**3.3. Greenhouse antagonism tests**

The highest development rate was 5.69 mm/day for *T. harzianum*, followed by *T*. *viridae* at 5.04 mm/day and *T*. *atroviridae* at 4.85 mm/day; with respect to strain of *F. oxysporum* Fo-A had a rate of 3.80 mm/day, where a significant difference (p = 0.023) occurs among strains of

**Table 3.** Macroscopic characterization of the colonies of Mexican strains *Trichoderma* spp. and *F. oxysporum* in culture

Cottony Regular Regular Green/yellow Radial 0.75 b 4.85c

Cottony Abundant Abundant Green/white Radial 1.25 a 5.69a

Cottony Abundant Abundant Green/white Radial 0.64 c 5.04b

Velvety Low Regular White/pink Radial 0.83 b 3.80d

**mycelium**

160 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

**Color Form Growth** 

**rate (cm/ day) \***

**Development rate(mm/day) \***

The results of the percentage of inhibition of *Trichoderma* spp. strains on *F. oxysporum* by the dual culture method are shown in **Figure 1**, the Mexican strains of *Trichoderma* spp. inhibited the growth of the pathogenic fungus, where they presented a percentage of inhibition of radial significant growth (PIRG) [p = 0.056] at the Fo-A strain of *F. oxysporum*, with *T. harzianum* which showed higher antagonistic activity, with an average value of 81.50% (PIRG), followed

**Figure 1.** Percentage of radial growth inhibition (PICR) in replicates of *Trichoderma* spp., on *F. oxysporum* in dual culture.

\*Different letters in the columns mean statistical differences in percent inhibition with Tukey's test (p < 0.05).

**3.2. Confrontation of** *Trichoderma spp.* **on** *F. oxysporum in vitro*

\*Different lowercase letters indicate significant differences with the Tukey test (P = 0.05)

*Trichoderma* spp. (**Table 3**).

**Code Texture Density Aerial** 

Tav-T7 *T. atroviridae*

Th-T4 *T. harzianum*

Tv-T3 *T. viridae*

(Fo-A) *F. oxysporum*

PDA.

The treatment inoculated with *F. oxysporum* showed symptoms of the disease in the root and aerial part (**Table 4**), presenting the highest values in incidence and severity, this in comparison with the other treatments evaluated in this study. These results coincide with that observed by Kim [40], who point out the damage caused by *Phytophthora* sp., at the root and crown of the stem of chile plants under greenhouse conditions, similar results in this research.

In the present investigation, the lowest incidence and severity was obtained in the treatment based on *T. harzianum* with 6%, presenting slight dry circular lesions in the root and without symptomatology in the aerial part. *T. harzianum* has the ability to produce enzymes such as cellulases, β-1,3-glucanase and chitinases, which degrade the cell wall of phytopathogens [41].

Treatments based on *T. harzianum*, *T*. *atroviridae* and *T*. *viridae*. used in this research work, present antagonistic efficacy against *F. oxysporum* with a survival ranging from 62.7 to 76.4% in comparison to the control treatment (**Figure 3**), which had a survival rate of 46%; while

**Figure 2.** Antagonistic capacity, according to the scale of Bell for *Trichoderma* spp. on *F. oxysporum*. (A and B = *T. harzianum* and *T*. *atroviridae* overlapping *F. oxysporum*, has a type II interaction, covers 2/3 of the surface of the medium, stops its growth and can overgrow it, and C = *T. viridae* presents an interaction of type I, where it covers the entire surface of the medium and stops its growth).


**Table 4.** Percentage of incidence and severity caused by *F. oxysporum* at 30 days after sowing in tomato plants (*L. esculentum* Mill).

tomato plants inoculated with only *F. oxysporum* achieved the lowest survival with 26.3%. Michel [38] performed antagonistic studies with native isolates obtained from tomato crops planted in Tlayacapan, Morelos-Mexico, with which it confronted *Trichoderma* spp. against *Alternaria solani* and *Phytophthora infestans* achieving a range of inhibition percentage from 16.3 to 85.5%, results that are similar to those obtained in this study.

variables, highlighting the treatment based on *T. harzianum*, which showed an average height of 22.64 cm/plant, an average stem thickness of 1.00 mm and an average dry biomass of 0.18 g; denoting a significant increase in comparison with the control, which showed an average height of 13.57 cm/plant, an average stem thickness of 0.20 cm and an average dry biomass of 0.02 g; while the treatment inoculated with the *F. oxysporum* strain Fo-A presented the lowest averages of height (12.66 cm/plant), 0.10 mm of stem thickness and 0.02 g in dry biomass (**Table 5**). Romo [42], mention that *Trichoderma* spp. has antifungal properties, thanks to the production of substances such as: trichodermine, dermadina, sequisterpeno, suzukacillina, alamethicina, trichotoxina, acetaldehyde, as well as extracellular enzymes such as β-1,3 glucanase, chitinase and cellulase that degrade The host cell walls and allow the penetration of

**Figure 4.** Significant statistic differences of each of the treatments in green weight. (a) *T. viridae,* (b) *T. harzianum,* (c) *T.* 

**Table 5.** Mean values of height, stem diameter and dry biomass of tomato plants (*L. esculentum* Mill) under greenhouse

the antagonist's hyphae, reducing its propagation in the root.

*atroviridae*, (d) *Perkins*-C21, (e) *Tricovel*-25, (f) control and (g) *F. Oxysporum*.

**Identification** 

conditions.

**Root (cm)**

**\* P. aerial (cm)**

**\* Total height (cm)**

\*Different lowercase letters indicate significant statistic differences with the Tukey test (P = 0.05)

Witness 4.53 c 9.04 d 13.57 b 0.20 d 0.19 b 0.02 b *F. oxysporum* 3.70 c 8.96 d 12.66 b 0.10 c 0.18 b 0.02 b *T. viridae* 9.25 ab 12.18 abc 21.43 a 0.80 a 0.62 b 0.05 b *T. atroviridae* 9.04 ab 12.38 abc 21.42 a 0.60 b 0.48 b 0.04 b *T. harzianum* 10.58 a 11.05 c 22.64 a 1.00 b 0.97 b 0.18 a Perkins-C21 8.30 b 13.81 a 20.12 a 0.30 a 0.83 b 0.07 b Tricovel- 25 7.81 b 12.73 b 20.54 a 0.30 d 0.66 a 0.06 b

**\* Stem diameter (mm)**

Biological Control of *Fusarium oxysporum* in Tomato Seedling Production with Mexican Strains…

**\* Weight (green)**

**\* Weight (dry)**

http://dx.doi.org/10.5772/intechopen.72878

*\**

163

**key**

For the variables height, stem thickness and dry biomass of each treatment, showed significant differences (p = 0.043) among the strains of *Trichoderma* spp.; being the treatments based on Mexican strains *Trichoderma* spp. those that presented better results in the evaluated

**Figure 3.** Percentage of mortality and survival caused by *F.* oxysporum, evaluated 30 days after tomato planting (*L. esculentum* Mill). \*Different lowercase letters indicate significant differences for % of mortality and survival with Tukey test (p < 0.05).

Biological Control of *Fusarium oxysporum* in Tomato Seedling Production with Mexican Strains… http://dx.doi.org/10.5772/intechopen.72878 163


**Table 5.** Mean values of height, stem diameter and dry biomass of tomato plants (*L. esculentum* Mill) under greenhouse conditions.

tomato plants inoculated with only *F. oxysporum* achieved the lowest survival with 26.3%. Michel [38] performed antagonistic studies with native isolates obtained from tomato crops planted in Tlayacapan, Morelos-Mexico, with which it confronted *Trichoderma* spp. against *Alternaria solani* and *Phytophthora infestans* achieving a range of inhibition percentage from

**Table 4.** Percentage of incidence and severity caused by *F. oxysporum* at 30 days after sowing in tomato plants

**Root damage Damage in aerial part**

For the variables height, stem thickness and dry biomass of each treatment, showed significant differences (p = 0.043) among the strains of *Trichoderma* spp.; being the treatments based on Mexican strains *Trichoderma* spp. those that presented better results in the evaluated

**Figure 3.** Percentage of mortality and survival caused by *F.* oxysporum, evaluated 30 days after tomato planting (*L. esculentum* Mill). \*Different lowercase letters indicate significant differences for % of mortality and survival with Tukey

test (p < 0.05).

(*L. esculentum* Mill).

16.3 to 85.5%, results that are similar to those obtained in this study.

**Treatment % incidence \*Severity scale % incidence**

162 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

Witness 28 e 11 *F. oxysporum* 70 h 58 *T. viridae* 13 b 10 *T. atroviridae* 15 b 13 *T. harzianum* 6 a 4 Perkins-C21 24 e 14 Tricovel-25 26 e 14

> variables, highlighting the treatment based on *T. harzianum*, which showed an average height of 22.64 cm/plant, an average stem thickness of 1.00 mm and an average dry biomass of 0.18 g; denoting a significant increase in comparison with the control, which showed an average height of 13.57 cm/plant, an average stem thickness of 0.20 cm and an average dry biomass of 0.02 g; while the treatment inoculated with the *F. oxysporum* strain Fo-A presented the lowest averages of height (12.66 cm/plant), 0.10 mm of stem thickness and 0.02 g in dry biomass (**Table 5**). Romo [42], mention that *Trichoderma* spp. has antifungal properties, thanks to the production of substances such as: trichodermine, dermadina, sequisterpeno, suzukacillina, alamethicina, trichotoxina, acetaldehyde, as well as extracellular enzymes such as β-1,3 glucanase, chitinase and cellulase that degrade The host cell walls and allow the penetration of the antagonist's hyphae, reducing its propagation in the root.

**Figure 4.** Significant statistic differences of each of the treatments in green weight. (a) *T. viridae,* (b) *T. harzianum,* (c) *T. atroviridae*, (d) *Perkins*-C21, (e) *Tricovel*-25, (f) control and (g) *F. Oxysporum*.

Harman [43], argues that *T. harzianum* stimulates the growth of plants by producing metabolites that promote developmental processes, which allow greater root development and absorbent hairs, which favors the mobilization of nutrients in the soil, thus improving nutrition and water absorption; also accelerates the decomposition of organic matter and minerals [44].

**Author details**

Omar Romero Arenas1

and Manuel Huerta Lara1

Puebla (BUAP), México

México

**References**

Dora Ma. Sangerman-Jarquín<sup>2</sup>

\*

DOI: 10.1007/s11274-012-1071-9

123. DOI: 10.1111/j.1365-3059.1995.tb02723.x

DOI: 10.1094/Phyto-85-469

Francis; 1998. pp. 153-172

org/reddy/reddyharzianum

bej.2007.05.012

\*Address all correspondence to: batprofesor@hotmail.com

, Jesús Francisco López Olguín1

, Conrado Parraguirre Lezama1

Biological Control of *Fusarium oxysporum* in Tomato Seedling Production with Mexican Strains…

1 Centro de Agroecología, Instituto de Ciencias, Benemérita Universidad Autónoma de

2 Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP), Campo Experimental Valle de México, Carretera Los Reyes-Texcoco, Texcoco, Estado de México,

[1] Santos-Villalobos S, Barrera-Galicia GC, Miranda-Salcedo MA, Peña-Cabriales JJ. Burkholderia cepacia XXVI siderophore with biocontrol capacity against *Colletotrichum gloeosporioides*. World Journal of Microbiology and Biotechnology. 2012;**28**:2615-2623.

[2] Howell CR, Stipanovic RD. Mechanisms in the biocontrol of *Rhizoctonia solani* induced cotton seedling disease by *Gliocladium virens*: *Antibiosis*. Phytopathology. 1995;**85**:469-472.

[3] Candela ME, Alcazár MD, Espín A, Egea-Gilabert C, Almela L. Soluble phenolic acids in *Capsicum annuum* stems infected with *Phytophthora capsici*. Plant Pathology. 1995;**44**:116-

[4] Verma M, Brar S, Tyagi R, Surampalli R, Valero J. Antagonistic fungi, *Trichoderma* spp., panoply of biological control. Biochemical Engineering. 2007;**37**:1-20. DOI: 10.1016/j.

[5] Cook RJ, Baker KF. The Nature and Practice of Biological Control of Plant Phatogens. St. Paul, Minnesota: American Phytopathological Society. 1989; 589 p. ISBN: 0890540535

[6] Chet I, Benhamou N, Haran S. Mycoparasitism and lytic enzymes. In: Harman GE, Kubicek CP, editors. Trichoderma and Gliocladium. Vol. 2. London, UK: Taylor and

[7] Sandoval VMC, ZMCI N. Producción de conidios de *Trichoderma harzianum rifai* en dos

[8] Ghisalbertí EL, Sivasithamparam K. Antifungal antibiotics produced by *Trichoderma* spp. Soil Biology and Biochemistry. 1991;**23**:1011-1020 http://www.bashanfoundation.

medios de multiplicación. Fitosanidad. 2011;**15**(4):215-221. ISSN:1562-3009

, Dionicio Juárez Ramón<sup>1</sup>

,

165

, Primo Sánchez Morales<sup>1</sup>

http://dx.doi.org/10.5772/intechopen.72878

The native strains of *Trichoderma* spp. presented higher biomass (green), emphasizing the treatment based on *T. harzianum*, which showed a root height of 10.58 cm and a green biomass of 0.97 g; denoting a significant increase in comparison with the control "**Figure 4**", which showed an average root height of 4.53 cm, and a green biomass of 0.19 g, while the treatment inoculated with the *F. oxysporum* strain Fo-A, presented the lowest root height averages, with 3.70 cm and 0.18 g in green biomass.
