Open access peer-reviewed chapter

Evaluation of the Efficiency of Some Antagonistic Trichoderma spp. in the Management of Plant Parasitic Nematodes

Written By

L. Bina Chanu, N. Mohilal and M. Manjur Shah

Submitted: November 13th, 2014 Reviewed: January 7th, 2015 Published: July 16th, 2015

DOI: 10.5772/60082

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Abstract

Plant parasitic nematodes cause great economic losses to agricultural crops worldwide. They along, with their hosts, are not isolated in the ecological system, but are strongly influenced by antagonists, parasites and pathogens. Though pesticides appear to be the most economical and efficacious means of controlling plant pathogens, toxicological, environmental and sociological concerns have led to drastic reductions in the availability of efficient commercial nematicides. These restrictions have forced farmers to look for an integral system that makes use of other means of disease control. Species of spiral nematodes, Helicotylenchus and Scutellonema, were among the most abundant plant parasitic nematodes of the mulberry plant. Eco-friendly control of the parasitic nematodes could be achieved by means of endoparasitic fungi (like Hirsutella, Meria, Nematophthora and Nematoctonus), trapping fungi (like Arthrobotrys and Duddingtonia) or parasitic fungi (like Paeceilomyces lilacinus). During the course of this present work, Trichoderma Pers. Ex. Fr. was found to be one of the most effective fungi in controlling the eggs and J2 of Meloidogyne javanica. The present study outlines the comparative efficacy of five Trichoderma species (T. viride, T. harzianum, T. longibrachiatum, T. koningii and T. hamatum) against Helicotylenchus sp. and Scutellonema sp. The study also outlines the effect of Trichoderma viride Persoon on Scutellonema spp. and Helicotylenchus sp., effect of Trichoderma harzianum Raifae on Scutellonema sp. and Helicotylenchus sp., effect of Trichoderma longibrachiatum Rifai on Scutellonema sp. and Helicotylenchus sp., effect of Trichoderma koningii Oudeom on Scutellonema sp. and Helicotylenchus sp., and lastly effect of Trichoderma hamatum (Bonord) Bainier on Scutellonema sp. and Helicotylenchus sp.

Keywords

  • Plant parasitic nematodes
  • mulberry plant
  • fungus
  • antagonistic Trichoderma spp
  • biocontrol

1. Introduction

Plant parasitic nematodes cause great economic losses to agricultural crops worldwide. They along, with their hosts, are not isolated in the ecological system, but are strongly influenced by antagonists, parasites and pathogens. Though pesticides appear to be the most economical and efficacious means of controlling plant pathogens, toxicological, environmental and sociological concerns have led to drastic reductions in the availability of efficient commercial nematicides. These restrictions have forced farmers to look for an integral system that makes use of other means of disease control. This imperative approach involves a mixture of agro-technical, biological, chemical and genetic (breeding) means of plant disease control [20, 24, 36]. Species of spiral nematodes, Helicotylenchus and Scutellonema, were among the most abundant plant parasitic nematodes of the mulberry plant. Reductions in length and weight of shoot, number and weight of leaves, and number of leaf buds were the characteristic symptoms of the infection of spiral nematodes [10]. Rao and Swarup [26] found stunting of the plants and reduction in fresh and dry weights of both shoot and root system in sugarcane due to Helicotylenchus dihystera. Besides chemicals, various researchers suggested other control measures in view of the need to replace highly toxic and potentially polluting chemicals used to control plant parasitic nematodes and fungi with less dangerous chemicals, or preferably with biological control agents and botanicals [21]). The discovery of new biocontrol agents and the demonstration of their value in reducing disease incidence and severity has opened promising new avenues for practical applications in agriculture as well as for promoting environmental safety [8]. Considerable efforts have been made by many researchers for the management of different plant parasitic nematodes with the use of Trichoderma harzianum [1 - 5, 23, 28, 33].

Eco-friendly control of the parasitic nematodes could be achieved by means of endoparasitic fungi (like Hirsutella, Meria, Nematophthora and Nematoctonus), trapping fungi (like Arthrobotrys and Duddingtonia) or parasitic fungi (like Paeceilomyces lilacinus). But there are problems in the culture of the fungi, such as unavailability of their host, and the generalist feeding nature of fungi that means they can become trapped on and digest beneficial as well as pest species of nematode. The general approach has been to go to locations where nematodes have reached high densities, and extract parasitized individuals from the soil. Then, the fungi were cultured and tested as parasites of the nematode pest. The mycoparasitic ability of Trichoderma sp. against soil-borne plant pathogens allows for the development of biocontrol strategies [11, 13, 14, 16, 24]). Windham et al. [40] reported reduced egg production in the root-knot nematode Meloidogyne arenaria following soil treatment with Trichoderma harzianum and T. koningii preparations. Combining T. harzianum with neem cakes reduced the population of citrus nematode, Tylenchulus semipenetrans [25]. Reduction of M. javanica infection with several isolates of Trichoderma lingnorum and T. harzianum has been reported [32]. Trichoderma may also promote plant growth [19].

During the course of this present work, Trichoderma Pers. Ex. Fr. was found to be one of the most effective fungi in controlling the eggs and J2 of Meloidogyne javanica. The fungi is characterized by rapidly growing colonies bearing tufted or postulate, repeatedly branched conidiophores with lageniform phialides and hyaline or green conidia borne in slimy heads. They can be cultured and isolated from any type of soil. Considering the importance of the fungal genus containing species that have the potential for economic impact, the present study was carried out to determine the comparative efficacy of five Trichoderma species (T. viride, T. harzianum, T. longibrachiatum, T. koningii and T. hamatum) against Helicotylenchus sp. and Scutellonema sp.

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2. Materials and Methods

2.1. Extraction of nematodes

Soil samples from around the rhizospheric regions of mulberry plants were collected and processed through Cobb’s sieving and decanting method followed by Baerman’s funnel technique [38]. The nematodes were observed under stereoscopic microscope and were counted using a Syracuse counting disc.

2.2. Isolation and enumeration of Trichoderma sp. from soil

The fungi were isolated through the serial soil dilution plate method [39]. Then, 10 g of oven dried fungi was added to a sterile Erlenmeyer flask with 90 ml sterile water, and the mixture was stirred with a magnetic stirrer for 20-30 minutes. A blender was used for blending the samples. While the suspension was in motion, 10 ml of solution was taken and added to 90 ml sterile water in a screw-cap flask or medicine bottle. It was shaken for one minute and 10 ml of the suspension was transferred to a 90 ml sterile water blank. The process was repeated until the desired dilution was obtained. Ten millilitres of soil solution was pipetted and mixed with 90 ml distilled water and marked to 10-3. From 10-2 and 10-3 test tubes about 5 ml solution was added to culture media contained in four petri dishes (two of each) and kept at laminar flow for 3-4 days.

To facilitate uniform spreading of the suspension over Czapek Dox agar surface at pH 5.5, the plate was placed on a turntable and the suspension spread with a flamed L-shaped rod with one hand, while rotating the turntable with the other. To obtain distinct colonies, plates were prepared 2-3 days before use or placed for a few hours at 35 to 40° C after pouring to ensure a dry agar when the suspension was added. A water film on the freshly poured plates caused excessive spreading of organisms. The plate was incubated for a few days at 24-30° C and colony counted.

The composition of the culture media was as follows:

  1. Sodium nitrate (Na2 NO3) - 1.0 g

  2. Magnesium sulphate (Mg SO4⋅7 H2O) - 0.5 g

  3. Potassium chloride (K Cl) - 0.25 g

  4. Potassium dihydrogen phosphate (KH2PO4) - 0.5 g

  5. Ferrous sulphate (FeSO4.7 H2O) - 0.5 g

  6. Sucrose - 15.0 g

  7. Agar agar - 10.0 g

  8. Distilled water – 500 ml

A broth media was made by mixing together the substances excepting the agar. It was kept for 24 hours to dissolve the substances completely. Five to six drops of the broth were removed with a dropper into different autoclaved cavity blocks.

2.3. Inoculation of nematode and fungi

Five to six drops of the broth were removed with a dropper into different autoclaved cavity blocks. Then, 0.1 ml of selected Trichoderma spp. was transferred into the cavity block containing the broth. Next, 200 female Scutellonema spp. and Helicotylenchus spp. each were also inoculated into the cavity blocks. The cavity blocks containing the whole mixture were incubated at room temperature covered upside down by autoclaved petri dishes. Uninoculated nematodes on the broth were also kept as control. Observation of the nematodes under stereoscopic binocular microscope to record their mobility and fungal infection was done at each 12-hour interval. Each treatment was replicated three times.

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3. Results and Discussion

The fungus attacked the nematodes though the production of conidia, sticky spores and mycelia, which on contact adhere to the cuticle and germinate, forming germ tubes that penetrated the nematodes. Then, they extended their hyphae inside the nematodes after penetration of the cuticle by conidia formation. These hyphae multiplied profusely. They inactivated the parasitic nematodes and immobilized them due to production of certain antibiotics and compounds. Observations of the immobility and parasitism of the nematodes due to the fungi were made every 12 hours. Each observation was replicated three times and the results are represented in tables 1–10. Photographs with graphs of parasitism are also provided in figures 1-32.

3.1. Effect of Trichoderma viride Persoon on Scutellonema spp. and Helicotylenchus sp. (table 1 and 2)

After 12 hours of inoculation, the fungus produced mycelium and conidia. The highest immobility was found at 108 hours of inoculation. The spores attached at the middle and anterior end of the body, and made the nematode immobile in the case of Scutellonema spp. After 24 hours of inoculation, many mycelium were found attached to the entire body of the nematode, due to which the body of the nematode was deformed and became shrunken, killing many of the nematodes. Body constrictions of nematodes might be due to the sucking of body contents by the fungus. Fifty percent of Scutellonema spp. out of 200 nematodes were immobilized by the fungus at 84 hours, and complete immobilization was observed at 108 hours of inoculation. In the case of Helicotylenchus spp., infection started within 24 hours of inoculation, and complete infestation occurred within 4-5 days of inoculation. The conidiophores of Trichoderma viride were less complicated; they formed aerial hyphae and coiled around the body of the nematode, producing smaller branches and ultimately forming a conifer-like branching system.

Figure 1.

Effect of fungal inoculums of Trichoderma viride on Scutellonema

Figure 2.

Effect of fungal inoculums of T. harzianum on Helicotylenchus

Observation Active female nematodes Non-active female nematodes % of non-active female nematode Control
Active Non-active
12 hours 184 14.33 7.16 200 0
24 hours 174.33 25.66 12.83 200 0
36 hours 165.33 32.00 15.83 200 0
48 hours 148.66 52.66 26.33 200 0
60 hours 134.33 62.66 31.26 200 0
72 hours 120.66 84.00 42.00 200 0
84 hours 94.00 109.33 54.66 200 0
96 hours 48.66 154.0 77.0 200 0
108 hours 0.0 200.0 100 200 0
SE m± 19.33 19.67 9.84 0.0 0
C.D. at 0.05* 3.52 60.66 1.71 0.0 0

Table 1.

Effect of fungal inoculums Trichoderma viride on the activities of Scutellonema spp.

* Significant at 0.05 level of significance.


Observation Active nematode Non-active female nematode % of non- active nematode Control
Active Non-active
12 hours 200 0 0 200 0
24 hours 182 17 9 200 0
36 hours 172 27.66 13.83 200 0
48 hours 166 34 17 200 0
60 hours 145 54.66 27.33 200 0
72 hours 133 66.66 33.33 200 0
84 hours 126 74 37 200 0
96 hours 97 103 51.5 200 0
108 hours 46.33 153.66 76.83 200 0
120 hours 13 187 93.5 200 0
132 hours 0 200 100 200 0
SEm± 19.87 19.87 9.67 0.0 0
C.D. at 0.05* 2.760 2.760 1.38 0.0 0

Table 2.

Effect of fungal inoculums Trichoderma viride on the activities of Helicotylenchus spp.

* Significant at 0.05 level of significance.


3.2. Effect of Trichoderma harzianum Raifae on Scutellonema sp. and Helicotylenchus sp. (table 3 and 4)

There was no infection after 24 hours of inoculation, but 3 % of the nematodes were immobile. Infection and parasitism of the nematode occurred after 48 hours of inoculation. The highest immobility was found at 108 hours of inoculation. Many mycelia grew over the body of the nematode. The conidiophores were seen to be multiple-branched, forming loose tufts which arose in distinct and continuous ring-like zones. The main branches, mostly in groups of two or three, stood at right angles, and the length increased with the distance from the tip of the main branch, giving a conical or pyramidal appearance. The body cuticle of the nematode was suppressed. The mycelia tip ran parallel to the nematode. There was rapid and excessive coiling on the target host. The mycelium coiled with its constricting networks of loops at the anterior region of the body and the head region, making constrictions that might be due to the sucking of body contents. After 96 hours of inoculation, there was complete immobilization of the nematodes. In the case of Helicotylenchus, the highest percentage of infection and immobility occurred during 96th hour of inoculation.

Figure 3.

Effect of fungal inoculums of T. harzianum on Helicotylenchus

Observation Active female nematodes Non-active female
nematodes
% of non-active female nematode Control
Active Non-active
12 hours 196.66 4.0 3.0 200 0
24 hours 198.66 5.0 3.5 200 0
36 hours 193.33 6.33 3.16 200 0
48 hours 161.33 36.0 17.66 200 0
60 hours 122.66 76.0 38.0 200 0
72 hours 104.0 90.0 45.0 200 0
84 hours 89.66 104.66 52.33 200 0
96 hours 00 200 100 200 0
SEm± 22.88 22.47 11.15 0.0 0
C.D. at 0.05* 3.72 3.99 2.99 0.0 0

Table 3.

Effect of fungal inoculums Trichoderma harzianum on the activities of Scutellonema spp.

* Significant at 0.05 level of significance.


Observation Active nematode Non-active nematode % of non-active nematode Control
Active Non-active
12 hours 197 3 1.5 200 0
24 hours 193 7 3.5 200 0
36 hours 184.66 15.33 7.66 200 0
48 hours 162 38 19 200 0
60 hours 125.66 74.33 37.16 200 0
72 hours 108 92 47 200 0
84 hours 93 107 54 200 0
96 hours 53 147 73.5 200 0
108 hours 0 200 100 200 0
SEm± 21.34 19.32 10.67 0.0 0
C.D. at 0.05* 2.33 2.33 0.36 0.0 0

Table 4.

Effect of fungal inoculums Trichoderma harzianum on the activities of Helicotylenchus spp.

* Significant at 0.05 level of significance.


3.3. Effect of Trichoderma longibrachiatum Rifai on Scutellonema sp. and Helicotylenchus sp. (table 5 and 6)

After 12 hours of inoculation, 4 % of the total nematode population was found to be immobile, with the highest immobility at 108 hours of inoculation. Infection started before 20 hours of inoculation. Hyphae of the fungus strain formed an appressorium-like structure in close contact with the nematode. They produced penetration holes in the cuticle of the nematode. The penetrated cuticle rapidly lost turgor and collapsed. At contact with the nematode cuticle, the hyphae branched dichotomously at the tip. The hyphae were not observed to coil around the nematode cuticle, and instead grew along the cuticle. However, penetration was not evident. Despite the absence of visible penetration, the nematode cuticle lost turgor pressure, wrinkled and collapsed. Finally, both the cuticle and body content of the nematode completely disintegrated. In the case of Helicotylenchus, the highest immobility was found at 60 hours of inoculation.

Figure 4.

Effect of fungal inoculums of T. longibranchiatum on Scutellonema, whole body

Figure 5.

Effect of fungal inoculums of T. longibranchiatum on Scutellonema, head region

Observation Active female nematodes Non-active female
nematodes
% of non-active female nematode Control
Active Non-active
12 hours 186.0 10.0 4 200 0
24 hours 180.0 18.0 9.0 200 0
36 hours 164 36 18.0 200 0
48 hours 154 46.33 22.33 200 0
60 hours 78.33 84.33 40.33 200 0
72 hours 83 121.33 60.13 200 0
84 hours 62 136.66 68.0 200 0
96 hours 27.33 173 86.53 200 0
108 hours 00 200 100 200 0
SEm± 21.70 21.81 10.97 0.0 0
C.D. at 0.05* 2.72 3.14 2.54 0.0 0

Table 5.

Effect of fungal inoculums Trichoderma longibrachiatum on the activities of Scutellonema spp.

* Significant at 0.05 level of significance.


Observation Active nematode Non-active nematode % of non-active nematode Control
Active Non-active
12 hours 193 7 3.5 200 0
24 hours 186 14 7 200 0
36 hours 174 26 13 200 0
48 hours 154 45 23 200 0
60 hours 83 117 58.5 200 0
72 hours 77 123 61.5 200 0
84 hours 52 147.66 83.5 200 0
96 hours 33 167 83.5 200 0
108 hours 12.66 187.33 93.66 200 0
120 hours 0 200 100 200 0
SEm± 22.24 22.05 11.02 0.0 0
C.D. at 0.05* 2.06 2.06 1.05 0.0 0

Table 6.

Effect of fungal inoculums Trichoderma longibrachiatum on the activities of Helicotylenchus spp.

* Significant at 0.05 level of significance.


3.4. Effect of Trichoderma koningii Oudeom on Scutellonema sp. and Helicotylenchus sp. (table 7 and 8)

There was no effect during the first 12 hours of inoculation in the case of Helicotylenchus spp., but 7 % of Scutellonema spp. were immobilized during that time. After 24 hours of exposure, conidia attachment of the nematode was found. The conidia stuck towards the cephalic region and stylet of the nematode. Maximum immobility in the case of Scutellonema occurred at 144 hours of nematode exposure to the fungus, while it occurred at 168 hours of exposure in the case of Helicotylenchus sp. At 48 hours of exposure, hyphae formation was found around the body of Helicotylenchus and at the anterior and posterior part of the body of Scutellonema spp. The hyphae penetrated towards the body cuticle of the nematode and sucked the body contents, affecting the nematode. This might be attributed to the fungus’s production of endo- and exochitinases by which hyphae penetration of the nematode cuticle was made possible.

Figure 6.

Effect of fungal inoculums of T. koningii on Scutellonema

Figure 7.

Effect of fungal inoculums of T. koningii on Helicotylenchus

Observation Active female nematodes Non-active female nematodes % of non-active female nematode Control
Active Non-active
12 hours 186.0 13.0 7 200 0
24 hours 176.0 23 12 200 0
36 hours 166 33 17 200 0
48 hours 135 65 33 200 0
60 hours 127.0 73.0 37 200 0
72 hours 113.0 87.0 44.0 200 0
84 hours 97.0 100 50 200 0
96 hours 77 122 60 200 0
108 hours 65 132 65 200 0
120 hours 37 161 80 200 0
132 hours 13 185 92 200 0
144 hours 00 200 100 200 0
SEm± 17.269 17.192 8.483 0.0 0
C.D. at 0.05* 6.97 2.17 92.10 0.0 0

Table 7.

Effect of fungal inoculum Trichoderma koningii on the activities of Scutellonema spp.

* Significant at 0.05 level of significance.


Observation Active female nematodes Non-active female nematodes % of non-active female nematode Control
Active Non-active
12 hours 200 00 00 200 0
24 hours 198 1.33 0.66 200 0
36 hours 194 4.66 2.33 200 0
48 hours 179.66 16 7 200 0
60 hours 165 30 14 200 0
72 hours 133 62 30 200 0
84 hours 1235.33 70.0 34 200 0
96 hours 111 85 43.3 200 0
108 hours 95 101 51.3 200 0
120 hours 73.33 122 61.8 200 0
132 hours 60 135 68.3 200 0
144 hours 44.66 150 75.93 200 0
156 hours 13 183.0 92 200 0
168 hours 00 200 100 200 0
SEm± 17.67 17.55 8.88 0.0 0
C.D. at 0.05* 26.088 3.089 1.891 0.0 0

Table 8.

Effect of fungal inoculum Trichoderma koningii on the activities of Helicotylenchus spp.

* Significant at 0.05 level of significance.


3.5. Effect of Trichoderma hamatum (Bonord) Bainier on Scutellonema sp. and Helicotylenchus sp. (table 9 and 10)

There was no effect on the nematode Scutellonema spp. during the first 60 hours of exposure to the fungus, or 72 hours in the case of Helicotylenchus sp. Immobilizations of a few Scutellonema sp. were found at 72 hours of exposure, while this occurred at 96 hours of exposure in the case of Helicotylenchus sp. Hundred percent immobility of Scutellonema sp. was found at 300 hours of inoculation, and in the case of Helicotylenchus, it was found at 444 hours of exposure. Infection of Scutellonema sp. started after 68 hours of exposure, and 80 hours in case of Helicotylenchus. Direct growths of the mycoparasite from the body of the nematodes were observed. There was spore formation inside the body of the nematode and shrinkage of body contents occurred. Trichoderma hamatum produced aspersoria-like structures attached to the host cell wall. Subsequently several different types of interaction occurred. The fungus either grew parallel to and along the host hyphae or coiled around the host. In Helicotylenchus sp., the parasite penetrated into and grew within the cuticle. The cuticle became vacuolated, shrank, collapsed and finally disintegrated. The oesophageal part of the nematode had shrunken and the tail region was disintegrated into two, as in a fork.

Figure 8.

Effect of fungal inoculums of T. hamatum on Scutellonema

Figure 9.

Effect of fungal inoculums of T. hamatum on Scutellonema, head region

Figure 10.

Effect of fungal inoculums of T. hamatum on Scutellonema, tail region

Figure 11.

Effect of fungal inoculums of T. hamatum on Helicotylenchus

Observation Active female nematodes Non-active female nematodes % of non-active female nematode Control
Active Non-active
12 hours 200 00 00 200 0
24 hours 200 00 00 200 0
36 hours 200 00 00 200 0
48 hours 200 00 00 200 0
60 hours 200 00 00 200 0
72 hours 195 5 2.5 200 0
84 hours 184.33 15.66 7.8 200 0
96 hours 173.66 26.33 13.16 200 0
108 hours 165 35 17 200 0
120 hours 152 48 24 200 0
132 hours 146.33 53.66 26.83 200 0
144 hours 135.33 64.66 32.3 200 0
156 hours 123 77 38.5 200 0
168 hours 117 83 41.5 200 0
180 hours 105.33 94.66 44 198 2
192 hours 91 109 54.4 196 4
204 hours 80.33 123.66 61.83 194 6
216 hours 68.66 134.66 61.83 193 7
228 hours 54 146 73 191 9
252 hours 37 163 81.5 189 11
264 hours 26 174 87.33 187 13
276 hours 16.33 183.66 95.83 185 15
288 hours 8.33 191.66 95.83 183 17
300 hours 00 200 100 181 19
SEm± 4.907 13.98 7.14 1.25 1.25
C.D. at 0.05* 3.57 3.27 2.55 0.0 0.0

Table 9.

Effect of fungal inoculum Trichoderma hamatum on the activities of Scutellonema spp.

* Significant at 0.05 level of significance.


Observation Active female nematodes Non-active female nematodes % of non-active female nematode Control
Active Non-active
24 hours 200 00 00 200 0
48 hours 200 00 00 200 0
72 hours 200 00 00 200 0
96 hours 191.33 6.0 3 200 0
120 hours 180 15.66 7.66 200 0
144 hours 174.66 24 12 200 0
156 hours 150.66 50.66 25.33 200 0
180 hours 134.66 62.33 31.16 200 0
204 hours 122 77 48.5 194 6
228 hours 106 93 46.5 193 7
252 hours 85.33 119 59.16 191 9
276 hours 77 125 62.5 189 11
300 hours 63 140.66 70.33 187 13
324 hours 47 153 77.33 185 15
348 hours 30 171 85.63 183 17
372 hours 28.66 177 88.5 181 19
396 hours 17.66 186.33 94.83 179 21
420 hours 8.66 195.0 94.16 177 23
444 hours 00 200 100 176 24
SEm± 16.13 16.519 8.229 2.00 2.0
C.D. at 0.05* 2.86 3.61 7.08 0.0 0.0

Table 10.

Effect of Trichoderma hamatum on the activities of Helicotylenchus spp.

* Significant at 5 % level of significance.


Several possible mechanisms have been suggested to be involved in Trichoderma antagonism, such as production of volatile or non-volatile antibiotics by the fungus [6], space- or nutrient- (carbon, nitrogen, iron, etc.) limiting factors that compete with the host [31], and direct mycoparasitism whereby the host cell wall is degraded by the lytic enzymes secreted by Trichoderma [9]. Trichoderma harzianum produced antibiotic 6-pentyl-α-pyrone, which had the dual effect of inhibiting pathogen growth and down-regulating genes for biosynthesis of trichothecenes, a class of mycotoxins with broad-spectrum antimicrobial activity [12]. Trichoderma longibrachiatum produced three main hydrolytic enzymes: protease, β-1, 3-glucanase and chitinase, which were involved in fungal cell wall degradation. Trichoderma koningii has also been found to produce cell wall degrading enzymes – chitinases, β-1, 3-glucanase and cellulose – which aid in the colonization of their host cells, while isonitrin, homothalin A, melanoxadin, trichodermin, ergokonin, viridian, viridio fungin A, B and C produced by the fungus act in antibiosis [22]. Sharon et al. [29] studied the mechanism involved in the attachment and parasitic processes with special emphasis on the important role of the nematode’s gelatinous matrix (gm) in direct nematode-fungus interactions, and suggested that carbohydrate-lectin-like interactions might be involved in the attachment of conidia to the nematodes. The authors also found that parasitism was one of the modes of action of Trichoderma species against Meloidogyne javanica. Trichoderma longibrachiatum produced nematotoxic concentrations of acetic acid. Secondary metabolites from fungi also contained compounds which were toxic to plant parasitic nematodes [17, 30]. Trichoderma are also known to produce toxins and antibiotics like malformin, hadacidine, gliotoxin, viridian and penicillin [37], which might contribute to the inactivity and immobility of the nematodes. Parasitic interactions between Trichoderma and nematodes might take place in soil, on root surfaces [29] and in the rhizosphere, sites that could be colonized by these opportunistic avirulent plant symbionts [18]. The improved attachment and parasitism observed in vitro could facilitate the development of new strategies to affect interactions between the nematode, plant and fungus for successful biocontrol.

The tested species of Trichoderma show a significant effect on the activity of nematodes. The results indicated that T. harzianum followed by T. longibrachiatum, T. viride, T. koningii and T. hamatum were effective in controlling the plant parasitic nematodes Helicotylenchus sp. and Scutellonema sp. Trichoderma sp. was considered imperfect filamentous (Deuetromycetes, Hyphomycetes, Phialasporace, Hyphales, Dematiaceae), and was the most common saprophytic fungi in the rhizosphere found in almost any soil.

Using beneficial fungi for control of plant disease is a useful and acceptable method for farmers. As such, the fungal microbe Trichoderma sp. can be used in biological control in the Integrated Pest Management (IPM) programme to achieve good success. Among the tested species of Trichoderma, Trichoderma harzianum is a potential candidate. These results are in agreement with [7, 15, 27, 34, 35].

Figure 12.

Effect of fungal inoculums of Aspergillus sp. on J2 M. javanica (8 days after inoculation) 10 X 10x

Figure 13.

Effect of fungal inoculums of Aspergillus sp. on egg and J2 M. javanica (39 days after inoculation) 10 X 10x

Figure 14.

Effect of fungal inoculums of Mucor sp. on egg and J2 M. javanica (39 days after inoculation) 10 X 10x

Figure 15.

Effect of fungal inoculums of Mucor sp. on J2 M. javanica (39 days after inoculation) 10 X 40x

Figure 16.

Effect of fungal inoculums of Paeceilomyces sp. on egg and J2 M. (39 days after inoculation) 10 X 10x

Figure 17.

Effect of fungal inoculums of Paeceilomyces sp. on egg of M. javanica javanica (39 days after inoculation) 10 X 40x

Figure 18.

Effect of fungal inoculums of Penicillium sp. on egg and J2 M.javanica (39 days after inoculation) 10 X 10x

Figure 19.

Effect of fungal inoculums of Penicillium sp. on egg of M. javanica (8 days after inoculation) 10 X 10x

Figure 20.

Effect of fungal inoculums of Trichoderma sp. on J2 of M. javanica (39 days after inoculation) 10 X 10x

Figure 21.

Effect of fungal inoculums of Trichoderma sp. on egg of M. javanica (39 days after inoculation) 10 X 10x

Figure 22.

Effect of different inoculums of J2 Meloidogyne javanica on mulberry plants (Var. S10)

Figure 23.

Galled roots of mulberry plant (Var.S10) due to M. javanica

Figure 24.

Graph showing absolute frequency, relative density, prominence value and importance value of soil and plant parasitic nematodes associated with mulberry plants at Govt. Silk Farm, Wangbal, Thoubal District, during the year 2006

Figure 25.

Graph showing absolute frequency, relative density, prominence value and importance value of soil and plant parasitic nematodes associated with mulberry plants at Govt. Silkfarm, Wangbal, Thoubal District during the year 2007

Figure 26.

Graph showing absolute frequency, relative density, prominence value and importancevalue of soil and plant parasitic nematodes associated with mulberry plants at Govt. Silk Farm, Wangbal, Thoubal District during the year 2008

Figure 27.

Graph showing absolute frequency, relative density, prominence value and importance value of soil and plant parasitic nematodes associated with mulberry plants at Govt. Silk Farm, Wangbal, Thoubal District during years the 2006–2008

Figure 28.

Pie-chart representation of the nematode genera during the year 2006

Figure 29.

Pie-chart representation of the nematode genera during the year 2007

Figure 30.

Pie-chart representation of the nematode genera during the year 2008

Figure 31.

Total nematode population of the family Hoplolaimidae at four different sites in valley districts of Manipur

Figure 32.

Relationship between the total nematode populations at four sites with their physical-chemical parameters

References

  1. 1. Abdel-Bari, N. A., Aboul-Eid, H. Z., Anter, E. A. and Noweer, E. A. Effects of different fungal filtrates on Meloidogyne incognita larvae in laboratory bioassay tests. Egyptian J. Agronematol. 4: 49–69.2000
  2. 2. Add-Elmoity, Riad, F. W. and El-Eraki, S. Effect of single mixture of antagonistic fungi on the control of root-knot nematode Meloidogyne incognita. Egyptian J. Agric. 71: 91–101.1993
  3. 3. Al-Fattah, A., Dababat, A. and Sikora, R. A. Use of Trichoderma harzianum and Trichoderma viride for the biological control of Meloidogyne incognita on tomato. Jordan J. Agric. Sci. 3 (3): 297–309.2007
  4. 4. Ali, A. H. H. and Barakat, M. I. E. Utilisation of Trichoderma harzianum as a biocontrol agent against root-knot nematode Meloidogyne incognita. Egyptian J. Biol. Pest Control. 4: 67–77.1994
  5. 5. Badr, S. T. A.: Effects of seven fungal filtrates, singly and combined with three nematicides on Meloidogyne javanica juveniles. Egyptian J. Agronematol. 5: 105–113.2001
  6. 6. Baker, R. and Griffin, G. J. Molecular strategies for biological control of fungal plant pathogens. In R. Reuveni (ed.), Novel Approaches to Integrated Pest Management, pp. 153–82. Boca Raton, FL, USA: Lewis Publisher. 1995
  7. 7. Bokhari, F. M. Efficacy of some Trichoderma species in the control of Rotylenchulus reniformis and Meloidogyne javanica. Archives Phytopathol. and Plant Prot. 42 (4): 361–369.2009
  8. 8. Boland, G. J. Biological control of plant diseases with fungal antagonists: Challenges and opportunities. Canadian J. Plant Pathol. 12: 295–299.1990
  9. 9. Chet, I. Trichoderma - application, mode of action and potential as biocontrol agent of soil borne plant pathogenic fungi. In: I Chet (ed.), Innovative Approaches to Plant Disease Control, pp. 137-160. New York: John Wiley & Sons. 1987
  10. 10. Deka, S. B. K. Studies on nematodes associated with mulberry, Morus alba L. M.Sc. Thesis, TNAU, Coimbatore, Tamil Nadu, India.1994
  11. 11. Dubey, S. C., Suresh, M. and Singh, B. Evaluation of Trichoderma species against Fusarium oxysporum fsp. circeris for integrated management of chick pea wilt. Biological Control. 40: 118–127.2007
  12. 12. Duffy, B. K., Simon, A. and Weller, D. M. Combination of Trichoderma koningii with fluorescent pseudomonads for control of take-all on wheat. Phytopathol. 86: 188-194.1996
  13. 13. Elad, Y., Zimmand, G., Zags, Y., Zuriel, S. and Chet, I. Use of Trichoderma harzianum in combination or alteration with fungicides to control cucumber grey mold (Botrytis cinerea) under commercial greenhouse condition. Plant Pathology. 42: 324–356.1993
  14. 14. Elad, Y. Biological control of foliar pathogens by means of Trichoderma harzianum and potential modes of action. Crop Prot. 19: 709–714.2000
  15. 15. Farouk, M. I., Rahman, M. L. and Bari, M.A. Management of root - knot nematode of tomato using Trichoderma harzianum and organic soil amendment. Bangladesh J. Plant Pathology. 18: 33–37.2002
  16. 16. Freeman, S., Minz, D., Kolesnik, I., Barbul, O., Zreibil, A., Maymon, M., Nitzani, Y., Kirshner, B., Rav-David, D., Bitu, A., Dag, A., Shafir, S. and Elad, Y. Trichoderma biocontrol of Collectotrichum acutatum and Botrytis cinerea and survival in strawberry. European J. Plant Pathology. 110: 361–370.2004
  17. 17. Hallmann, J. and Sikora, R. A. Toxicity of fungal endophyte secondary metabolites to plant parasitic nematodes and soil-borne plant pathogenic fungi. European J. Pl. Pathol. 102: 155–162.1996
  18. 18. Harman, E.G. (1991): Seed treatments for biological control of plant diseases. Crop Protection. 10: 166 – 171.
  19. 19. Inbar, J., Abramsky, M., Cohen, D. and Chet, I. Plant growth enhancement and disease control by Trichoderma harzianum in vegetable seedlings grown under commercial conditions. European J. Plant Pathol. 100: 337–346.1994
  20. 20. Kerry, B. R. Biological control. In R. H. Brown and B. R. Kerry (eds.), Principles and Practices of Nematode Control in Crops, pp. 233–263. New York: Academic Press.1987
  21. 21. Oostendrop, M. and Sikora, R. A. Utilisation of antagonistic rhizobacteria as a seed treatment for the biological control of Heterodera schachtii in sugarbeet. Revue de Nematol. 12: 77–83.1989
  22. 22. Orole, O. O. and Adejumo, T. O. Activity of fungal endophytes against four maize wilt pathogens. African J. Microbiol. 3(1): 969–973.2009
  23. 23. Pandey, G., Pandey, R. K. and Paul, H. Efficacy of different levels of Trichoderma viride against root-knot nematode in chickpea (Cicer arientinum L.). Annu Plant Protect. 26: 971–977.2003
  24. 24. Papavizas, G. C. Trichoderma and Gliocladium biology, ecology, and potential for biocontrol. Annual Review Phytopathol. 23: 23–54.1985
  25. 25. Parvatha, R. P., Rao, M. S. and Nagesh, M. Management of citrus nematode, Tylenchulus semipenetrans by integration of Trichoderma harzianum with oil cakes. Nematol. Medit. 24: 265-267.1996
  26. 26. Rao, V. R. and Swarup, G. Pathogenicity of the spiral nematode, Helicotylenchus dihystera to sugarcane. Indian J. Nematol. 4 (2):160–166.1974
  27. 27. Reddy, P. P., Rao M. S. and Nagesh, M. Management of the citrus nematode, Tylenchulus semipenetrans by integration of Trichoderma harzianum with oil cakes. Nematologia Mediterranea. 24: 265–267.1996
  28. 28. Sharon, E., Bar-Eyal, M., Chet, I., Herrera-Etrella, A., Kleifeld, O. and Spiegel, Y. Biological control of the root-knot nematode Meloidogyne javanica by Trichoderma harzianum. Phytopathology. 91 (7): 687–693.2001
  29. 29. Sharon, E., Chet, I., Viterbo, A., Bar-Eyal, M., Nagan, H. and Samuels, G. J. Parasitism of Trichoderma on Meloidogyne javanica and role of the gelatinous matrix. European J. Plant Pathol. 118: 247–258.2007
  30. 30. Sikora, R. A., Niere, B. and Kimenju, J. Endophytic microbial biodiversity and plant nematode management in African agriculture. In P. Neuenschwander, C. Borgermeister and J. Langewalder (eds.), Biological Control in IPM Systems in Africa, pp. 179–192.2003
  31. 31. Sivan, A. and Chet, I. Microbial control of plant diseases. In R. Mitchell (ed.), New Concepts in Environmental Microbiology, pp. 335-354. New York: Wiley-Liss Inc.1992
  32. 32. Spiegel, Y. and Chet, I. Evaluation of Trichoderma spp. as a biocontrol agent against soil-borne fungi and plant-parasitic nematodes in Israel. Integrated Pest Management Reviews. 3: 169–175.1998
  33. 33. Stephen, Z. A., Hassoon, I. K. and Antoon, B. G. Use of biocontrol agents and nematicides in control of Meloidogyne javanica root-knot nematode on tomato and eggplant. Pakistan J. Nematol. 16: 151–155.1998
  34. 34. Stephen, Z.A., El-Behadli, A.H., Al-Zahroon, H.H., Antoon, B.G., Georgees, S.S.H. (1996): Control of root – knot - wilt disease complex on tomato plants. Dirasat Agric. Sci. 23: 13 – 16.
  35. 35. Stephen, Z.A., Hassan, M.S. and Hasoon, I.K. (2002): Efficacy of fenamiphos, Trichoderma harzianum, Paeceilomyces lilacinus and some organic soil amendments in the control of root-knot root-rot wilt disease complex of eggplant. Arabian J. Plant Protection. 20: 115.
  36. 36. Stirling, G. R. Biological Control of Plant Parasitic Nematodes. CAB International, Wallington, UK. 282.1991
  37. 37. Subramanian, C.V. (1964): Predatory observations on host parasitic relationships on plant disease. Indian Phytopath.Soc.Bull. 2: 5 – 17.
  38. 38. Thorne, G. Principles of Nematology. New York, Toronto & London: McGraw-Hill Book Company.1961
  39. 39. Waksman, S. A. and Fred, B. A tentative outline of the plate method for determining the number of micro-organisms in the soil. Soil Sciences. 14: 27–28.1922
  40. 40. Windham, G. L., Windham, M. T. and Williams, W. P. Effects of Trichoderma spp. on maize growth and Meloidogyne arenaria reproduction. Plant Des. 73: 493–494.1989

Written By

L. Bina Chanu, N. Mohilal and M. Manjur Shah

Submitted: November 13th, 2014 Reviewed: January 7th, 2015 Published: July 16th, 2015