Tomato Plant Protection

#### **Chapter 5**

## Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact, Challenges, and Management

*Indhravathi Chintapalli and Usha Rayalcheruvu*

#### **Abstract**

Insect-borne plant viruses cause huge yield loss in the world's most important crops. Understanding viral transmission mechanisms involves defining plant virus receptors inside their insect vectors. Tomato leaf curl virus (ToLCV) is the most devastating virus for worldwide tomato production. Understanding the biology of ToLCV and devising management techniques are critical in combating this global threat. Researchers are looking into using advanced technologies to detect plant viruses quickly and handle them properly for long-term agriculture. This review's main goal is to highlight management solutions for effectively combating ToLCV outbreaks and worldwide spread. Resistance genes for plant viruses in agriculture have been identified using morphological, biochemical, and molecular markers from the ancient to the present era. Such techniques are extremely basic. Traditional virus identification methodologies should be integrated with current and advanced tools for efficient virus improvement in crops. This review's main goal is to highlight management solutions for effectively combating ToLCV outbreaks and worldwide spread. For this aim, we focus on the impact of ToLCV on the world's agriculture and the significance of recent advances in our comprehension of its interactions with its host and vector. Another important topic is the role of mutations and recombination in shaping the ToLCV genome's evolution and regional distribution.

**Keywords:** plant viruses, crop, yield, significant impact, challenges, molecular techniques

#### **1. Introduction**

In both tropics and subtropics of the world, tomato cultivation [*Solanum lycopersicon* L.] is significant and widespread. Tomato production has received greater attention recently because it is not only regarded as a dietary supply of the vitamins C, potassium, folate, and K, but also as a source of revenue and a significant factor in ensuring food security. China, India, the United States, Italy, Turkey, and Egypt are the world's top tomato-producing nations. The total area under tomato cultivation worldwide is 4.582 Mha, with a yield of 150.51 mt. India is expected to have produced 21 million metric tons of tomatoes for the fiscal year 2021. India, which is second on the list of countries producing tomatoes during the measured period, accounts for 10.51 percent of the world's total tomato production cultivated 781,000 hectares or more. Production: 243,367 hg/ha. It is the second most significant vegetable. The major states in India are Andhra Pradesh, Karnataka, Orissa, Maharashtra, West Bengal, Bihar, Gujarat, Chhattisgarh, Tamil Nadu, and Jharkhand. The highest tomato producer in India is Andhra Pradesh, which produces 5962.21 thousand tons of tomatoes annually (from FY 2015 to FY 2020). In comparison to the prior fiscal year, the cultivated area increased. Tomato production has received greater attention recently, and tomatoes are often regarded as protective foods due to their high lycopene content, which aids in the prevention of various cancers. However, there are numerous obstacles to the production of tomatoes.

Plant viruses are considered to be predominantly damaging to their cultivated crop hosts' lives. In the majority of instances analyzed, virus-cultivated agricultural plant interactions have a detrimental impact on host morphology and physiology, resulting in disease [1, 2]. Viral diseases impact a lot of vegetable crops. The world's food supply is seriously threatened by crop diseases brought on by pathogenic microbes. Viruses, viroids, phytoplasma, bacteria, fungi, and nematodes are some of the pathogens that cause infectious plant diseases. Viral diseases pose a serious threat to sustainable and productive agriculture globally, causing annual economic losses. Plant pathogen infections are one of the main factors limiting crop productivity globally, and any destructive issues are caused by the wide range of viral isolates with highly variable degrees of virulence. They are immovable and often pass from one plant to another via a live thing called a vector or carrier. Since they have piercing-sucking mouthparts that enable them to reach and feed on the contents of plant cells, aphids, whiteflies, thrips, and leafhoppers are the most frequent carriers of plant viruses. Viruses can also be spread by other insects, mites, nematodes, fungi, contaminated seeds, pollen, vegetative propagation material, plant-to-plant contact, and other pests [3]. Emerging diseases, which are characterized by a rapid rise in disease incidence, geographic scope, and/or pathogenicity, have the most impact. Although the source of plant viruses is unknown, various suggestions have been put forth that suggest a possible insect vector as a possible explanation for the similarities between some plant and animal viruses. Plant viruses are challenging to control because they are widespread throughout the world and are effectively transmitted to their host plants by vectors. Although the length and specificity of the interactions between viruses and vectors vary, some recurring motifs in vector transmission have emerged: Plant viruses bind to specific sites in or on vectors and are retained there until they are transmitted to their plant hosts; viruses bind to specific sites in or on vectors and are retained there until transmission to their plant hosts; and viruses determine the virion's structural proteins, which are essential for transmission, as well as additional nonstructural helper proteins in some cases [4]. Entry, encapsulation, translation, replication, cell-to-cell movement, encapsidation, vascular transport, and plant-to-plant transmission—which can be horizontal through vectors or mechanical wounds, or vertical through seeds and pollen—are the basic steps in the successful infection of a plant by a virus. Viruses must combat host defenses such as RNA silencing and protein-mediated general and targeted immunity [5]. The majority of plant pathogenic viruses have an essential component to their infection cycle: acquisition and dissemination by an insect vector. Sap-sucking insects spread the virus in two ways: persistent transmission and nonpersistent transmission, which refer to how long it

#### *Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

takes an insect to acquire and transmit the virus, and circulative transmission; in some cases, it then involves virus replication in the cells of the insect host. Plant viruses can interact with their insect host in a variety of ways. Replicating viruses can also cause the insect host to mount general and targeted defenses. A recurring character is a need for specific molecular interactions between the virus and host, frequently via proteins, for the virus to interact with its insect host or carrier. By preventing virus absorption and transmission, plant protection strategies can be supported by knowledge of the interactions between plant viruses and the insects that serve as their hosts. Here, we offer a perspective centered on identifying existing and novel strategies with research directions to facilitate control of plant viruses by better understanding and focusing on virus-insect molecular interactions with these interactions in insect vectors of plant viruses, and we consider technical advancements for their control that may be more broadly applicable to plant virus vectors [6].

The increase of publications published on the topic during the past 15 years demonstrates the resurgence of interest in plant virus evolution. In the past 5 years, several new viruses have been described, some of which have novel genetic characteristics that have prompted the suggestion of the formation of new genera and the revision of the virus taxonomy status. There is a need for work aimed at understanding the processes involved in plant viral evolution, because contemporary plant virus evolution research has been regarded from a molecular, rather than populational, approach. Plant viruses create a significant amount of genetic variation that is present both within and between species using a variety of ways. Plant RNA viruses and pararetroviruses most likely have replication processes that are very error-prone, leading to a lot of mutations and a quasispecies nature. Although the origin of the diversity in the plant DNA viruses is not entirely apparent, it does exist. Recombination and reassortment are commonly used by plant viruses as evolutionary forces, as are occasionally other methods including gene duplication and hyperinflation [7].

Even though there is no proof of variation in the mutation rate, the amount of variety detected in different species of plant viruses is extremely varied. Plant viruses are thought to result in significant annual losses across the globe. Recent climate change events may have made this problem worse, and climate change will likely have an impact on how diseases spread in the future, which may affect how plant viruses spread. Increases in temperature, atmospheric carbon dioxide concentration, water availability, and the frequency of extreme weather events will all have a direct and indirect impact on plant viruses by affecting their hosts and vectors. Climate change may have an impact on plant viruses' virulence and pathogenicity, which will increase the frequency and scope of disease outbreaks. The natural defensive process of plants, known as autophagy, has become crucial in the interactions between plants and pathogens. In plants, it serves as an antiviral defense mechanism [8]. The virus alteration demonstrates how plant viruses can control, subvert, or even employ the autophagy system for pathogenicity. However, accumulating evidence from virus modification shows that: (1) high mutation rates are not necessarily adaptive, as a significant portion of the mutations is deleterious or lethal; (2) despite having a high potential for genetic variation, populations of plant viruses are not highly variable, and genetic stability is the norm rather than the exception; and (3) the degree of genetic variation constriction in virus-encoded proteins is comparable to that in their eukaryotic hosts and vectors [9].

Although it is difficult to trace the evolutionary history of viruses and practically impossible to regulate virus disease over the long term, their propensity for fast adaptation makes them a great model system for research on the broad mechanisms

underlying molecular evolution. More exemplary research was done in the second half of the twentieth century, demonstrating the infectiousness of RNA alone, (ii) the resolution of RNA-protein interactions in the structure determined by X-ray fiber diffraction, (iii) the existence of a distinct region on the virus for the start of encapsidation, (iv) the definition of the virus sequence and open reading frames (ORFs); (v) open reading frames and the definition of the viral sequence (ORFs); and (vi) cDNA clone that is biologically active [10]. This substantially contributed to our comprehension of reproduction and transmission, resulting in a new understanding of viruses among scientists in a subsequent generation. In addition to improving our knowledge of the local ecology and fitness of mechanically transmitted viruses, this upcoming research must expand our understanding of virus structure and transporters of small molecules. The process of developing effective host-virus interactions, including how different species move through a vector in different ways. The top nine virus list is shown in **Table 1**, in descending order [19].

#### **1.1 Viruses, crops affected, and damage caused**

Hence, there is an urgent need to improve its productivity with the help of modern technological implementation to shield the tomato plants against various biotic and abiotic factors.

One of the main biotic limitations is virus-associated. One of the most significant factors restricting its cultivation and productivity is tomato leaf curl disease (ToLCD). It frequently suffers from a range of infections, which causes significant yield losses. Infections caused by fungi, bacteria, and phytoplasma are only a few of the many viral diseases that affect it. The most significant and damaging viral pathogen in many regions of the world is the tomato leaf curl virus (ToLCV), a geminivirus, which is responsible for all documented viral diseases in tomatoes [20–27]. Based on the genome organization, host range, phylogenetic relationships, and insect vectors, geminiviruses have been classified into nine genera: Becurtovirus (two species), Begomovirus (>320 species), Curtovirus (three species), Mastrevirus (>30 species), Eragrovirus (one species), Topocuvirus (one species), and Turncurtovirus (one species). Begomovirus is the largest genus in the Geminiviridae family, and it contains multiple notable species, including ToLCV, which infects tomato cultures in Asia and Australia. ToLCV is the name given to a group of vector-transmitted geminivirus genus [28]. Geminiviruses are made up of one or two circular ssDNA genomic components of 2500–3000 nucleotides encapsulated in paired icosahedra or geminate particles (called geminiviruses), as we know them now. Inside the host cell, their ssDNA genome is converted into a dsDNA intermediate and rapidly replicated (**Figure 1**). The introduction of high-yielding tomato varieties has been accompanied by ToLCV infection [29–31]. In India, in Andhra Pradesh, the disease is widespread in tomatoes during the summer season in southern parts and autumn in northern parts and causes yield losses ranging from 27 to 100% [32–34].

#### **1.2 Genome organization**

Plant viruses belonging to the Geminiviridae family have circular genomes made of single-stranded (ss) DNA, which encodes their genetic material. The genomes of all geminiviruses are identical, as is DNA-A, the section of bipartite begomoviruses that encodes the proteins necessary for replication, control of gene expression, getting past host defenses, encapsidation, and insect transmission. Two proteins that enable


#### *Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*


## **Table**

 **1.** *The top 10 viruses list in rank order.*

*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

#### **Figure 1.**

*Å structure of a plant geminivirus—Nanoviruses have* T *= 1 icosahedral capsids of 18nm in diameter.*

intracellular and intercellular movement in host plants are encoded by the second component, DNA-B. The unknown is the origin of the DNA-B component. This work aimed to investigate the relationship between the bipartite begomovirus DNA-A and DNA-B components to unravel their evolutionary histories and gain insight into the potential origins of the DNA-B component [35].

One of the genomes recognized are begomovirus species, and they have all Old World origins (OW). Monopartite viruses have only one molecule of nucleic acid [36]. The majority of dsDNA viruses exist in monopartite forms. It has been demonstrated that monopartite begomoviruses interact with betasatellites, which are ssDNA satellites. In contrast to monopartite begomoviruses, which are phloem confined and only cause stunting and leaf curl symptoms, several bipartite begomoviruses infect both phloem and other tissues, causing leaf curling, crumpling, and mosaic/mottling symptoms, and are sap transmissible [37–40].

#### *1.2.1 Gene functions*

The six ORFs that make up DNA-genome A's are used to encode the signals for the six most significant viral proteins, which range in size from 11 to 40 kDa. Both viruses detect ORF. Monopartite begomoviruses have complementary-sense ORFs (C1–C4) and virion-sense ORFs (V1 and V2), respectively. Together with the V2 protein, the virion-sense coat protein (CP) promotes viral mobility in plants and is responsible for insect transmission. The sole viral protein necessary for viral DNA replication is the geminiviral replication-associated protein (Rep). The ToLCV intergenic region has a 120-bp segment that Rep particularly binds to. Two significant movement proteins, BV1 and BC1, are encoded by DNA-B and are in charge of long-distance mobility between cells. Globally, both economically significant and weed crops suffer enormous economic losses due to begomoviruses and their satellites. Monopartite begomoviruses typically develop a virus/satellite complex with a betasatellite and cause severe leaf curl in tomatoes Betasatellites linked with geminivirus play a variety of roles in pathogenesis [41]. In India, the papaya leaf curl virus (PaLCuV) associated with aster betasatellite yellow vein disease was found. In Pakistan, PeLCV infection of Pedilanthus tithymaloides plants was connected to the betasatellite for tobacco leaf curl [42, 43]. Recently, the alphasatellite, a 1.4-kb additional satellite DNA, has been

connected to the tomato leaf curl virus (TYLCV). Although they multiply autonomously inside their hosts, they depend on the helper virus for encapsulation, movement inside the plant, and vector transmission. They can multiply independently within their hosts, but they, like betasatellites, depend on the helper virus for encapsulation, mobility inside the plant, and vector transmission. The earliest satellites discovered in relationship with the ToLCV were deltasatellites, an Australian example (ToLCV). Previously known as ToLCV-Sat, is currently called tomato leaf curl deltasatellite (ToLCD). The ToLCD (682 nt long) and the helper virus have no significant sequence similarities other than the origin of replication [44].

#### *1.2.2 Geminiviruses replication*

Geminiviruses employ their insect vectors to transfer their encapsidated DNA to plants. Due to the lack of a model system for eukaryotic replication, details of geminivirus replication remain unknown. Other research indicates that the replication initiator protein (Rep) is the viral protein most crucial to viral DNA replication. Yellow mungbean mosaic mutations in CR, Rep, AC5, and other replication-related components and viral proteins, as well as a lack of host factors, inhibited the replication of the India virus. When injected into protoplasts, DNA-A can replicate itself and contain the genes required for viral replication [45]. In mature plant cells, geminiviruses promote the expansion of the DNA replication apparatus to create a favorable environment for replication. On the other hand, a lot is still unknown about the process of C4-induced cell division. The physiological advantage of promoting cell division might produce a situation where DNA viruses can multiply easily. There is proof that ToLCV can be replicated within the vector, that is, whiteflies. A fivefold increase in viral accumulation was observed in the early stages of the insect's life cycle, followed by a decline in the later stages, according to the first study to demonstrate this phenomenon, which was published in 2015.

V1, V2, and C3 displayed increased expression after nonviruliferous flies fed on TYLCV-infected tomato plants for 1 to 3 days. The midgut epithelial cells were also found to have complementary viral DNA, a sign of viruses that replicate [5, 46]. This suggests that these cells served as viral replication hubs. Another study found that insects that consumed diseased tomato plants accumulated 10 times more viral gene transcripts. In the later stages of the investigation, the reduction in virus titers was also attributed to autophagy [47]. Contrarily, experimental evidence supports the notion that TYLCV cannot replicate within its vector. The quantity of viral transcripts increases only during the acquisition phase and thereafter remains constant, according to other research, indicating that replication is not occurring [48, 49].

Many DNA viruses depend on their hosts' transcription and replication systems. Geminiviruses only employ a small number of proteins, and they rely on host enzymes to carry out their functions. These genes are transcribed by begomoviruses using their bidirectional promoters. It produces several overlapping RNA species, some of which are polycistronic. The TATA box initiates transcription, and the RNA produced by the virus is polyadenylated, showing that the host transcription machinery aids in the geminivirus' transcription. One complementary-sense transcript (BC1) is produced for DNA-B.

#### **1.3 Tomato leaf curl virus**

The Food and Agriculture Organization of the United Nations estimates that the tomato is the most widely grown tomato crop in the world, producing about 180

#### *Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

million tons annually [50]. Growing for the processing industry accounts for one-fourth of the 160 million tons. The annual processing capacity of factories operated by significant food corporations is about 39 million tons of tomatoes. In all tropical locations of the world, ToLCV is one of the most virulent viruses that can cause catastrophic illnesses in vegetable crops [51]. Since ToLCV was originally identified in solanaceous crops, numerous instances of harm to other crops have surfaced all across the world (**Table 2**). It has a devastating impact on the development and production of several plant groups with significant agricultural economic importance. In 1948, Vasudeva and Samraj reported the first ToLCV case in India. ToLCNDV has expanded to other vegetable and fiber crops, according to several studies from the last 10 years [52]. Tomato leaf curl New Delhi virus with mosaic and leaf curl disease has just been discovered in chrysanthemum. This virus is attributed to diseases in a wide variety of plant species, including fruit crops and ornamental plants [53]. These begomoviruses may act as reservoirs for crops that are crucial to the economy [2, 11, 50]. Because it causes one of the most prevalent and economically significant tomato diseases in the world, ToLCNDV significantly hinders tomato output in general [54]. As it has spread across the Indian subcontinent, ToLCNDV's host range has significantly increased. India and Pakistan have reported cases of ToLCV linked to cotton leaf curl disease between 2013 and 2015 [55, 56]. The insects harm the plant directly by sucking phloem sap (**Figure 2**), which stunts growth, causes early wilting, prematures defoliation, and ultimately results in yield loss. They also damage the plant indirectly by excreting honeydew, which promotes the growth of fungi on the surfaces of leaves and fruit. When a peach plant was infected with the leaf curl virus, the leaves had very little chlorophyll and performed little or no photosynthesis. Previous research has suggested that ToLCV causes increased reactive oxygen species (ROS) production in tomato leaves infected with it [57].

#### **1.4 Occurrence and yield loss**

According to geographic distribution, the ToLCD is expanding quickly. It affects tomatoes and significantly reduces agricultural yields in the Southeast United States and around the world [58]. If infected, susceptible tomato types could lose up to 100% of their production (142,251). The tobacco leaf curl virus on tomato first appeared naturally in India in 1942 according to Pruthi and Samuel, who were followed by Vasudeva and Samraj in 1948. Later, Andhra Pradesh, Hyderabad [59, 60], and Tamil Nadu [61] first reported the detailed characterization of ToLCV from Karnataka by Govindu and his coworkers in 1964. The prevalence of ToLCD in South India rapidly increases from 27 to 90 percent in susceptible cultivars, leading to yield losses of up to 90 percent [62]. The virus's extreme invasiveness and a dearth of efficient control methods allowed it to spread globally and cause a serious pandemic. Nine different ToLCV isolates have been found in India [11]. In the United States, the TYLCV-like ToLCV has been identified [63, 64]. The tomato leaf curl viral illness, which has a significantly high disease incidence in both the Rabi and Summer seasons, 96.80 percent and 98.43 percent, respectively, according to Khandare et al. [65], produces severe leaf curl disease. However, the first reports of tomato yellow leaf curd disease and the connection between radish leaf curl virus (RaLCV) isolates and a tobacco disease were both made in 2012 by Singh and his colleagues [66]. The majority of Indian isolates of ToLCVs are caused by a monopartite tomato leaf curl Joydebpur virus (ToLCJoV) that causes severe leaf curl. Numerous plants, including natural fibers and chillies, have been discovered to be infected by monopartite ToLCJoVs [67].

#### *Tomato - From Cultivation to Processing Technology*


*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*


#### *Tomato - From Cultivation to Processing Technology*


#### **Table 2.**

*There were reports of huge crop devastation all across the world due to ToLCV.*

Tomato leaf curl Palampur virus (ToLCPalV) strains/variants' importance is an increasing hazard to cucurbit output in India. Because of the recombination breakpoint of the viral genome, these valuable crops are now being destroyed and impacted by ToLCV. ToLCV has emerged as a major limiting factor and problem for farmers and scientists alike, with a special focus on management initiatives for preventing the spread of ToLCD. Because of the economic importance of LCV, efforts have been made to understand LCV pathophysiology and generate tolerant plants using breeding and transgenic techniques [61].

*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

#### **Figure 2.**

*Schematic representation of Bemisia tabaci [insect vector]-mediated transmission of plant viruses in leaf—Spreads begomoviruses by sucking the sap from the phloem of leaf curl virus-infected plants (red geminate particles). As a result of the infection, the plants show characteristic begomovirus symptoms such as vein yellowing, foliar yellow mosaics, and leaf curling.*

#### **1.5 Geographical distribution**

Cucumber mosaic virus (CMV), tomato spotted wilt virus (TSWV), tomato aspermy virus (TAV), tobacco mosaic virus (TMV), tomato bushy stunt virus (TBSV), potato Y virus (PVY), and ToLCV are among the viruses that infect tomatoes [68–70]. Additionally, mixed infections frequently contain these viruses [71]. In the southern region of India, a monopartite ToLCV is the most prevalent of the various ToLCVs [72]. There are several species of begomoviruses, viz. bipartite tomato leaf curl Palampur virus (ToLCPalV) and tomato leaf curl New Delhi virus (ToLCNDV), monopartite tomato leaf curl Kerala virus (ToLCKeV), tomato leaf curl Patna virus (ToLCPaV), tomato leaf curl Ranchi virus (ToLCRnV), tomato leaf curl Rajasthan virus (ToLCRaV), tomato leaf curl Pune virus (ToLCPV), tomato leaf curl Bangalore virus (ToLCBaV), tomato leaf curl Lucknow virus (ToLCLuV), tomato leaf curl Karnataka virus (ToLCKaV), tomato leaf curl Gujarat virus (ToLCGuV), and tomato leaf curl Joydebpur virus (ToLCJoV). The begomovirus, ToLCGuV, that exists in both mono and bipartite forms has been reported from Varanasi [73]. Both of these categories are common in India [74, 75]. ToLCNDV is a rare Old World bipartite begomovirus. It is ubiquitous on the Indian subcontinent but found elsewhere in the Far East, Middle East, North Africa, and Europe [76, 77]. According to Sahu et al., the occurrence of Guar leaf curl alphasatellite (GLCuA) could be attributed to whitefly migration from Pakistan [78, 79]. ToLCV has been linked to infections in a variety of hosts between 2000 and 2010 [80]. Only the DNA-A component appears to be present in the monopartite genomes of the tomato leaf curl Bangalore virus from Bangalore

[81, 82] and tomato leaf curl Karnataka virus from Karnataka [83]. The PaLCuV has been found in many different crops across the globe [84]. Numerous begomoviruses may pick papaya plants to survive in a variety of uncertain environments. The presence of chili leaf curl virus and its related tomato leaf curl betasatellite in the *Cucurbita maxima* host reveals the virus's potential harm to this crop [85]. There is a chance that this virus complex could speed up the spread to other crops. There is a risk that the virus could spread to other countries via air, sea, and possibly through borders, particularly among countries that share borders. It is critical to avoid any chance of the virus spreading to new hosts in other nations by enacting effective quarantine legislation.

#### **1.6 Virus-vector interaction**

There are often two components to a vector-transmitted pathogen: host-pathogen interaction and vector-virus contact (**Figure 3**). In the past, research on plant-virus interactions has focused on viral mobility, replication, symptom development, and the plant's reaction to infection. Vector-virus interaction, on the other hand, has been an insufficient investigation.

One of the most invasive creatures is the Bemisiatabaci [Hemiptera: Aleyrodidae], often known as the whitefly as a vector for ToLCV. It has been ranked among the top 100 worst invasive alien species in the world. It is a cryptic species that includes at least 39 Hemiptera species and is found naturally throughout the world's tropical and subtropical regions [86]. In the last 20 years, *B. tabaci* has spread around the globe, likely as a result of the transportation of agricultural products [87]. It has become one of the most destructive agricultural pests. The bug damages the plant directly by sucking phloem sap, which results in stunted development, early wilting, premature defoliation, and ultimately a loss of production, as well as indirectly by excreting

**Figure 3.**

*The infection cycle starts when the vector comes into contact with the virus in the plant and acquires it. The virus must then survive long enough in or on the vector to be transmitted to a new host and released into the plant cell.*

#### *Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

honeydew, which encourages fungal growth on the surfaces of leaves and fruits [88]. Over 600 plant species are targeted by Bemisiatabaci, and viruliferous whiteflies creating a feeding site directly contributes to leaf curl virus transmission to plant hosts [89]. Alfalfa leaf curl virus (ALCV), which is spread by Aphids craccivora [90], is one of two Capulavirus species that are spread by aphids. Aphid transmission was only found in 2015, even though the Aphididae family contains most species described as plant virus vectors [91, 92]. An unclassified geminivirus from the genus Capulavirus can spread ALCV [93]. Being polyphagous explains B. tobaci wide host range and ability to spread a lot of viruses to more than 300 plant species from 63 families [94]. Forty-nine begomovirus species have been related to ToLCD, while 17 have been linked to TYLCD [95, 96]. The two primary tomato geminiviruses that infect tomatoes and significantly reduce output are ToLCV and TYLCV [97].

The key features of ToLCV acquisition, retention, and egestion by the vector have been the focus of investigations into how viruses interact with their host's machinery fluids. The maxillary stylet is made up of three stylets: a mandibular stylet and the salivary canal. Individual whiteflies may acquire varying amounts of viral particles in their exoskeletons even when fed on the same tissue for the same significant period. In insects, the precibarium and cibarium taste organs control whether virus particles move through circulatory or noncirculatory channels. Circulating persistent viruses circulate and stay in the body of the insect for the majority of its life, whereas noncirculative viruses never breach the gut barrier [77]. Any alteration to the protein that supports virus retention and transmission (CP) interferes with both processes [98]. ToLCV uses endosymbionts from whiteflies that release proteins to stop damage in the open circulatory system. ToLCV's CP interacts with GroEL, a molecular chaperone, and is shielded from deterioration. It has been demonstrated that Hsp70 interacts with CP and blocks transmission. The continuous cycle between the host and vector due to their circulative pathway affects prospective agricultural output all over the world. ToLCV particles enter the salivary glands of viruliferous flies while they feed on phloem sap and are then discharged into plants [99].

#### **1.7 Plant-virus interaction**

Geminiviruses are intracellular parasites that must successfully influence plant cell activities to multiply, block antiviral defenses, and spread throughout the plant [44, 100]. They inject viral DNA into the host cell during infection, disrupting the host gene silencing system. During viral infection, the production of ROS in the host cells is raised to prevent systemic virus migration up to specific cells [63]. To counteract the self-damage produced by ROS, the host creates glutathione peroxidases (GPXs), which lower ROS levels in the cells. The competence of a virus to co-opt and alter processes in a particular host plant will influence how the virus-plant relationship turns out. To hijack the molecular machinery of the host cell, geminiviruses produce a small number (between 4 and 8) of tiny, rapid evolving, multifunctional proteins, encoded by bidirectional and partially overlapping ORFs [101]. ToLCV is a whiteflytransmitted vector-borne disease (**Figure 4**), and this is the first time ToLCNDV has been identified as a seed-transmissible virus in zucchini squash plants in Italy. The leaf curl virus was found in early seedlings sprouted organically from fallen fruits [18]. After being injected into the host, virus particles multiply and travel to other areas of the body, causing symptoms [102]. Geminiviruses manipulate E2F transcription factor activity to produce an S-phase environment [103]. Beet curly top virus (BCTV) infection causes cell expansion (hypertrophy) and division (hyperplasia) [104].

**Figure 4.**

*Plant-pathogen interactions-global environmental change, study the environmental parameters (biotic and abiotic) influencing the biology of plant viruses and their transmission by vectors.*

DNA virus replication in differentiated plant cells requires the induction of cell division. To achieve efficient replication, DNA viruses encode the C4 protein, which encourages cell proliferation. The viral protein may interact with yeast and alter its growth and development, since the expression of BCTV C4 caused a 100-fold decrease in transformation efficiency [105, 106]. The glycogen synthesis kinase-3 (GSK3) homolog in N. benthamiana, NbSK, is hijacked by the C4 protein of the tomato leaf curl Yunnan virus (TLCYnV). According to Mei and his coworkers in 2018 [107], SK is a target of geminivirus C4, and activation of cell division is necessary for DNA virus replication in differentiated plant cells. According to Chandan et al., infected tomato plants exhibited increased expression of LeCTR1, one of the ethylene signaling pathway's negative regulators. LeCTr1 gene silencing caused by the tobacco rattle virus (TRV) increased ToLCJoV infection tolerance [101].

#### **1.8 ToLCV recombination, mutation, and evolution**

Begomovirus disease complexes are quickly developing to increase their host range and overcome resistance sources through recombination, component capture, and mutations.

#### *1.8.1 Mutation*

Genetic variation in the tomato yellow leaf curl Sardinia virus [TYLCSV] is a result of interspecific recombination as well as mutation, natural selection, and genetic drift [108]. RNA viruses have a high mutation rate as a result of error-prone replication mediated by viral RNA-dependent RNA polymerases (RDRP). Recombination that occurs during mixed infections and mutations is what drives viral evolution. DNA viruses like TYLCV, whose replication is facilitated by plant DNA polymerases, should have modest mutation rates as a result of proofreading activity [109]. However,

studies disagree with these assumptions, and considerable variability has been reported. Serious outbreaks have been triggered by recombinant TYLCVs with resistance-breaking capabilities [110, 111].

#### *1.8.2 Recombination and evolution*

Mixed infections and high viral replication levels are two conditions that may promote recombination [103]. As recombination hotspots, the IR, V1, V2, and C1 regions of the genome have been identified [112–118]. Studies from the Punjab regions of Pakistan and India recently suggest that CLCuMuV has reemerged in the Indian subcontinent, which is consistent with observations from earlier studies. Its dominance and frequent emergence in North Indian regions have been demonstrated by recent studies [55]. Researchers examined and studied recombination events in the viruses to see if they had any consequence on viral transmission due to the high diversity of begomoviruses in North India. Geminiviruses have evolved and appeared as a result of recombination. The Cotton Leaf Curl Multan betasatellite [CLCuMBC1]'s satellite conserved region (SCR) and A-rich portions were particularly prone to recombination [67]. Additionally, recombination was found in the Rep and A-rich regions of the GLCuA, supporting earlier findings that suggested these sites were recombination hotspots [11]. As shown by DNA-A components of ToLCNDV isolates [119], recombinations involving genomic sequences from other begomoviruses or sequences of unknown origin are frequent in this genus.

Recombination happened in 458 occurrences among begomoviruses related to cotton leaf curl disease during 459 mixed infections [120]. If these viruses travel to other locations, 460, where they are common, new viruses may emerge [4]. The Recombination Detection Program version 4 [RDP4] was used to identify possible recombination events in sequences [121]. SeqMan and GenBankBLASTn were used to put together sequence reads (**Table 3**). MEGA7 was used to perform pairwise nucleotide sequence analysis and build phylogenetic trees [139]. This sequence analysis is useful to determine the threshold level at 91 percent sequence identified demarcation for begomovirus classification that has been set [140].

#### **2. Plant viruses management and detection**

Human activity is creating conditions that encourage the spread of begomoviruses. To avoid ToLCV outbreaks and agricultural losses, administrative, legal, and technological procedures should be implemented. Insect vector biological management may also be explored to minimize insect vector infestation and disease dissemination. We discuss recent developments in the identification, characterization, and detection of plant viruses and virus-like compounds using nano-coupled molecular method approaches in this review article [141]. The objective of this essay was to accurately identify the most significant plant viruses as reported by Molecular Techniques contributors. We are well aware that between disciplines and regions, importance and priority might differ locally (**Table 4**). In the past, biological assays were a crucial tool for determining a plant's health condition as well as for the identification and characterization of a specific virus. The labor- and time-intensive biological indexing has few practical uses nowadays, though it is still important for plant virus research. Instead, molecular diagnostics is the primary method used for the precise and sensitive detection of the majority of viruses and virus-like diseases [151]. Some recent developments



*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*


*Tomato - From Cultivation to Processing Technology*


*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

> **Table3.**

 *Some plant viruses are shown in the table, along with their Genbank accession numbers.*


*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*


#### **Table 4.**

*For the identification of plant viruses and virus-like pathogens, nanotechnology integrated molecular method techniques.*

in point-of-care (POC) nucleic acid extraction technology are summarized in this study. Emerging bacterial, viral, fungal, and other pathogen-caused human and plant diseases present a persistent threat to global health and food security [152]. Although there are numerous pipelines for finding plant viruses, they all have a similar structure. In POC diagnostics, plant samples are examined right away at the sampling site for disease screening. Rapid point-of-care (POC) molecular diagnostics of plant diseases is becoming increasingly important for disease control and agricultural protection. The identification of the disease-causing pathogens and their pathogenesis is revealed at the genomic level by nucleic acid-based molecular diagnostics. One of the most important and efficient steps in creating control strategies for plant viral infections is still the development of reliable and early detection technologies [117].

#### **2.1 Conventional measures before, during, and after vegetation**

The virus itself is frequently not the main problem, but rather the vector that it travels on. To reduce the population of *B. tabaci*, insecticides including imidacloprid, acetamiprid, dinotefuran, and thiamethoxam are frequently applied. Whitefly infestation in nurseries may be avoided by using netting. Eretmocerus eremicus and other biocontrol agents could be very effective control agents. One of the main challenges in managing pathogens is keeping a pathogen alive in many hosts. Alternate hosts serve as a repository for inoculum both during the growing season and during periods when there is no crop. Recent research indicates that the effectiveness of using chitosan as a biocontrol agent against ToLCV has increased when combined with pseudomonas. Because begomoviruses are so common, breeders should take them into account when making selections for resistance.

#### **2.2 Biotechnological approaches for viral disease diagnosis**

The family Geminiviridae (genus Begomovirus) has more than 100 different viruses. Because of improvements in cloning and low-cost sequencing, the number of accessible genome sequences has significantly increased. Tomatoes have a small DNA genome that is simple to clone. Numerous tomato viruses can be identified, as well as new or emerging viruses and viroids, using general virus detection technologies like enzyme-linked immunosorbent assay (ELISA) or polymerase chain reaction (PCR). For the accurate identification of well-known and novel viruses and viroids, a bioinformatics pipeline based on the alignment and assembly of sRNA or DNA sequences is necessary. Key viral genes called ToLCNDV miRs affect the fundamental processes involved in virus emergence. RNAi-based viral gene silencing and sense/antisense technology have been used to create transgenic resistance. The use of next-generation sequencing [NGS] technology to quickly, precisely, and affordably detect miRs has become commonplace. The generation of markers for marker-assisted selection (MAS) of resistant genotypes may now be done swiftly because to the development of NGS technology and high-throughput genotyping platforms [153]. The use of transgenic methods to control viral infections is extremely successful. The public hostility to genetically modified organisms (GMOs) in developing countries like India has inhibited their implementation. Infection is reduced by 75% in transgenic tomato plants that produce dsRNA-containing sequences from the IR, V1, and V2 regions of TYLCV-Oman. The identification of VSR may lead to the development of disease resistance strategies and other biotechnological applications. RNA silencing sometimes referred to as RNA interference or RNAi shields plants from viroids and invading viruses [111]. The AC4/C4 protein is a pathogenicity protein that impacts plant development and can be used as a useful tool for research into cell cycle regulation, hormone signaling, cell differentiation, and plant development. It has been demonstrated that transgenic lines of AC4 viruses led to abnormal phenotypes and developmental patterns in several different host plants. Disease-resistant tomato varieties can assist with gene-pivoting and resistance-breeding [154].

Elite tomato breeding lines were chosen using a mix of phenotypic and molecular screening techniques for ToLCD, late blight, and RKN. To develop fresh market tomato lines that are begomovirus-resistant, AVRDC, The World Vegetable Center, initiated a campaign. A lipid transfer protein, nucleotide-binding site, and leucinerich repeat (NBS-LRR) proteins, posttranscriptional gene silencing machinery and other defense genes are expressed specifically in the host in response to ToLCNDV infection. To build resistance against various ToLCV strains, six resistance loci have been employed in tomato plants. Of the six resistance/tolerance loci, Ty-1, Ty-2, and Ty-3 are the ones that are used most frequently. These results suggest that these Ty loci increase host defense via a different mechanism from the R gene-mediated hypersensitive response (HR). Increased expression of genes associated with the lignin and SA biosynthesis pathways has been related to improved plant virus defense, according to reports. A global miR profiling study has shown that a large number of miRs were differently changed in ToLCV-ND-infected Pusa Ruby tomato leaf tissue. The study also identified miRs that demonstrated differential expression between the sensitive and tolerant cultivars. ToLCD resistance is conferred by the expression of artificial microRNA targeting the ATP-binding region of AC1 in transgenic tomatoes without affecting tomato output. The antisera's usefulness in begomovirus identification in field samples and reservoir hosts is demonstrated by polyclonal antibodies generated using purified intact virus and rCP of tomato leaf curl Bangalore virus [155]. From tomato and N. benthamiana leaves that had been treated with the virus, ToLCBV was successfully purified. Different Indian isolates of the begomovirus in plant and weed species might be recognized using monoclonal antibodies. ToLCBVC viral infections were caused by biologist Devaraja in tomato samples as well as other

*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

crop and weed plant types. Total viral resistance is provided by transgenic plants that produce Cas9 and dual gRNAs that target different regions of the cotton leaf curl Multan virus (CLCuMuV) single-stranded DNA genome, offering a special method for developing geminivirus resistance. Clustered, regularly interspaced short palindromic repeats/CRISPR-associated proteins are the foundation of the genome-editing technique CRISPR/Cas. It originated from the bacterial and archaeal adaptive immune systems that resist viruses [156, 157].

#### **2.3 Nanotechnology-based nucleic acid/viral particle detection**

High-throughput sequencing (HTS) has enabled virologists to identify an unprecedented number of viruses, advancing our understanding of the variety of viruses in nature and, in particular, revealing the virome of many crops. The gaps in our knowledge of virus biology have, however, frequently become wider as a result of these new virus discoveries. Enzyme-linked immunosorbent assays (ELISA) and direct tissue blot immunoassays are two immunological methods now utilized to detect infections in plants (DTBIA). For the identification and detection of pathogens, DNA-based techniques such as polymerase chain reaction (PCR), real-time PCR (RTPCR), and dot blot hybridization have also been proposed [158].

The sensitivity and specificity of virus nucleic acid sequence identity are predicted to be improved by using nanotechnology-based techniques, which are thought to be more effective, safer, and target-specific. Compounds made of nanoparticles (NPs) can simulate ligand and receptor binding to particular target-specific plant diseases, such as the interaction between an antigen and an antibody. The gold standard for plant disease diagnostics uses molecular assays based on nucleic acids and antibodies. Being one of the most fascinating and dynamic fields of research, nanotechnology has a significant impact on a wide range of fields, including science, engineering, medicine, and agriculture. Nanomaterials, whose diameters typically vary from 1 to 100 nm, can offer enhanced surface-to-volume ratios as well as special chemical, optical, and electrical properties, making them excellent candidates for the analysis of plant diseases. For the quick detection of a variety of human and plant illnesses, lateral flow assays (LFA) and electrochemical sensors have been used as some nano-based approaches (**Table 5**). Fast identification of plant diseases is now possible because of portable imaging equipment (such as cellphones) backed by nanostructures. Due to the extraordinary biosecurity of designed molecular recognitions at the nanoscale, which has seen exceptional development in the past decade, nanoscale materials are promising possibilities for plant disease detection. Overall, thanks to recent advancements in rapid plant DNA extraction technology made possible by microneedles, tiny DNA sequencing chips, and smartphone-based volatile organic (VOC) sensors, many traditional laboratory tests, like nucleic acid amplification, sequencing, and volatile organic compound (VOC) analysis, may now be performed directly in the crop field in a much faster and more affordable manner [163].

#### *2.3.1 Challenges in nanotechnology*

The environmental impact and toxicity of engineered nanomaterials, the quickness of data sharing and disease forecasting, and long-term sensor stability in extreme conditions like extreme cold or heat, prolonged sun exposure, and heavy wear are the three main challenges that currently exist for plant diagnostic tools. The first issue is that safety issues must be resolved before any nanosensors can be commercialized and


#### **Table 5.**

*Only a few methods use nanotechnology-based electrochemical nucleic acid sensing in disease diagnosis.*

used in the field because some nanoparticles, like QDs, may be hazardous. More thorough toxicity testing and regulation are required for nanosensors that will be used on living plants or consumable agriculture and food items, because dangerous

*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

nanomaterial residues may infiltrate the food chain and be consumed by end users. Regarding the second issue, the new generation of nanosensors is anticipated to be more wirelessly connected and capable of providing measurement in close to real time as the primary requirement for disease diagnosis is consistently the timely report and forecast of infection events on-site. Finally, before any sensors can be deployed to the actual yield, more resilient and robust sensors that can endure a wide range of environmental factors (such as temperature, humidity, air pollution, etc.) in the agricultural yield are predicted [164].

#### **2.4 Need to resolve**

#### *2.4.1 Several outstanding questions and future directions are highlighted*

	- What role in the pathophysiology of TYLCV do endogenous noncoding RNAs, such as sRNAs and lncRNAs, play?
	- Is there another way to safeguard against the virus? It is imperative to hunt for new sources of resistance given the virus's quick global spread.
	- Investigations focusing on this area may help to fix the problem, because there is still debate on whether TYLCV can reproduce inside the whitefly.
	- The effect of suppressor TYLCV proteins in dampening the host RNAsilencing machinery has been investigated extensively. It is yet unclear if they can inhibit proteins involved in other processes, such as the ubiquitinproteasome system or autophagy.
	- By pyramiding various poisons, can we develop broadspectrum resistance against chewing and sap-sucking insects?
	- Tma12 kills *B. tabaci* in two ways.
	- Will phloem-specific promoters improve the performance of dsRNA and Tam12?
	- Will the poisons cause *B. tabaci* to become resistant?
	- Can the simultaneous expression of many toxins and/or dsRNAs prevent resistance breaking?

### **3. Concluding remarks**

One of the most widely investigated plants viral diseases is the ToLCV. On the Indian subcontinent, ToLCV is the most hazardous bipartite begomovirus, and it has quickly spread to other regions of the world. The implementation of innovative management techniques is dependent on the availability of information regarding

*Leaf Curl Disease a Significant Constraint in the Production of Tomato: Impact… DOI: http://dx.doi.org/10.5772/intechopen.106733*

ToLCV-associated viruses and their epidemics, which is lacking. ToLCNDV has an incredibly diverse population, with mutations that have different host ranges and some that are better adapted to infecting particular host plants. The invention of novel techniques to defend plants from infection will be facilitated by an understanding of the fundamental mechanisms behind such host adaptation. GM techniques based on gene silencing have presented exceptionally significant options for plant viral resistance tactics, and their long-term promise should not be overlooked. Conventional control procedures alone are insufficient for ToLCV control. However, combining many of these approaches following suggestions based on an understanding of the disease's epidemiology may make managing ToLCNDV outbreaks easier. As a result, it is suggested that integrated management techniques integrating numerous control practices be used. However, more research about the epidemiology and ecology of this multifaceted disease is needed to develop efficient management strategies. Because this virus does seem to have a quicker response time than available controls, we should be able to predict the nature and diversity of ToLCV outbreaks in a more dynamic environment, which will have drastic effects on virus vectors. The discovery of more potent and long-lasting strategies to prevent epidemics also depends on a detailed awareness of ToLCV polymorphism and the factors that influence the growth of its inhabitants.

### **Conflicts of interest**

The authors declare no conflicts of interest.

#### **Author details**

Indhravathi Chintapalli and Usha Rayalcheruvu\* Sri Padmavati Mahila Visvavidyalayam (Women's University), Tirupati, India

\*Address all correspondence to: ushatirupathi2020@gmail.com

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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### **Chapter 6**

## Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato Cultivation

*Refik Bozbuga, Songul Yalcin Ates, Pakize Gok Guler, Hatice Nilufer Yildiz, Pınar Aridici Kara, Bekir Bulent Arpaci and Mustafa Imren*

#### **Abstract**

Several pathogens and pests damage tomato plants, and only one and/or more pathogens and pests can coexist in the same plant at the same time. As several numerous pathogens are found in the same plant, the damage to the tomato plants is higher. Pathogens such as nematodes, viruses, viroids, bacteria, and insects adversely affect the growth and development of tomato plants. They may infect roots or upper part of the plant and can cause not only slow down the growth of plants, but also crop losses and their death. Damaging of plant caused by pathogens and pests reduces the market value of plant products. Those pathogens and pests are also called biotic stress agents. The damage, mode of infection, and the mechanism of infection in each tomato plant and pathogens might be different. This situation is crucially important to understand plant pathogen relationship in detail in terms of controlling pests and pathogen. The effect of each pest/pathogen on tomato plants during the cultivation, the type of damage, and new developments and perspectives on morphological and molecular aspects in tomato-pathogen interactions will be discussed in this chapter.

**Keywords:** nematode, viroid, bacteria, virus, insects, pathogens, resistance, pest, biotic stress

#### **1. Introduction**

Tomato (*Solanum lycopersicum* L.), member of the family Solanaceae, is a cultivated plant with a very large cultivation area in the world. According to 2021 FAO data, the amount of tomatoes produced in the worldwide is 187 million tonnes. The highest production amount is in China, followed by Turkey in the third place [1]. Tomato (*S. lycopersicum* L., family Solanaceae) is one of the most produced crops worldwide, and Turkey is placed in top five countries in terms of the production of Solanaceae family [2, 3]. At least 12% of the world's agricultural products are lost every year due to plant

diseases caused by some pathogenic microorganisms and 20% due to some insect pests. Disease factors, pest organisms, and weeds in agricultural products can cause significant economic losses and damage. If the necessary controls against these factors are not made, crop losses can reach from 35% to 100%. The 60–75% of the diseases observed in plants are caused by fungal and bacterial diseases, 10–15% by viral disease (virus and viroids), and 10% by other pathogens and some environmental stress factors [4].

Viruses are commonly encountered in the living ecosystem. Since it does not have a complete cellular structure, it interacts with prokaryotic and eukaryotic organisms and maintains its own existence [5, 6]. In recent years, plant viruses and their mechanisms of action have been widely studied due to the loss of agricultural products and their effects on fruit-vegetable quality. Plant viruses have either single-stranded RNA (ssRNA) or double-stranded RNA or DNA nucleotides [7].

Nematodes are one of the most abundant multicellular organisms on the earth. They may live as plant and animal parasites and/or free living. Parasitic nematodes may infect humans, plants, and animals [8]. Among nematodes, about 4100 nematode species have been identified as plant-parasitic nematodes [8]. They cause significant crop losses on tomato plants.

Bacterial, viroid diseases, and insect pests give also significant crop losses affecting tomato production in many regions in the world.

In this chapter, the effect of each pest/pathogen (virus, nematode, viroid, bacteria, and pests) on tomato plants during the cultivation, the type of damage, and new developments and perspectives on morphological and molecular aspects in tomatopathogen interactions are given.

#### **2. Viruses disease**

Tomato viruses are transmitted by vector insects, plant material, and seeds [9]. Transmission of tomato viruses is important to determine the plant material used in the diagnosis, to choose the method of diagnosis, to prevent the spread of the virus, and to develop a method of struggle against the virus. In this part, we examine under two subtitles that some viral diseases, the main host of which is tomato, are transmitted only by plant materials including seeds and are transmitted by vector insects and/or plant material together. In addition, in this section, the general information and classification of viruses, their genetic characteristics, symptoms and damage in tomato plants, and preventing against the viruses have been briefly explained.

#### **2.1 Tomato viruses transmitted by plant parts including seeds**

#### *2.1.1 Tomato brown rugose fruit virus*

Tomato brown rugose fruit virus (ToBRFV) was first reported in tomato in Jordan [10]. ToBRFV belongs the family Virgaviridae and genus *Tobamovirus*, has rod-shaped particles with encapsidating a positive-sense single-stranded RNA (ssRNA) [11, 12]. ToBRFV is basically transmitted by mechanical ways as plant-plant contact, workers, tools, equipment, and irrigation water. The virus is also effectively transmitted by seeds [10]. In addition, bumblebees transmit the virus on tomatoes [13]. The virus has severe symptoms as mosaic blotch, narrowing on leaves and brown rugose, yellowing spots on fruits. Moreover, the virus reduces the quality of the fruit and causes the

#### *Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

fruit to be unmarketable [14]. ToBRFV is detected by enzyme-linked immunosorbent analysis (ELISA), polymerase chain reaction (PCR)-based analysis by specific primers, and genome sequencing, NGS (next-generation sequencing) [10, 14–16].

#### *2.1.2 Pepino mosaic virus*

*Pepino mosaic virus* (PepMV) was originally identified in pepino (*Solanum muricatum*) in Peru, in 1974 [17]. Following pepino, the virus was firstly detected in tomato, in Netherlands [18]. PepMV belongs to the family Flexiviridae and genus *Potexvirus*, has a positive-sense ssRNA genome with non-enveloped, flexible, rod-shaped particles [17]. Although PepMV isolates show a high genomic similarity, they differ from the original source isolate that causes disease in tomato [9]. Observing leaf symptoms are yellow and mosaic spots, scorching, and deformations [9]. The common transmission way of PepMV is mechanical basis such as plant sap, contaminated tools, and surfaces [9]. The virus has been also transmitted by recirculating hydroponic system, bumblebees, and the root-infecting fungus *Olpidium virulentus* between tomato plants [19–21]. In addition, conventional polymerase chain reaction (PCR), quantitative PCR (qPCR) methods as TaqMan assays and restriction fragment length polymorphism (RFLP) are also have been used for detection of virus and identification of different genotypes [19, 22].

#### *2.1.3 Tobacco mosaic virus*

*Tobacco mosaic virus* (TMV) was the first virus detected [23], belongs the family Virgaviridae and genus *Tobamovirus* [24, 25]. TMV has rod-shaped and encapsulating particles with a single-stranded RNA (ssRNA) [26–28]. The first viral protein structure sequenced belongs to TMV [29, 30]. TMV is transmitted by mechanically including workers, tools, and propagating materials [31]. Because the virus has oldest genomic information, it has widespread host plants including tomato [32]. TMV has characteristic symptoms on the leaves such as light and dark green spots and malformation. Moreover, TMV infections have also caused necrotic rings, browning, and number and size reducing on fruits [33]. In addition to the serological analysis method for TMV, numerous molecular detection methods and diagnostic studies have been carried out [34]. In general, virus-free seeds, plantlets, and hygienic measures have to be used to prevent from virus like other tobamoviruses.

#### *2.1.4 Tomato mosaic virus*

*Tomato mosaic virus* (ToMV) belongs the family Virgaviridae and genus *Tobamovirus* [12, 35]. The particles of virus are rod-shaped and encapsulating with a genome single-stranded RNA (ssRNA) [26]. ToMV has high rate of infectivity, effective seed transmission, and mechanic transmission easily by working hands, tools, soil, and plants parts [12, 36]. Like as other tobamoviruses, ToMV causes malformation, spotting and clearing on tomato leaves, and malformation on fruit and reducing the yields [36]. As with other tobamoviruses, virus-free seeds, plantlets, and hygiene measures should generally be used to prevent the virus.

#### *2.1.5 Tomato mottle mosaic virus*

Tomato mottle mosaic virus (ToMMV) was firstly identified in Mexico in 2013, belongs the family Virgaviridae and genus *Tobamovirus*, has four open reading frames (ORFs) including the movement protein (MP) and coat protein (CP) in genome [37]. As other tobamoviruses, ToMMV is inclined to mechanical transmission including contacts, hands, tools, the greenhouse structure, and bumblebees. Moreover, seed transmission is also possible with infected seeds [12]. ToMMV causes the mosaic symptoms, chlorosis, and leaf deformation on tomato plants [38]. The virus can be detected by using polymerase chain reaction (PCR) basis methods [39]. Management of the ToMMV is possible by using virus-free seeds and plantlets and using hygienic measures [40].

#### **3. Plant-parasitic nematodes**

Plant-parasitic nematodes are significant pests and cause crop losses, with an estimated yearly loss of USD 173 billion [41]. It is likely that 10% of world crop production is lost as a result of plant-parasitic nematode damage [42]. Most of the plant-parasitic nematodes feed on roots and decrease the uptake of water and nutrients [43]. Stylets of the plant-parasitic nematodes are important apparatus used to puncture plant cells and uptaking nutrient contents. The main signs shown by plants affected by nematodes are stunted development, wilting, and susceptibility to contamination by other plant pathogens [44]. Although there are many plant-parasitic nematodes, the most vital plant-parasitic nematodes in the USA are *Heterodera glycines*, *Meloidogyne fallax*, *Meloidogyne chitwoodi Globodera pallida*, *Ditylenchus dipsaci*, *Litylenchus crenatae*, *Globodera rostochiensis*, *Meloidogyne enterolobii*, *Pratylenchus fallax* and *Bursaphelenchus xylophilus* [45]. Similarly, *Meloidogyne* spp., *Aphelenchoides besseyi*, *Nacobbus aberrans*, *Pratylenchus* spp., *B. xylophilus*, *Heterodera* and *Globodera* spp, *Xiphinema index*, *Radopholus similis*, *D. dipsaci*, and *Rotylenchulus reniformis* are most important nematodes in terms of plant pathology [46]. Root-knot nematodes: The nematodes belonging to the *Meloidogyne* genus termed root-knot nematodes are polyphagous plant pathogens [47]. They may be found worldwide and parasitize the species of higher plants [47]. Root-knot nematodes, *Meloidogyne* genus, which are obligate plant parasites, are economically important and damage plants. They are found in many parts of the world and have the ability to parasitize any high plants [47]. They disrupt plant physiology and decrease crop quality and yield [9, 48]. Root-knot nematodes have 106 species [47]. *M. hapla*, *M. incognita*, *M. arenaria,* and *M. javanica* are major species; however, *M. fallax*, *M. minor*, *M. chitwoodi*, *M. exigua*, *M. paranaensi,* and *M. enterolobii (=M. mayaguensis*) are minor root-knot nematode species [41].

The genus of *Meloidogyne* compromises more than 100 species in the world [46]. Root-knot nematodes are named because of their characteristic features, as they typically cause root galls. While young plants may not survive high infection by a nematode, mature plants often show low yield and growth retardation. Among the root-knot nematodes, *M. graminicola* may cause damage to cereals in South Africa, the USA, Australia, and Mexico [44]. *M. arenaria*, *M. incognita*, and *M. javanica* are good hosts of some cereal cultivars such as rye, barley, oat, and wheat under greenhouse conditions [49]. *M. hapla* is distributed in temperate regions, and yield losses caused by some root-knot nematode species are valued at approximately \$10 billion [50]. Root-knot nematodes cause damage and induce a unique feeding site structure termed giant cells within the plant roots. Cell wall molecular architecture of nematode feeding site is changed [51]. *M. javanica, M. arenaria, M. graminicola, M. incognita*, and *M. hapla* are some of the most damaging species; some species cause more damage to their host than other species. For instance, *M. graminicola* is one of the main

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

#### **Figure 1.**

*The root-knot nematode, M. incognita, induced root galls in tomato plants (left) and control-uninfected healthy tomato plant roots (right). The nematode cause galls in tomato roots (right).*

problems in rice fields that develop special hook-like knots on the roots of rice plant roots [52]. Root-knot nematodes induce feeding cells and become sedentary within approximately 48 hours after nematode infection [53]. The second stage juveniles of root-knot nematodes can infect the plant roots. More than one species of root-knot nematodes in the same plant tissues can be found. The nematode causes galls in the root system (**Figure 1**), disrupts the vascular tissues, and restricts the exchange of water and nutrients. Growth slows down, wilting, stunting, and yellowing of leaves are seen. During a severe infection, the plant may completely dry out. The secondary damage of root-knot nematodes is that soil-borne pathogens may enter nematodeinduced wounds in plants [54].

#### **4. Tomato pests and their control**

#### **4.1** *Tuta absoluta* **(Meyrick) (Lepidoptera: Gelechiidae)**

Tuta absoluta is the main pest in open field and greenhouse tomato cultivation. Adult butterflies are active at night. They lay their eggs, usually under the leaves, in the lower part of the sepals of buds and immature fruits. Its larvae damage all parts of the tomato plant except the root and in each period. The larva feeds by opening

galleries between the two epidermes on the leaves of the tomato. The plant may dry out completely due to the galleries opened in the green part of the plant. The pest enters under the sepals of immature tomato fruits. The damaged fruit loses its market value, and rots occur when secondary microorganisms settle in the galleries opened in the fruit [55]. As a biotechnical method, pheromone + water trap or pheromone + light + water trap can be used in greenhouse tomato cultivation for mass trapping against tomato moth [55].

#### **4.2** *Bemisia tabaci* **(Genn.) and** *Trialeurodes vaporariorum* **(Westw.) (Hemiptera: Aleyrodidae)**

The damage of these pests is important in tomatoes, cucumbers, peppers, beans, and eggplant [56]. Whitefly adults use the underside of leaves for feeding, laying eggs and resting. Larvae and adults feed by sucking plant sap. As a result of suction, yellowing occurs in the form of spots on the leaf. In addition, the pest secretes a sweet substance during feeding, with the development of fumagine fungi on this substance, a black layer forms on the leaves, and these parts cannot assimilate. For this reason, the plant weakens, plant growth is adversely affected, yield and quality decrease. Whiteflies give an average of 9–10 offspring per year, depending on the temperature, and a female lays an average of 200-300 eggs. Whitefly adults also play an important role in the transmission of some viral diseases. Especially *Tomato yellow leaf curl virus* (TYLCV) is carried by Tobacco whitefly [55].

#### **4.3** *Liriomyza trifolii* **(Burgess),** *L. bryoniae* **(Kalt.),** *L. huidobrensis* **(Blanchard) (Dip.: Agromyzidae)]**

Especially tomato, cucumber, and beans are among the important hosts of leaf fly, which is a polyphagous pest. Adults and larvae of the pest cause damage to the plant. Adults lay their eggs between the two epidermes of the leaf [55]. Larvae emerging from the egg feed on the parenchyma tissue between the two epidermes in the leaf, and as a result, galleries are formed. In the following periods, these areas turn yellow, dry, and fall off. It indirectly causes loss of product and value by delaying development in young seedlings and plants [55]. A female can lay about 400 eggs in her lifetime at 30°C. It can give about 10 offspring under greenhouse conditions. In order to obtain healthy plants in the cultural struggle, precautions should be taken against pests, especially during the seedling period, For this purpose, ventilation openings must be covered with gauze. Weeds around and inside the greenhouse must be destroyed. Contaminated plant residues must be destroyed. The soil must be kept moist and the pupae must rot from moisture by mulching, and larvae should be prevented from becoming pupae by passing into the soil. Entry-exit and ventilation openings in greenhouses should be covered with gauze or fine-hole wire to prevent the entry of adults. Yellow sticky traps are used in biotechnical control since planting seedlings. One of the most important parasitoids is Diglyphus isaea Walker (Hym.: Eulophidae). In case of 10 larvae per leaf in tomato, chemical control is decided [55].

#### **4.4 Aphids [***Myzus persicae* **(Sulz.),** *Aphis gossypii* **Glov.,** *A. fabae* **Scop.,**  *Macrosiphum euphorbiae* **(Thomas) (Hem.: Aphididae)]**

Aphids are particularly damaging to tomatoes, peppers, eggplants, cucumbers, and zucchini. Aphids cause damage by sucking plant sap. Due to the suction, the

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

leaves take a shrivelled, curled appearance. As a result of this suction, the plant weakens, development stops, the yield and quality of the product deteriorate. The sweet substances they secrete cover the plant surface by causing fumagine, and damage occurs as a result of the plant's obstruction to assimilation and respiration. It is also the vector of viral diseases. It is known that only *M. persicae* is the vector of 50 different viruses [55]. Contaminated plants and weeds should be cleaned from inside the greenhouse. Among the predators, especially the species belonging to the Coccinellidae, Chrysopidae, and Syrphidae families and the parasitoids *Aphidius* species are very important in terms of biological control. For chemical control against Aphids in tomato, it is decided to apply if 20 individuals are seen per leaf [55].

#### **4.5** *Tetranychus urticae* **Koch. (Acarina: Tetranychidae)**

As a polyphagous pest, *T. urticae* is particularly damaging to tomatoes, beans, cucumbers, eggplant, peppers, and zucchini [83]. The females lay their eggs on the underside of the leaves, between the webs they weave along the leaf veins. The larva that emerges from the egg becomes adult by passing the protonymph and deutonymph stages. Larvae change three shirts until they reach adulthood [55]. A female can lay 100–200 eggs. Depending on the climatic conditions and the host, it can produce 10–12 offspring per year in greenhouses [56]. As a cultural precaution in the fight against spider mites, plant residues contaminated with the pest should be removed from the environment. Soil cultivation should be done, and weeds should be combated. In its biological control, especially Phytoseids, Coccinellids, and predatory thrips are the first preferred natural enemies. If five nymphs + adults per leaf are determined in chemical control against spider mites in tomato, the application is decided [55].

#### **4.6 Thrips [***Thrips tabaci* **Lind.,** *Frankliniella occidentalis* **Pergande. (Thys.: Thripidae)]**

Thrips particularly give damage to tomatoes, cucumbers, peppers, eggplants, and beans. Adults and larvae injure the epidermis layer of leaves, stems, and fruits of plants and feed by absorbing the sap. The cells in the area where the thripsin is fed die and white silvery spots appear. As a result, the assimilation capacity of the leaves decreases and the leaf edges curl. As a result of feeding on fruit or capsules, silvery spots appear, and deformities occur. *T. tabaci* lay 70–100 eggs during their lifetime. It completes one offspring in an average of 14–30 days. It gives 3–10 offspring per year. *F. occidentalis* lays 150–300 eggs during its lifetime. It gives a maximum of 15 offspring per year. As a cultural precaution, plant residues contaminated with pest should be destroyed. Of the natural enemies, especially *Orius* spp., it is important for biological control. In the chemical control of thrips, if 20 nymphs per leaf or three nymphs + adults (adults-larvae) are determined per flower, the application is decided [55].

#### **5. Tomato bacterial diseases**

#### **5.1 Bacterial canker** *Clavibacter michiganensis* **subsp.** *michiganensis* **(Smith)**

*Clavibacter michiganensis* subsp. *michiganensis* (CMM) is a xylem-inhabiting bacteria [57]. Optimal growth conditions are at 24–38°C and 7 and 8 P. But it found to grow

#### **Figure 2.**

*Vascular color change of tomato plant by Clavibacter michiganensis subsp. michiganensis (CMM). The bacteria inhabit in the xylem. The color of the plant vascular tissues is cream-yellow to brown.*

in plant xylem at pH 5 [57, 58]. The disease is seed-borne, and bacteria may survive in or on the seed coat. Contaminated soil equipment and other materials serve as inoculum sources for short periods. Infected plant materials and soils with infected plant debris are important inoculum sources by providing long life periods of bacteria. After the plant is infected, bacteria invade xylem vessels, and it moves systemically throughout a plant. Disease causes weak and stunted plants. Infected seedlings may be quickly collapsed. Bacterial canker caused vascular (systemic) and parenchymal (superficial) symptoms. The early symptoms are wilting, curling browning, and wilting of the leaves, especially along one side of the plant. Wilting of the lower leaves can be seen toward the flowering stage. The wilting may progress upward of the plant. The wilted parts can dry out in a short time. As a result of the superficial infections, necrotic or slightly raised spots may appear on the surfaces of leaves, on the stems, and on petioles. In infected plant, cream-yellow to brown coloring of the vascular tissues can be seen (**Figure 2**).

#### **5.2 Bacterial pith necrosis**

Bacterial pith necrosis disease is caused by several pathogenic bacteria, *Pseudomonas corrugata* (Scarlett et al.) Roberts and Scarlett, *P. cichorii* (Swingle) Stapp, *P. mediterranea* Catara et al., *P. viridiflava* (Burkholder) Dowson, *P. fluorescens*, *Pseudomonas marginalis* Brown (Stewens), *Dickeya chrysanthemi*, *Pectobacterium carotovorum* subsp. *carotovorum* [59–61]. The disease affects tomato plants (*S. lycopersicum*), especially in greenhouse production. The disease was first described in Britain in 1970 by Scarlett et al. [62]. Disease-causing agents are generally opportunistic bacteria to cause disease when the plant is under stressful conditions. High humidity, high N fertilizer, and low night temperatures encourage rapid plant growth, and the formation of the juicy structure is a disease favorable condition [63]. The major entry place for bacteria is the wounds caused after secondary sprout removal, which is a common practice in staked tomato fields. Disease agents generally survived in seeds, soil, and infected plant debris for 6–8 months [64]. The disease may occur in

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

#### **Figure 3.**

*Bacterial pith necrosis: general wilting and stem necrosis by tomato pith necrosis and stem necrosis and vascular coloring of tomato plants caused by tomato pith necrosis. The brown discoloration is seen.*

the field and covered greenhouse crops, especially during winter in greenhouse crops. The symptoms are similar to the infections caused by the pathogens *P. viridiflava*, *P. corrugata*, *P. mediterranea*, *P. carotovorum*, or *Pectobacterium atrosepticum* [65–67]. Typical symptoms of pith necrosis on tomato plants consisted of general plant wilting, yellowing, and brown to black spots or lesions developing on the stem, petiole, and fruit stalk (**Figure 3**). Internally, pith tissues developed water-soaking, brown discoloration, hollowing, and soft rotting. In some cases, browning also occurs in the vascular tissues (**Figure 3**).

#### **5.3 Bacterial speck disease** *Pseudomonas syringae* **pv.** *tomato* **(Okabe) Young, Dye, Wilkie**

Bacterial speck of tomato is a serious problem in many greenhouse and field production areas. Disease can occur at every growing stage of tomato, but it causes severe infections at cool, moist conditions. The optimal growth temperature of the bacteria is 24–30°C Disease development stops in hot weather conditions. The disease is ubiquitous [68], Bacteria can survive epiphytically on weed hosts [69]. Bacteria can maintain the viability for 1–2 years as saprophytically on diseased plant residues in the soil [70].

The disease is seed-borne. Infection may begin with soil with contaminated seeds or plant debris. Secondary contamination occurs from wounds or natural openings. Water droplets play an important role in the spread of the disease. During the seedling period, brown-black spots sometimes surrounded by chlorotic margin are seen on the leaves and stems of the seedlings, and sometimes these spots spread and cause drying of the seedling. The spots on the leaves are small, round, dark in color, and unlimited. A yellow halo is usually seen around these spots, which are 1–3 mm in diameter. The spots coalesce over time and form large necrotic areas that lead to deformation and drying of the leaf. Superficial large brown spots are seen on the main stem and branches, leaves, and flower stalks (**Figure 4**) [71].

#### **Figure 4.**

*The symptoms of bacterial speck disease P. syringae pv. Tomato (Pst). Large spots on tomato stems (left), flower spots (middle), spots on fruit stalks and fruits (right) by Pst.*

#### **5.4 Bacterial spot of tomato** *Xanthomonas vesicatoria* **Vauterin et al.,** *Xanthomonas euvesicatoria* **(Jones et al.);** *Xanthomonas perforans* **(Jones et al.).**

Bacterial spot of tomato is a worldwide disease. *X. vesicatoria* Vauterin et al., *X. euvesicatoria X. perforans* have been identified to cause bacterial spot disease on tomato. The disease was firstly discovered in South Africa in 1914 [72]. High relative humidity and overhead irrigation are optimal conditions for disease development. The optimum growth temperature of these bacteria is 29°C. 20–35°C temperature conditions promote disease development, while night temperatures lower than 16°C suppress disease development. Infected seeds may serve as a major inoculum source. The agent can survive on or in the seed for a year or more. *Xanthomonads* may also survive epiphytically in the tomato phyllosphere. Under favorable conditions, epiphytic populations can cause severe infections or outbreaks, especially in transplants [73]. Tomato bacterial spot caused necrotic lesions on the leaves, stems, petals, and flowers, and fruit [74]. Circular water-soaked lesions appear on seedlings (**Figure 5**). They later dry and turn dark brown to black [75]. Sometimes, halos are present around the spots. Primary lesions coalesce, resulting in extensive necrosis and a blighted appearance (**Figure 5**).

#### **5.5 Bacterial wilt of tomato** *Ralstonia solanacearum*

Bacterial wilt (BW) is the most important disease affecting tomato production in many regions [76]. It causes severe wilting of economically important crops such as tomato, potato, eggplant, chili, and non-solanaceous crops such as banana and groundnut. *R. solanacearum* is an aerobic obligate organism. It was classified as four races and five biovars. Race 1 has a very wide host range mainly flowering crops. Race 2 attacks bananas, race 3 has worldwide effects on tomatoes, potatoes, and other *Solanaceae* plants, and race 4 infects ginger [77]. *R. solanacearum* can survive on weeds and alternative non-host plants epiphytically. Infected soil and crop residues may serve as important inoculum sources [78]. The pathogen is carried in tomato seeds [79].

Initial symptom of bacteria in tomato is wilting of upper leaves (**Figure 6**). Complete wilting of the plants is observed in a short time. Brown discoloration of the infected vascular tissues and visible white or yellowish bacterial ooze can be seen [80]. *Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

#### **Figure 5.**

*The symptoms of bacterial spot of tomato Xanthomonas spp. Water-soaked lesions of the disease on seedlings (left), leaf spots of X. euvesicatoria in greenhouse grown tomatoes (right).N. YILDIZ.*

#### **Figure 6.**

*The symptoms of BW of tomato R. solanacearum. Wilting caused by R. solanacearum is seen on the leaves of tomato plant.*

#### **6. Tomato viroids**

Some viroids are pathogenic, some can continue to multiply asymptomatically in susceptible plant species. Viroids are classified in two families, Avsunviroidae

and Pospiviroidae. It has been reported that there are eight species in the family Pospiviroidae, which cause symptoms to occur intensely in tomatoes, especially in the Solanaceae family [81].

Common symptoms of viroid infection depending on viroid species and variant (species and strain), variety, temperature, and light conditions include chlorosis, tanning, leaf deformation, reduced plant growth, severe yield loss, and non-marketable fruit symptoms in tomato plants [82].

#### **6.1 Potato spindle tuber viroid**

The genus *Pospiviroid* of the family Pospiviroidae; *Potato gothic virus*, *Potato spindle tuber pospiviroid* (PSTVd), *Potato spindle tuber virus*, *Tomato bunchy top viroid* has been named under different names. The PSTVd factor is included in the EPPO A2 list. PSTVd was the first to be identified as a new viroid and is quite different from bacteria and viruses [83]. PSTVd is located in the family Pospiviroidae of the *Pospiviroid* genus [84]. While the main host is potato (*Solanum tuberosum* and other *Solanum* spp.), tomato (*S. lycopersicum*), pepper (*Capsicum* spp.), and other vegetables and ornamental plants and weeds from the Solanaceae family also constitute the host series. Infections in ornamental plants and weeds are generally asymptomatic. It has been determined that many species in the Solanaceae family and a few species in other families can be transmitted experimentally [85].

The type and severity of PSTVd symptoms vary depending on the viroid strain, host species and variety, and environmental conditions. PSTVd infections can be asymptomatic or produce symptoms ranging from mild to severe. PSTVd may cause more severe symptoms at higher temperatures [86]. In tomato, early in infection, infected plants show slow growth and chlorosis in the upper part of the plant, while in advanced stages the growth reduction may become more severe and leaves may turn red and/or purple and become more fragile (**Figure 7**). At this stage, flowering and fruiting may stop. In advanced stages, plants may die or partially recover.

#### **6.2** *Citrus exocortis viroid* **(***Citrus exocortis pospiviroid***) (Indian tomato bunchy top viroid)**

The disease agent was observed for the first time with the symptom of bark scaling on the three-leaf rootstock of citrus fruits, and it was revealed that it was transmitted

#### **Figure 7.**

*Potato spindle tuber pospiviroid (PSTVd) induced plant symptoms on tomato plants. PSTVd symptoms of tomato plant (Money maker cv. (left) and H5656 cv. Standard cultivar (right)). Control plant represents the uninfected plants.*

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

#### **Figure 8.**

*The symptoms of CEVd (Citrus exocortis pospiviroid) (Indian tomato bunchy top viroid). CEVd symptoms of Gynura aurantiaca indicator plant and tomato plant (H5656 cv. Standart cultivar). Control plant represents the uninfected plants.*

#### **Figure 9.**

*The symptoms of PSTVd (A) and CEVd (B) in S. lycopersicum L. (Hünkar cv.) and C. annuum L. (Sunam F1 cv.) plants.*

by the bud [87]. In 1972, this factor was determined to be a viroid [88]. The agent is classified as a *Citrus exocortis viroid* (CEVd) species in the *Pospiviroid* genus of the family Pospiviroidae. CEVd is one of the best characterized viroids today. Exocortis disease is called citrus dwarfing viroid disease in our country. CEVd can cause scaling in the bark tissue of citrus trees, peeling and general stunting of the plant [89, 90]. Decreased growth, stunting may occur, chlorosis in leaves may become more severe, turning into reddening, bruising, and/or necrosis (**Figures 8** and **9**).

#### **6.3** *Columnea latent viroid*

*Columnea latent viroid* (CLVd) agent was first detected in the *Columnea erythrophaea* plant in the US state of Maryland in 1989, and it was stated that the agent was present asymptomatically in this plant [91] but it was later determined that it

#### **Figure 10.**

*The symptoms of CLVd in pepper plants. CLVd (A) and Mexican papita viroid (MPVd) (B) symptoms of S. lycopersicum L. (Hünkar cv.) and C. annuum L. (Sunam F1 cv.) plant.*

produced PSTVd-like symptoms in potatoes and tomatoes [92]. The agent is *Brunfelsia* spp., *Columnea* spp., *Gloxina* spp. and *Nematanthus* species are generally asymptomatic (latent) in ornamental plants [93, 94]. Both PSTVd and MPVd were found naturally in wild Solanum species [95]. In tomatoes, CLVd can cause general stunting, deterioration of leaf structure, formation of thin-stemmed plants, tanning of leaves, chlorosis and leaf epinasticity, as well as necrosis of leaves, stems, and petioles (**Figure 10A**).

#### **6.4** *Mexican papita viroid*

The MPVd agent was first identified in 1996 in the plant *Solanum cardiophyllum*, a wild solanum species in Mexico [95]. The symptom caused by MPVd in plants is observed as a general stunting and the formation of chlorotic and purplish spots on the leaves (**Figure 10B**). Depending on the severity of the infection, the fruit size decreases and/or no fruit is formed. There are uncertainties about how the agent is transported. The sequence of MPVd was determined to be very similar to that of TPMVd (93%) and PSTVd [95].

#### **6.5 Tomato apical stunt**

*Tomato apical stunt* (TASVd) causes severe symptoms in tomato plants shortening of the internodes, leaf deformation and yellowing, shrinkage, and less coloration of fruits (**Figure 11A**). TASVd has been reported in the Ivory Coast, Tunisia [96], Senegal [97]. TASVd has also been detected asymptomatically in some ornamental plants (e.g., *Brugmansia*, *Cestrum*, *Solanum jasminoides*, *S. rantonetii*, *Streptosolen jamesonii*). TASVd is transported by seed, by plant sap during mechanical processes (during pruning, etc.). While it is not carried by pests such as *M. persicae* and *B. tabaci*, it is carried with pollen with the help of bumblebees during pollination. There is insufficient data on the geographical distribution, host range and epidemiology of TASVd, and control of viroids is difficult in practice [98].

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

#### **Figure 11.**

*The symptoms of TASVd in plants. TASVd (A) and ToCDVd (B) symptoms of S. lycopersicum L. (Hünkar cv.) and C. annuum L. (Sunam F1 cv.) plant.*

#### **6.6 Tomato chlorotic dwarf viroid**

Tomato chlorotic dwarf viroid (TCDVd) agent was first detected in 1996 in a tomato greenhouse in Manitoba, Canada [99]. As the hosts of the agent; *Brugmansia* spp. and hybrids, *Petunia* spp., *Solanum melongena*, *Verbena* spp., and *Vinca minor* plants have been reported. The agent has been found in Arizona and Hawaii [100, 101], India [102], Slovenia [103]. It has caused disease in tomatoes grown in greenhouses in [104]. General stunting, curling of leaves, chlorosis that may turn bronze or purple in later periods (**Figure 11B**), necrosis in petioles and veins, leaf epinasticity, apical bunching, small It causes losses in total yield with the appearance of cracked fruit formations [105].

#### **6.7** *Tomato planta macho viroid*

*Tomato planta macho viroid* (TPMVd) agent was first detected in the tomato state of Morelos, Mexico, in 1982 [106]. Seven species in the Solanaceae family have been reported as natural hosts of TPMVd to date. Since the fruits of the infected plants are in the size of balls and they are completely unmarketable, great commercial losses have been experienced. Although this factor was initially thought to be a viral disease, it was later determined to have a viroid etiology [107–109]. In infected tomato plants, the first symptoms begin 10–15 days after the infection as growth cessation. Chlorosis, epinasty, wrinkling, wrinkling are seen on the leaves and the leaves become brittle. Later, the leaves shrink and turn yellow and stand upright. Although excessive and early fruit formation is seen, the fruits remain small. No seeds are formed in the fruit or fruits with very few seeds are formed. In general, severe stunting is observed in the plant and the fruits may lose their market value. The main symptom occurring within the cells is necrosis caused by the collapse of the phloem [106]. TPMVd affects plant growth (**Figure 12**). It has been reported that the agent can be transmitted mechanically and by the vector *M. persicae*, but there is no conclusive evidence of seed transmission [110].

#### **Figure 12.**

*The effect of TPMVd on plants. TPMVd (A (60 days), B (21 days)) symptoms of S. lycopersicum L. (Hünkar cv.) and C. annuum L. (Sunam F1 cv.) plant.*

#### **7. Plant resistance to pathogens**

Many devastating diseases widely distribute throughout the world in tomatogrowing areas and tomato hosts more than 200 species of pests and pathogens [111]. Bacterial canker caused by seed-borne organism *Clavibacter michiganensis* subsp. michiganensis (CMM) is a destructive disease in both field and protected cultivation of tomato crops. *S. hirsutum*, *S. peruvianum*, *S. pimpinellifolium,* and *S. chilense* are the wild relatives to improve resistance source of *S. lycopersicum* [112–115]. Inheritance of the resistance was controlled by four-gene model [116]. Inheritance of the CMM resistance in wild relatives has been explained by at least four genes [117] and quantitative trait loci (QTL) associated with resistance in interspecific cross [118]. Two major loci Rcm 2.0 and Rcm 5.1 introgressed from LA407 (*S. hirsutum*) have been identified on second and fifth chromosome and explained epistatically 68% of the variation [119].

Whitefly transmitted tomato yellow leaf curl virus (TYLCV Genus *Begomovirus*, Family Geminiviridae) has been threatened to tomato production throughout the temperate regions of the world since 1930s [120]. TYLCV and/or TYLCV-like viruses have many strains and genomic recombinants causing similar symptoms [121]. TYLCV-resistant tomato breeding program was initiated in Israel where first symptoms were observed in the world [122]. TY-20 has been improved as the first hybrid variety resistant to TYLCV from *S. peruvianum* (line M-60) and *S. lycopersicum* (line 10) [123]. Cucumber mosaic virus (CMV) has been divided into subgroups (I and II) and generates stunting, filiform leaves, and necrosis. A single dominant resistance gene Cmr derived from chromosome 12 of *S. chilense* accession (LA458) contributes complete or partial resistance to cultivars [124]. Potato virus Y (PVY) and tobacco etch virus (TEV) are two of main viruses belonging potyviridae transmitted by many species of aphids infect to tomato plants. The recessive gene pot-1 sourced from PI 247087 contributes resistance by single recessive genes both TEV and PVY [125, 126]. ToMV and TMV are named synonymously vice versa. Three dominant resistance genes Tm-1, Tm2, and Tm22 are used to improve resistant varieties derived from PI 235673 (*S. lycopersicum*) [127], PI 126926 (*S. peruvianum*) [128], and PI 128650 (*S. peruvianum*) [129], respectively. *S. peruvianum* is the wild relative used as genetic resource for resistance to *Meloidogyne* spp. Resistance is conferred by a single eight

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

Mi-1 to Mi-8 dominant gene located on chromosome 6 and 12, controls *M. incognita*, *M. arenaria*, and *M. javanica* [130]. Resistance sources to *Meloidogyne* spp. are PI128657 (Mi or Mi-1), PI270435-2R2 (Mi-2) PI126443-1MH (Mi-3), LA1708-1 (Mi-4) PI126443-1MH (Mi-5), PI270435-3MH (Mi-6 and Mi-7) PI270435-2R2 (Mi-8). Mi-3, Mi-7 and Mi-8 genes confer resistance to virulent strain *M. incognita* 557R. Nematode resistance is heat-sensitive in tomato. Mi-4, Mi-5, and Mi-6 genes contribute resistance over 30°C. LA2884 (*S. chilense*) line has heat stable resistance [131]. Potato spindle tuber viroid (PSTVd), tomato chlorotic dwarf viroid, citrus exocortis viroid, Columnea latent viroid, TASVd, tomato planta macho viroid (including Mexican papita viroid), and pepper chat fruit viroid have been identified as causal agents of pospiviroids in tomato. Ther is no commercial variety resisting to pospiviroids [132]. Potato spindle tuber viroid (PSTVd) causes yield loss, plant stunting, leaf chlorosis, smaller fruits. It is one of the most prevalent viroid species attacked to tomato plants. Four accessions belonging *S. chilense* and *S. habrochaites* have been reported less than 50% of PSTVd infection [133]. *S. pimpinellifolium* (LA0373, LA0411) and *S. chmielewskii* (LA1028) plants reported highly tolerant to PSTVd [134].

#### **8. Conclusions**

Plant-pathogens and pests are significantly important and cause an immense amount of crop losses worldwide. Plant-parasitic nematodes, insects, bacteria, viroid, and viruses damage crops at a high rate. Some groups of those diseases and pests parasitize the specific host plant, while others are polyphagous. Identification of plant-parasitic nematodes, insects, bacteria, viroid, and viruses and determination of the parasitism mode of action are important in terms of controlling pests and disease. Plant pathogens and pests show very different symptoms in plants, for example, root knot nematodes cause galls, bacteria cause color changes in plant stems and roots, viruses and viroids cause color changes and deformities in plants. The species of some insects that cause not only their own damage, but also secondary damages due to the fact that some of them carry viruses (for instance *M. persicae* is the vector of numerous viruses). Therefore, in order to grow disease-free plants, it has to be protected of healthy plants from plan-pathogens and pests. In controlling diseases and pests, it is important to have a deep understanding of the host-parasite interactions using cutting-edge technology and techniques. It is also crucially significant for future studies to fully understand host parasite interactions at morphological, molecular, and genetics level.

#### **Conflict of interest**

The authors declare no conflict of interest.

### **Author details**

Refik Bozbuga1 \*, Songul Yalcin Ates2†, Pakize Gok Guler3†, Hatice Nilufer Yildiz3†, Pınar Aridici Kara3†, Bekir Bulent Arpaci4† and Mustafa Imren5†

1 Faculty of Agriculture, Department of Plant Protection, Eskisehir Osmangazi University, Eskisehir, Turkey

2 İzmir Directorate of Agricultural Quarantine, İzmir, Turkey

3 Biological Control Research Institute, Yuregir, Adana, Turkey

4 Faculty of Agriculture, Department of Horticulture, Cukurova University, Saricam, Adana, Turkey

5 Faculty of Agriculture, Department of Plant Protection, Abant Izzet Baysal University, Bolu, Turkey

\*Address all correspondence to: refik.bozbuga@ogu.edu.tr

† Authors contributed equally to this work. Plant parasitic nematodes (by R Bozbuga and M Imren), insects (by PA Kara), viroid diseases (by PG Guler), bacterial diseases (by HN Yildiz), plant resistance to pathogens (by BB Arpaci), virus diseases (by SY Ates) in tomato plants are written in this book chapter.

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Host-Pathogen and Pest Interactions: Virus, Nematode, Viroid, Bacteria, and Pests in Tomato… DOI: http://dx.doi.org/10.5772/intechopen.106064*

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### **Chapter 7**

## Viral Diseases of Tomato – Origins, Impact, and Future Prospects with a Focus on Tomato Spotted Wilt Virus and Tomato Yellow Leaf Curl Virus

*Stephen F. Hanson*

#### **Abstract**

Tomatoes are affected by a number of viruses, with tomato spotted wilt virus (TSWV) and tomato yellow leaf curl virus (TYLCV) being two of the most damaging. TSWV and TYLCV have severely impacted tomato production worldwide for the past several decades at levels that led to both of these viruses being included in the list of top ten most important plant viruses. While they were first described in the early 1900s, both of these viruses emerged in the 1980s to become the severe and persistent problems they are today. The emergence of both viruses was facilitated in part by the emergence and expansion of more efficient insect vectors. Natural sources of resistance, especially from wild relatives of tomato, have provided some measure of control for both viruses to date. This chapter summarizes the origins, emergence, and impacts of these viruses, along with current approaches and future prospects for control, including both natural and engineered resistance.

**Keywords:** tomato spotted wilt virus, TSWV, tomato yellow leaf curl virus, TYLCV, RNAi, SIGS, spray-induced gene silencing, RNA interference

#### **1. Introduction**

Tomato (*Solanum lycopersicum*) is a member of the Solanacea family of plants that also includes potato, chili and bell peppers, and eggplant. Tomato is a ubiquitous crop produced worldwide for a variety of uses ranging from high value fresh fruit to use in a variety of processed products including ketchup, pastes, soups and stews, and canned pasta sauces. Tomatoes are grown under a variety of conditions including open fields, plastic or green houses, screenhouses, and indoor growth rooms.

Tomatoes are one of the most important vegetable crops in the world, valued for both their flavor and nutritional qualities including being rich in vitamins A and C as well as minerals like calcium, potassium, and phosphorus. According to FAO statistics, tomatoes are the most widely produced vegetable, with production levels

of ~170 million tons annually and accounting for ~16% of all vegetable production worldwide [1] coming from ~5 million cultivated hectares. Tomato production has been steadily increasing over recent decades, with China, the US, and India being the largest producers.

Tomato was likely domesticated by indigenous peoples in Mexico and became an important vegetable crop in Central America prior to the arrival of Europeans. Tomatoes were first introduced to Europe by conquistadors returning from the Americas then spread across Europe and the Spanish empire. Tomatoes spread quickly around the globe and even reached China during the 16th century [2].

Tomatoes are affected by many diseases, like all domesticated crops that have been extensively bred and grown in high-density monoculture. Diseases affecting tomato include those caused by bacteria, fungi, viruses, and nematodes. Viruses cause some of the most consistent and severe losses of tomato crops (reviewed in [3–5]). This chapter will focus on two viruses that have caused serious problems in tomato production for several decades, tomato spotted wilt virus (TSWV) and tomato yellow leaf curl (TYLCV). Both of these viruses were included in the top ten most damaging plant viruses, with TSWV and TYLCV occupying the second and third spots on the list, respectively [6].

TSWV and TYLCV provide interesting contrasts on a number of levels including genome structure (RNA for TSWV and DNA for TYLCV), the origin of TSWV appearing to have been disseminated around the globe along with tomatoes and/ or peppers, while TYLCV has emerged more recently and its spread has been partly facilitated by humans; TSWV has an extremely broad host range that includes plants and animals, while the host range of TYLCV is much more limited. These two viruses also share some common themes including the role of insect transmission in their emergence as leading pathogens, the strong potential for natural resistance to play a role in controlling damage, and the potential for biotech/genetic engineering solutions to reduce damage caused by these viruses. This chapter will examine some of the commonalities and differences between TSWV and TYLCV as well as current and potential future prospects for control of these highly damaging pathogens.

#### **2. Tomato spotted wilt virus background**

TSWV causes severe losses in tomato and many other crops worldwide. Symptoms of TSWV in tomato include spotting, often ring spots, and uneven ripening that renders the fruit unmarketable, along with bronzing and wilting of vegetative tissue (**Figure 1**). The first known report of spotted wilt disease on tomatoes was in 1915 in Australia [7]. This spotted wilt disease was shown to be thrips transmitted in 1927 [8] and attributed to a virus in 1930 [9]. TSWV was subsequently reported in various regions around the glove, including Hawaii and Europe, where it appeared sporadically for several decades until emerging as a more regular and profound problem in the 1980s. Since that time, TSWV has become one of the most damaging plant viruses in the world, being cited for regularly causing over \$1 billion in annual crop losses worldwide since the mid-1990s [10] and being recognized as the second most damaging plant virus in the world [6].

TSWV is a member of the Tospovirus genus within the family Bunyaviridae. TSWV virions are pleomorphic pseudo-spherical, with a diameter ranging from ~70 to 120 nm, and are enveloped in a host-derived membrane [11]. The RNA genome segments inside the envelope are encapsidated in N protein [12]. The virions also

*Viral Diseases of Tomato – Origins, Impact, and Future Prospects with a Focus on Tomato… DOI: http://dx.doi.org/10.5772/intechopen.108608*

#### **Figure 1.**

*Tomato spotted wilt on tomato and chili pepper fruit. Typical symptoms of TSWV, including uneven ripening and spotting of fruit on tomato (left) and chili pepper (right).*

contain the L protein, which is the viral RNA-dependent polymerase [13]. TSWV is mechanically transmissible to most plant species it infects, and plants can be infected with either virions or membrane-free ribonucleoprotein complexes that contain the N protein-encapsidated genome segments [14].

Tospoviruses have a tripartite negative sense (or ambi-sense) genome (**Figure 2**). The three genomic RNAs are designated by size as large (L), medium (M), and small (S) RNAs. The L RNA have an entirely negative sense, while the M and S RNAs have ambi-sense and encode genes in both the viral genome sense and viral complement senses [15]. The TSWV genome codes for five proteins overall [16]. The L protein is coded on the viral or negative sense on the L RNA and is the viral RNA-dependent polymerase [13, 17]. The M RNA has ambi-sense and codes for the NSm protein in the genome sense and the polyprotein that is processed into the two structural glycoproteins in the genome complement sense. The non-structural protein NSm has been shown to promote cell to cell and long-distance movement during infection [16, 18]. The glycoproteins were formerly referred to as G1 and G2 but are now denoted as Gn and Gc, indicating their N- or C-terminal location in the precursor polyprotein. The glycoproteins decorate the surface of the virions and are required for thrips transmission [19, 20]. The ambi-sense S RNA codes for the nonstructural protein NSs in the genome sense and the N protein in the genome complement sense. The NSs protein promotes thrips transmission and also functions as a suppressor of RNA silencing

#### **Figure 2.**

*Genome structure of TSWV. Cartoon representation of the tripartite TSWV genome showing the L, M, and S RNAs approximately to scale. Yellow boxes show positions of open reading frames in the genomic (L, NSm, NSs) and genome complement (glycoprotein precursor and N) senses.*

[21, 22], while the N protein is the nucleocapsid that encapsidates viral RNA to form RNPs [23]. The N protein is also required for local spread, suggesting that RNPs may be the functional viral unit involved in local spread [24].

Reverse genetic systems have been a valuable tool that enabled in vitro infection from cloned cDNA and DNA copies of plant virus genomes, mutational analysis of virus genes, evaluation of chimeric viruses, and more. Unfortunately, reverse genetics systems have been unavailable or difficult to develop for viruses with negative or ambi-sense genomes, including TSWV. The recently reported rescue of TSWV from cloned cDNAs is an exciting step forward that will enable reverse genetic analysis of TSWV to TSWV researchers [25].

TSWV has an extremely broad host range and is a rare case of a virus that infects hosts in two different kingdoms as it replicates in both plants and its thrips vector [26]. This observation led to the speculation that TSWV may be a thrips-infecting virus that evolved to also infect plants, which may partially explain its severity as a plant virus. For plants, the host range of TSWV includes over 1000 different plant species in 82 botanical families encompassing both monocotyledonous and dicotyledonous plants [27]. This extremely broad host range likely contributes to TSWV disease persistence since there is a high likelihood that alternate hosts will be present even when susceptible crops are not being grown.

TSWV is transmitted by at least 10 different species of thrips with *Frankliniella occidentalis*, commonly known as the wester flower thrips, being the most efficient vector species [28, 29]. Transmission is circulative and propagative [30, 31]. While adult thrips can acquire TSWV, they are unable to transmit it, and transmission only occurs when thrips acquire TSWV as first- or second-stage larva [29, 32, 33]. While adult thrips can acquire TSWV, they are unable to transmit; thus, the acquisition of TSWV by adult thrips is a dead end for TSWV. Thrips larvae can acquire TSWV with acquisition access periods as short as 15 min although transmission efficiency increases with longer acquisition access periods, and an acquisition period of 4 days was reported to result in 74% of emerging adult thrips being competent for TSWV transmission [34]. Thrips that acquire TSWV remain infected and able to transmit TSWV for life due to the circulative propagative nature of transmission.

TSWV is thought to be acquired by thrips via an animal virus-like receptor-mediated interaction that is rare among plant viruses. The demonstration that a truncated soluble form of the TSWV glycoprotein Gn interferes with thrips transmission of TSWV, presumably by blocking TSWV receptors in the thrips midgut, suggests that the glycoproteins are the viral proteins that mediate virion acquisition [35]. Identification of thrips receptors for TSWV has been an area of interest since it may lead to strategies for blocking thrips transmission of TSWV. While early reports of thrips proteins that interact with TSWV [36] generated some excitement, these initial leads appear to have been dead ends (S. Hanson, unpublished). More recent work has identified different thrips proteins that interact with TSWV virions or glycoproteins and are therefore promising candidates for receptors that mediate TSWV acquisition in thrips [37].

TSWV was described as occurring in many different parts of the world going back to the mid-1900s. This worldwide distribution as a minor pathogen before emergence as one of the most damaging agricultural viruses suggests that TSWV may have spread around the world with host plants like tomato and pepper as they were brought back from meso-America and subsequently spread around the globe. Molecular phylogeny studies that have shown that TSWV often exists as a stable populations in geographically isolated regions and may have spread around the world *Viral Diseases of Tomato – Origins, Impact, and Future Prospects with a Focus on Tomato… DOI: http://dx.doi.org/10.5772/intechopen.108608*

with tomatoes and/or peppers when these plants were introduced to Europe and beyond by Spanish explorers returning from the Americas [38]. The emergence of TSWV as a more widespread and damaging disease started in the 1980s, likely due to the spread of the more efficient western flower thrips vector into areas that were already infested with TSWV.

#### **3. TYLCV background**

Serious outbreaks of tomato yellow leaf curl disease were reported in the late 1920s in the Jordan Valley [39]. Typical symptoms of TYLCD include mosaic chlorosis and stunting of affected plants (**Figure 3**). Since then, numerous outbreaks of TYLCD happened around the Mediterranean in the 1960s. From there, it spread throughout the Middle East to Central Asia, Africa, and the Americas. TYLCV is now considered to be ubiquitous across tropical, subtropical, and temperate regions [40]. During the 1980s, outbreaks of TYLCV became more common and widespread, with some being noted as causing up to 100% loss in affected areas of Italy and the Dominican Republic [41, 42]. All of this led to TYLCV being recognized as one of the most severe viral pathogens of tomato worldwide [43, 44] and to TYLCV being ranked the third most important plant virus in the world [6].

TYLCV is a member of the geminiviridae family, characterized as having single stranded genomes that replicate via a rolling circle type of mechanism and unique twinned icosahedral capsids (reviewed in [45]). There are nine recognized genera within the geminiviridae, and TYLCV is part of the begomovirus genus, which is characterized as being transmitted by whiteflies and infecting dicot plants [46]. The large number of individual viruses within the begomovirus genus has led to several revisions for how groupings are determined and individual viruses are named within this group [47, 48]. The begomovirus genus contains numerous distinct tomatoinfecting members, with the TYLCV subgroup being recognized as one of the most damaging to agriculture [47]. With so many closely related members, the TYLCV subgroup is often treated as a complex of closely related strains that are individually identified by including the location where the strain was recognized in the name, such as for tomato yellow leaf curl sardinia virus denoted as TYLCSV (recent listing in table 1 of [47]). In addition to the large number of strains identified to date, mixed infections that produce recombinant/chimeric variants are believed to happen frequently [49].

#### **Figure 3.**

*TYLCV symptoms. Typical symptoms of TYLCV on tomato, including stunted plants (left) and mosaic chlorosis (right).*

TYLCV was the first member of the begomovirus genus with a monopartite genome, with most begomoviruses having bipartite genomes (**Figure 4**). The genome of TYLCV is ~2.7 Kb and codes for genes in both the viral and complementary senses [50]. The relatively small and simple nature of geminivirus genomes has facilitated extensive reverse genetic analysis via infectious DNA clones that have been obtained for many geminiviruses including TYLCV.

The viral sense codes for two open reading frames (ORFs), with V1 encoding the capsid protein and V2 coding for a multifunctional protein that functions to both facilitate movement and suppress RNA silencing [51, 52]. The genome complementary sense strand encodes four overlapping ORFs that have broad functions in viral replication, transcription, and host interactions. The C1 ORF encodes the replicationassociated protein that contains ATPase and DNA nicking domains [53]. The C1 protein promotes rolling circle replication directly by initiating and terminating rolling circle replication via DNA nicking and ligase activities and indirectly by recruiting host factors involved in viral DNA replication. The C2 ORF codes for a transcriptional activator protein (TrAP) that regulates early and late gene expression. The C3 ORF codes for a replication enhancer protein (Ren). The C4 ORF is involved in symptom development and movement [54]. Like all geminviruses, TYLCV contains a large intergenic region that facilitates bidirectional transcription and contains the origin of replication, including a requisite stem-loop sequence, where rolling circle replication begins and ends.

TYLCV, like all begomoviruses, is transmitted by whiteflies (*Bemisia tabaci*) in a circulative manner (reviewed in [55]). Acquisition and inoculation can both

#### **Figure 4.**

*TYLCV genome. Cartoon representation of the circular ssDNA genome of TYLCV. Open reading frames are shown in yellow. The large intergenic region containing the origin of replication and bidirectional promoters is shown in red.*

*Viral Diseases of Tomato – Origins, Impact, and Future Prospects with a Focus on Tomato… DOI: http://dx.doi.org/10.5772/intechopen.108608*

be as quick as 15 minutes. While several *B. tabaci* biotypes are able to transmit geminiviruses, the emergence and spread of the B biotype that is highly efficient at transmitting geminiviruses played a key role in increasing the spread and severity of geminivirus diseases, including TYLCV, that started in the 1980s.

#### **4. Management of TSWV and TYLCV**

In spite of many differences in virus biology, the factors that lead to the emergence of these viruses and measures for control share a lot of commonalities. A great deal of work has gone into reducing losses caused by TSWV and TYLCV over the past few decades, with some promising advances, although much remains to be done as both viruses still cause extensive losses in tomatoes at present. Standard IPM-based practices, especially those that limit insect vectors, are widely used for controlling both TSWV and TYLCV [56–58]. Although these IPM-based approaches can produce modest reductions in disease, they are not able to prevent all diseases. Breeding for disease resistance has shown some success for both TSWV and TYLCV, and thus, resistance breeding programs are likely to continue as a focus into the future. While not broadly adopted at present, genetic engineering (GE) has shown great potential for controlling both TSWV and TYLCV. The high cost of developing GE lines, extensive regulatory requirements, and concerns about consumer acceptance of GE crops have severely limited the adoption of these approaches for control of diseases in agricultural crops (reviewed in [59]). Thus, GE-based approaches hold great promise for controlling diseases if GE crops become more widely accepted for use in agriculture.

Since the emergence and expansion of more efficient vector species was a major driver in increasing damage caused by these viruses, a number of approaches, especially integrated disease management approaches, have focused on reducing populations of insect vectors or managing production aspects like time of planting to reduce exposure of plants to viruliferous insect populations [58, 60]. Strategies based on insect vector control remain challenging for several reasons, including the lack of effective insecticides, the rapid evolution of insecticide resistance, the fact that both thrips and whiteflies are successful on a number of alternate hosts, and very quick transmission when viruliferous insects enter agricultural fields.

#### **5. Genetic resistance**

Natural resistance has been a highly successful and long-relied-upon strategy for controlling many plant pathogens. Often, wild relatives are found to contain sources of resistance that can be introgressed back into domesticated lines where resistance has been lost.

Several resistance genes have been described for TSWV. These include the single dominant R genes like the Tsw gene from *Capsicum chinense* and the Sw-5 from *Lycopersicon peruvianum* that have provided commercially useful resistance to TSWV resistance in tomato [61–63]. Both of these genes confer a typical hypersensitive response (HR)-based resistance that usually prevents systemic infection by stopping pathogens at the site of inoculation [64]. Molecular studies on TSWV strains with re-assorted genomes were used to determine that the NSm gene is the avirulence determinant recognized by the Sw-5 gene [65].

Natural resistance genes have also been described for several geminiviruses, with many of the resistance genes coming from non-domesticated relatives (reviewed in [66]). This is especially true for TYLCV [66]. The tomato relative *Solanum chilense* is noted as the most common source of TYLCV resistance genes identified to date [66]. At least twelve different sources of resistance to TYLCV were described as of 2020 (summarized in [67]). The Ty-2 gene appears to be a canonical R gene with typical nucleotide binding (NB) and leucine-rich repeat (LRR) regions [68], while others are clearly not classical R genes, but are rather genes involved in RNA metabolism, basic metabolism, cell status sensing, or signaling. The Ty-1 and Ty-3 resistance genes appear to be alleles of a gene [69] that encodes for RNA-dependent polymerase and cause increased cytosine methylation in replicated genomes [70]. Members of the WRKY group III transcription factors have been shown to play a role in TYLCV defense signaling [71]. Still other genes involve in hexose transport or other metabolic processes [72].

Unfortunately, single dominant R genes tend to have limited durability and are often overcome as pathogens evolve to escape the resistance. This is the case for many of the resistance genes described above. Resistance breaking strains of TSWV that overcome the Sw-5 genes have emerged several times independently in different areas including Europe, the US, and Australia [73–75]. Multiple independent cases of resistance breaking TSWV variants have also been reported for the Tsw genes [61, 76]. Resistance breaking has also been observed for several of the described TYLCV genes. Ty-2 mediated resistance was reported to be overcome by TYLCV-Sardinia [77] and an isolate of the mild strain of TYLCV [78]. The Ty-1 gene has been shown to be overcome occasionally under high disease pressure [79].

The generation of resistance breaking strains does not mean that R genes are not useful for control of TSWV and TYLCV. On the contrary, genetic resistance has proven to be one of the most effective tools for limiting TSWV and TYLCV losses to date. And the generation of resistance breaking strains is both typical and expected for any single dominant R gene against any evolving pathogen. For R genes to provide long-term utility, they need to be cycled through, with tomorrow's R genes being discovered while today's are in use. Fortunately, wild relatives of tomato appear to be a robust source for the discovery of new R genes that may be able to supply novel sources of genetic resistance to these viruses well into the future. This is evidenced by one recent study that has evaluated ~700 accessions derived from 13 wild tomato species, where ~140 of the lines were symptom free after inoculation with TYLCV [66]. Based on this, it is likely that wild species will continue to be a robust source of natural resistance genes that will help in reducing TSWV- and TYLCV-caused losses for the foreseeable future.

It should also be noted that while R genes are the most common form of resistance gene found historically, single dominant R genes are not the only type of genetic resistance to pathogens. There are several examples of multigenic resistance and tolerance that provide long-term stable reductions in pathogen losses. One current example is a multigenic field resistance that appears to be providing long-term durable control of TSWV in peanuts [80]. Sequence-level population analysis of multiple TSWV genes did not detect any resistance-related selection in TSWV populations, indicating that this multigenic resistance is likely to be durable. While this resistance appears to be based on high-level tolerance, it provides commercially useful control of TSWV in peanuts, a crop that suffered serious losses from TSWV prior to deployment of this

resistance. Future work using marker-assisted breeding and similar approaches may be useful for developing tomato lines with similar multigenic resistance to TSWV and TYLCV in the future.

#### **6. Engineered resistance**

Genetic engineering is an approach that has proven useful for developing resistance to many plant pathogens including many plant viruses. This is true for TSWV and TYLCV, where numerous approaches for creating engineered resistance have been reported over the past several decades. While several approaches have been described, gene silencing/RNAi approaches (reviewed in [81]) are the most common. Despite promising research results, genetically engineered virus resistance has not been widely adopted due to several barriers, including the high costs for the development of commercial lines approved for human consumption and public resistance to GMO crops (reviewed in [59]).

The first description of engineered resistance to TSWV was described in 1991 [82]. Since that time, several additional examples of engineered resistance to TSWV have been reported, including the use of chimeric RNAi-inducing genes that confer broad spectrum tospovirus resistance [83, 84]. Despite these promising results, engineered resistance to TSWV has yet to be deployed in commercial crops.

Numerous examples of engineered resistance have also been described for geminiviruses in general and TYLCV in particular (reviewed in [85]). Similar to TSWV, the first reports of engineered geminivirus resistance also date back to the 1990s, with many of these attempts using virus-derived resistance targeting the viral genes involved in replication, movement, or encapsidation [86–88]. Examples also include numerous descriptions of anti-sense RNA- and RNAi-based resistance. There are also some interesting examples of non-pathogen-derived resistance, including the use of peptide aptamers that interfere with the function of geminivirus replication-associated proteins that were found to confer high-level tolerance to several diverse begomoviruses, including TYLCV and tomato mottle virus [89]. Still other approaches have targeted host functions like those involved in modulating host defenses [90, 91]. Approaches that modulate host resistance responses have also shown promising results.

Geminiviruses are one rare example where engineered resistance has been approved and deployed in crops produced for human consumption [92]. In this case, common beans engineered to express an RNAi construct targeting the Rep gene of bean golden mosaic virus (BGMV) proved to be highly resistant to begomoviruses affecting bean production in Brazil [93]. The lack of natural resistance sources for BGMV, in spite of decades of screening, made engineered resistance an attractive alternative for BGMV. Extensive multi-year field testing showed that this gene effectively protected common beans from BGMV-caused losses, which had previously reduced yields by 40–100% [94]. So far, this resistance is only approved for use in Brazil. The effectiveness of this approach for controlling BGMV-caused losses, and similar levels of conservation among the Rep genes of TYLCV isolates, suggests that this approach has strong potential for controlling TYLCV-caused losses. While genetic engineering holds great promise for controlling TYLCV, the substantial barriers associated with development costs, regulatory approval, and consumer acceptance must still be overcome before engineered resistance approaches can be broadly utilized.

The time, cost, and consumer acceptance barriers to deploying genetically engineered resistance in crop plants intended for human consumption have spurred innovation aimed at producing similar resistance mechanisms without using transgenic plants. Promising approaches in this area include the use of exogenous doublestranded RNAs that are sprayed on plants to induce an RNAi response in a process referred to as spray-induced gene silencing (SIGS; [95]). SIGS has shown promise against several viral pathogens including TSWV [96]. Another similar approach uses endophytic bacteria engineered to express dsRNAs that can induce and RNAi response in plants. This bacterial-mediated RNAi, sometimes referred to as transkingdom RNAi, has shown promise in reducing infection by fungal and viral plant pathogens [97]. It will be interesting to see if SIGS or transkingdom RNAi evolve into useful technologies that provide control of plant pathogens while successfully skirting the barriers that have prevented more widespread adoption of genetically engineered approaches for control of plant pathogens like TSWV and TYLCV.

#### **7. Summary**

Tomatoes are the most widely produced vegetable on earth, and viruses have been a persistent problem in tomato production for as long as tomato has been cultivated as a crop. TSWV and TYLCV have been serious yield-limiting constraints on tomato production for the past several decades. Tried and true practices like traditional resistance breeding and integrated disease management have allowed continued production of tomatoes in spite of the severe losses these viruses can cause. It is likely that both of these viruses will be better controlled in the future based on the rich body of knowledge developed to date for these viruses. In particular, the abundance of natural resistance sources that are known to be present in wild relatives will continue to be a valuable source of natural resistance genes. Biotech is also likely to play a bigger role in the future on several levels. Marker-assisted breeding and other related approaches will speed introgression of natural resistance resources into commercial cultivars. And if (or when) the cost and societal acceptance barriers are reduced, approaches like engineered resistance and technologies like SIGS are certain to reduce virus caused losses.

#### **Author details**

Stephen F. Hanson Department of Entomology, Plant Pathology, and Weed Science, New Mexico State University, New Mexico, USA

\*Address all correspondence to: shanon@nmsu.edu

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Viral Diseases of Tomato – Origins, Impact, and Future Prospects with a Focus on Tomato… DOI: http://dx.doi.org/10.5772/intechopen.108608*

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#### **Chapter 8**

## Management of Branched Broomrape in Field Processing Tomato Crop

*Francesco Lops, Laura Frabboni, Antonia Carlucci, Annalisa Tarantino, Maria Luisa Raimondo and Grazia Disciglio*

#### **Abstract**

In recent years, there has been a considerable increase in land area used for tomato (*Lycopersicon esculentum* Mill.) in many countries around the world. The essential role is played by Italy at a worldwide level as the country with the third biggest production of tomatoes for processing. *Phelipanche ramosa* (L) Pomel, commonly known as branched broomrape, is a root holoparasitic weed for many crops, particularly for the processing tomato. Due to its physical and metabolic overlap with the crop, its underground parasitism, and hardly destructible seed bank, the control of this parasite in the field is difficult. Results of research studies, many of them on environmentalfriendly methods such as preventive, agronomic, and biological carried out in southern Italy, are discussed and summarized. The results can constitute a relevant basis for further experimental studies.

**Keywords:** orobanche, *Phelipanche ramosa*, control methods, processing tomato crop, cultural practices

#### **1. Introduction**

Tomato (*Lycopersicon esculentum* Mill.) is the vegetable crop with the highest demand and the greatest economic value in the world. Tomato trade and production have particular importance in tropical, subtropical, and mild regions of the world, for both fresh and processing markets [1]. In recent years, there has been a considerable increase in the world land area used for tomato production. The essential role is played by Italy at a worldwide level as the country with the third biggest production of tomatoes for processing after the United States and China. The 2021 tomato processing campaign in Italy closed with a production of just over 6 million tons of processed product, up 17% compared with 2020. Italy's production is 13% of the world's and 53% of Europe [2].

In Italy, as in other areas of the world [3, 4], and especially in the Mediterranean basin, the tomato crop and other species (broccoli, fennel, parsley, celery, and chamomile) are undergoing increased attack of a holoparasitic plant with obligate

**Figure 1.** *A summarized life cycle of a branched broomrape (from Osipitan et al., 2021) [6].*

root belonging to the *Orobanchaceae* family, the *Phelipanche ramosa* (L.) Pomel (syn. *Orobanche ramosa* L.), commonly known as the branched broomrape. Tomato is highly vulnerable also to similar species, as the *Phelipanche aegyptiaca* Pomel (syn. *O. aegypt*iaca) and *O. cernua* Loefl., which are known to cause damage and yield reductions in this crop [5]. The broomrape seeds only germinate in response to specific chemicals (strigolactones) released by the host plant, and the plant spends most of its life cycle underground (**Figure 1**) [7, 8].

Following germination, the seedlings attach to the host roots by the production haustoria that penetrate the host tissues until they reach the vascular system for uptake of water and nutrients, assimilate, and grow at the expense of the host plant's resources [5]. *P. ramosa* attacks tomato roots early in the growing season, within 14–28 days after transplanting (DAT), depending on the temperature conditions, and the shoot usually emerges within 35–56 DAT [9]. Once connected to a host plant, broomrape grows rapidly, forming a tubercle (a storage organ for nutrients and water extracted from the host) underground. Multiple shoots (up to about 20) develop from the tubercle and emerge above the soil surface, and then grow to stalks from 15 cm to 30 cm in height (**Figures 2** and **3**). Flowering begins within 3–7 days after a broomrape shoot emerges above the soil surface. A mature broomrape plant can release more than 500,000 seeds (from 0.2 to 0.4 mm), which can remain dormant and viable for many years (> 20) in soil [5]. The number of emerged shoots per surface unit, and/or number and dry weight of parasitic plants per host plant, can be used as indicator to monitor *Phelipanche infestation* [10].

The air and soil temperature are the main factors that influence the dynamic of host/ parasite interaction and development. Moreover, the optimum temperature for maximum germination of *Orobanche* seeds decreases as the level of their water stress increased [11].

*Management of Branched Broomrape in Field Processing Tomato Crop DOI: http://dx.doi.org/10.5772/intechopen.106057*

**Figure 2.** *Branched* P. ramosa *plant (F. Lops).*

**Figure 3.** P. ramosa *infestation in tomato (F. Lops).*

The presence of the parasite causes a significant reduction in the photosynthetic capacity of tomatoes, as shown by the higher SPAD chlorophyll indices detected on the leaves of infested tomato crop compared with the non-infected one (**Figure 4**).

**Figure 4.**

*Average SPAD values ± SD of parasitized and non-parasitized tomato plants, measured at 53 days after transplanting. Different letters indicate significant differences at P <0.05 according to Tukey's test [12].*

**Figure 5.**

*Relationship between tomato marketable yield and number of emerged branch shoots of P. ramosa detected at the end of tomato cycle (harvesting time) [14].*

This generates a loss of biomass of their aerial organs [13] and a significant decrease in crop yield (**Figure 5**), mesocarp thickness, fruit color, compactness, content of soluble solids, of ashes, and of ascorbic acid [15].

#### **2. Management of** *P. ramosa* **in the field**

Effective control of *P. ramosa* is difficult because, as already mentioned, most of its life cycle occurs below the soil surface. Thus, the effective management of this parasitic weed will require a long-term and an integrated approach. Measures to

*Management of Branched Broomrape in Field Processing Tomato Crop DOI: http://dx.doi.org/10.5772/intechopen.106057*

successfully contain the problems due to *P. ramosa* need to be targeted at: i) reduction of the existing *P. ramosa* seed bank in the soil; ii) prevention of further seed production; and iii) prevention of seed dissemination. These objectives are mutually dependent. Practices to control this parasite include several methods (preventive, chemical, agronomic, and biological), which help to avoid germination, infection, or strong reproduction of the weed [16, 17].

#### **2.1 Prevention methods**

Preventing the movement of parasitic weed from infested into un-infested areas or its spread in recently infested fields is a crucial component of control. Principal measures are to remove the *Orobanche* prior to flower opening; the quarantine for a period of at least 2 years, and in subsequent years only rotational crops may be cultivated (e.g., in California, these crops are those approved by the local agricultural commissioner); clean and disinfect all equipment used in a field with broomrape infestation [6, 17]. As for seed eradication on farm equipment, quaternary ammonium compounds have been found effective in *Phelipanche* and *Orobanche* spp. [18].

#### **2.2 Chemical methods**

Herbicides that currently are in use for parasitic weed broomrape control in various crops are sulfonylurea and imidazolinones. Sulfonylurea herbicides are absorbed through the host plant foliage and roots with rapid acropetal and basipetal translocation. Imidazolinone herbicides are absorbed and translocated through the host to the meristematic tissues. The most successful method to the parasite control in processing tomato is to apply sulfonylurea herbicides, on foliage and by injection through the drip irrigation system in preplanting, or post-emergence, or post-planting [19]. Soil herbigation (saturating the soil with sulfonylureas) effectively controls pre-attached stages of broomrapes [20], but this is hardly compatible with other agricultural cropping practices, as detrimental for many crop seedlings for several weeks or months. Applying sulfosulfuron to the soil three times, at 200, 400, and 600 growing degree days, followed by two applications of imazapic to the tomato foliage late in the season, effective Egyptian broomrape control has been achieved [21, 22]. In the conditions of southern Italy, the best parasite control and tomato yield performances were obtained with sulfonylureas (rimsulfuron and chlorsulfuron) applied through drip irrigation in pretransplant at 25.0 and 5.0 g a.i. ha−1, and in post-transplant at 75.0 and 15.0 g a.i. ha−1, respectively [23].

#### **2.3 Agronomic methods**

In order to integrate the use of chemical methods, there has been an increased effort to research suitable methods (fertilization, soil solarization, long-term rotation, soil management, sowing, or transplanting date) for the control of this parasitic weed, even because there is an increasing market for organically grown tomatoes, where the use of chemical pesticides is not an option [24].

#### *2.3.1 Fertilization*

Broomrape infestations occur mainly in soils poor nitrogen (0.2 and 1.8 ‰) and organic matter (1–2%) such as many soils of southern Italy [25], where the Italian

research studies related in this chapter were carried out. Also, phosphate in deficient soil showed a suppressive effect of *P. ramosa* parasitism [26]. Therefore, soil fertility management can contribute to the management of this parasite. Phosphorous and nitrogen have been described to downregulate strigolactones exudation in some crop species [27–29].

Direct contact with fertilizer, such as urea and ammonium, may be toxic to broomrape, inhibiting seed germination and seedling growth [30]. Urea fertilizer, due to hydrolysis in soil, produces ammonium ion, which probably exerts the toxic effect on the parasite [31].

Nitrogen fertilizer (80 kg ha−1 N) or sulfur (8 t ha−1 S) applied prior to the tomato seedling transplant showed a suppressive effect on the seed germination of *Phelipanche* [32]. Also, the mixtures of chicken manure and sulfur significantly reduced the dry weight of *Orobanche* and increased eggplant and potato yield compared with the control [33].

Organic compounds are widely used in cropping systems to increase soil organic matter, structural stability, water holding and cation exchange capacities, and as a source of nutrients [34].

Recently, in the olive production and/or processing areas, as those of southern Italy, the use of oil mill wastewater (OMW) has been proposed as a suitable method for the containment of *P. ramosa.* In this regard, several trials dealing with the OMW distributed on the heavy infested soils at the dose of 80 m3 ha−1, 40 days prior to tomato seedling transplant (**Figure 6**), and incorporated into the soil later, revealed a significant reduction (between 34 and 76%) of emerged *P. ramosa* plants with respect to the untreated control (**Figure 7**), limiting the additional seed production of this parasite [35]. This could be due to the organic and mineral compounds, as nitrogen, phosphorus, and potassium contained in the OMW, which could improve the nutrient status of the tomato plants in addition to the effects of phenols present in the OMW that could produce a reduction of *P. ramosa* seed germination [36–38]. Therefore, the tomato marketable yield showed a significantly higher value in the OMW treatment than the untreated control. No significant differences for the fruit qualitative characteristics were observed [35].

**Figure 6.** *Mechanical distribution of OMW on the soil (F. Lops).*

*Management of Branched Broomrape in Field Processing Tomato Crop DOI: http://dx.doi.org/10.5772/intechopen.106057*

#### **Figure 7.**

*Average number per m−2 of P. ramosa for OMW and control at the time of the tomato harvest in the different trials. Different letters indicate significant differences at P < 0.05 according to Tukey's test [35].*

Furthermore, in recent years, the use of organic fertilizers or "plant biostimulant" compounds has encountered increasing interest in agriculture because they play roles in various soil and plant functions [39]. Some of these compounds of natural origin, such as natural amino acids, were also suggested for use in *P. ramosa* management strategies being able to inhibit seed germination [40, 41]. Experimental results in Italy indicated that using the commercial product "Radicon®" (a suspension-solution containing humic substances), at the time of transplanting (immersing the root of the seedlings in a 1.5% solution), and incorporating it into the soil in the first 3 irrigation interventions, produced a reductions of 68.1% of emerged shoots in comparison with the untreated control. These substances introduced into the soil rhizosphere can cause severe physiological disorders of the germinating *P. ramosa* seeds, thus reducing the number of developing tubercles of the parasite [42].

#### *2.3.2 Soil solarization*

Solarization is used in many warm climate countries, as pre-tomato planting treatment. Its consists of heating the soil through sun energy achieving temperatures above 45°C, by covering a wet soil with transparent polyethylene sheets for a period of 4–8 weeks during the warmest season [43]. This method for the high cost per surface unit is not readily applicable at large scale [44]. Solarization may be more effective if combined with added nitrogen fertilizers as chicken manure [45].

#### *2.3.3 Rotation*

Decreasing the frequency of tomato cultivation prevents *P. ramosa* seed bank increases, maintaining the seed bank dormant and reducing the rate of seed bank replenishing. However, it is a long-term strategy due to the long viability of seed bank [16], which requires at least a nine-course rotation in order to prevent broomrape seed bank increases [46]. Its efficacy for broomrape cultural control can be increased including trap and/or catch crops as components in the rotation [16].

The trap crops are species (e.g., *Medicago sativa*, *Vigna unguiculata*, *Pisum sativum,* and *Linum usitatissimum*) whose root exudates induce broomrape seed germination,

but these species do not allow attachment or support broomrape seedling growth and survival [47].

Catch crops are host plants that support normal parasitism, but they are harvested as green vegetables after the parasite seeds germinated and before the flowering and seed dispersal stages of the parasite itself. For instance, *Brassica campestris* when managed properly as a catch crop can result in up to a 30% reduction in the size of broomrape seed bank [48].

#### *2.3.4 Soil management*

The soil tillage management must aim at reducing the seed bank, while minimizing the production of new seeds. In this regard, inversion plowing results in burial of a large proportion of seed in the tillage layer, carrying them at a depth from which they cannot germinate, although they remain viable in the deep soil for a long period of time [49, 50]. Deep plowing has been suggested to bring seeds of parasitic weeds to a depth with less oxygen availability and therefore a reduction in its germination capacity [51, 52]. Eizenberg et al., 2007 observed that the deep plowing ≥ 12 cm strongly reduced broomrape infection severity in terms of number of parasites, total parasitic biomass, delayed broomrape emergence and prevention of flower initiation, and seed set. Results of another study [53], carried out in two heavily infested fields in southern Italy, showed significant lower parasite attachments on tomato roots, the lower dry weight of emerged and underground-branched shoots per host plant in 50 cm deep plowing compared with 30-cm-deep plowing (**Table 1**).

#### *2.3.5 Sowing or transplanting date*

The air and soil temperature are the main factors influencing the dynamic of host/ parasite interaction and development. Temperature is strongly connected with the climatic conditions, which are themselves related to the periods for crops seedling into the field. Delayed sowing is consistently reported to reduce infection of winter crops such as oilseed rape [30]. Also, in spring-summer crops such as sunflower, modified planting dates provided the indirect effect of temperature on Orobanche parasitism [54]. In this regard, a study by Kebreab et al., 1999 [55] reports that at supra-optimal temperatures for germination of *O. crenata* seeds (i.e., above 25°C), they will not


#### **Table 1.**

*Mean value ± SD of total attachments, dry weight of emerged shoots, and tubercles per tomato plant of 30-cm-deep plowing compared with 50 cm one. Different letters in each column of each field and plowing treatment are differing significantly at P* ≤ *0.05, according to Tukey's test.*

#### *Management of Branched Broomrape in Field Processing Tomato Crop DOI: http://dx.doi.org/10.5772/intechopen.106057*

germinate. In a research carried out in southern Italy [14], a delay in seedling transplanting date from April to the hottest May reduced the *P. ramosa* infestation by 77%. Indeed, the daily maximum temperature was almost always below 25°C from April to mid-May, the period corresponding to the first stage of the tomato cycle for the early crop (transplant in April), while it increased to the threshold values always higher than 25°C starting from mid-May. This technique would give the host plant a time advantage over the *P. ramosa* and thereby make the tomato crop more competitive against this parasitic weed.

#### **2.4 Biological methods**

#### *2.4.1 Bioherbicide*

Biological agents such as pathogens *Fusarium* spp. (e.g., *Fusarium oxysporum* and *Fusarium arthrosporiodes*) or *Ulocladium botrytis,* incorporated into the soil by drip irrigation in field, are able to infect the pre-attached broomrape stages, and efficacy in reducing number and weight of emerging broomrapes [56, 57]. Due to the parasitic plant life cycle, multiple applications of *Fusarium* at the soil level would be necessary [58]. Conidial suspension of two *F. oxisporum* isolated reduced *O. crenata* and *P. ramosa* germination *in vitro* by 76–80%, in root chambers by 46–50%, and in polyethylene by 40–55% [59]. Fungi can be applied in the field together with solid growth media (such as wheat, corn, or rice grains) or in granules containing the biocontrol agent nutrients [60]. Compost activated by *Fusarium* was efficient in reducing the infection, by minimizing the number of parasitic spikes on the host tomato plant. This might be due to the additive effects on the seed germination of the parasite of the organic compound along with the soilborne fungi [61, 62]. Both granular soil applications and conidial suspensions of *Fusarium sp*. caused extensive mortality of *P. ramosa* in pot experiments. On the contrary, in field experiments, results were inconsistent as reduction *P. ramosa* shoot number and biomass [63, 64]. The main obstacle to the use and development of biocontrol agents is the poor field efficacy of the known pathogens. Soil-active biocontrol agents for *Phelipanche* must be able to contend with soil microorganisms without negatively affecting the host crop [65].

#### *2.4.2 Resistant varieties*

Cultivation of resistant varieties is another sustainable method to control *Phelipanche* [66, 67]. In addition, it is a useful component of an integrated approach, because easy to combine with other measures such as soil fertility amendments, land preparation, or soil tillage. Several mechanisms underlying the resistance of plants to the *P. ramosa* parasite have been described [68]. These include low stimulation of broomrape seed germination, pre-haustorial resistance, phytoalexin induction, high levels of peroxidase activities, lignification of host endodermis and xylem vessels, cell wall deposition, development of an encapsulation layer in the cortical parenchyma, induction of pathogenesis-related proteins, and sealing of host xylem vessels by deposition of mucilage [69]. Considered that this parasite requires stimulants exuded by the host roots, in order to germinate and reach the host root, varieties that exude stimulants at low levels or secrete inhibitors, they could be suitable for reducing parasite infection [70, 71]. An example of tomato cultivars resistant to *P. ramosa* infestation was reported by Qasem and Kasraw, 1995 [72]. The low germination stimulant phenotype of tomato has been reported in mutants owing to reduced exudation of strigolactones [73]. A successful screening program in a heavily

broomrape-infested field, to locate a resistant tomato line from a fast neutron-mutagenized M2 tomato population, was reported in Israel [74]. However, at present there are no commercial varieties for the broomrape control in tomato [6]. Research is needed in this regard to select from the wide range of varieties resistant to this parasite.

#### **2.5 Integrated method**

The single control practices described above are often only partially effective and sometimes inconsistent. Therefore, the most feasible way of coping with the weedy root parasites is *via* the integration different preventive measures and control instruments on a long-term basis into the given farming system [75]. The real challenge is to integrate practices that obtain optimum efficiency in terms of reduction of existing seed banks, prevention of seed production, and avoidance of seed dissemination with affordable costs. A computer simulation on integrated approach with a selection of appropriate cultural methods such as hand weeding, trap/catch cropping, delayed planting, resistant cultivars, and solarization demonstrates the importance of preventing new seeds entering the soil seed bank [76]. Resistant crop varieties and delayed transplant, for instance, are generally considered the useful components of an integrated approach that are usually easy to combine with other measures such as rotation, soil fertility amendments, and land preparation or soil tillage, and suitable to promote tomato plant growth and to reduce the *P. ramosa* infestation. Advantages of these sustainable approaches are no chemical applications that are known to cause damage to the environment.

#### **3. Conclusion**

The spread of branched broomrape is of great concern in tomato and other susceptible crop production systems in many countries around the world. This review summarizes the main control measures for the weedy root parasites *Phelipanche* and *Orobanche* in processing tomato, namely prevention, chemical, agronomic, and biological control. Some of these methods are commercially widely used by farmers (herbicidal control), some are in the final stages of development toward commercialization (resistant varieties), and some still require further development and improvement before commercial implementation (bioherbicide control). As for chemical control of broomrape, it should take the environment into consideration by encouraging reductions of herbicides, by carefully calibration of doses and timing of treatments depending on the underground phenology of broomrape determined by local conditions. One of the most promising directions is the precision agriculture approach of site-specific weed management. In this approach, herbicide is applied only in the infested area according to the spatial variation of parasite infestation in the field. Furthermore, it is desirable to improve the environmentally friendly, sustainable, and practical parasite control methods and use them in an integrated way. Therefore, future efforts must aim at improving these parasite control methods in accordance with new cultivation technologies suitable for the development of the processing tomato.

*Management of Branched Broomrape in Field Processing Tomato Crop DOI: http://dx.doi.org/10.5772/intechopen.106057*

### **Author details**

Francesco Lops\*, Laura Frabboni, Antonia Carlucci, Annalisa Tarantino, Maria Luisa Raimondo and Grazia Disciglio Department Agriculture Food Natural Science Engineering, University of Foggia, Foggia, Italy

\*Address all correspondence to: francesco.lops@unifg.it

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Section 3

## Tomato Processing Applications

#### **Chapter 9**

## Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their Application as Antimicrobial Agent

*Aistė Balčiūnaitienė, Jonas Viškelis, Dalia Urbonavičienė and Pranas Viškelis*

#### **Abstract**

Silver nanoparticles (AgNPs) biosynthesized using by-products of tomatoes extracts as reducing and capping agents show multiple possibilities for solving various biological problems. The aim of this study was to expand the boundaries on AgNPs using novel low toxicity and production cost phytochemical method for the biosynthesis of nanoparticles from tomatoes aqueous extracts. Biosynthesized AgNPs were characterized by various methods (SEM, EDS). Determined antioxidative and antimicrobial activity of plant extracts was compared with the activity of the AgNPs. TEM results show mainly spherical-shaped AgNPs, size distribution of which depends on the plant leaf extract type; the smaller AgNPs were obtained with tomatoes extract (6–45 nm AgNPs). Besides, AgNPs show strong antimicrobial activity against broad spectrum of Gram-negative and Gram-positive bacteria strains and fungi.

**Keywords:** silver nanoparticles, green synthesis, tomatoes by-products, antimicrobial activity

#### **1. Introduction**

There are various bacterial, fungal, viral, and other microscopic life forms in our environment. Microorganisms make up 80–90% of the earth's total biomass, and even under "clean" conditions, several thousand fungal spores can be inhaled per day. Many microorganisms are harmless or even beneficial, but others can be extremely dangerous or even deadly. The current way of life creates favorable conditions for the spread of infections (food from distant lands, work in air-conditioned rooms, frequent trips to foreign countries, visits to hospitals, etc.). The human body is not sterile, and it is colonized by many microorganisms that are part of the normal microflora and live like harmless commandants.

Microorganisms living under normal conditions on the skin, nasopharynx, and intestine play an important protective role, as they prevent the growth of pathogenic microorganisms in these places. As the bodies condition changes (weakened immunity, disease, or trauma), the so-called non-pathogenic bacteria can become pathogenic and cause infections. Wounds are susceptible to contamination by microorganisms both externally and from internal sources in the body, such as the nasopharynx, skin, and gastrointestinal tract. Infection is the result of a constantly changing interaction between microorganisms, the human being as their host, and the environment around them. Exposure of the Gram-positive and Gram-negative bacteria strains to the host's defense capacity interferes with wound healing and potentially dangerous changes in the body due to infection.

According to research, more than 23,000 people die each year in Europe from invasive (or systemic) infections caused by *Staphylococcus aureus* (*S. aureus*) and *Escherichia coli* (*E. coli*). It has also been observed that these infections are increasing rapidly due to the progressive, excessive use of antimicrobials, which allows pathogenic microorganisms to evolve and acquire multiple antibiotic resistance. Therefore, scientists around the world are constantly looking for new ways and materials to combat the colonization of pathogenic microorganisms [1]. Thus, the problems associated with unwanted bacterial adhesion to the surfaces of medical equipment, as well as the colonization of surgical equipment, implants, and other health-related products, pose a significant risk to public health. The formation of biofilms also directly affects many industrial processes: food processing and storage, water treatment processes, maritime transport and management. Various antimicrobial agents are commonly used against biofilms and their infections, but microbiological control of the process is hampered by the ability of pathogenic microorganisms to attenuate or acquire full resistance to antimicrobial compounds, including antibiotics. Despite ongoing efforts by scientists to avoid bio-contamination and additional control measures implemented by industry, there is still no effective solution to protect the surface of equipment from colonization by pathogenic microorganisms. For these reasons, the need for antimicrobials is greater than ever before.

#### **2. Antimicrobial agents**

The increasing level of pollution by microorganisms and infections creates the need for new antimicrobial agents. Therefore, the research on the development and application of polymer composites with antimicrobial activity is of great interest.

Plant-mediated synthesis imparts several advantages to metal nanoparticles (MNPs) technology for the development of alternative products against infectious diseases. Indeed, most of green MNPs from plant-derived materials are highly effective and nonspecific antimicrobial agents, showing remarkable activities against the growth of a broad spectrum of bacterial and fungal species, in both planktonic and biofilm forms, including nosocomial and multidrug-resistant strains [2, 3].

Materials with antimicrobial activity are abundant. One of the largest groups is natural or synthetic antibiotics, which inhibits the appropriate stage of synthesis of the microorganism's cellular proteins. However, excessive use of antibiotics has led to the emergence of strains of bacteria that are resistant to most antibiotics, posing a significant risk to public health. As a result, other substances with antimicrobial activity are increasingly being used. Their nature can be very different. These are, in particular, substances of natural origin: vegetable (various essential oils, medicinal plant extracts, etc.) and animal (e.g., lysozyme, lactoferrin), microbes (nisin, natamycin, etc.), as well as inorganic and organic synthetic and hybrid derivatives of a

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

#### **Figure 1.**

*The choice of antimicrobial modification method.*

**Figure 2.**

*The main methods of antimicrobial modification of polymers [4].*

nature. Polymeric materials that are resistant to the colonization of microorganisms and the spread and multiplication of pathogenic microorganisms are also one of the groups of antimicrobials. They usually consist of a polymeric matrix and an embedded antimicrobial component.

The choice of antimicrobial modification methods depends on many factors (**Figure 1**).

The main methods of antimicrobial modification of polymers are as presented in **Figure 2**.

#### **3. Antimicrobial activity of metal nanoparticles**

One of the most abundant groups of substances with antimicrobial activity suitable for polymer modification is inorganic compounds and metal nanoparticles. This group consists of metals (Ag, Au, Cu, etc.), metal oxides (ZnO, TiO2, etc.) [5], nonmetallic oxides (SiO2). In most cases, the size of antimicrobial nanomaterials ranges from 1 to 100 nm. They can be of organic or inorganic origin, but inorganic substances are most commonly used. Nanoparticles are the most widely used because they have broad-spectrum antibacterial properties against both Gram-negative and Gram-positive bacterial strains [6]. The main reason why nanoparticles are an alternative to antibiotics is their ability to inhibit multiresistant microorganisms in some cases. Nanoparticles have a large surface area that increases interaction with microorganisms, resulting in strong antimicrobial activity. Nanoparticles with a smaller size and a higher surface area to weight ratio are more efficient at breaking biofilms. The particle shape also has a significant effect on the degradation efficiency of biofilms (e.g., rod-shaped particles are much more efficient than spherical forms). There are various methods for the synthesis of nanoparticles, which can be divided into two main classes: (1) bottom-up, and (2) from top to bottom [7].

In general, the chemical, physical, mechanical, and antimicrobial properties of nanoparticles depend on their chosen precursor. Nanoparticle microorganisms act in different ways, and the mechanism of their action depends on the origin of the nanoparticle [8]. Nanoparticles have antibacterial (inhibits DNA replication, enzyme functions, etc.), antiviral (blocks the attachment of viruses to the cell wall), antifungal (breaks down the cell membrane), and other effects.

Nanotechnology is the science, engineering, and technology that studies matter at the atomic, molecular, or supramolecular levels to yield nanometric materials and nanosystems with improved properties such as high surface-to-volume ratio and high dispersion in solution. With size typically ranging between 1 and 100 nm, these nanomaterials and nanosystems can be synthesized by chemical, physical. and/or biological methods [9]. In comparison with chemical and physical methods that involve costly and toxic chemicals, the biological synthesis pathway based on the usage of biological sources (plants, bacteria, fungi, and algae) is hoisted as a real rescue route. In spite of that, the biological methods do not envisage the use of toxic catalysts and reagents, dealing exceptionally with the intracellularly or extracellularly produced metabolites within fermentation routes, and this method requires big input of costly materials, well-developed protocols and guidelines, and microbiological hands-on experience to ensure cell culture and nanoparticles purification under highly aseptic conditions.

In contrast, the use of plant-derived extracts, juice, hydrolysates, etc., for the biosynthesis of metal nanoparticles (MNPs) seems to be an environmentally friendly, cost-effective, robust, sustainable alternative with moderate reaction conditions [10]. The plant-mediated synthesis of nanoparticles is also biocompatible, clinically adaptable, and easily up-scalable for industrial production [11]. Plants could represent

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

continuous source of natural antioxidants and antimicrobials (polyphenols, flavonoids, tannins, terpenoids, alkaloids, essential oils, etc.) suitable for green synthesis of nanoparticles with desirable properties. Under proper extraction conditions dealing with nontoxic organic solvents, diverse spectrum of non-deleterious reducing agents could be acquired [12].

Recent evidence in the field of nanotechnology revealed that the morphological parameters of nanoparticles (e.g., size and shape) can be modulated by varying the concentrations of bioactive compounds and reaction conditions (e.g., temperature and pH). Due to multiple therapeutic and biological activities such as antioxidative, antimicrobial, anti-inflammatory, anticancer, eugenol as a representative of phenylpropanoids received tremendous interest among researchers. The crude extracts recovered from such herbal plants as Lamiaceae, Lauraceae, Myrtaceae, and Myristicaceae, the major compound of which was eugenol, have been investigated in terms of reducing ability for nanoparticles synthesis. However, less explored are other sources of this unique molecule, especially by-products that also could provide adequate quantities of eugenol. Considering the evidence on the presence of eugenol, a principal component of lignin in cereals and their by-products (bran), and already established protocol for lignocellulose biomass hydrolysis, it is speculated that the process of biorefining could represent a sustainable and reliable way of bran utilization for the production of eugenol-based nanoparticles, thereby contributing to waste reduction. Additionally, using different sources of metals (salts or oxide) in combination with plants, the biological reduction method allows the synthesis of a large number of green MNPs, including silver (Ag), gold (Au), zinc oxide (ZnO), platinum (Pt), palladium (Pd), copper (Cu), iron oxide (Fe2O3 and Fe3O4), nickel oxide (NiO), magnesium oxide (MgO), titanium dioxide (TiO2), and indium oxide (In2O3).

Considering the above, exploration of the plant systems as potential bio-factories for MNPs has gained considerable attention, especially for researchers working in the field of phytonanotechnology, pharmaceutical, and clinical microbiology as well as medicine [7]. Indeed, due to the surging popularity of green methods, more than 2000 research papers and reviews related to antibacterial, antifungal, and antibiofilm properties of MNPs have been published. Noteworthy, most of the reviews and research articles published so far focused mainly on predicting the antimicrobial mechanisms of MNPs and parameters that may influence their antibacterial, antifungal, and activities such as.

**Figure 3.** *Mechanisms of antimicrobial action of nanoparticles on bacterial cells.*

Unfortunately, it appears from these reviews that the methods used for assessing the antibacterial, antifungal, and antibiofilm efficiency of MNPs are only partially elaborated in terms of standardization process; therefore, it is hard to correlate or compare data from different studies to pinpoint the high values of antimicrobial nanoparticles. Moreover, such methodologies and models are usually hard to extrapolate to real products.

Metal nanoparticles can affect the bacterium even in several ways, causing extremely strong antimicrobial activity (**Figure 3**). The most common and widely used silver nanoparticles, elemental silver has been widely used as an antimicrobial agent since ancient times. To improve their antibacterial activity and reduce their toxicity, silver ions can be transformed into metallic silver nanoparticles through biological and biomimetic methods of synthesis. Green AgNPs have demonstrated the ability to reduce microbial infections in the skin and burn wounds and prevent bacterial colonization on the surface of various medical devices such as catheters and prostheses. Acting as capping agents, different multifunctional phytochemicals contribute efficiently to these antimicrobial activities [8]. Moreover, AgNPs can express synergism with standard antibiotics such as gentamycin and streptomycin [13]. Hence, these combinations can effectively be used against antibiotic-resistant pathogens. Additionally, antifungal activities of AgNPs have extensively been studied and demonstrated in the literature [14]. In the frame of the fight against antibiotic resistance, green synthesized AgNPs may be used as vehicles to transport oligonucleotide-based antimicrobial. Their synthesis can be performed by physical, chemical, or biological methods. Particle size, morphology, and antimicrobial activity differ according to the chosen method [15].

Numerous studies have shown that silver nanoparticles, in both colloidal and ionic forms, have a broader spectrum of antibacterial activity than most other nanoparticles. Due to their unique optical, electrical, and chemical properties, silver nanostructures are widely used in a variety of industries. However, they are most commonly used in health care and medicine due to their strong antimicrobial activity against many pathogenic microorganisms—Gram-positive, Gram-negative, and antibiotic-resistant bacterial species, fungi, and viruses.

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

#### **Figure 4.**

*Factors determining the antimicrobial activity of silver nanoparticles.*

Rapid wound healing is due to decreased matrix metalloproteinase and increased neutrophil apoptosis in the wound induced by silver compounds. Matrix metalloproteinase is thought to be able to initiate inflammation and thus slow wound healing, so its regulation is very important [16]. Silver nanoparticles are also used in bone cement in various disinfectants [13] as antimicrobial agents. Beside this, their effect depends on several factors presented in the figure (**Figure 4**).

Antimicrobial activity of silver ions is obtained by reacting with the main components of the bacterium:


Due to the small size and very large specific surface area, silver nanoparticles adhere firmly to the surface of the bacterium. Silver ions, by interacting with the bacterial cell membrane and the sulfur compounds present in its proteins, impair its functionality. Further, silver nanoparticles penetrate the cell and damage DNA. Silver ions react with phosphorus compounds present in DNA, disrupting the process of DNA replication, which inhibits bacterial proliferation. They also degrade bacterial proteins, especially enzymes that catalyze metabolic reactions and other vital cellular processes. In addition, nanoparticles lead to the formation of reactive oxygen species, which are active and unstable molecules that can damage cellular DNA, protein structures, and cell membranes [17].

The antimicrobial activity of silver ions in Gram-positive and Gram-negative bacterial cultures may be different due to differences in bacterial cell structure. The cell wall of Gram-positive and Gram-negative bacteria has a complex, semi-rigid structure. The structure of the wall is very important because it determines the ability of the bacteria to cause disease and resistance to certain antibiotics. The wall thickness of the bacterial cells is unequal. The cell wall of Gram-positive prokaryotes is composed of a network of macromolecules called peptidoglycan or murein,

polysaccharides, lipids, and proteins. The wall thickness is much higher (20–80 nm) than that of Gram-negative bacteria. Their prokaryotic cell wall is composed of several layers: The inner dense electron layer (2–3 nm) is composed of peptidoglycan, two dense electron bands separated by an electron-conducting cavity, a space separated by the periplasmic cavity of the cytoplasmic membrane. The cell wall of Grampositive microorganisms adheres closely to the cytoplasmic membrane [18]. These differences between bacterial species lead to unequal interactions between antimicrobial compounds. It is clear that metal nanoparticles are promising as antimicrobial agents and therapeutic agents due to their biological, physical, and chemical properties. They can solve many problems in the field of nanomedicine. However, there is a lack of knowledge about the long-term effects of nanoparticles on human health and the environment. Nanoparticles are stable and can accumulate in the environment; they have a tendency to agglomerate and can therefore change their dimensions. Toxicity studies of nanoparticles have shown that metal nanoparticles can act at the organ, tissue, cell, muscle, and protein levels. Nanoparticles are extremely small in size and can easily spread through air or water and adversely affect the skin, lungs, and brain (especially nanoparticles with dimensions 10 nm).

Therefore, the search for other substances with antimicrobial activity, such as the use of plant-derived substances to obtain antimicrobial compounds, is intensifying [19].

#### **4. Morphology and antimicrobial activity of tomatoes by-products and green AgNPs**

The aim was to compare the morphological differences of tomato pulp by variety. From **Figure 5**, the micrographs presented by SEM can show that the tomato particles are irregular in shape, with an uneven, layered surface, and that the particles appear to be composed of discrete slender shapes without any visible particles on the surface. The average particle diameter is very uneven as it was not fractionated, but the particle size could be harmonized by choosing milling techniques and conditions. Also, the particle size may vary depending on the desired properties.

The aim is to obtain stable and externally resistant colloidal solutions of silver nanoparticles and to investigate the antimicrobial efficacy of synthesized nanoparticles. Silver nanoparticles (AgNPs) were obtained by crude metal synthesis by reducing and stabilizing silver nitrate in extracts from bioactive compounds.

The morphology of lyophilized AgNPs of biologically active tomato pulp used in the work was investigated by SEM methods. From **Figure 6**, the microstructures of tomato by-products AgNPs can be concluded from the irregularly shaped particles but do not form agglomerations, which will have a positive effect on antimicrobial

**200 Figure 5.** *SEM images of tomato by-products particles*

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

**Figure 6.** *Tomatoes "Vilina" by-products TEM micrographs, surface EDS spectra, and elemental analysis*

activity. From the photos provided by TEM, we can see a clearer morphology of the particles. The particles are spherical and do not form agglomerates (**Figure 6**).

In this case, individual agglomerations can already be observed. Scanning of metal particles at selected locations where AgNPs are suspected shows peaks in the 3.0 keV region of the EDS spectra that can be attributed to silver binding energy, and this can be detected at first and third samples. In second, sample AgNPs could not be found, but the biomatrices had antimicrobial activity. Therefore, we can say that the particles formed. With the help of TEM microscopy, we can see that the particles obtained are particles with a clear spherical shape, but in individual cases we can observe the formation of agglomerates (**Figure 7**).

**201** The TEM images show an uneven surface with AgNPs. A high silver content in the biomatrix was identified (**Figure 8**). The particles remain irregular in shape and do not

**Figure 7.** *Tomatoes "Laukiai" by-products TEM micrographs, surface EDS spectra, and elemental analysis*

tend to form large aggregates, which is likely to have a positive effect on antimicrobial activity. From the presented photos, we can see the particle shape, size distribution, and agglomeration tendency of AgNPs. In this case, the largest particles are obtained. Also in their form the resulting spheres. The particles obtained have a relatively high polydispersity, which is likely to have a positive effect on antimicrobial activity.

The antibacterial activity of organic colloidal solutions of AgNPs was tested for both Gram-negative and Gram-positive bacterial strains and fungi. From the results *Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

**Figure 8.** *Tomatoes by-products mix TEM micrographs, surface EDS spectra, and elemental analysis*

presented in **Table 1**, it can be concluded that silver nanoparticles in organic media actively interact with the bacterial membrane and disrupt their functions.

The results of the antifungal efficacy studies of AgNPs are presented in **Table 1**. Two different fungal cultures were selected: *Candida albicans* (*C. albicans*) and *Rhodotorula glutinis* (*R. glutinis*). In humans, *C. albicans* can cause external infections and life-threatening systemic infections, and *R. glutinis* is an opportunistic pathogen that can cause infection in


#### **Table 1.**

*Antimicrobial activity of the greenly synthesized AgNPs.*

a weakened immune system. From the results presented in the table, it can be concluded that AgNPs obtained using different Russian tomatoes with different syntheses inhibited the growth of *C. albicans* and *R. glutinis* colonies. Meanwhile, extracts without particles did not show this effect.

It is clear that metal nanoparticles are promising as antimicrobial agents and therapeutic agents due to their biological, physical, and chemical properties. They can solve many problems in the field of nanomedicine. However, there is a lack of knowledge about the long-term effects of nanoparticles on human health and the environment. Nanoparticles are stable and can accumulate in the environment, and they have a tendency to agglomerate and can therefore change their dimensions. Toxicity studies of nanoparticles have shown that metal nanoparticles can act on the organ, tissue, cell, muscle, and protein levels. Nanoparticles are extremely small in size and can easily spread through air or water and adversely affect the skin, lungs, and brain (especially nanoparticles with dimensions ≤10 nm). Therefore, the search for other substances with antimicrobial activity, such as the use of plant-derived substances to obtain antimicrobial compounds, is intensifying.

#### **5. Conclusions**

Green nanoparticles obtained by green synthesis methods, which have a wide range of antibacterial properties against both Gram-negative and Gram-positive bacterial strains and fungi, expand their applications in orthopedics, biomedicine, and medicine, as well as in other industries. Recently, the range of substances resistant to microbial colonization and multiplication of pathogenic microorganisms are increasing due to the increasing use of extracts of medicinal plants and plant by-products, which are strong antioxidants with anticancer, antibacterial, anti-inflammatory, antiallergic, antiviral, hepatoprotective effects. One of the most important antioxidants accumulated in plants is phenolic compounds, the

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

mechanism of action of which is related to their ability to neutralize free radicals, protect against diseases caused by oxidative stresses, and reduce various forms of reactive oxygen species. It can be assumed that the modification of green nanoparticles with multifunctional hybrid particles can increase and expand their scope. Such antimicrobial and functional biomatrices are obtained using secondary by-products and Ag.

Stable colloidal solutions of AgNPs with high antibacterial activity in organic media have been obtained, which completely inhibit various bacterial cultures.

#### **Acknowledgements**

This study was financed by the Lithuanian Research Centre for Agriculture and Forestry and attributed to the long-term research program "Horticulture: agrobiological foundations and technologies."

#### **Conflict of interest**

The authors declare no conflict of interest.

#### **Appendices and nomenclature**


### **Author details**

Aistė Balčiūnaitienė\*, Jonas Viškelis, Dalia Urbonavičienė and Pranas Viškelis Lithuanian Research Centre for Agriculture and Forestry, Institute of Horticulture, Babtai, Lithuania

\*Address all correspondence to: aiste.balciunaitiene@lammc.lt

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Tomatoes By-products Extracts Mediated Green Synthesis of Silver Nanoparticles and Their… DOI: http://dx.doi.org/10.5772/intechopen.105976*

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## Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa

*Hans Adu-Dapaah, Michael Kwabena Osei, Joseph Adjebeng-Danquah, Stella Owusu Nketia, Augustine Antwi-Boasiako, Osuman Alimatu Sadia, Peter Ofori Amoako and Richard Agyare*

#### **Abstract**

Tomato production in Africa has increased due to increased population, rising consumer demands for nutritious and healthy food and potential use of improved technologies. Demand-led' plant breeding puts producers and consumers at the heart of research and development involving stakeholders even before the research starts. These 'stakeholders' are not only farmers but key actors along the tomato value chain. They influence how the tomato is traded as: fresh food and processing product. This chapter focuses on different approaches to fast-track tomato breeding so as to contribute to the transformation of African agriculture by enabling small scale farmers to compete in local and regional markets, by increasing the availability and adoption of high performing tomato varieties that meet market demands. It further outlines development of varieties that meet farmer needs, consumer preferences, and market demand in Africa. These new varieties are designed to meet client needs by connecting plant breeders with crop value chains, seed distribution organizations, and encouraging enterprise and entrepreneurship in transforming agriculture in Africa. Lastly, it outlines the prospects and challenges associated with demand-led breeding of tomato and offers suggestions to increase food security in Africa.

**Keywords:** demand-led, tomato, breeding, emerging markets, consumers, producers

#### **1. Introduction**

The production and utilization of tomato has increased over the years in Africa [1]. However, demand for the crop exceeds supply owing to its economic importance and increasing popularity as a result of processing, value addition and consumption [2]. According to ref. [3], open field cultivation dominates production of tomato in Africa hence exposes the crop to biotic and abiotic stresses, resulting in yield reduction and poor fruit quality [4, 5]. Increasing population, reduced natural resources as well as extreme climate change has further worsened the glitches of food security. The focus

of most plant breeding programmes is to develop and make available to end-users, crop varieties that meet their needs and capable of solving their everyday problems. Therefore, to facilitate the adoption and utilization of the end product, there is the need to take into consideration the expectations or demands of the prevailing market. Plant breeding approaches that consider active participation of farmers in the identification of their challenges, mitigation approaches and preferences in new varieties is crucial in the adoption of the resultant varieties [6].

Conventional methods of crop improvement require longer period, are time-consuming, costly, and restrict supply of genetic diversity. These challenges are exacerbated by the growth of human population (7.8 billion) in 2020, projected to nearly 10 billion by 2050 [7]. The need for strategies to increase the genetic advance in foods crops including tomatoes [8] for achieving food and nutrition security especially in Africa cannot be overemphasized. Speed breeding (SB) provides an opportunity to fast-track the breeding cycle, a key component of breeder's equation [9]. In diverse and marginal environments like Africa [10], appropriate plant breeding strategies should be developed to enhance the adoption and utilization of new varieties by farmers, consumers and industry.

The demand-led breeding approach will not only increase development of new tomato varieties but meet the needs of changing market preferences. Demand-led breeding combines the best practices in market led new variety design with innovative plant breeding methods and integrates both of these with the best practices in business as a new way of breeding crops to deliver benefits. The approach puts stakeholders especially producers and consumers at the heart of research and development by involving stakeholders even before the research starts. These 'stakeholders' are not only farmers but include other actors along the tomato value chain. They influence how the tomato is traded either as fresh food or processed product. According to ref. [11], demand-led breeding takes an integrated approach to new variety development and requires a comprehensive analysis that takes into consideration the target clients, their needs and how these may change.

This chapter focuses on different approaches to fast-track demand-led tomato breeding so as to contribute to the transformation of African agriculture, by empowering small-scale farmers to better compete in local and regional markets. It stresses on demand-led breeding and strategies to hasten breeding of tomato varieties that will be appropriate for consumers as well as other relevant stakeholders. It further discusses seed system in the sub-Sahara Africa and projects seed security as paramount in food security. It outlines the development of varieties that meet farmer needs, consumer preferences, and market demand in Africa. Finally, it outlines the challenges and prospects associated with demand-led breeding of tomato and offers suggestions to enhance food security in Africa.

#### **2. Tomato breeding objectives in Africa**

Tomato production in Africa has increased [1], but demand for the vegetable crop continues to surpass supply due to its economic relevance and growing popularity as a result of processing, value addition, and consumption [2, 5]. In Africa, less than 5% of areas dedicated to vegetable crop production including tomato are subjected to protected or controlled environments such as greenhouses [12]. Open field cultivation is paramount in tomato production in Africa [3, 13], exposing the crop to numerous biotic and abiotic stresses, causing reduction in yield and fruit quality [4, 5, 14].

Approaches to improve tomato resilience as well as productivity include breeding for resistance/tolerance to biotic and abiotic stresses and improvement of fruit quality to meet consumer/industry preferences.

#### **2.1 Breeding for resistance/tolerance to biotic stresses**

In Africa, one of the limiting factors impeding tomato production is biotic stress. This is induced by the living components of the environment [15] such as weeds, insect pests and diseases [2, 16–18]. In an attempt to address this situation, the indiscriminate use of chemicals such as pesticides which is harmful to human health [19] and the environment [18, 20] has made the demand-led breeding approach environmentally safe. Plant breeding is one of the most cost-effective and environmentally friendly methods of controlling biotic stresses in tomato production. In Africa, a number of breeding programmes have been undertaken to enable breeders develop tomato cultivars that are resistant to diseases and insect-pests [21]. In the tropics, major diseases affecting tomato production include bacterial wilt caused by *Ralstonia solanacearum*, late blight caused by *Phytophthora infestans, Stemphyllium spp* and Fusarium wilt caused by *Fusarium oxysporum* [22, 23]. Major pests of tomato include nematodes, thrips, aphid, cotton bollworm and mites [24]. Outbreaks of the tomato leaf miner (*Tuta absoluta*) have caused substantial damages in some West African countries [25, 26].

The pests and pathogens causing the above-mentioned diseases are genetically diverse with vast potential to generate new forms and hence difficult to control [16]. In view of this, conventional breeding together with marker-assisted selection (MAS) [21, 23], Quantitative Trait Loci analysis, and Hybridization [27] have been adopted for improving tomato resistance to biotic stresses. For instance, in recent decades extensive breeding programmes via the use of a series of in-region trials and collection of germplasm have been used to screen and develop varieties that are commercially acceptable and resistant to diseases [24]. Currently, the Crops Research Institute under the Council for Scientific and Industrial Research (CSIR), Ghana and West African Centre for Crop Improvement (WACCI), University of Ghana have released tomato varieties that have shown reasonable tolerance to late blight, Fusarium wilt and nematodes [28, 29].

#### **2.2 Breeding for tolerance to abiotic stress**

Similarly, abiotic factors tend to cause a myriad of considerable qualitative and quantitative crop losses to tomato production, especially in open field production. Due to threats posed as a result of climate change, tomato-producing environments are bedevilled with abiotic stresses such as heat, drought, water logging and salinity [24, 30]. Water deficit [31] and heat stress [32] are the most predominant abiotic factors that threaten tomato production in sub-Saharan Africa. Abiotic stresses can cause up to 70% yield losses [33]. Though considerable efforts have been made to ameliorate the continental menace through the use of agronomic approaches such as increased irrigation and temperature regulation, some of these efforts have proven futile over recent decades. An alternative promising approach to address these stresses is the development of tolerant tomato cultivars/lines through breeding [30]. One of the major constraints affecting tomato production is heat stress [34]. This is due to the increasing day and night temperatures as a result of climate change [35]. Breeding for heat tolerance has become one of the primary objectives of breeding programmes in

Africa [5]. In Ghana, Nkansah, King 5, DV2962 have been identified as heat-tolerant tomato cultivars [21, 32]. Currently, the CSIR-Crops Research Institute is developing heat tolerant tomato varieties with funding from the Korea government through Korea Africa Food and Agriculture Co-operation Initiative (KAFACI).

Most farmers in Africa grow tomatoes under open field conditions and as such rely solely on rainfall. Under such conditions, drought stress which is one of major constraints in open field tomato production [36] occurs due to erratic rainfall pattern. This consequently affects plant growth and development, resulting from reduced nutrient uptake [37] leading to increase in flower abscission, low percentage of fruit set, reduction in yield as well as fruit quality [38, 39]. There is therefore the need to breed and develop improved drought-tolerant varieties [38]. However, developing tomato cultivars tolerant to drought stress has been a neglected objective in many tomatoes breeding programmes [40], since the breeding objectives tend to focus much more on biotic stresses, prolonging shelf life, and determination of genetic variability among continental accessions. Although advances in molecular research and plant breeding have resulted in the introduction of drought-tolerant tomato cultivars in most developed countries, breeding efforts in sub-Saharan Africa (SSA) have focused on yield as the primary selection criteria, with little attention for drought tolerance [5, 40]. Nonetheless, a few screening trials have been conducted in countries such as Kenya to evaluate the susceptibility and tolerance of tropical cultivars derived from the AVDRC-The World Vegetable Center and the National Gene Bank of Kenya to drought stress [31].

#### **2.3 Breeding to improve tomato fruit quality to meet consumer/industry preferences**

Breeding for fruit quality is one of the major objectives of the tomato breeding programmes in Africa [41]. Due to the economic and nutritional importance of this perishable crop, breeders over the decades have put in great efforts to prolong its shelf life and organoleptic quality. Extensive studies have been reported on fruit quality traits such as the size, shape, total soluble solids, pH, colour, firmness, ripening, nutritional content, and flavour [42–44]. Therefore, tomato breeding initiatives have focused on boosting fruit quality and understanding its genetic and molecular diversity [44]. Recently, fruit colour is becoming increasingly important in the fresh market due to the awareness of the health benefits of carotenoids in the tomato fruit. Regarding processing of the tomato fruit in the industries, content of total soluble solids has also received lot of attention [44]. For instance, technologies such as pure line selection, hybridization, irradiation-induced mutation, and the crossing of local cultivars with exotic ones are ongoing breeding schemes in African countries such as Ghana to improve the fruit quality and shelf life of tomato [5, 14]. In addition, [45] reported studies on the utilization of single nucleotide polymorphism (SNPs) to evaluate the shelf life and fruit quality of F1 tomato progenies.

#### **3. Demand-led tomato breeding**

#### **3.1 Overview of demand-led breeding**

To facilitate the adoption and utilization of the end product, there is the need to take into consideration the expectations or demands of the prevailing market.

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

According to [6, 46, 47] plant breeding approaches that consider active participation of farmers in the identification of their challenges, mitigation approaches and preferences in new varieties is crucial in the adoption of the resultant varieties. The concept of demand-led breeding encompasses the approach whereby the situation in the prevailing market for new crop varieties considers the type of traits to incorporate into new varieties that will meet the expectations and satisfy the consumer or end-user needs. Whereas, participatory plant breeding or variety selection considers farmers involvement at different stages of the breeding programme, demand-led breeding encompasses various considerations from different actors such as processors, aggregators, marketers and consumers [48].

#### **3.2 Demand-led principles and approaches for tomato breeding**

Unlike participatory breeding that is more localized with limited scope, demand-led breeding involves more global focus. It takes into consideration a broader range of tools such as market research, value addition and modern product promotion strategies. It focuses more on the demands of the market rather than adoption for cultivation thereby producing a product that would be in high demand once released for cultivation. Demand-led approaches focus on the use of market information and intelligence to develop indices that are used to rank traits based on the monetary value and preferences from all potential end-users of the final product [49]. According to [50] demand-led variety design is based on six core principles; client needs and preferences, value chain analysis, market research, market trends and drivers, public and private sector linkages and multidisciplinary teams.

Demand-led or client-oriented breeding should consider client needs and preferences. This is crucial in considering the breeding objectives whether for industrial processing or home consumption. In Ghana, tomato production is either for processing into paste or direct consumption by consumers who purchase from the open market [51]. A demand-led programme should consider value chain analysis and innovation systems that involve all the actors in the value chain of the crop. For instance, in the value chain, common actors include farmers, aggregators or middlemen, transporters, traders, processors and consumers. Another consideration for demand-led breeding is market research. This allows the breeder to define the standard and priority for the traits or client preferences and validate the key assumptions at every stage of the breeding process. As a result, the breeder is kept abreast with the demands of the market in order to provide a product which will be readily adopted by the producers, marketers and consumers alike.

Demand-led breeding is also based on market trends and drivers which normally influence farmers' choices of crop varieties to adopt for cultivation. Prevailing circumstances and future occurrences such as climate change, national policies regarding certain commodities can all influence the kind of varieties that would be needed for cultivation [50]. For instance, a government initiative on tomato production or establishment of tomato processing factory may change the focus of variety design towards home consumption to varieties with good processing attributes. Another key principle guiding demand-led breeding is integration or linkage between private and public sector. It focuses on fostering cordial relationships between breeders and other actors in the value chain such as seed producers and distributors as well as other actors in the value chain. All these actors are involved in the identification and priority setting of client needs that result from the market research. Through this approach, breeders know what to breed for, the farmers also know what to cultivate

to meet the market requirements. Farmers are also linked to ready market. This approach promotes synergies between the various actors and culminates in benefits that far exceed what can be achieved with the different actors acting independently [51]. Demand-led breeding relies on multidisciplinary teams to achieve its objective. Demand-led breeding follows an innovative approach that utilizes a broad range of expertise and competencies of different actors with specific roles towards the design and development of the proposed variety. It is expected that the different experts will contribute to the development of the ideal product profile which possesses all the desired attributes and is responsive to the needs of the target group irrespective of the gender [52].

#### **3.3 New variety design and product profile for tomato breeding: Ghana as a case study**

To facilitate large scale adoption and commercialization of a new crop variety, such variety must meet the needs and expectations of the intended end-users [53]. Therefore, product profiles are developed for such a desired variety. A product profile encompasses a number of traits in a new variety that farmers would prefer compared to the variety they are already cultivating [54]. Product profiles are developed based on the array of clients that are targeted by the breeding programme following market research or broadscale stakeholder consultation. Several factors may influence the choice of crops to cultivate or the variety of a particular crop a farmer may cultivate and these have implications on the overall product design. A study by ref. [55, 56] revealed that the number of attributes or traits preferred by tomato farmers in the Wenchi municipal was positively influenced by the gender, education level, access to credit, household size, level of education, contact with extension staff, membership of farmer-based organization, farm size and off-farm income. This implies that different varieties are likely to be adopted by farmers in the different categories. As a result, these factors need to be considered by breeders and breeding programmes in designing and developing product profiles of new tomato varieties to meet the needs of the different clientele. For this reason, a particular breeding programme can have several product profiles that will define the type of varieties that would be developed [48].

Another survey carried out in seven regions of Ghana (Bono, Ahafo, Bono East, Ashanti, Greater Accra, Eastern and Upper East regions) involving 12 tomato growing communities found that tomato farmers in these areas prefer tomato varieties with large fruit size, high rounded shaped and red in colour [51]. A similar study by ref. [55, 56], indicated that majority of the farmers interviewed prefer firmness and extended shelf-life in their new tomato varieties. Though past plant breeding efforts in tomato have focused on morphological and molecular diversity studies, screening against biotic and abiotic stresses [5], breeding objectives must target other traits that may be of benefits to a wide array of end-users. Current efforts have targeted breeding for extended shelf-life through incorporation of genes from wild relatives [21, 45, 55, 56]. Development of an early maturing tomato varieties was achieved through hybridization of cherry tomato and Pectomech, a popular commercial variety [27].

In order to meet the current market demands and changing climate, there is the need to design and develop new tomato varieties that meet the requirements of different clients. As must-have traits, all new tomato varieties must be resistant to common pests and diseases such as whiteflies (*Bemisia tabaci*), tomato leaf miner (*Tuta absoluta*), bacterial wilt, Fusarium wilt as well as resilient to the prevailing

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

environmental conditions such as heat and drought. For home consumption, the new tomato varieties must be rounded in shape with large fruit sizes and red in colour. In order to meet the industrial market, there is the need to develop new varieties with high pulp content and/or brix for good paste production. To facilitate rapid adoption by farmers, the new varieties need to be resistant to most biotic stresses that prevail in most of the growing ecologies.

#### **4. Fast track/speed breeding for demand-led tomato varieties**

Speed Breeding involves manipulation of light, temperature, plant population and application of single seed descent method to identify major traits [57, 58]. Various methods which have been used to improve the cycle of turnover and are extensively classified as speed breeding (SB). SB is exploited in several tomato breeding programmes involving population generation, pyramiding traits, phenotyping, assessment of agronomic traits, genomic selection and genomic editing [58, 59]. Majority of the SB approaches target improvement in the tomatoes for fruit quality, fruit yield, and tolerant to stresses. Tomato is a model crop of the Solanaceae family that supports SB due to it short maturity period, diploid genome, convenience of Agrobacteriummediated transformation thus supporting mutation breeding, CRISPR-Cas9 application [60, 61]. SB strategies used in developing demand-led tomato varieties such as Flavr Savr in America as the first engineered tomato by biotechnology method [62]. SB strategies such as marker-assisted selection, participatory plant breeding, mutation, and clustered regularly interspaced short palindromic repeat (CRISPR-/Cas9) system could be used by Africans in developing demand-led tomatoes. There is little information as to a tomato variety developed in Africa via the SB strategies. This implies that African tomato breeders must take advantage of current breeding tools.

#### **4.1 Marker assisted breeding**

Marker-assisted selection (MAS) and marker assisted breeding (MAB) progresses the effectiveness of crop improvement via accurate transfer of genomic sections of significance and hasten the recovery of the recurrent parent genome. The application of MAS and MAB support genomic selection which rely on molecular markers in assisting crop breeding. MAS in tomato improvement is traced around the 1930s [63]. MAS have been used to improve the traits that are related to disease, morphological, and physiological in most crops [59]. Specifically, in tomatoes, integration of SB into MAS results into transfer of beneficial alleles. For instance, SB and genomic led to purify tomato hybrids [64]; identify heat tolerant tomatoes [20, 65, 66].

The achievement of MAS is highly dependent on several critical issues including the number of target genes to be transferred, the distance between the target gene and the flanking markers, number of genotypes selected in each breeding generation, the nature of germplasm and the technical options available at the marker level. The power and efficacy of genotyping are anticipated to develop with the advent of markers like single nucleotide polymorphisms (SNP).

#### **4.2 Participatory plant breeding**

When farmers and other actors are involved in a breeding programme to either to participate or collaborate with scientists in every stage of the breeding programme is

termed as a participatory plant breeding (PPB) [67]. PPB is recommended as a one of the breeding approaches that enhance crop improvement, empower and promote farmers right, and increases the acceptance rate when new varieties and or technologies are introduced to farmers [68–70].

Tomato is an essential crop to Africans, hence there is continual increase in demand. Therefore, PPB serves to intensify farmers access and openness to breeding of new tomatoes in Africa. Thus, PPB will aid in rapid improvement and delivery of farmers and customers preferred tomato variety [71]. In Africa, the smallholder farmers serve as the foundation for the food system [72] PPB is observed to be useful to the smallholder farmers, especially in Africa. Application of PPB strategies in Tanzania assisted in evaluating and releasing a tomato variety resistant to late blight resistant [73] PPB has the potential to improve farmers preferred tomato but it has not been fully utilized. Hence, the various actors in the tomato breeding must take full advantage of PPB.

#### **4.3 Mutation**

Mutation is among the efficient approaches for enhancing crop traits without changing the well-optimized genomic background of the crop. Mutation is used to study variation and traits of interest for improving fruit quality, male fertility and disease resistance [74, 75].

For instance, tomato variety M82 through mutation developed mutant line with a variant of eIF4E 1 showed resistance to the potyvirus strains [76]. Tomato fruits are known to be influenced by *rin* (ripening inhibitor) and PL gene that relates to fruit softening. Mutating of *rin* and silencing of PL gene results in delaying ripening. However, *rin* reduced nutritional components, flavour and colour of the fruits but PL gene had recorded otherwise [77, 78]. Tomato mutant iaa9-3 line is capable of developing parthenocarpic variety that would be seedless and of high quality [79]. Mutation resulted in generating heat-tolerant tomatoes which exhibited high pollen fertility and fruit set [80, 81]. These are evidence which support the contribution of mutation breeding to the tomato industry around the globe. Tomato breeding in Africa has not witnessed the anticipated progress hence breeders should be encouraged to use mutation.

#### **4.4 Clustered regularly interspaced short palindromic repeat (CRISPR-/Cas9) system**

Gene editing technologies (zinc finger nuclease—ZFN; transcription activatorlike effector nucleases—TALENs; and clustered regularly interspaced short palindromic repeat—CRISPR/Cas) applications have witnessed some successes in some crops within the Solanaceae family [82]. These technologies aim at modifying genome by generating novel desirable alleles that will foster the improvement and subsequent release of a new varieties and/or augmenting the genetic pool of desired alleles. The CRISPR/cas9 is the desired technology due to its high specificity and low cost [83]. Gene editing in tomatoes has been successful due the availability of its genome sequence (https://solgenomics.net/organism) and its annotation [84]. Thus, resulting in several research relating to improvement to stresses (biotic & abiotic), fruit quality (nutritional value, shelf life, & colour), and plant architecture [85].

CRSPR-Cas9 was first used in tomatoes in generating the first needle-leaf mutant in 2014 by knocking out *Argonaute 7* [86]. Most products produced by *Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

gene editing have no commercial value [87, 88] but tomato is an exception [61]. Application of gene editing in tomato by Mlo1 mutant showed resistance to powdery mildew. Then again, through selfing at T0, a mutant of mlo1 T-DNA was achieved [89]. Tomato is more sensitive to fusarium wilt disease by knocking out Solyc08g075770 by CRISPR-Cas9 [90]. Similarly, tomato bzr1 mutant via CRISPR reduced the production of hydrogen peroxide (H2O2) which improved heat tolerance in tomato [91]. Tomato plant's response to drought has been improved through gene editing technologies by manipulating CBF1 (C-repeat binding factor 1) and MAPK3 [92, 93]. CRISPR-Cas9 application have resulted in obtaining herbicideresistant tomatoes plants. A study by ref. [94] resulted in over 70% edited tomatoes plants exhibiting resistant to the pesticide chlorsulfuron. Similarly, CRISPR-Cas9 mutated carotenoid dioxygenase 8 (*CCD8*) and more Axillary Growth1 (*MAX1*) involved in the promotion of strigolactone synthesis, a key component required for the germination of *Phelipanche aegyptiaca* seeds thereby resulting in producing *Podalirius aegyptiaca*-resistant tomato plants [95, 96]. Fruit set in tomatoes is influenced by pollination and fertilization, a CRISPR/Cas9 via mutation developed parthenocarpic tomato which is attractive to farmers as it reduces labour cost of fruit setting [97].

There is numerous evidence that support that, CRISPR-/Cas9 system has the potential to facilitate SB in tomatoes. However, there is little evidence that show it application in Africa. Other researchers confirm that, agricultural production is low in Africa compared to other continents [98]. A study by ref. [14] recommended that Africa should embrace technologies such as CRISPER to develop novel crops such as tomato genotypes to sustain its production. It is time that Africa embrace modern breeding technologies that support SB for tomato industry to be sustained due to increase demand.

#### **5. Tomato seed system in sub-Saharan Africa**

Achieving food security is dependent on seed security as well as timely availability of quality seed in adequate quantity at the right price and time. This is very fundamental in increasing production and productivity. Population increase, depleted natural resources and extreme climate variability has worsened the problems of food security to help mitigate this problem, a functional seed system is needed. Seed systems include interrelated institutions that develop new cultivar, produce, test, certify and market the seed. An effective seed system has the potential to increase productivity in a marginal way as good quality seed alone has the potential of increasing yield of crop by up to 20–30% [99]. Tomato is a food security and high value crop that improves the livelihood and income among smallholder farmers in the sub-Saharan Africa. Farmer's access to high quality tomato seeds is the surest way to ensure a resilient tomato industry in Africa.

#### **5.1 Types of tomato seed system**

In Africa, farmers have different ways of obtaining their quantity and quality of seed which they need for production. Tomato seed system varies greatly depending on the locality, market availability and farmers knowledge on seed system and supply and can be basically grouped into formal, informal or the combination of the formal and the informal.

#### *5.1.1 Formal seed system*

Formal seed system in Africa is usually government supported with the active involvement of the public institutions (**Figure 1**). It is a holistic approach involving evaluations of genetic resources, breeding and the development of new materials, certification and distribution of the planting materials to farmers. The tomato seed system is not formalized in most parts of Africa but a form of semi-formal seed system where seed companies are involved in the distribution of imported tomato seed to farmers. In sub-Saharan Africa tomato seed is still imported from outside the continent, while local companies continue to produce seed of open-pollinated varieties. Countries like South Africa and Tanzania have a formal tomato seed system where both the government and the private sector (Seed companies) are involved in

**Figure 1.** *Schematic distribution of the formal seed system.*

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

the development and distribution of tomato varieties [100]. The first private seed company in Tanzania (Alpha seed) was established almost three decades ago. It was into the sale of open-pollinated tomato varieties developed by the World Vegetable Centre [100].

Currently, Tanzania can boast of about 25 vegetable seed companies involved in tomato seed production and is expected to grow from 25 million USD in 2018 to 65 million USD by 2023 [101]. In most African countries for example., Ghana and Nigeria, seed companies are involved in on-farm trials of breeding and participatory cultivar selection, seed- related research, seed multiplication, seed conditioning and quality assurance, repackaging of the seeds, storage and distribution to final end users in this case farmers [102]. Currently, in Ghana, a formal tomato seed system is about springing up due to the release of the first official tomato varieties by CSIR-Crops Research Institute, Kumasi, Ghana and West Africa Centre for Crop Improvement (WACCI) at the University of Ghana.

#### *5.1.2 Informal tomato seed systems*

The informal tomato seed system, otherwise termed as "local or farmers saved-seed system" account for about 80% of the seed stock [103] supply to the farmers. It is an unorganized system that includes identification, seed saving, seed exchange, production and distribution by the farmer according to his/her knowledge of the plant and it is highly localized (**Figure 2**). The informal tomato seed system in Africa apart from being farmer or community-based practices; also includes different local level seed production initiatives organized by either farmer group, non-governmental organization or both, working outside the formal regime of the organized seed sector. Other characteristics of the informal tomato seed sector within the sub-region includes the non-law regulatory system, farmer to farmer seed exchange and this deals with individual community with small seed quantities usually demanded by farmers [104].

In Africa, countries like Tanzania, Ethiopia, Mali, Burkina Faso, Nigeria, Kenya, and South Africa although have some kind of formal seed system for some vegetables but the tomato seed system is largely informal [24]. Farmers within these countries purchase the imported hybrid or open pollinated tomato seed from agro-dealers and seed companies to produce their own tomato seed. These seeds once they are produced are easily marketed and exchanged from farmer to farmer by irregular means for many seed generations. On the other hand, farmers after acquiring some seed production skills from the extension officers often develop their own seeds from hybrid tomatoes i.e., they advance it to the F2 themselves for local marketing. This is usually as a result of high cost of the F1 hybrids [104].

#### **5.2 Tomato seed value chain**

A seed value chain is a series of actors or stakeholders. The value chain involves input suppliers, producers and processors, to exporters and buyers engaged in the activities required to bring an agricultural product from production to end consumer through various actors as shown in **Figure 3**. A value chain, therefore, incorporates productive transformation and value addition at each stage of any value chain. At each stage in the tomato value chain, the product changes hands through chain actors, transportation costs are incurred, and generally, some form of value is added. Tomato value chain results from diverse activities including input

#### **Figure 2.**

*Schematic distribution of the informal tomato seed system.*

supply, production, transportation, marketing, processing, distributions, retailing, and consumption.

#### *5.2.1 Input suppliers*

Input suppliers are the producers of agricultural inputs such as seeds, pesticides, fertilizers, mulching sheets, etc. needed for the production of tomatoes. Through company owned, and other company dealers they sell their products to the farmers. Moreover, they also provide technical guidance on inputs usage and timely supply of inputs to the tomato farmers. They do maintain good relationships with the farmers and act as one of the informal sources of finance. Regarding the delivery of inputs like *Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

**Figure 3.** *Linkages and flow of tomato value chain in Africa.*

improved seed, herbicides and pesticides, and credit among others, public and private extension services provide extension services to the farmers.

#### *5.2.2 Producers*

They are the initial link in tomato value chain. Producers decide what to produce, how much to produce when to grow and sell. Three types of the production system can be observed viz., subsistence production, small-scale commercial production, and largescale commercial production. Subsistence production is carried out for

household consumption and produced in small quantities. The produce from the first category of farmers generally does not enter the market or enters in a very limited quantity especially in the local marketplace. Small- and large-scale commercial farmers sell most of their products to various market intermediaries. The producers generally deal with traders and wholesalers. In most cases, farmers depend on village level traders for price information.

#### *5.2.3 Marketing*

The main aggregators usually buy the initial tomato from the main farmers to a special location within the village where traders buy them and transport to desired markets. Such collection and transporting activities are carried out either by the local trader, or an outside trader regularly visiting the location.

#### *5.2.4 Wholesalers*

They usually depend on the various intermediate sized loads and put the tomatoes into large uniform units. These activities all contribute to price determination Wholesalers are market participants who buy large quantities of tomato and resell to other traders. Wholesalers often buy the tomatoes at the farm gate and other road site.

#### *5.2.5 Processors*

Processors are the secondary processing industries. The tomato processed products manufactured by the sample processors include tomato paste, sauce and ketchup. They usually collect fresh tomatoes from wholesalers and other sellers in major tomato production areas during peak season and glut in the market at cheaper prices.

#### *5.2.6 Distributors*

The distributors normally buy processed tomato products from processors and supply to small grocery stores and supermarkets. They generally sell products of different companies in different formats of retailers.

#### *5.2.7 Retailers*

They are the middlemen that include the supermarket and another large-scale retailer who divide large shipments of produce and sell it to consumers in small units. The basic function they provide is bulk breaking. Retailers are the sellers of tomatoes to the ultimate consumers through multiple channels such as small grocery stores, exclusive fruits and vegetable shops, supermarkets and exporters. They normally buy from wholesalers and sell both fresh tomatoes and other tomato processed products in smaller quantities with a higher profit margin. The retailers are the final connection that deliver tomato to consumers. They are many as compared to others and their function is selling tomato to consumers in small volumes after receiving large volumes from producers.

#### *5.2.8 Consumers*

It is the last link in the tomato market value chain. The consumers always make production meaningful and they usually set the pace for production. From the

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

consumers' perspective, the shorter the value chain the more likely the retail price going to be cheaper and affordable. The consumers are mostly classified into individual/household consumers and larger consumers like the restaurants, hotels and local food joints [105].

#### **5.3 Constraints in tomato seed production**

In Africa, the production and supply of most agricultural commodities is seasonal and tomato is not an exception [106]. This has contributed to the price adjustment of tomatoes based on the trend of supply [107] which follows the normal curve of demand and supply, as the demand increase the supply decreases and the reverse is true. In Ghana, Nigeria and most African countries where tomatoes production is considered as the game changer in both nutritional and in the economic sense, but yet cannot meet their production demand has for the past decades seen fluctuation in the rural wholesale price by a marginable percentage [108, 109].

This trend in price fluctuation of tomatoes in Africa has not only affected the quantity and quality of the tomatoes [110] but also affected the tomato seed production. The following are some of the challenges faced by tomatoes producers in the sub-region;


#### **6. Challenges and prospects of demand-led tomato breeding**

The demand-led breeding approach will not only increase development of new tomato varieties but rather meet the needs of changing market preferences. Below are few highlighted prospects and challenges of demand-led tomato breeding methods in Africa.

#### **6.1 Challenges**

The major challenge for demand-led tomato breeding is lack of adequate funding covering aspects of research, training of researchers and technical officers, establishment of tomato breeding infrastructures, etc. [111]. Researchers, governments and private investors should partner to strategize and examine the potential benefits, implementation and how to sustain demand-led tomato breeding.

In addition, there is inadequate genetic resources to meet demand-led tomato breeding programmes in Africa [112]. To improve future tomato breeding

programmes, countries in Africa should prioritize the collection and conservation of local landraces which possess useful agronomic genes to sustain future breeding programmes [113]. Overreliance of improved exotic tomato lines/cultivar in most African countries, however, makes demand-led tomato breeding programmes unsustainable. This is because, the imported tomato variety may not meet the actual needs of the actors in the tomato value chain.

Last but not least, well- resourced laboratories to explore modern techniques in crop improvement are lacking in Africa. Most African countries are lagging behind regarding the utilization of basic to advanced biotechnology techniques such as marker assisted selection, genetic engineering and genome editing to facilitate plant breeding programmes [14]. Demand-led tomato breeding programmes would require most of these above-mentioned techniques to reduce the time for variety development.

#### **6.2 Prospects**

Tomato breeding based on demand has the potential to gather and understand information about tomato value chain actors' preferences for single or multiple traits. In order to accomplish this, more comprehensive quantitative research methodologies will be required to identify well-informed preferred traits by tomato value chain actors. To develop excellent demand-driven tomato breeding, scouting for traits insights along the value chain is a key [49]. Furthermore, using market intelligence, a comprehensive quantitative breeding index to rank the preferred traits for demandled tomato breeding schemes can be developed. The "monetary values, preferences from all potential breeding clients, from the farm to the consumer's table" [49] may be the basis for trait ranking. As a result, this analysis will aid in determining trait improvement priorities and maximizing not only genetic gains, but also actual variety adoption, while ensuring that released varieties have traits that are preferred by all key value chain actors and stakeholders. This strategy will hasten the adoption of new tomato varieties.

Demand-driven tomato breeding can facilitate and harness the adoption of a customer- and data-driven approach that adds value to released tomato variety, whiles meeting the actual needs of a diverse range of customers and breeding clients [114]. Thus, involving various actors along the value chain especially in the early breeding process will promote early adoption, acceptance and consumption of newly released tomato varieties. As such, this breeding method will contribute enormously in achieving broader societal goals including increased food and nutrition securities concomitant with improved health, poverty reduction, climate resilience and environmental conservation.

Again, demand-led tomato breeding can help increase the economic value for targeted tomato traits. Since, traits of significant importance to various actors in the tomato value chain are known and ranked, economic value for specific or highly ranked traits could be improved. For instance, a change in dietary preferences and requirements has caused a shift for tomato fruits with improved quality such as fruits with high lycopene content [115] and sugar level [116]. A situation where consumers' preferred traits are successfully introgressed into a newly released tomato varieties, they will be willing to pay premium prices for these tomato fruits. Thus, the market value can be increased and then improve the livelihoods of various actors in the tomato value chain.

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

#### **7. Conclusion**

Tomato breeding programmes have focused on breeding for resistance to biotic and abiotic constraints which cause severe yield reduction. In addition to yield, tomato varieties are also bred for quality traits such as colour, firmness, flavour and extended shelf-life to meet consumer or industry preferences. Demand-led breeding which targets consumer and market preferences is based on six core principles which need to be considered in designing new tomato varieties Attributes desired by Ghanaian and tomato producers from other African countries considered in new variety designs include pests and diseases resistance, tolerance to environmental conditions as well as for quality traits and attributes such as shape, colour, and high pulp content.

Various fast-track breeding approaches are employed for rapid progress in tomato breeding. Speed breeding techniques reduce the long breeding cycle compared to conventional tomato breeding. Examples of such approaches include marker assisted breeding (MAB), participatory plant breeding (PPB), mutation breeding and clustered regularly interspaced short palindromic repeat (CRISPR-/Cas9) system which have been used to accelerate the breeding process in tomato. Two key seed systems are common in the tomato seed value chain. These are the formal system which is regulated, and the informal system which mainly operates in the rural areas. The tomato seed system is faced with challenges such as: inadequate government support, absence of seed laws and enabling environment, under-developed private sector among others. Challenges facing demand-led tomato breeding include lack of involvement of all stakeholders in the tomato value chain. Inadequate research infrastructure, few trained personnel and inadequate genetic resources. These have limited to scope of most tomato breeding programmes to meet consumer needs.

Finally, demand-led tomato breeding has the potential to gather relevant information on the preferences of tomato value chain actors to inform breeders on what product profile to develop. Comprehensive quantitative research methodologies will help identify the requisite traits that would be preferred by current and future markets. Demand-driven tomato breeding can help breeders to design products that have the potential of satisfy consumer needs and facilitate rapid adoption by farmers and other end-users. Adequate funding, governmental support and active collaboration between researchers, private investors and farmers, and education on the potential benefits demand-led breeding are needed to implement and sustain demand-led tomato breeding.

### **Author details**

Hans Adu-Dapaah1 \*, Michael Kwabena Osei<sup>2</sup> , Joseph Adjebeng-Danquah3 , Stella Owusu Nketia4 , Augustine Antwi-Boasiako2 , Osuman Alimatu Sadia2 , Peter Ofori Amoako4 and Richard Agyare3

1 CCST-CSIR College of Science and Technology, Kumasi, Ghana

2 CSIR-Crops Research Institute, Kumasi, Ghana

3 CSIR-Savannah Agricultural Research Institute, Nyankpala, Ghana

4 College of Basic and Applied Sciences (CBAS), University of Ghana, Legon, Ghana

\*Address all correspondence to: hadapaah@yahoo.com

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

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*Perspective Chapter: Accelerating Demand-Led Tomato Breeding for Emerging Markets in Africa DOI: http://dx.doi.org/10.5772/intechopen.106737*

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#### **Chapter 11**

## Recovery and Valorization of Tomato By-products in R&D EU-Funded Projects

*Marcello Casa and Michele Miccio*

#### **Abstract**

In the last years, the European Commission has been funding numerous projects regarding the valorization of food wastes. Tomato by-products received great attention especially in Spain, Italy, Greece, and Portugal due to high volumes and high concentration of valuable compounds. Among 40 funded projects about the management of tomato wastes in general, 14 projects are strictly connected to the valorization and exploitation of the tomato residues/by-products after processing and are of great interest for their scientific, technical, and economical outcomes. They received an overall budget of around 37 M€ over 35 years, involving 20 European and 4 non-European countries, with project coordinators located in Germany, the Netherlands, and Italy in most of the cases. This chapter delivers general information about these projects, assessing and reporting scientific and technical results. Moreover, the interconnection is highlighted among them by focusing on the contribution they gave to the European know-how, the management of the by-products and the progress they reached in waste minimization and valorization. Finally, the industrial and environmental outcomes of these projects have been reported by highlighting issues and problems that are still to be overcome.

**Keywords:** tomato by-products, waste valorization, European Union, funded projects

#### **1. Introduction**

In the last years, the European Commission has been funding projects regarding the valorization of food wastes. Tomato by-products received great attention especially in Spain, Italy, Greece, and Portugal due to high volumes and high concentration of valuable compounds. Among 40 funded projects about the management of tomato wastes in general, 14 projects are strictly connected to the valorization and exploitation of the tomato residues/by-products after processing and are of great interest for their scientific, technical, and economical outcomes. They received an overall budget of around 37 M€ over 35 years, involving 20 European and 4 non-European countries, with project coordinators located in Germany, the Netherlands, and Italy in most of the cases. This chapter delivers general information about these projects, assessing and reporting scientific and technical results. Moreover, the interconnection is highlighted among them by focusing on the contribution they gave to the European expertise, the

management of the by-products and the progress they reached in waste minimization and valorization. Finally, the industrial and environmental outcomes of these projects have been reported by highlighting issues and problems that are still to be overcome.

#### **2. Funded projects**

The Community Research and Development Information Service (CORDIS) [1], namely the European Commission's primary source of results from the projects funded by the EU's framework programs for research and innovation, was used to gather all information such as project factsheets, participants, reports, deliverables, and links to open-access publications about tomato by-products valorization. In the first instance, from research in this database, it came up that on 352 funded projects including the keyword "TOMATO" only 10% take into consideration wastes or by-products produced by harvesting, transformation, and use of this vegetable. In particular, the research on CORDIS with "TOMATO" and "WASTE" as keywords gives forty projects as a result. Other searches with other keywords were conducted with less significant results: for example, "TOMATO" and "VALORIZATION" give 9 projects as a result, or "TOMATO" and "RESIDUE" return 23 projects as a result. As it is possible to see from **Figure 1** the number of funded projects in this field of application had a strong increase in the last 5 years, probably due to the growing interest, shown by academia and industries, in waste reduction, valorization of materials so far considered as undesirable by-products, and exploitation of the high-value compounds contained in these waste streams.

Then, these forty projects were deeply studied, and it was possible to divide them into eight categories regarding the topic:


• Production of biogas from residues

#### **Figure 1.**

*Distribution during last years of funded research projects on tomato waste.*

*Recovery and Valorization of Tomato By-products in R&D EU-Funded Projects DOI: http://dx.doi.org/10.5772/intechopen.106768*


**Figure 2** reports a bar chart of the number of projects per field of application. Among these, only fourteen projects are strictly connected to the valorization and exploitation of the tomato residues/by-products after transformation processes. In **Table 1**, the main information is reported about these projects of interest, sorted by topic.

#### **Figure 2.**

*Number of projects per field of application.*



#### **Table 1.**

*Main information about funded European projects on valorization and exploitation of tomato wastes.*

The information reported in the previous table was analyzed and summarized in the next chart to synthetically show the distribution of budget and participants among the considered application categories (**Figure 3**).

The overall budget is around 40 M€ involving 20 European and 4 non-European countries, with project coordinators located in Germany, the Netherlands, and Italy in most cases. It is worth notice that the field of biorefining, the one in which this thesis is involved, even if it is not the one with the highest number of the funded project, exhibits the highest budget and is the one with more partners involved. It is so probably because, even if the application of the biorefinery concept to tomato residual

#### **Figure 3.**

*Distribution of budget and participants among the considered application categories.*

by-products is quite new, the European Commission believes that research in this field could strongly increase the EU technological level. In the next paragraph, the outcome of these projects will be reported and briefly discussed.

#### **2.1 Early projects**

Projects funded before 2001 lack results reports, for different reporting policies of the European Commission. Anyway, the project QLK1-CT-2000-2041,137 had likely as an outcome a patent EP1676888B1 entitled Method of obtaining lycopene from tomato skins and seeds [2], assigned to *Conservas Vegetales de Extremadura* SA, which was the coordinator of the project. The patent refers to a process for obtaining lycopene from tomato skins and seeds. The carotenoid is obtained after a series of steps of dehydration, seed separation, pelletization, extraction, distillation, and crystallization. The extraction solvent is hexane and the purity of the lycopene obtained is between 65% and 85%, depending on the raw material.

#### **2.2 TOM**

The title of the project was "Development of new food additives extracted from the solid residue of the tomato processing industry for the application in functional foods." Partners of the TOM project had developed and optimized an extraction process whereby lycopene is extracted in tomato seed oil from tomato plant processing residue. This can then be used in functional food products and cosmetics. The carried-out process involves the use of supercritical carbon dioxide (CO2) [3]. The yield in tomato seed oil is 3–6%. The lycopene yield depends on raw material and ranges between 15 and 180 ppm, which is very low considering the extraction yield nowadays.

#### **2.3 Bioactive-net**

The title of the project was "Cultivation and processing of tomato, olive, and grape are the main agricultural businesses in the South European countries. Production of tomato paste, olive oil, and grape" and the main objectives of the project were:


Remarkable was the study on the best available technologies (BATs) to separate vitamins, antioxidants, essential oils, and other valuable compounds from the processing residues. In *Guida pratica sui COMPOSTI BIOATTIVI ottenibili dai SOTTOPRODOTTI della TRASFORMAZIONE DEL POMODORO* they reported the main technologies available for: residues drying, lycopene extraction, and lycopene purification. Moreover, an economic assessment that compares solvent and supercritical extraction for this compound was reported [4]. The report clearly shows from an economic and technological point of view that supercritical CO2 is rarely favorable, while solvent extraction is profitable only when a high amount of tomato by-products is processed.

#### **2.4 Lycosol**

The title of this 2019 project is "Feasibility Analysis on the Extraction of Lycopene from Tomato Peel through Organic Synthesis." LycoSOL project proposes an environmentally friendly solution based on natural ingredients. The method involves extracting and processing healthy ingredients from the waste from food processing. The project aims to develop the process of extraction and encapsulation from plant waste, targeting production from tomato peels. No results reports or scientific papers have been already disseminated.

#### **2.5 Pro-enrich**

The title of this 2018 project is "Development of novel functional proteins and bioactive ingredients from rapeseed, olive, tomato and citrus fruit side streams for applications in food, cosmetics, pet food." Pro-Enrich was aimed at optimizing existing biomass fractionation technologies and validating novel extraction approaches beyond the current state of the art with reference to the Technology Readiness Level (TRL) assessment system (i.e., from TRL2 through to TRL 4/5) to isolate and purify proteins, polyphenols, and dietary fibers and pigments. The products being targeted are food ingredients, pet food, cosmetics, and adhesives. These were to be developed through an iterative process of feedstock mapping, laboratory process development, functionality/performance testing of samples by upscaling to pilot plant and industry level. Rapeseed, tomato peels and citrus waste were studied in the project. First, a review paper on waste composition and edible protein extraction for the selected feedstock was published [5]; then, a first pilot plant for protein production from rapeseed was started [6]; finally, the following bioactive ingredients were successfully extracted, and are waiting for Scale-up to demonstration scale:


*Recovery and Valorization of Tomato By-products in R&D EU-Funded Projects DOI: http://dx.doi.org/10.5772/intechopen.106768*

Food: animal-based protein. Pet food: animal-based protein. Adhesives: petrochemically derived phenolics up to 40%.

#### **2.6 BIOCOPAC**

The title of the project is "Development of bio-based coating from tomato processing wastes intended for metal packaging." BIOCOPAC initiative looked at tomato by-products to satisfy some of these needs. The goal was to develop a natural lacquer liner for tins that are made from the cutin raw material contained in discarded tomato skins. The coating was aimed to be applied to internal and external surfaces of food tins to ensure consumer health and safety. The next step was to develop the bio-resin and the lacquer. Scientists developed two different formulas to produce the lacquer, one specifically designed for tinplate and a generic one for all types of metal can. BIOCOPAC produced canned goods using these lacquers, demonstrating that the lacquer performs as well as current products. An interesting outcome of the project is a Life Cycle Assessment (LCA) conducted using the SimaPro software, version 7.1. The analyses compared the LCA of a conventional epoxy-based lacquer to a bio-lacquer, tomato cutin based, obtained from tomato processing waste. The results showed clear environmental benefits of the "Bio-lacquer." The benefit of the cutin lacquer lies in the saving of natural resources and the recovery of part of the skins. This can lead to lower consumption of fossil fuels and lower CO2 emissions.

BIOCOPAC project merged with the BIOCOPAC+ project, funded under LIFE+ Environment Policy and Governance project application (Grant Agreement No. LIFE13 ENV/IT/000590). The project was started on the 1st of June 2014 and lasted for 36 months. The project was industry-driven and focused on demonstration activities aimed to prove the technical feasibility and effectiveness of the cutin extraction and production systems currently developed at a laboratory scale. Its outcomes were a prototype pilot plant for cutin extraction, installed at *Azienda Agricola Virginio CHIESA* (IT) and a cutin-based lacquer production site in SALCHI (IT) plant [7].

#### **2.7 BIOPROTO**

The project title is "Bioplastic production from tomato peel residues." The team investigated the possibility of creating a bioplastic film from discarded tomato skins. The idea proved feasible, yielding scalable and biodegradable options for food packaging. Results yielded a new set of films and coatings taken from the lipid portion of plant cuticles, reported in **Figure 4**. The outcome also represented a potentially scalable and cheap process for the manufacture of bioplastics intended for use in food packaging. BIOPROTO's new plastic was biodegradable, with minimal environmental impact [8].

#### **Figure 4.** *Photographs of bioplastic made by tomato cuticle during the BIOPROTO project.*

#### **2.8 ECOFUNCO**

The tile project is "ECO sustainable multi-FUNctional biobased COatings with enhanced performance and end-of-life options." The overall objective of project ECOFUNCO [9] was to select, extract and functionalize molecules (proteins, polysaccharides, cutin) from highly available, low valorized biomass such as tomato, legumes, sunflower, etc. for the development of new bio-based coating materials to be applied on two different substrates (i.e., cellulosic and plastic-based), with improved performances compared to currently available products and at the same time with the more sustainable end of life options. The products to be developed in the project were in particular:


The ECOFUNCO project final event has been taken on June 17–18, 2022 in the form of "1st Conference on Green Chemistry and Sustainable Coatings."

The event confirmed that the ECOFUNCO project developed sustainable biobased and compostable coatings to be applied on bioplastics and cellulose substrates to reach the same properties as fossil-based packaging materials. Also, the use of nanofibrils to add antimicrobial properties to tissues has been demonstrated.

The ECOFUNCO coordination and management demonstrated a great mind opening and a forward-looking sensitivity as they dedicated a session of the final conference to other related EU-funded projects, that is, FISH4FISH, PRESERVE, RECOVER, PROLIFIC, and Agrimax, in order to foster the cooperation between European projects.

#### **2.9 Refresh**

The title of the project is "Resource Efficient Food and dRink for the Entire Supply cHain." The overall aim of the REFRESH project was to significantly contribute toward the objective of reducing food waste across the EU by 30% by 2025 and maximizing the value from unavoidable food waste and packaging materials. The project aims to gather information about the main and most present food waste in the European countries, find the known way to exploit these by-products, and create a simplified tool to help the decision-maker to valorize at best these side streams, both in terms of economic feasibility and environmental impact. Tomato by-products are one of the considered waste streams. The project outcomes are 6 scientific publications regarding food waste, from their management to their reduction, a website and a software tool [10]. One of the main outcomes of the REFRESH project is a deliverable with the TOP20 waste streams in Europe, carefully reporting


#### **Figure 5.**

*Tomato pomace is one of the TOP20 food wastes in Europe.*

their current management and the reason for selection. Tomato by-products are in the list (see **Figure 5**).

Another main outcome is FORKLIFT, a spreadsheet learning tool that applies a partial lifecycle greenhouse gas impact and costing calculation approach for six key examples of unpreventable food processing co-products, by-products, or wastes (collectively referred to as *side flows*):


FORKLIFT allows users to interpret the results of the effects of intervention while making it possible to compare the results with alternative products available on the market [11]. For tomato pomace conventional solutions for its exploitation were selected and modeled in the FORKLIFT® tool, allowing for evaluation via LCA and LCC, cost and CO2 emission for different scenarios of valorization, and to easily compare them as a support to decision making. **Figure 6** shows the interface of the tool. In the analysis of tomato pomace, the following valorization routes were considered:


#### **Figure 6.**

#### **Figure 7.**

*FORKLIFT output for lycopene production.*

For example, with this spreadsheet is possible to compare lycopene production cost and emission with carotenoid production from microalgae (**Figure 7**).

#### **2.10 AGRIMAX**

The project title is "Agri and food waste valorization co-ops based on flexible multi-feedstocks biorefinery processing technologies for new high added value applications." The goal of the project was to extract the significant amounts of valuable compounds contained in food industry wastes, AgriMax [12] combined affordable and flexible processing technologies for the valorization of side streams from the horticultural culture and food processing industry to be used in a cooperative approach by local stakeholders. The project merged previous knowledge and outcome of other European projects, such as cutin extraction and exploitation studied in the BIOCOPAC project. LCA and LCC studied the best approach to minimize the environmental impact of the new value chains. Moreover, a pilot multi-feedstock bio-refinery process was set up at two demonstration sites in Spain (Pilot Plant at Indulleida S.A.) and Italy Pilot Plant (at Chiesa Virginio EC). Currently, the Italian pilot plant is valorizing the tomato by-products, producing cutin bioplastic, a small amount of lycopene and compost. The pilot plant flowsheet is reported in **Figure 8**.

*Recovery and Valorization of Tomato By-products in R&D EU-Funded Projects DOI: http://dx.doi.org/10.5772/intechopen.106768*

**Figure 8.**

*Flowsheet of Italian pilot plant located in the factory of Azienda Agricola Virginio Chiesa, Canneto Sull'Oglio (MN), Italy.*

#### **3. Conclusion**

In conclusion, 11% of funded European projects having tomato as a topic are dealing with tomato wastes and by-products. Forty projects were found when searching CORDIS with "tomato" and "waste" as keywords; 14 regard by-products valorization, categorizable in the following topics: production of bioplastic or biofilm, extraction of high-value compounds, preparation of food additives or fodder, biogas production via fermentation and biorefining of tomato by-products. The overall budget, that European Commission furnished to the participants, has been around 40 M€ in about 35 years. These projects involved 130 participants coming from all over the world. Extraction of compounds is the topic of most projects, but the highest budget has been awarded to biorefining. This is also the main focus of the research activities first explored and then directly pursued by the authors [13, 14] of this chapter. Projects on extraction technology development had as an outcome the optimization of commercial techniques, leading to patents; moreover, some studies showed that supercritical CO2 is never economically feasible for lycopene extraction. PRO-ENRICH is the only project about food additives that were recently found, to start a pilot plant for protein production from different waste streams, including tomato pomace. In the last years, bioplastic production from tomato by-products received great attention and funding, leading a pilot plant in Italy to produce metal packaging cover with a biofilm obtained from tomato peels. Recent projects (AGRIMAX and REFRESH) aim to best exploit food waste, making recourse to a biorefining approach. Main problems remain in the tomato by-products valorization: the high economic or environmental cost of lycopene extraction, as also underlined [15] by the authors of this chapter; the absence of a 'green' alternative for cutin extraction, and the difficulty in finding a biomass

similar to tomato pomace in order to overcome the seasonality issue. Moreover, a lack of data, studies, and projects on energy recovery from tomato by-products was evidenced by the present survey.

#### **Acknowledgements**

This work has been carried out in the frame of the "Training on-the-job" initiative (*Dottorati di ricerca con caratterizzazione industriale*) funded by Regione Campania (DGR No. 156 21/03/2017 - POR Campania FSE 2014/2020 - *obiettivo specifico 14 azione* 10.4.5) in Italy.

#### **Author details**

Marcello Casa and Michele Miccio\* Department of Industrial Engineering, University of Salerno, Fisciano, Italy

\*Address all correspondence to: mmiccio@unisa.it

© 2022 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*Recovery and Valorization of Tomato By-products in R&D EU-Funded Projects DOI: http://dx.doi.org/10.5772/intechopen.106768*

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[2] García JE. Method of obtaining lycopene from tomato skins and seeds, EP1676888B1, Oct. 24, 2012. Accessed: Oct. 14, 2020. Online: https://patents. google.com/patent/EP1676888B1/en

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[6] Pro-Enrich project. Available from: https://www.pro-enrich.eu. [Accessed: July 21, 2022]

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[15] Casa M, Miccio M. Lycopenecontaining tablets production from tomato peels by environment-friendly extraction: Simulation and discussion. Chemical Engineering Transactions [Internet]. Jun 30, 2022;**92**:583-588. Available from: https://www.cetjournal. it/index.php/cet/article/view/ CET2292098 [cited 6 Sep. 2022]

### *Edited by Pranas Viškelis, Dalia Urbonavičienė and Jonas Viškelis*

The tomato is a valuable vegetable, popular all over the world. This book covers interesting research topics including tomato plant nutrition, production and chemical composition, tomato plant protection, and sustainable tomato processing technologies. This book will be of value to researchers, academics, and students in the field of agronomy, food, pharmacy, and other sectors.

Published in London, UK © 2022 IntechOpen © zahra solgi / iStock

Tomato - From Cultivation to Processing Technology

Tomato

From Cultivation to Processing Technology

*Edited by Pranas Viškelis,* 

*Dalia Urbonavičienė and Jonas Viškelis*