Biological control of root-knot nematode,Meloidogyne incognita infesting tomato

Talc based formulations of two antagonistic fungi, Acremonium strictum W. Gams and Aspergillus terreus Thom were tested separately and together for their ability to suppress the development of root-knot disease of tomato caused by the root-knot nematode, Meloidogyne incognita Kofoid & White in two consecutive trials (2007–08). Tomato seedlings were each inoculated with M. incognita at 2 infective second stage juveniles /g of soil. M. incognita caused up to 48% reduction in plant growth parameters compared to un-inoculated control. Control efficacy achieved by combined soil application of both fungi, in terms of galls/root system and soil population/50 ml of soil, was 66 and 69% respectively at 60 days of inoculation compared to control. Soil application by individual fungus did not achieve as much effectiveness as the biocontrol agents applied together. The combined treatment was found to have antagonistic effect on M. incognita development and increased plant vigor. Incorporation of fine powder of chickpea pod waste with talc powder was beneficial in providing additional nutrients to both plant and biocontrol agents and increased the activity of the nematophagous fungi in soil. A. strictum and A. terreus were successfully established in the rhizosphere of tomato plants up to the termination of the experiment.     

Effects of some plant extracts on larval hatch of the root-knot nematode,Meloidogyne incognita

Abstract

The inhibitory effect of water extract of seed, leaf and bark of five plants, viz., Tamarindus indica, Cassia siamea, Isoberlinia doka, Dolnix regia and Cassia sieberiana was evaluated on larval hatch of Meloidogyne incognita in the laboratory. All the plant parts inhibited larval hatch of M. incoginta Percentage inhibition was higher in the seeds followed by the leaves and bark. Degree of inhibition observed, was directly related to the concentration of the extract. The standard suspensions inhibited hatching by about 97% while dilutions of S/100 inhibited larval hatch by 3%. Nematicidal activity of the plant parts of the five plants showed that C. siamea was the most effective followed by C. sieberiana, I. doka, T. indica and D. regia.     

Biocontrol of root-knot nematode Meloidogyne incognita by the isolates of Bacillus on tomato

Fifteen isolates of Bacillus, isolated from the root-knot nematode suppressive soils, were used for the biocontrol of Meloidogyne incognita on tomato. Bacillus isolates B1, B4, B5 and B11 caused greater inhibitory effect on hatching of M. incognita than caused by other isolates. In addition, these isolates (B1, B4, B5 and B11) caused greater colonisation of tomato roots and also caused greater increase in the growth of tomato seedling than caused by other isolates. All the isolates of Bacillus were able to increase growth of tomato and caused reduction in galling and nematode multiplication in green house tests. Isolates B1, B4, B5 and B11 caused a greater increase in growth of tomato and higher reduction in galling and nematode multiplication than other isolates in a green house test. These isolates were also tested for hydrogen cyanide (HCN) and indole acetic acid productions. Only one isolate (B13) produced HCN out of 15 tested. On the other hand, isolates B5, B11, B4 and B1 showed greater production of IAA than the other 11 isolates tested. This study suggests that Bacillus isolates B5, B11, B4 and B1 may be used for the biocontrol of M. incognita on tomato.     

Biocontrol of root-knot nematode Meloidogyne incognita by the isolates of Pseudomonas on tomato

Biocontrol of root-knot nematode Meloidogyne incognita was studied on tomato using 15 isolates of fluorescent Pseudomonads isolated from pathogen suppressive soils. Pseudomonas aeruginosa (isolates Pa8, Pa9 and Pa3) caused greater inhibitory effect on hatching of M. incognita than other isolates. In addition, isolates Pa8, Pa9 and Pa3 caused greater colonisation of tomato roots and also caused a greater increase in the growth of tomato seedlings. These isolates also caused a greater increase in growth of tomato and higher reduction in galling and nematode multiplication in a green house test than is caused by other isolates. Isolates Pf1, Pf5, Pf6 and Pa13 were unable to increase growth of tomato and caused less reduction in galling and nematode multiplication compared to other isolates. Only 10 isolates produced siderophores on chromo-azurol sulfonate (CAS) agar medium and isolate Pa12 showed greater production of siderophore followed by Pa11, Pa9, Pf10, Pa3 and Pf5. Similarly, isolates Pa14, Pa12, Pf10, Pa9, Pa8, Pa7 and Pa6 produced greater amount of HCN than the other isolates tested. Isolates Pa8 and Pa9 showed greater production of IAA than the other 13 isolates tested. This study suggests that P. aeruginosa isolates Pa8 and Pa9 may be used for the biocontrol of M. incognita on tomato.     

Response of five lettuce cultivars to root-knot nematode, Meloidogyne incognita

The root-knot nematode, Meloidogyne incognito (Kofoid et White) Chitwood is an important pathogen of vegetables. Five commercial cultivars of lettuce (Lactuca sativa L.) were evaluated under greenhouse conditions for resistance to Meloidogyne incognita, Benguet population. Plants were inoculated with 1000 eggs collected from 'Apollo' tomato (Lycopersicon esculentum) roots. The degree of galling and number of egg masses were assessed 4 and 8 weeks after inoculation. Host plant response was classified as immune, highly resistant, resistant, moderately resistant, intermediate, moderately susceptible, and highly susceptible based on the resistance index of Kouamè et at., 1998 [RI = (gall2 + egg2)]. Inoculation of 1000 eggs/plant significantly affected the growth and yield of the five lettuce cultivars 4 and 8 weeks after inoculation. A significant interaction was observed between treatment and cultivar during the two evaluation periods in terms of marketable and non-marketable yield, plant height, root weight, number of galls and number of egg masses. A reduction in growth and yield was observed in the cultivars Ballon, Lollo Rosa and Red Wave. Significant differences were noted in the number of galls and egg masses among the different cultivars tested. The highest average number of galls was obtained from the cultivars Red Wave, Ballon and Lollo Rosa. Cultivar Ballon had the highest average number of recovered nematode while Gilaben had the lowest with 15 and 4 per roots, respectively after 4 weeks inoculation. After 8 weeks, nematode was highest in cultivar Red Wave (615) and lowest in Great Lakes (70). Based on the host response, cultivars Great Lakes and Gilaben were rated highly resistant and resistant, respectively, while Red Wave, Ballon and Lollo Rosa were rated intermediate.     

Interactions between the root-knot nematode Meloidogyne incognita and Glomus mosseae in banana

The effects of the interaction between the arbuscular mycorrhizal fungus Glomus mosseae and the root-knot nematode Meloidogyne incognita on growth and nutrition of micropropagated ;Grand Naine banana (Musa AAA) cultivar was studied under greenhouse conditions. Inoculation with two G. mosseae isolates significantly increased growth of plants in relation to non-mycorrhizal plants. Response to mycorrhizae was as effective as with an optimum P fertilization in promoting plant development for most growth parameters. Meloidogyne incognita had no apparent effect on the percentage of root colonization in mycorrhizal plants. In contrast, G. mosseae suppressed root galling and nematode buildup in the roots. The percentage of mycorrhizal colonization was high (over 80%) in low P fertilized plants, but optimum P rates for bananas (four times higher than low P) significantly reduced mycorrhizal colonization. Most elements were within sufficiency levels for banana with exception of N which was low for all treatments. Mycorrhizal plants fertilized with a low P rate showed higher N, P, K, Ca, and Mg contents as compared to non-mycorrhizal plants low in P with or without the nematode. Inoculation with G. mosseae favours growth of banana plants by enhancing plant nutrition and by suppressing nematode reproduction and galling during the early stages of plant development.     

Pathogenic potential of root-knot nematode Meloidogyne incognita and root-rot fungus Fusarium solani on chilli (Capsicum annuum L.)

Damaging threshold levels of root-knot nematode Meloidogyne incognita and root-rot fungus Fusarium solani on plant growth parameters, viz., plant length, fresh and dry weight of chilli were determined by conducting their pathogenicity trials in pot experiments. The results revealed a significant reduction in the plant growth parameters at and above the inoculum level of about 1000?J₂ per plant of M. incognita and the highest reduction was recorded at 8000?J₂ per plant. Significant reduction in plant growth parameters was recorded at 1.00?g mycelial mat of F. solani per plant, while the highest reduction was observed at 8.00?g mycelial mat per plant. The damaging threshold level was 1000?J₂ per plant of M. incognita and 1.00?g mycelial mat of F. solani.     

Pathogenicity of root-knot nematode,Meloidogyne incognita on Lens culinaris (Medik.)

Effects of the root-knot nematode (Meloidogyne incognita) on lentil (Lens culinaris) were studied under greenhouse conditions. The plants were inoculated with 250, 500, 1000, 2000 and 4000 J₂ per plant. Plant growth, yield, nodulation, seed weight, chlorophyll, nitrogen, phosphorus and potassium, (NPK) contents, as compared to control, were found decreased in all the nematode infected plants. The extent of reduction increased with an increase in inoculum levels. The reductions were significant at 500 J₂ and at higher inoculum levels, i.e. 1000, 2000 and 4000 J₂ per pot over the control. An increase in inoculum level caused enhancement in galling, egg mass production and nematode population. At higher inoculum levels, the population of the nematode in the root as well as in the soil increased to a greater magnitude than at lower inoculum levels. On the contrary, reproduction factor (RF) and rate of population increase (RPI) decreased with increasing inoculum levels.     

Bio-activity of fungal culture filtrates against root-knot nematode egg hatch and juvenile motility and their effects on growth of mung bean (Vigna radiata L. Wilczek) infected with the root-knot nematode,Meloidogyne incognita

Culture filtrates of selected soil fungi, namely Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Fusarium oxysporum, Penicillium vermiculatum and Rhizopus nigricans exhibited variable response to egg hatching and mortality of the root-knot nematode, Meloidogyne incognita. Higher concentrations of the culture filtrates of all the fungi inhibited egg hatching and proved to be toxic to the juveniles of M. incognita. In addition, development of the gall and multiplication of M. incognita were also found adversely affected in varying degrees on all the plants of Vigna radiata treated with the filtrates. The culture filtrate of A. niger showed highest toxicity to the nematode than those of any other fungus tested. Soil drench application of the culture filtrates gave better seedling growth and least nematode multiplication in comparison to seed soaking treatment.     

In vitro studies of antagonistic fungi against the root-knot nematode,Meloidogyne incognita

The present study was carried out in vitro to determine the efficacy of indigenous fungi isolated from egg masses of root-knot nematode, Meloidogyne incognita on egg parasitism, egg hatching, mobility and mortality against root-knot nematode, M. incognita. The tested fungi were Acremonium strictum, Aspergillus terreus, A. nidulans, A. niger, Chetomium aubense, Chladosporium oxysporum, Fusarium chlamydosporium, F. dimarum, F. oxysporum, F. solani, Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma viride and T. harzianum. All tested fungi showed varied effects against the nematodes. Culture filtrates of A. strictum was very effective against the nematode in regards to egg parasitism (53%), egg hatching inhibition (86%) and mortality (68%) compared to controls. A. strictum was found to have an advantage over P. lilacinus, P. chlamydosporia, T. viride and T. harzianum in that it caused greater mortality of the second stage juveniles (J₂). A. terreus did not show egg parasitism but was found to be highly toxic against second stage juveniles (J₂) causing high mortality (around 68%). Thus, A. strictum and A. terreus showed good biocontrol potential against root-knot nematode, M. incognita under in vitro conditions.     

Isolation and characterization of a cDNA encoding a serine proteinase from the root-knot nematode Meloidogyne incognita

This report describes the first serine proteinase gene isolated from the sedentary nematode Meloidogyne incognita. Using degenerate primers, a 1372bp cDNA encoding a chymotrypsin-like serine proteinase (Mi-ser1) was amplified from total RNA of adult females by RT-PCR and 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of Mi-ser1 encoded a putative signal peptide and a prodomain of 22 and 33 amino acids, respectively, and a mature proteinase of 341 amino acids with a predicted molecular mass of 37,680Da. Sequence identity with the top serine proteinases matches from the databases ranged from 23 to 27%, including sequences from insects, mammals, and other nematodes. Southern blot analysis suggested that Mi-ser1 is encoded by a single or few gene copies. The pattern of developmental expression analyzed by Northern blot and RT-PCR indicated that Mi-ser1 was transcribed mainly in females. The domain architecture composed of a single chymotrypsin-like catalytic domain and the detection of a putative signal peptide suggested a digestive role for Mi-ser1.     

Evaluation of oil dispersion formulation of nematophagus fungus,Pochonia chlamydosporia against root-knot nematode,Meloidogyne incognita in cucumber

Root-knot nematode, Meloidogyne incognita is considered as one of the major non-insect pests of crops. The management of these root feeders becomes highly challenging due to a strong host-parasitic relationship. Pochonia chlamydosporia is a nematophagus fungus that colonizes eggs of nematodes. This study aimed to test the efficacy of P. chlamydosporia (NAIMCC-SF0039) against M. incognita. An oil dispersion formulation of P. chlamydosporia was prepared using emulsifiers and vegetable oil. This formulation had a shelf-life of 90 days (3.3 × 10⁸ CFU/mL) at room temperature (28 ± 1 °C). The inhibitory effect of oil formulation was tested against M. incognita by inoculating it on the egg mass. We found that colonization of the gelatinous matrix occurred on the third day of inoculation followed by complete egg parasitization on the seventh day. A greenhouse trial was laid out to evaluate the biocontrol potential of P. chlamydosporia in cucumber (Cucumis sativus). The results showed that the application of talc formulation of P. chlamydosporia at the rate of 1 kg per acre during planting, followed by delivery of 1 L of oil dispersion formulation through drip lines at 30-day intervals caused the highest reduction of nematode infestation. This treatment recorded 67.9 and 57.5% reduction in egg masses and soil nematode population respectively than that of control.     

Effect of certain organic materials in controlling Meloidogyne incognita root-knot nematode infesting banana

Efficacy of certain plant wastes as onion bulb envelope, dry leaves of sugar beet, fleabane and jojoba, filter cake or mud as sugar cane industrial residue and nile fertile mineral bio-fertilisers were studied under field conditions for managing Meloidogyne incognita on banana Cv. Williams. All the tested treatments significantly (P ≤ 0.05 and 0.01) proved to be effective in reducing the studied nematode criteria during the growing season of banana. The highest percentage reductions of 87.5 and 85.5% were recorded in the number of second-stage juveniles caused by fleabane at vegetative and harvest stages, respectively. As for galls, the highest percentage reductions of 80.4 and 79.6% were achieved at harvest stage by sugar beet waste and filter cake residue, respectively. Also, sugar beet waste was the best at increasing banana fruit yield per feddan (77.0%), followed by jojoba (53.1%) and fleabane (50.4%). The number of fingers and hands per bunch increased by the different materials at various degrees.     

Integrated management of root-knot nematode, Meloidogyne incognita infestation in tomato and grapevine

An integrated approach with the obligate bacterial parasite, Pasteuria penetrans and nematicides was assessed for the management of the root-knot nematode, Meloidogyne incognita infestation in tomato and grapevine. Seedlings of tomato cv. Co3 were transplanted into pots filled with sterilized soil and inoculated with nematodes (5000 juveniles/pot). The root powder of P. penetrans at 10 mg/pot was applied alone and in combination with carbofuran at 6 mg/pot. Application of P. penetrans along with carbofuran recorded lowest nematode infestation (107 nematodes/200 g soil) compared to control (325 nematodes/200 g soil). The rate of parasitization was 83.1% in the carbofuran and P. penetrans combination treatment as against 61.0% in the P. penetrans treatment only. The plant growth was also higher in the combination treatment compared to all other treatments. A field trial was carried out to assess the efficacy of P. penetrans and nematicides viz., carbofuran and phorate in the management of root-knot nematode, M. incognita infestation of grapevine cv. Muscat Hamburg. A nematode and P. penetrans infested grapevine field was selected and treatments either with carbofuran or phorate at 1 g a.i/vine was given. The observations were recorded at monthly interval. The results showed that the soil nematode population was reduced in nematicide treated plots. Suppression of nematodes was higher under phorate (117 nematodes/200 g soil) than under carbofuran (126.7 nematodes/200 g soil) treatment. The number of juveniles parasitized was also influenced by nematicides and spore load carried/juvenile with phorate being superior and the increase being 17.0 and 29.0% respectively over the control. The results of these experiment confirmed the compatibility of P. penetrans with nematicides and its biological control potential against the root-knot nematode.