Mixed Infection and Natural Spread of ‘Candidatus Phytoplasma asteris’ and Mungbean Yellow Mosaic India Virus Affecting Soya Bean Crop in India

Suspected phytoplasma and virus‐like symptoms of little leaf, yellow mosaic and witches’ broom were recorded on soya bean and two weed species (Digitaria sanguinalis and Parthenium hysterophorus), at experimental fields of Indian Agricultural Research Institute, New Delhi, India, in August–September 2013. The phytoplasma aetiology was confirmed in symptomatic soya bean and both the weed species by direct and nested PCR assays with phytoplasma‐specific universal primer pairs (P1/P6 and R16F2n/R16R2n). One major leafhopper species viz. Empoasca motti Pruthi feeding on symptomatic soya bean plants was also found phytoplasma positive in nested PCR assays. Sequencing BLASTn search analysis and phylogenetic analysis revealed that 16Sr DNA sequences of phytoplasma isolates of soya bean, weeds and leafhoppers had 99% sequence identity among themselves and were related to strains of ‘Candidatus Phytoplasma asteris’. PCR assays with Mungbean yellow mosaic India virus (MYMIV) coat‐protein‐specific primers yielded an amplicon of approximately 770 bp both from symptomatic soya bean and from whiteflies (Bemisia tabaci) feeding on soya bean, confirmed the presence of MYMIV in soya bean and whitefly. Hence, this study suggested the mixed infection of MYMIV and ‘Ca. P. asteris’ with soya bean yellow leaf and witches’ broom syndrome. The two weed species (D. sanguinalis and P. hysterophorus) were recorded as putative alternative hosts for ‘Ca. P. asteris’ soya bean Indian strain. However, the leafhopper E. motti was recorded as putative vector for the identified soya bean phytoplasma isolate, and the whitefly (B. tabaci) was identified as vector of MYMIV which belonged to Asia‐II‐1 genotype.     

CRISPR/Cas9‐mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean

Flowering is an indication of the transition from vegetative growth to reproductive growth and has considerable effects on the life cycle of soya bean (Glycine max). In this study, we employed the CRISPR/Cas9 system to specifically induce targeted mutagenesis of GmFT2a, an integrator in the photoperiod flowering pathway in soya bean. The soya bean cultivar Jack was transformed with three sgRNA/Cas9 vectors targeting different sites of endogenous GmFT2a via Agrobacterium tumefaciens‐mediated transformation. Site‐directed mutations were observed at all targeted sites by DNA sequencing analysis. T1‐generation soya bean plants homozygous for null alleles of GmFT2a frameshift mutated by a 1‐bp insertion or short deletion exhibited late flowering under natural conditions (summer) in Beijing, China (N39°58′, E116°20′). We also found that the targeted mutagenesis was stably heritable in the following T2 generation, and the homozygous GmFT2a mutants exhibited late flowering under both long‐day and short‐day conditions. We identified some ‘transgene‐clean’ soya bean plants that were homozygous for null alleles of endogenous GmFT2a and without any transgenic element from the T1 and T2 generations. These ‘transgene‐clean’ mutants of GmFT2a may provide materials for more in‐depth research of GmFT2a functions and the molecular mechanism of photoperiod responses in soya bean. They will also contribute to soya bean breeding and regional introduction.     

Rapid Diagnosis of Soya Bean Root Rot Caused by Fusarium culmorum Using a Loop‐Mediated Isothermal Amplification Assay

We report a rapid diagnosis of soya bean (Glycine max L.) root rot caused by Fusarium culmorum, using a loop‐mediated isothermal amplification (LAMP) assay. We used the CYP51C gene sequence to design LAMP assay primers specific for F. culmorum. The LAMP assay amplified the target gene efficiently in 60 min at 63°C. The sensitivity of the assay was 100 pg/μl of genomic DNA. Among the tested soya bean pathogens, a positive colour (sky blue) was only observed in the presence of F. culmorum with the addition of hydroxynaphthol blue (HNB) dye prior to amplification, whereas other species isolates showed no colour change. Suspected diseased soya bean samples collected in the field from Jiangsu, Shandong and Anhui provinces and Beijing were diagnosed successfully using the LAMP assay reported here. This study provides a new and readily available method for rapid diagnosis of soya bean root rot caused by F. culmorum.     

In vitro digestion enhances anti-adhesion effect of tempe and tofu against Escherichia coli

Aims: Enterotoxigenic Escherichia coli is one of the main pathogenic bacteria causing diarrhoea. Earlier studies have shown that tempe—a fungal fermented soya food—has anti‐adhesive activity against E. coli in vitro. Our aims were to challenge the anti‐adhesive activity under gastro‐intestinal conditions and to assess the activity of the nonfermented soya product tofu. Methods and results: In this study, we compared the anti‐adhesive activity of two major soya bean products, tempe and tofu, and their ileum efflux after transit through a dynamic gastrointestinal system simulating digestion in the human stomach and small intestine. The results showed that both tempe and tofu have an anti‐adhesive activity against E. coli in vitro. Tempe and tofu, after digestion through the stomach and small intestine, have even higher anti‐E. coli adhesive activity. Conclusions: In addition to the proven in‐vivo activity of tempe, this confirms the potential antidiarrhoeal effect of both the soya products tempe and tofu. Significance and Impact of the Study: As tofu has a much greater circle of consumers, this finding is relevant for the health of a large part of the world’s population.     

Effects of floral resources on the efficacy of a primary parasitoid and a facultative hyperparasitoid

Resources added to agroecosystems to enhance biological control are potentially available to multiple members of the resident insect community—not only the biological control agents for which the resources are intended. Many studies have examined the effects of sugar feeding on the efficacy of biological control agents. However, such information is lacking for other, interacting species such as facultative hyperparasitoids, which may contribute to pest suppression but can also interfere with introduced biological control agents. Under greenhouse conditions, we tested the direct effects of sugar and nectar provisioning on the longevity, host‐killing impact and offspring production of two pupal parasitoids associated with leek moth, Acrolepiopsis assectella: the introduced biological control agent, Diadromus pulchellus, and the native facultative hyperparasitoid, Conura albifrons. Adding sucrose, buckwheat or a combination of buckwheat and common vetch to a sugar‐deprived system (potted leek plants in cages) increased parasitoid longevity and resulted in higher leek moth parasitism and mortality compared to water or common vetch treatments. However, the two parasitoid species exhibited a distinct temporal response to the treatments, likely influenced by differences in their life histories. This study provides insight into how integrating conservation biological control techniques could affect the success of a classical biological control programme.     

Can plants mediate the humoral immunity and biochemical metabolism of entomopathogen‐infected caterpillars?

Plant‐insect herbivore‐entomopathogen interactions are one of the hot topics in biological control and humoral immunity, and biochemical metabolism are important responses of herbivores to pathogen infection. Entomopathogens are key biocontrol agents of caterpillars, but how plants affect the responses of caterpillars to these organisms is not well understood. We studied hormonal immunity (lysozyme and phenoloxidase activities) and biochemical metabolism (total protein and lipid contents) of Beauveria bassiana‐infected beet armyworm (Spodoptera exigua) larvae that feed on five different host plants (soya bean, Chinese cabbage, edible amaranth, water convolvulus and pepper). Results indicated that plant species differentially affected lysozyme and phenoloxidase activity and lipid content, but had no effect on protein content of pathogen‐infected caterpillars. Both lysozyme and phenoloxidase activities were generally higher in entomopathogen‐infected larvae that feed on edible amaranth or water convolvulus compared with the other three plants from days 1 to 5 after treatment. Plant species did not affect in regular changes during the 5 days in the lipid content of infected or non‐infected caterpillars. Our study reveals that plants fail to affect the biochemical metabolism but plants can mediate the humoral immunity of caterpillars to defend against pathogens. This study provides insight into plant‐mediated effects on the response of herbivores to pathogens.     

Towards the development of a sustainable soya bean‐based feedstock for aquaculture

Soya bean (Glycine max (L.) Merr.) is sought after for both its oil and protein components. Genetic approaches to add value to either component are ongoing efforts in soya bean breeding and molecular biology programmes. The former is the primary vegetable oil consumed in the world. Hence, its primary usage is in direct human consumption. As a means to increase its utility in feed applications, thereby expanding the market of soya bean coproducts, we investigated the simultaneous displacement of marine ingredients in aquafeeds with soya bean‐based protein and a high Omega‐3 fatty acid soya bean oil, enriched with alpha‐linolenic and stearidonic acids, in both steelhead trout (Oncorhynchus mykiss) and Kampachi (Seriola rivoliana). Communicated herein are aquafeed formulations with major reduction in marine ingredients that translates to more total Omega‐3 fatty acids in harvested flesh. Building off of these findings, subsequent efforts were directed towards a genetic strategy that would translate to a prototype design of an optimal identity‐preserved soya bean‐based feedstock for aquaculture, whereby a multigene stack approach for the targeted synthesis of two value‐added output traits, eicosapentaenoic acid and the ketocarotenoid, astaxanthin, were introduced into the crop. To this end, the systematic introduction of seven transgenic cassettes into soya bean, and the molecular and phenotypic evaluation of the derived novel events are described.     

CRISPR/Cas9 and TALENs generate heritable mutations for genes involved in small RNA processing of Glycine max and Medicago truncatula

Processing of double‐stranded RNA precursors into small RNAs is an essential regulator of gene expression in plant development and stress response. Small RNA processing requires the combined activity of a functionally diverse group of molecular components. However, in most of the plant species, there are insufficient mutant resources to functionally characterize each encoding gene. Here, mutations in loci encoding protein machinery involved in small RNA processing in soya bean and Medicago truncatula were generated using the CRISPR/Cas9 and TAL‐effector nuclease (TALEN) mutagenesis platforms. An efficient CRISPR/Cas9 reagent was used to create a bi‐allelic double mutant for the two soya bean paralogous Double‐stranded RNA‐binding2 (GmDrb2a and GmDrb2b) genes. These mutations, along with a CRISPR/Cas9‐generated mutation of the M. truncatula Hua enhancer1 (MtHen1) gene, were determined to be germ‐line transmissible. Furthermore, TALENs were used to generate a mutation within the soya bean Dicer‐like2 gene. CRISPR/Cas9 mutagenesis of the soya bean Dicer‐like3 gene and the GmHen1a gene was observed in the T₀ generation, but these mutations failed to transmit to the T₁ generation. The irregular transmission of induced mutations and the corresponding transgenes was investigated by whole‐genome sequencing to reveal a spectrum of non‐germ‐line‐targeted mutations and multiple transgene insertion events. Finally, a suite of combinatorial mutant plants were generated by combining the previously reported Gmdcl1a, Gmdcl1b and Gmdcl4b mutants with the Gmdrb2ab double mutant. Altogether, this study demonstrates the synergistic use of different genome engineering platforms to generate a collection of useful mutant plant lines for future study of small RNA processing in legume crops.     

Improving nutritional quality and fungal tolerance in soya bean and grass pea by expressing an oxalate decarboxylase

Soya bean (Glycine max) and grass pea (Lathyrus sativus) seeds are important sources of dietary proteins; however, they also contain antinutritional metabolite oxalic acid (OA). Excess dietary intake of OA leads to nephrolithiasis due to the formation of calcium oxalate crystals in kidneys. Besides, OA is also a known precursor of β‐N‐oxalyl‐L ‐α,β‐diaminopropionic acid (β‐ODAP), a neurotoxin found in grass pea. Here, we report the reduction in OA level in soya bean (up to 73%) and grass pea (up to 75%) seeds by constitutive and/or seed‐specific expression of an oxalate‐degrading enzyme, oxalate decarboxylase (FvOXDC) of Flammulina velutipes. In addition, β‐ODAP level of grass pea seeds was also reduced up to 73%. Reduced OA content was interrelated with the associated increase in seeds micronutrients such as calcium, iron and zinc. Moreover, constitutive expression of FvOXDC led to improved tolerance to the fungal pathogen Sclerotinia sclerotiorum that requires OA during host colonization. Importantly, FvOXDC‐expressing soya bean and grass pea plants were similar to the wild type with respect to the morphology and photosynthetic rates, and seed protein pool remained unaltered as revealed by the comparative proteomic analysis. Taken together, these results demonstrated improved seed quality and tolerance to the fungal pathogen in two important legume crops, by the expression of an oxalate‐degrading enzyme.     

Evaluation of indigenous bacterial strains for biocontrol of the frogeye leaf spot of soya bean caused by Cercospora sojina

Aims: Assessment of biological control of Cercospora sojina, causal agent of frogeye leaf spot (FLS) of soya bean, using three indigenous bacterial strains, BNM297 (Pseudomonas fluorescens), BNM340 and BNM122 (Bacillus amyloliquefaciens). Methods and Results: From cultures of each bacterial strain, cell suspensions and cell‐free supernatants were obtained and assayed to determine their antifungal activity against C. sojina. Both mycelial growth and spore germination in vitro were more strongly inhibited by bacterial cell suspensions than by cell‐free supernatants. The Bacillus strains BNM122 and BNM340 inhibited the fungal growth to a similar degree (I～52–53%), while cells from P. fluorescens BNM297 caused a lesser reduction (I～32–34%) in the fungus colony diameter. The foliar application of the two Bacillus strains on soya bean seedlings, under greenhouse conditions, significantly reduced the disease severity with respect to control soya bean seedlings and those sprayed with BNM297. This last bacterial strain was not effective in controlling FLS in vivo. Conclusions: Our data demonstrate that the application of antagonistic bacteria may be a promising and environmentally friendly alternative to control the FLS of soya bean. Significance and Impact of the Study: To our knowledge, this is the first report of biological control of C. sojina by using native Bacillus strains.     

Optimization of Nutrient and Fermentation Parameters for Antifungal Activity by Streptomyces lavendulae Xjy and Its Biocontrol Efficacies against Fulvia fulva and Botryosphaeria dothidea

This study has been proved that statistical experimental designs offer efficient and feasible approaches for determination of culture medium and fermentation conditions of Streptomyces lavendulae Xjy. Nutrition and cultural conditions have high influence on the antibiotic production of S. lavendulae Xjy. Soya bean meal, NaCl and (NH₄)₂SO₄ were identified as the best ingredients for high antifungal activity of Xjy based on single variable experiments. Using response surface methodology, the best concentrations of medium ingredients were 1.46% soya bean meal, 2.25% NaCl and 1.26% (NH₄)₂SO₄. The potential components were reduced from seven to three, and the antibiotic activity was increased by 10%. The optimized conditions for Xjy in a 250‐ml Erlenmeyer flask were initial pH 5, medium volume 25 ml/250 ml and inoculation volume 10% at 30°C with 200 rpm by response surface methodology. Validation experiments carried out to check the accuracy of the models indicated that the predicted values agreed with the experimental values. The extension of necrosis lesion of Botryosphaeria dothidea was impeded by the application of Xjy in apple fruit. In tomato plants, Xjy showed the 43.7–51.7% curative and protective effects against Fulvia fulva. The optimal medium not only gave a 20% increase in antifungal activity, but it provided a simpler formulation. The results will set a basis for further study on large‐scale fermentation volumes using S. lavendulae Xjy and be useful for the development of more advanced control strategies on plant diseases.     

The Role of Cotyledons in Vegetative Growth and Nitrogen Assimilation by Soya Bean

The removal of both cotyledons from soya bean seedlings 10 daysafter sowing, when the primary leaves were unfolded, reducedtheir stem height, branching, leaf production and dry weightat flowering by a similar proportion whether they were nodulatedor nitrate-dependent. Nitrogen assimilation by the shoots ofnitrate-dependent plants was increased by the removal of onecotyledon and reduced by the removal of both cotyledons althoughthese effects were not significant. Both these treatments significantlyincreased the amount of nitrogen in the shoots of nodulatedplants at flowering, mainly by more than doubling the nitrogencontent of their leaves. In contrast, the proportion of thetotal plant nitrogen in the leaves of nitrate-dependent plantswas almost constant. These results suggest that the cotyledonsmarkedly inhibit nitrogen assimilation by nodulated plants butdo not appreciably affect nitrogen assimilation by plants dependentsolely on inorganic nitrogen for their nitrogen supply. Glycine mux (L) Merr., soya bean, cotyledons, nitrogen assimilation, growth     

Begomovirus and Associated Satellite Components Infecting Cluster Bean (Cyamopsis tetragonoloba) in Pakistan

Cluster bean (Cyamopsis tetragonoloba) is a legume that is grown widely on the Indian subcontinent. Leaf curl symptoms of cluster bean plants collected in the Punjab, Pakistan, were shown to be associated with the begomovirus Papaya leaf curl virus; the first time this virus has been identified infecting cluster bean in Pakistan. The virus was shown to be associated with Tomato leaf curl betasatellite. Additionally, some cluster bean plants were shown to also harbour Cotton leaf curl Multan alphasatellite. The significance of these findings is discussed.     

Potassium phosphites in the protection of common bean plants against anthracnose and biochemical defence responses

The aim of this study was to investigate the effectiveness of potassium phosphites for the control of anthracnose and the mode of action of these products on common bean plants against Colletotrichum lindemuthianum, comparing it with the standard resistance inducer acibenzolar‐S‐methyl. The protection of plants against anthracnose was evaluated in greenhouse after treatment with potassium phosphites (Phosphite A and B, 5.0 ml/L), acibenzolar‐S‐methyl (0.25 g/L), or no treatment (control). Two sprayings of the treatments were performed, respectively, at V4 stage (three trifoliate leaves) and at the R5 stage (flower buds present). The inoculation with C. lindemuthianum was performed 5 days after the first spraying. Phosphite formulations A and B reduced the severity of anthracnose by 68.7% and 55.6%, respectively, and the presence of phosphites in the leaf tissues were detected at concentrations between 1 and 3 mm by 7 days after spraying. These same concentrations of phosphites reduced the mycelial growth of C. lindemuthianum in vitro by 15.0% to 25.7%. In addition, the activities of defence enzymes and the levels of phenolic compounds and lignin were assessed. Phosphite treatments enhanced the activity of various enzymes, including superoxide dismutase, peroxidase, chitinase, and β‐1,3‐glucanase, and increased the lignin and a small increase in the levels of soluble phenolics. This study provides evidence that phosphite treatments control anthracnose by acting directly on C. lindemuthianum and by inducing the production of defence responses.     

Floral resources to enhance the potential of the parasitoid Aphidius colemani for biological control of the aphid Myzus persicae

Flowering plants have been widely used to enhance biological control. However, this approach has been limited to some extent by the lack of suitable flowering plant species of wide applicability, particularly for global pests. A key example is the green peach aphid, Myzus persicae (GPA). It is commonly attacked by the polyphagous koinobiont aphidiidae, Aphidius colemani, which is also of global occurrence. Here, eight flowering plants were evaluated for the potential enhancement of GPA biological control using A. colemani under laboratory conditions. These included buckwheat (Fagopyrum esculentum), alyssum (Lobularia maritima), white rocket (Diplotaxis erucoides.), wild mustard (Sinapis arvensis), lavender (Lavandula angustifolia), wild marjoram (Origanum vulgare), thyme (Origanum marjorana) and pepper mint (Mentha piperita). The effects of access to these flowers on the longevity (days), potential fecundity (number of dissected eggs) and parasitism rate for A. colemani compared with the control treatment (water) were studied. Longevity of A. colemani which had access to buckwheat was 4–5 times longer than the control and 2–3 times longer than it was in the other plant treatments; the latter did not differ significantly between each other. Potential fecundity of A. colemani was the highest when it had been provided with buckwheat flowers. Exposing A. colemani to flowering plants for longer time intervals (12 hr and 24 hr) increased the number of eggs produced compared with 6 hr. The number of parasitized aphids/female A. colemani with buckwheat flowers was the highest of all treatments; it ranged from 14 in the control to 219 with buckwheat. Further studies should be carried out under field conditions to determine the effect of a range of flowering plants on A. colemani. For example, although buckwheat was highly effective, in many climates it may be a useful component in mixtures comprising other, more robust species.     

Rhizosphere bacteria antagonistic to soybean cyst (Heterodera glycines) and root-knot (Meloidogyne incognita) nematodes: Identification by fatty acid analysis and frequency of biological control activity

Rhizosphere bacteria were isolated from roots of young and mature plants with known antagonism to phytopathogenic nematodes, including velvet bean (Mucuna deeringiana), castor bean (Ricinus communis), sword bean (Cannavalia ensiformis), and Abruzzi rye (Secale cereale). Isolates from antagonistic plants were compared to soybean isolates for the frequency of antagonism to the root-knot (Meloidogyne incognita) and soybean cyst (Heterodera schachtii) nematodes in a disease assay with soybean. Bacterial isolates were identified using fatty acid analysis, and isolates which exhibited a significant reduction in incidence of soybean damage from both nematodes were characterized physiologically. The bacterial taxa associated with antagonistic plants were markedly different from soybean bacteria. Isolates from soybean were predominantly Bacillus spp., while those from antagonistic plants included more coryneform and Gram-negative genera. Pseudomonas cepacia and Pseudomonas gladioli were predominant among Gram-negative bacteria on antagonistic plants but were not isolated from soybean. Four to six times the number of bacteria from antagonistic plants, compared to soybean, significantly reduced disease incidence of both nematodes. No single pattern of physiological reactions was common among all these bacteria, suggesting that multiple mechanisms accounted for the observed biological control. The results suggest that rhizospheres of antagonistic plants may be useful sources of potential biological control agents for phytopathogenic nematodes.     

Integration of Pythium nunn and Trichoderma harzianum isolate T-95 for the biological control of Pythium damping-off of cucumber

Two biological control agents, Pythium nunn and Trichoderma harzianum isolate T-95, were combined to reduce Pythium damping-off of cucumber in greenhouse experiments lasting 3–4 weeks. T. harzianum T-95, a rhizosphere competent mutant, was applied to seeds and P. nunn was applied to pasteurized and raw soils naturally and artificially infested with Pythium ultimum. Some treatments were also amended with bean leaves to enhance the activity of P. nunn. The biological control of Pythium damping-off was evaluated in a Colorado soil (Nunn sandy loam) and an Oregon soil mix, which were replanted twice after 2 and 3 months. Interactions between P. nunn and T-95 were detected in the Colorado but not the Oregon soil. No consistent evidence of antagonism between P. nunn and T. harzianum was seen, and significant interactions were detected in the Colorado, but not the Oregon soil. In the first planting of some treatments, the combination of P. nunn and T. harzianum gave greater control of damping-off than either applied alone. P. nunn was most effective in soils that were pasteurized or amended with bean leaves. T. harzianum controlled Pythium damping-off in the Colorado, but not the Oregon soil. In both soils, disease declined over time in treatments amended with bean leaves but without P. nunn or T. harzianum added. This suppression was greater in the Colorado soil, which contained an indigenous population of P. nunn. This work demonstrates that two compatible biological control agents can be combined to give additional control of a soil-borne plant pathogen.     

Effect of rhizobia inoculation,N and P supply on Orobanche foetida parasitising faba bean (Vicia faba minor) under field conditions

Orobanche foetida is a chlorophyll-lacking holoparasite that subsists on the roots of plants and causes significant damage to the culture of some leguminous plants particularly faba bean. To evaluate the effect of rhizobia as a biological agent and the effect of nitrogen (N) and/or phosphorus (P) to control O. foetida parasitism in faba bean (Vicia faba minor), a trial was conducted in infested and non-infested fields with O. foetida in the Oued Beja Agricultural Experimental Unit, Tunisia. This field trial was performed during two consecutive cropping seasons using the susceptible cv. Badï, two selected rhizobia strains (Bj1 and Mat) and N combined or not with P. Mat strain showed an antagonistic effect which displayed a parasitism index two-fold lower than the control and carried two times less of emerged parasite spikes at crop maturity. Seed-yield losses caused by O. foetida infestation were very high and reached 95% for the control. Despite this high reduction, faba bean inoculated with Mat strain showed an average yield three-fold higher than the control. Broomrape parasitism did not significantly affect the protein/starch ratio of seeds for all treatments. The Mat strain is a potential candidate for developing an integrate method to control O. foetida parasitism on faba bean.     

Penetration and Reproduction of Meloidogyne javanica on Leguminous Crops

The use of antagonist plants is one of the most effective methods in the management of root–knot nematodes, and several plants recommended for this purpose are nitrogen‐fixing plants that act as green manure because of the amount of mass produced. The mode of action of some species has already been investigated, but this has yet to be elucidated for many plants researched as antagonists. Thus, this study aimed to assess the penetration and reproduction of Meloidogyne javanica on the root system of three species of crotalaria, as well as Mucuna deeringiana, Cajanus cajan, Canavalia ensiformis, Macrotyloma axillare and Stylosanthes capitata, with soya bean used as control treatment. Thus, plants inoculated with the nematode were assessed 5, 10, 15 and 20 days after inoculation (DAI), for nematode penetration and development. After 60 days of inoculation, the nematode reproduction factor (RF) was assessed. The plants did not prevent M. javanica penetration, which differed at varying degrees, according to the time of assessment and the plant species investigated. However, the nematode reproduction was impaired in all the leguminous crops assessed, with (RF)<1 in all the plants, while in soya bean the RF values were 2.85 and 2.56, in the respective experiments.