*2.1.2 Initial plant growth of common bean (Phaseolus vulgaris L.) variety Quivicán plants*

The common bean (*Phaseolus vulgaris* L.) is one of the edible legumes of greater consumption worldwide, providing an important source of proteins, vitamins and minerals to the diet of populations in developing countries in the Americas [37]. For this reason, research development has focused on improving crop response variables to increase their agricultural production. Thus, the objective of this study was to assess homoeopathic medicines to promote the overall performance and productivity of *P. vulgaris* L. (white-tinted Quivicán variety) during the stages of initial plant growth in pots with substrate, within cultivation houses with shadow mesh at Centro de Investigaciones Biológicas del Noroeste (CIBNOR), La Paz, Mexico.

**Test 1.** Certified seeds of this Quivicán variety (Seed Company, Villa Clara, Cuba) were used; a completely randomised design was used with three HOM treatments, T1 (MgM-31CH), T2 (MgMnP-3CH) and T3 (MgM-31CH + MgMnP-3CH), and a control with distilled water (DW), each one with four replicates. This

**163**

*Agricultural Homoeopathy: A New Insight into Organics DOI: http://dx.doi.org/10.5772/intechopen.84482*

**Table 1.**

*(Phaseolus vulgaris L.) variety Quivicán.*

experiment was developed to evaluate initial growth phase. Seeds were planted (five replications per treatment) in plastic pots (3 seeds/pot) with 5 kg of commercial substrate (Sogemix PM®). Emergence evaluation was done daily; when 50% + 1 of the seeds emerged, 1 ml of the respective HOM treatment and water for the control (NT) was applied around the stems of the plants on alternate days. After 35 days, we proceeded to measure stem length (SL), root length (RL), fresh biomass (g)

*Effect of HOM treatments on morphometric parameters during growth stage of the common bean* 

(SD) (mm) and number of leaves (L No). The yields corresponding to dry biomass (DRB, DLB, DSB) were also measured. The most relevant result of the study was highly favourable and statistically significant effect of HOM treatment T3 with

The cause-effect results in the plant model demonstrated a synergistic effect of the homoeopathic medicines included in HOM treatment T3, which evidently could not have been a placebo effect because it clearly favoured the growth of *P. vulgaris* variety Quivicán. Magnesium and manganese were components of T3; both are essential for the growth of any living cell and necessary secondary macronutrients for plant growth and development. Around 75% of foliar magnesium was involved in protein synthesis, and 15–20% of total magnesium was associated with pigments, a constituent element of the chlorophyll molecule and extremely important in photosynthesis [38]. Magnesium acts mainly as a cofactor of several enzymes involved in photosynthetic carbon fixation and also in basic metabolism [39]. Both magnesium and manganese play an important role in plant general nutrition and enhance or reinforce their resistance to diseases [40]. The results obtained in this study suggest that agricultural homoeopathy has application in the cultivation of the common bean (*Phaseolus vulgaris* L.) variety Quivicán, during the stages of initial plant growth.

Currently, numerous studies have focused on elucidating the negative effects of abiotic stress on agricultural crops. Saline stress associated with high temperatures and solar irradiation is the most important environmental process that stops cultivated plant growth, development and survival, decreasing productivity [41]. Worldwide, millions of hectares have a high degree of aridity and the presence of salts [42]. In Mexico, arid and semiarid regions constitute more than 50% of the national territory [43], so the study of these factors is important due to the negative impact they have on the agricultural sector. Salinity has environmental, social and economic consequences because sustainability and yield of the cultivated species decrease in the affected areas. Salinity affects plant metabolism and growth, causing a decrease in biomass production [44]. Global losses due to salt stress are estimated at 12 billion dollars per year and affect a fifth of the farmland [45]. Among the main harmful effects, decrease in water absorption, ion assimilation

), stem diameter

of root, stem and leaves (FRB, FSB, FLB), foliar area (FA) (cm2

respect to the untreated control group (**Table 1**).

**2.2 Attenuation of the effects of abiotic stress**


### **Table 1.**

*Multifunctionality and Impacts of Organic and Conventional Agriculture*

(≤2 and ≤23%, respectively). In general, most of the HOM treatments had a stimulating effect during the emergence stage (**Figure 2**) since significant differences were recorded in the morphometric variables evaluated. During germination, the length of stem (SL) was greater using T3 (5.5 cm) with respect to the control (4.3 cm). With HOM treatment T2, the highest root development was obtained with a length of 8.2 cm compared to the control (5.6 cm). Significant differences were observed in dry biomass of stem and root obtaining higher dry stem biomass with T3 and T5 (0.01 g) than that obtained with control and other HOM treatments (0.0027 g). When T1 was applied, a higher dry root biomass (0.002 g) was observed with respect to control (0.0005 g), and other HOM treatments (≤0.0013 g) were assessed. During the emergence stage, significant differences were found regarding stem length with HOM treatment T2 (6.6 cm), and with T3 a similar growth was obtained (5.9 cm). The length of the stem was smaller with the rest of the treatments (≤5.8 cm) but, even so, higher than the control (4.6 cm). Regarding root length, no significant differences were found; however, the seedlings treated with T4 and T7 had the highest root growth (4.5 and 4.6 cm, respectively) beyond the control (3.3 cm). Finally, with regard to biomass, no significant differences were

*Effect of HOM treatments on tomato (Solanum lycopersicum) emergence (% and rate).*

*2.1.2 Initial plant growth of common bean (Phaseolus vulgaris L.) variety Quivicán* 

The common bean (*Phaseolus vulgaris* L.) is one of the edible legumes of greater consumption worldwide, providing an important source of proteins, vitamins and minerals to the diet of populations in developing countries in the Americas [37]. For this reason, research development has focused on improving crop response variables to increase their agricultural production. Thus, the objective of this study was to assess homoeopathic medicines to promote the overall performance and productivity of *P. vulgaris* L. (white-tinted Quivicán variety) during the stages of initial plant growth in pots with substrate, within cultivation houses with shadow mesh at Centro de Investigaciones Biológicas del Noroeste (CIBNOR), La Paz, Mexico. **Test 1.** Certified seeds of this Quivicán variety (Seed Company, Villa Clara, Cuba) were used; a completely randomised design was used with three HOM treatments, T1 (MgM-31CH), T2 (MgMnP-3CH) and T3 (MgM-31CH + MgMnP-3CH), and a control with distilled water (DW), each one with four replicates. This

observed between HOM treatments and the control.

**162**

*plants*

**Figure 2.**

*Effect of HOM treatments on morphometric parameters during growth stage of the common bean (Phaseolus vulgaris L.) variety Quivicán.*

experiment was developed to evaluate initial growth phase. Seeds were planted (five replications per treatment) in plastic pots (3 seeds/pot) with 5 kg of commercial substrate (Sogemix PM®). Emergence evaluation was done daily; when 50% + 1 of the seeds emerged, 1 ml of the respective HOM treatment and water for the control (NT) was applied around the stems of the plants on alternate days. After 35 days, we proceeded to measure stem length (SL), root length (RL), fresh biomass (g) of root, stem and leaves (FRB, FSB, FLB), foliar area (FA) (cm2 ), stem diameter (SD) (mm) and number of leaves (L No). The yields corresponding to dry biomass (DRB, DLB, DSB) were also measured. The most relevant result of the study was highly favourable and statistically significant effect of HOM treatment T3 with respect to the untreated control group (**Table 1**).

The cause-effect results in the plant model demonstrated a synergistic effect of the homoeopathic medicines included in HOM treatment T3, which evidently could not have been a placebo effect because it clearly favoured the growth of *P. vulgaris* variety Quivicán. Magnesium and manganese were components of T3; both are essential for the growth of any living cell and necessary secondary macronutrients for plant growth and development. Around 75% of foliar magnesium was involved in protein synthesis, and 15–20% of total magnesium was associated with pigments, a constituent element of the chlorophyll molecule and extremely important in photosynthesis [38]. Magnesium acts mainly as a cofactor of several enzymes involved in photosynthetic carbon fixation and also in basic metabolism [39]. Both magnesium and manganese play an important role in plant general nutrition and enhance or reinforce their resistance to diseases [40]. The results obtained in this study suggest that agricultural homoeopathy has application in the cultivation of the common bean (*Phaseolus vulgaris* L.) variety Quivicán, during the stages of initial plant growth.

## **2.2 Attenuation of the effects of abiotic stress**

Currently, numerous studies have focused on elucidating the negative effects of abiotic stress on agricultural crops. Saline stress associated with high temperatures and solar irradiation is the most important environmental process that stops cultivated plant growth, development and survival, decreasing productivity [41]. Worldwide, millions of hectares have a high degree of aridity and the presence of salts [42]. In Mexico, arid and semiarid regions constitute more than 50% of the national territory [43], so the study of these factors is important due to the negative impact they have on the agricultural sector. Salinity has environmental, social and economic consequences because sustainability and yield of the cultivated species decrease in the affected areas. Salinity affects plant metabolism and growth, causing a decrease in biomass production [44]. Global losses due to salt stress are estimated at 12 billion dollars per year and affect a fifth of the farmland [45]. Among the main harmful effects, decrease in water absorption, ion assimilation

that can cause toxicity and nutritional imbalance and physiological changes, such as the reduction of the photosynthetic rate due to a lower leaf area, which reduces crop viability [46] are included.

Several alternatives have been studied to mitigate the effect of salinity in agriculture, such as genetic improvement, selection of tolerant varieties and physical treatments to seeds to induce tolerance or agrochemicals to stimulate plant growth. In these sense, HOM treatments imply the application of ultra-diluted substances, which are an eco-friendly and economic variant that could be used effectively in any condition and circumstance [47]. Unfortunately, this organic-like alternative has been little studied worldwide.

## *2.2.1 Salinity stress by NaCl in the bean crop (Phaseolus vulgaris L.)*

The common bean *Phaseolus vulgaris* L. is a key product in world food security [48], but the adverse environmental conditions, mainly drought and salinity in soils, affect their general performance reducing productivity and harvest [49]. Saline stress occurs due to high concentrations of sodium (Na+ ) and chlorine (Cl<sup>−</sup>) that seriously alter the plant metabolism, affecting its growth and development [50]. Water deficit is an osmotic stressing agent, specifically associated with salinisation, which reduces the rate of fixation of carbon dioxide (CO2) and affects processes associated with photosynthesis [51]. Abscisic acid (ABA) is a key hormone that regulates the responses of plants to abiotic stress [52] and initiates the activation of stomatal closure when facing salt stress. When it happens, CO2 levels decrease and consequently photosynthesis, causing oxidative stress [53]. The objective of this study was to evaluate HOM treatment NaM (7CH) as attenuator of the harmful effect of salinity induced by NaCl application. The experiment was developed at CIBNOR to assess the initial growth stage of the species and study the expression of the genes associated with physiological response of *P. vulgaris* against HOM treatments.

A completely randomised design was applied with HOM treatment T1 (NaM 7CH) and control (DW) each one with five replicates and two concentrations of NaCl (0 and 75 mM). Seeds of the variety white-tinted Quivicán (Empresa de Semillas, Villa Clara, Cuba) were used.

**Test 1.** The seeds were disinfected, then imbibed for 30 min in HOM treatment NaM 7CH or in DW (control treatment), planted in 5 kg plastic pots (3 seeds/ pot) with commercial substrate (Sogemix PM®) and grown for 35 days. Once the seeds germinated and the plants emerged, NaM 7CH or DW (15 ml) was applied on alternate days near the stem of each plant. The addition of NaCl began 15 days after germination and is applied gradually until reaching 75 mM to avoid osmotic shock.

The photosynthesis rate (A, μmol m<sup>−</sup><sup>2</sup> s<sup>−</sup><sup>1</sup> ) was measured with a LCpro-SD portable computer with a wide-blade camera (ADC, Hoddesdon, Herts, United Kingdom). Three measurements were made during the fourth week of the trial, on healthy leaves and on completely sunny days. At the begining of frlowering, a sample was taken to perform the expression analysis of the 9-cis-epoxycarotenoid gene (PvNCDE1) [54]. Total RNA isolation, cDNA synthesis and real-time PCR amplification were performed following the methodology reported by Morelos et al. [55], and the relative expression of PvNCDE1 was estimated following the model proposed by Hellemans et al. [56]. One-way ANOVA followed by Tukey's exact test was performed using STATISTICA 8.0 (StatSoft, Inc.). A *p*-value <0.05 was considered significant.

At the end of the study, photosynthesis showed a considerable decrease in the control (DW) when the plants underwent saline stress (**Figure 3**). In these stressed plants, stomatal closure was produced by ABA, generating a direct blockage of photosynthesis due to the limited uptake of CO2, an indispensable substrate for the Calvin cycle. When it happens, a blockage of the photo phase occurs due to the null

**165**

**Figure 3.**

*(p ≤ 0.05).*

**Figure 4.**

*Agricultural Homoeopathy: A New Insight into Organics DOI: http://dx.doi.org/10.5772/intechopen.84482*

demand of the reducing power and ATP, resulting in oxygen production with consequent damage to the chloroplast membranes and their subsequent disintegration [57]. Contrary to the above, in HOM-treated plants that were subjected in parallel to salt stress, photosynthesis increased and no significant difference *(p >* 0.05) was observed with respect to plants not subjected to salt stress by the addition of NaCl. This result suggests that plants receiving HOM treatment did not become stressed,

*Expression of the pvNCED1 gene of HOM-treated (NaM-7CH) bean Phaseolus vulgaris L., exposed to NaCl stress. Average values with different literals correspond to statistically different treatments (p ≤ 0.05).*

*Photosynthetic rate is recorded in Phaseolus vulgaris treated and non-treated with NaM-7CH and subjected to 0 and 75 mM NaCl. Average values with different literals in the same treatment or control differ statistically* 

The 9-cis-epoxycarotenoid gene (NCED) is known to be overexpressed under drought stress conditions in *P. vulgaris* bean, and it is considered fundamental for the regulation of ABA biosynthesis [58]. The increase of ABA under stress conditions causes a change in gene expression and adaptive physiological responses of plants [59]. As a result of this study, the relative expression of the pvNCED1 gene (**Figure 4**) was higher (*p* < 0.05) in the plants without HOM treatment of the control group (DW) than that were exposed to 75 mM of NaCl with respect to those not exposed to NaCl. The response of the HOM-treated plants did not show significant differences (*p* > 0.05) under normal conditions or in high salinity (75 mM NaCl), which means

despite having been exposed to saline conditions (**Figure 3**).

*Agricultural Homoeopathy: A New Insight into Organics DOI: http://dx.doi.org/10.5772/intechopen.84482*

#### **Figure 3.**

*Multifunctionality and Impacts of Organic and Conventional Agriculture*

*2.2.1 Salinity stress by NaCl in the bean crop (Phaseolus vulgaris L.)*

with physiological response of *P. vulgaris* against HOM treatments.

occurs due to high concentrations of sodium (Na+

Semillas, Villa Clara, Cuba) were used.

The photosynthesis rate (A, μmol m<sup>−</sup><sup>2</sup>

viability [46] are included.

has been little studied worldwide.

that can cause toxicity and nutritional imbalance and physiological changes, such as the reduction of the photosynthetic rate due to a lower leaf area, which reduces crop

Several alternatives have been studied to mitigate the effect of salinity in agriculture, such as genetic improvement, selection of tolerant varieties and physical treatments to seeds to induce tolerance or agrochemicals to stimulate plant growth. In these sense, HOM treatments imply the application of ultra-diluted substances, which are an eco-friendly and economic variant that could be used effectively in any condition and circumstance [47]. Unfortunately, this organic-like alternative

The common bean *Phaseolus vulgaris* L. is a key product in world food security [48], but the adverse environmental conditions, mainly drought and salinity in soils, affect their general performance reducing productivity and harvest [49]. Saline stress

ously alter the plant metabolism, affecting its growth and development [50]. Water deficit is an osmotic stressing agent, specifically associated with salinisation, which reduces the rate of fixation of carbon dioxide (CO2) and affects processes associated with photosynthesis [51]. Abscisic acid (ABA) is a key hormone that regulates the responses of plants to abiotic stress [52] and initiates the activation of stomatal closure when facing salt stress. When it happens, CO2 levels decrease and consequently photosynthesis, causing oxidative stress [53]. The objective of this study was to evaluate HOM treatment NaM (7CH) as attenuator of the harmful effect of salinity induced by NaCl application. The experiment was developed at CIBNOR to assess the initial growth stage of the species and study the expression of the genes associated

A completely randomised design was applied with HOM treatment T1 (NaM 7CH) and control (DW) each one with five replicates and two concentrations of NaCl (0 and 75 mM). Seeds of the variety white-tinted Quivicán (Empresa de

**Test 1.** The seeds were disinfected, then imbibed for 30 min in HOM treatment

s<sup>−</sup><sup>1</sup>

NaM 7CH or in DW (control treatment), planted in 5 kg plastic pots (3 seeds/ pot) with commercial substrate (Sogemix PM®) and grown for 35 days. Once the seeds germinated and the plants emerged, NaM 7CH or DW (15 ml) was applied on alternate days near the stem of each plant. The addition of NaCl began 15 days after germination and is applied gradually until reaching 75 mM to avoid osmotic shock.

portable computer with a wide-blade camera (ADC, Hoddesdon, Herts, United Kingdom). Three measurements were made during the fourth week of the trial, on healthy leaves and on completely sunny days. At the begining of frlowering, a sample was taken to perform the expression analysis of the 9-cis-epoxycarotenoid gene (PvNCDE1) [54]. Total RNA isolation, cDNA synthesis and real-time PCR amplification were performed following the methodology reported by Morelos et al. [55], and the relative expression of PvNCDE1 was estimated following the model proposed by Hellemans et al. [56]. One-way ANOVA followed by Tukey's exact test was performed using STATISTICA 8.0 (StatSoft, Inc.). A *p*-value <0.05 was considered significant. At the end of the study, photosynthesis showed a considerable decrease in the control (DW) when the plants underwent saline stress (**Figure 3**). In these stressed plants, stomatal closure was produced by ABA, generating a direct blockage of photosynthesis due to the limited uptake of CO2, an indispensable substrate for the Calvin cycle. When it happens, a blockage of the photo phase occurs due to the null

) and chlorine (Cl<sup>−</sup>) that seri-

) was measured with a LCpro-SD

**164**

*Photosynthetic rate is recorded in Phaseolus vulgaris treated and non-treated with NaM-7CH and subjected to 0 and 75 mM NaCl. Average values with different literals in the same treatment or control differ statistically (p ≤ 0.05).*

#### **Figure 4.**

*Expression of the pvNCED1 gene of HOM-treated (NaM-7CH) bean Phaseolus vulgaris L., exposed to NaCl stress. Average values with different literals correspond to statistically different treatments (p ≤ 0.05).*

demand of the reducing power and ATP, resulting in oxygen production with consequent damage to the chloroplast membranes and their subsequent disintegration [57]. Contrary to the above, in HOM-treated plants that were subjected in parallel to salt stress, photosynthesis increased and no significant difference *(p >* 0.05) was observed with respect to plants not subjected to salt stress by the addition of NaCl. This result suggests that plants receiving HOM treatment did not become stressed, despite having been exposed to saline conditions (**Figure 3**).

The 9-cis-epoxycarotenoid gene (NCED) is known to be overexpressed under drought stress conditions in *P. vulgaris* bean, and it is considered fundamental for the regulation of ABA biosynthesis [58]. The increase of ABA under stress conditions causes a change in gene expression and adaptive physiological responses of plants [59]. As a result of this study, the relative expression of the pvNCED1 gene (**Figure 4**) was higher (*p* < 0.05) in the plants without HOM treatment of the control group (DW) than that were exposed to 75 mM of NaCl with respect to those not exposed to NaCl.

The response of the HOM-treated plants did not show significant differences (*p* > 0.05) under normal conditions or in high salinity (75 mM NaCl), which means that the plants receiving homoeopathy were not stressed despite also receiving 75 mM NaCl since a lower expression of the pvNCED1 gene was found in the gene expression study. This result confirms a cause-effect relationship of T1 and the impossibility of a placebo effect in a plant model investigation where there is no possibility of suggestion of the treated individual.

### *2.2.2 Salinity stress by NaCl in basil (Ocimum basilicum L.)*

Basil is an important aromatic species for its use as flavouring and dry and fresh seasoning in the food industry; as stimulant, antispasmodic and antialopecic in pharmacy; and as aromatising cosmetics in the perfume industry [60]. The growing interest of consumers for products of natural origin stimulates the market of aromatic plants, making them a viable option for the organic agricultural sector and the possibility of exporting them fresh or processed in extracts, essences and oils used in culinary industries, cosmetics and pharmaceuticals [61]. This work aimed to assess the effect of NaM as an attenuator of NaCl stress in plant photosynthesis and biomass production of two varieties of basil, grown under hydroponic system.

**Test 2.** This experiment was developed at CIBNOR, and seedlings were obtained from certified organic seeds of basil *O. basilicum* L., Emily and Napoletano varieties (Vis Seed Company, USA). A completely randomised experimental design with factorial arrangement (2A × 2B × 3C) was used, considering the Napoletano and Emily as factor A, the concentration of NaCl (0 and 75 mM) as factor B and HOM treatments NaM 7CH and NaM 13CH as factor C. The study included a total of 12 treatments each with 4 replications. The bioassay was carried out in expanded polyurethane boxes of 69 × 38.5 × 25 cm and 38 L capacity, gauged with potable water (electrical conductivity 0.22 dS m<sup>−</sup><sup>1</sup> ). Six plastic pots (150 ml) with an experimental plant inside were fixed above each box letting the roots inside the box with liquid-enriched media pass through 1 inch holes. The plants received a nutritious solution adapted for basil according to Samperio [62].

The application of the HOM treatments and control (DW) began after a period of acclimatisation 7 days after transplant, spraying the aerial part of the plants with 150 ml plant<sup>−</sup><sup>1</sup> on alternate days. After 15 days, the application of saline treatments began gradually to avoid osmotic shock until a concentration of 75 mM was reached.

The photosynthetic rate (A, μmol m<sup>−</sup><sup>2</sup> s<sup>−</sup><sup>1</sup> ) was measured with (IRGA) LCpro-SD portable photosynthesis system equipment with a broad-leaf leaf chamber (ADC, Hoddesdon, Herts, UK) in a completely turgid and healthy leaf on completely sunny days (three measurements, 1 week before cutting); 45 days after transplant, the leaf area was determined (LA, cm2 ) by integrating leaf area metre (Li-Cor®, model-LI-3000A, series PAM 1701) and biomass using fresh weight of the aerial part (BFAP, g); an analytical balance was used (Mettler Toledo®, model AG204).

The cause-effect results of this study revealed significant differences between varieties × NaCl × HOM treatment for A (F2,60 = 4.14, *p* ≤ 0.020), FA (F2,36 = 2.87; *p* ≤ 0.01) and BFAP (F2,36 = 11.1; *p* ≤ 0.0001). Napoletano showed highest fresh weight of aerial part without the addition of NaCl and the HOM treatment NaM 7CH (**Table 2**). When both basil varieties were subjected to 75 mM NaCl, the photosynthetic rate decreased 53.6 and 63.6% with respect to the control in Napoletano and Emily, respectively. However, this result was reversed with the application of HOM treatments (NaM 7CH or NaM 13CH) in all variables and both varieties. At the end of the study, increases greater than 50% were obtained with respect to the plants that received 75 mM NaCl, but they did not receive a 'similar' HOM treatment.

These results suggest that *O. basilicum* plants naturally and positively responded to saline stress with both HOM treatments without the possibility of a placebo effect, both at cellular level and in the tissues as a whole, increasing growth. According to Zhu [63], stress agents offer environmental signals that are perceived and recognised by plants,

**167**

**Table 2.**

*Agricultural Homoeopathy: A New Insight into Organics DOI: http://dx.doi.org/10.5772/intechopen.84482*

**2.3 Attenuation of the effects of biotic stress**

*(O. basilicum) subjected to saline stress (NaCl).*

influences crop growth and development [34].

transduced into cells and re-transmitted, which generate a cascade of biochemical, physiological and genetic responses that allow the plant to adapt to change when it is gradual, but this response depends on each species and variety. It is possible to hypothesise that the presence of magnesium nanoparticles initially bioavailable in sea salt (NaM origin) has contributed to the formation of chlorophyll molecules and photosynthesis, which is the main process of plant biomass production. Additionally, Magnesium has a predominant role in enzymatic activity related to carbohydrate metabolism [64]. The results of this study suggest that agricultural homoeopathy besides being an organic-like treatment can increase general overall performance and productivity of *O. basilicum*, strengthening its ability to tolerate saline stress conditions without the need of using agrochemicals.

*Effect of the interaction of varieties × NaCl × NaM in the variables evaluated in two varieties of basil* 

With the indiscriminate use of pesticides in agriculture, the resistance of pathogenic microorganisms to the chemicals used has increased, which has negatively affected the environment and therefore the consumption of products with high levels of toxicity. Therefore, technological alternatives have been generated to replace conventional ones. Recently the application of safe products has increased in agriculture, many of which are framed in novel agricultural homoeopathy. Agricultural homoeopathy is an alternative for agricultural farmers, compatible with traditional, organic, ecological, biodynamic and even conventional agriculture, capable of influencing the biological processes of plants by controlling health problems caused by fungi, viruses and bacteria. It contributes to pest control and

**Test 1**. The experiment was developed at UTEQ, Ecuador, to assess in vitro activity of homoeopathic medicines against *Fusarium oxysporum* f. sp. lycopersici. One of the most important diseases affecting tomato crop is vascular wilt caused by *Fusarium oxysporum* f. sp. lycopersici (Fol) (Sacc.) Snyder and Hansen [65]. The control of fungal diseases in agriculture is controlled by agrochemicals including toxic pesticides whose excessive and indiscriminate use has caused the decrease or loss of the fertile

*Agricultural Homoeopathy: A New Insight into Organics DOI: http://dx.doi.org/10.5772/intechopen.84482*


#### **Table 2.**

*Multifunctionality and Impacts of Organic and Conventional Agriculture*

possibility of suggestion of the treated individual.

*2.2.2 Salinity stress by NaCl in basil (Ocimum basilicum L.)*

that the plants receiving homoeopathy were not stressed despite also receiving 75 mM NaCl since a lower expression of the pvNCED1 gene was found in the gene expression study. This result confirms a cause-effect relationship of T1 and the impossibility of a placebo effect in a plant model investigation where there is no

Basil is an important aromatic species for its use as flavouring and dry and fresh seasoning in the food industry; as stimulant, antispasmodic and antialopecic in pharmacy; and as aromatising cosmetics in the perfume industry [60]. The growing interest of consumers for products of natural origin stimulates the market of aromatic plants, making them a viable option for the organic agricultural sector and the possibility of exporting them fresh or processed in extracts, essences and oils used in culinary industries, cosmetics and pharmaceuticals [61]. This work aimed to assess the effect of NaM as an attenuator of NaCl stress in plant photosynthesis and biomass production of two varieties of basil, grown under hydroponic system.

**Test 2.** This experiment was developed at CIBNOR, and seedlings were obtained from certified organic seeds of basil *O. basilicum* L., Emily and Napoletano varieties (Vis Seed Company, USA). A completely randomised experimental design with factorial arrangement (2A × 2B × 3C) was used, considering the Napoletano and Emily as factor A, the concentration of NaCl (0 and 75 mM) as factor B and HOM treatments NaM 7CH and NaM 13CH as factor C. The study included a total of 12 treatments each with 4 replications. The bioassay was carried out in expanded polyurethane boxes of 69 × 38.5 × 25 cm and 38 L capacity, gauged with potable water (electrical conductivity 0.22 dS m<sup>−</sup><sup>1</sup>

plastic pots (150 ml) with an experimental plant inside were fixed above each box letting the roots inside the box with liquid-enriched media pass through 1 inch holes. The plants

The application of the HOM treatments and control (DW) began after a period of acclimatisation 7 days after transplant, spraying the aerial part of the plants with

began gradually to avoid osmotic shock until a concentration of 75 mM was reached.

s<sup>−</sup><sup>1</sup>

3000A, series PAM 1701) and biomass using fresh weight of the aerial part (BFAP, g);

The cause-effect results of this study revealed significant differences between varieties × NaCl × HOM treatment for A (F2,60 = 4.14, *p* ≤ 0.020), FA (F2,36 = 2.87; *p* ≤ 0.01) and BFAP (F2,36 = 11.1; *p* ≤ 0.0001). Napoletano showed highest fresh weight of aerial part without the addition of NaCl and the HOM treatment NaM 7CH (**Table 2**). When both basil varieties were subjected to 75 mM NaCl, the photosynthetic rate decreased 53.6 and 63.6% with respect to the control in Napoletano and Emily, respectively. However, this result was reversed with the application of HOM treatments (NaM 7CH or NaM 13CH) in all variables and both varieties. At the end of the study, increases greater than 50% were obtained with respect to the plants that

These results suggest that *O. basilicum* plants naturally and positively responded to saline stress with both HOM treatments without the possibility of a placebo effect, both at cellular level and in the tissues as a whole, increasing growth. According to Zhu [63], stress agents offer environmental signals that are perceived and recognised by plants,

portable photosynthesis system equipment with a broad-leaf leaf chamber (ADC, Hoddesdon, Herts, UK) in a completely turgid and healthy leaf on completely sunny days (three measurements, 1 week before cutting); 45 days after transplant, the leaf

on alternate days. After 15 days, the application of saline treatments

) was measured with (IRGA) LCpro-SD

) by integrating leaf area metre (Li-Cor®, model-LI-

received a nutritious solution adapted for basil according to Samperio [62].

an analytical balance was used (Mettler Toledo®, model AG204).

received 75 mM NaCl, but they did not receive a 'similar' HOM treatment.

The photosynthetic rate (A, μmol m<sup>−</sup><sup>2</sup>

area was determined (LA, cm2

). Six

**166**

150 ml plant<sup>−</sup><sup>1</sup>

*Effect of the interaction of varieties × NaCl × NaM in the variables evaluated in two varieties of basil (O. basilicum) subjected to saline stress (NaCl).*

transduced into cells and re-transmitted, which generate a cascade of biochemical, physiological and genetic responses that allow the plant to adapt to change when it is gradual, but this response depends on each species and variety. It is possible to hypothesise that the presence of magnesium nanoparticles initially bioavailable in sea salt (NaM origin) has contributed to the formation of chlorophyll molecules and photosynthesis, which is the main process of plant biomass production. Additionally, Magnesium has a predominant role in enzymatic activity related to carbohydrate metabolism [64]. The results of this study suggest that agricultural homoeopathy besides being an organic-like treatment can increase general overall performance and productivity of *O. basilicum*, strengthening its ability to tolerate saline stress conditions without the need of using agrochemicals.

#### **2.3 Attenuation of the effects of biotic stress**

With the indiscriminate use of pesticides in agriculture, the resistance of pathogenic microorganisms to the chemicals used has increased, which has negatively affected the environment and therefore the consumption of products with high levels of toxicity. Therefore, technological alternatives have been generated to replace conventional ones. Recently the application of safe products has increased in agriculture, many of which are framed in novel agricultural homoeopathy. Agricultural homoeopathy is an alternative for agricultural farmers, compatible with traditional, organic, ecological, biodynamic and even conventional agriculture, capable of influencing the biological processes of plants by controlling health problems caused by fungi, viruses and bacteria. It contributes to pest control and influences crop growth and development [34].

**Test 1**. The experiment was developed at UTEQ, Ecuador, to assess in vitro activity of homoeopathic medicines against *Fusarium oxysporum* f. sp. lycopersici. One of the most important diseases affecting tomato crop is vascular wilt caused by *Fusarium oxysporum* f. sp. lycopersici (Fol) (Sacc.) Snyder and Hansen [65]. The control of fungal diseases in agriculture is controlled by agrochemicals including toxic pesticides whose excessive and indiscriminate use has caused the decrease or loss of the fertile

soil layer and death of microorganisms in soil. Additionally, the pathogens have developed resistance to the active ingredient of the agrochemical that generally has a high cost [34]. Therefore, it is necessary to search for ecological and less polluting alternatives for the control of pests and diseases in agriculture, ensuring the safety and future of the agro-food industry. The objective of this study was to evaluate the in vitro effect of homoeopathic medicines on the pathogenic fungus *Fusarium oxysporum* f. sp. lycopersici, highly damaging various agricultural crops [66].

An experimental design which is completely randomised was applied with six homoeopathic medicines, each in two dynamisations (7 CH and 13 CH, Similia® CDMX, Mexico): T1 (MaP 7CH), T2 (ZiP 7CH), T3 (PhA 7CH), T4 (SiT 7CH), T5 (NaM 7CH), T6 (ArA 7CH), T7 (MaP 13CH), T8 (ZiP 13CH), T9 (PhA 13CH), T10 (SiT 13CH), T11 (NaM 13CH) and T12 (ArA 13CH) and one control (DW) to measure antifungal activity against *F. oxysporum*, using the method of the poisoned medium [67]. The diameter of the mycelium was measured daily, and the percentage of growth inhibition was determined by the formula [%inhibition = mycelial growth of the control − mycelial growth of the treatment/mycelial growth of the control × 100].

No significant differences were observed in radial growth of the phytopathogenic fungus in the HOM treatments with respect to the untreated control (NT). On the other hand, significant differences were observed between HOM treatments and their dynamisations (7 CH and 13 CH) with respect to NT. The dynamisation 13 CH increased the percentage of inhibition of the phytopathogenic fungus. These results confirmed the variability of the response induced by homoeopathic medicines in plant model, whose response depends on the dynamisation used [68]. These results are in agreement with those reported by Narváez-Martínez et al. [28] who used a homoeopathic treatment developed from a pathogen (nosode); when they applied different dynamics in tomato *Solanum quitoense* Lam, they found different effects against a pest caused by *Neoleucinodes elegantalis*. HOM treatments T2, T7 and T12 offered a greater inhibition percentage against the pathogen (70, 65 and 51%, respectively). These results agree with Tichavsky [69], who stated that *Phosphorus* homoeopathic medicine helped to control diseases caused by fungi, and two of these treatments contained phosphorus. According to Casali et al. [70], this result was due to the production of secondary metabolites (essential oils). However, during this study, HOM treatments T3 and T9 favoured the growth and reproduction of the fungus. Our results coincide with Damin et al. [71] who evaluated nine homoeopathic medicines against the pathogen *Metarhizium anisopliae* and obtained stimulation in the production of conidia by this fungus. It is necessary to conceptualise that homoeopathy acts on living beings; therefore, fungus can also be favoured with a HOM treatment.

The attained results revealed that the HOM treatments showed activity against *F. oxysporum*, highlighting *Zincum phosphoricum* and *Magnesium phosphoricum*. These cause-effect results in the plant model demonstrated that a placebo effect is not only absent in homoeopathic medicine but also supports agricultural homoeopathy. These results contribute to search for alternatives to control diseases caused by this phytopathogen in tomato plants, by using effective, innocuous and more eco-friendly tools to substitute the use of agrochemicals.
