Control of nematode disease of egg plant (Solanum aethiopicum) using manure

Pot experiment was conducted in the year 2010 and repeated in 2011 to examine the effects of organic manure (poultry, cow dung and domestic waste) and inorganic manure (NPK 15:15:15) on the yield, soil and root population of Meloidogyne incognita-infected Ethiopian egg plant Solanum aethiopicum in a greenhouse at Kabba college of agriculture, Ahmadu Bello University, Kabba, Nigeria. Each of the organic manure was applied as soil amendment at the rate of 5t/ha and the inorganic fertiliser (NPK) was applied at the rate of 200?kg/ha, while there was an untreated control that acted as standard check. The experimental design was a completely randomised design comprising of five treatments including control and each of the treatments was replicated four times. The result of the experiment showed that all the organic manures considered and NPK fertilisers were effective in suppressing the nematode’s negative effects on the plant, as shown by the improved yield, reduced soil and root population as well as reduced gall index of the organic and inorganic manure-treated plant compared with the control. The mean fruit yield of the manure-treated plant was of the range 18?±?1 fruits and NPK fertiliser had an average of fruit number of 17, while the untreated control recorded an average fruit number of 6.5. The organic and inorganic manure-treated plants recorded bigger fruit size compared with control, and are significantly different from the control. The soil and root population as well as root gall index are reduced in all the manure treatments compared with the control and are significantly different from the control. The result of this experiment confirmed that organic manure can be utilised to manage nematode in soil endemic with root-knot nematode M. incognita.     

Effect of poultry manure on the growth,yield and root-knot nematode (Meloidogyne spp.) infestation of carrot (Daucus carota L.)

Currently, synthetic pesticides which are highly detrimental to man and the environment are the principal means of nematode control. However, the use of organic amendments might provide a sustainable control option as nematicidal properties have been identified in many animal wastes. A field experiment was carried out to determine the effectiveness of different levels of poultry manure (PM) applications on root-knot nematode infestation of carrot. There were four treatments (0, 2, 4 and 6?t/ha of PM) laid out in a randomised complete block design with four replications. The various levels of PM were worked into the soil on raised beds before carrot seeds were sown. Data were taken on growth performance (plant height, number of leaves and canopy spread), yield characteristics (root weight, root length and root girth) and infestation (root-galling index and nematode population in the soil). PM significantly reduced both root galling and nematode population with 4?t/ha significantly increasing yield characters of carrot. Since PM significantly reduced (p?<?0.05) nematode infestation and promoted growth and yield of carrot, farmers are encouraged to use it preferably at 4?t/ha.     

Interaction of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato

The effects of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato were tested in a greenhouse experiment. Chicken layer manure was used as a carrier substrate for the inoculum of P. lilacinus. The following parameters were used: gall index, average number of galls per root system, plant height, shoot and root weights. Inoculation of tomato plants with G. mosseae did not markedly increase the growth of infected plants with M. javanica. Inoculation of plants with G. mosseae and P. lilacinus together or separately resulted in similar shoots and plant heights. The highest root development was achieved when mycorrhizal plants were inoculated with P. lilacinus to control root-knot nematode. Inoculation of tomato plants with G. mosseae suppressed gall index and the average number of galls per root system by 52% and 66%, respectively, compared with seedlings inoculated with M. javanica alone. Biological control with both G. mosseae and P. lilacinus together or separately in the presence of layer manure completely inhibited root infection with M. javanica. Mycorrhizal colonization was not affected by the layer manure treatment or by root inoculation with P. lilacinus. Addition of layer manure had a beneficial effect on plant growth and reduced M. javanica infection.     

Macrophomina phaseolina – Meloidogyne javanica disease complex in balsam (Impatiens balsamina): a new report

Balsam seedlings were inoculated with root-knot nematode Meloidogyne javanica Race-2 and Macrophomina phaseolina either individually or concomitantly, as well as sequentially with an interval of 15?days between the nematode or fungal inoculations to determine whether the interaction was concomitant or sequential. The greater reduction in plant growth characters was observed in the plants inoculated with M. javanica and M. phaseolina, either concomitantly or sequentially as compared to their individual inoculation. However, the highest reduction in plant growth characters were recorded in the plants inoculated with M. javanica Race-2 15?days prior to M. phaseolina followed by concomitant-inoculated M. javanica Race-2 and M. phaseolina, and M. phaseolina 15?days prior to M. javanica. The number of galls/root system and the reproduction factor of the root-knot nematode was reduced in the presence of root-rot fungus. The intensity of root-rot caused by M. phaseolina increased in the presence of root-knot nematode M. javanica as compared to when M. phaseolina was inoculated individually. Moreover, stem and collar-rot symptoms caused by M. phaseolina appeared only in the presence of root-knot nematode.     

Effect of root-rot fungus (Macrophomina phaseolina) on the life cycle of root-knot nematode (Meloidogyne javanica) Race-2 on balsam

The penetration of second stage juveniles of Meloidogyne javanica started within 12 hours after inoculation and the rate of penetration gradually increased with the passage of time up to the fifth day in the plants inoculated with root-knot nematode alone and up to the sixth day when plants were infected with root-knot nematode and root-rot fungus. Mostly, the penetration of second stage juveniles of Meloidogyne javanica took place in the meristematic region but in some cases the juveniles also penetrated into the root tips and oriented themselves near the stellar region almost parallel to the longitudinal axis of the roots. The life cycle of Meloidogyne javanica on balsam was completed within 25 days, whereas the duration of the life cycle and fecundity of females was adversely affected in the presence of fungus (Macrophomina phaseolina) and it took about 33 days to complete the life cycle, i.e. the presence of Macrophomina phaseolina delayed the life cycle of the root-knot nematode (Meloidogyne javanica) by eight days.     

Biological control of root-knot nematode (Meloidogyne javanica) disease by Pseudomonas fluorescens (Chao)

Plant growth-promoting Rhizobacteria is currently developed as an biocontrol agent against many plant pathogens. In this research, biological control of root-knot nematode (Meloidogyne javanica) by Pseudomonas fluorescens was investigated in greenhouse and laboratory experiments. Results showed that 10⁹?(CFU/ml) of P. fluorescens decreased nematode infection and other parameters significantly, compared to the control. P. fluorescens was able to cause destruction of nematode egg mass matrix and significantly decreased nematode egg hatching level. Specific activities of resistance-related enzymes, namely peroxidase (POX) and phenylalanine ammonia lyase (PAL), increased significantly in P. fluorescens-inoculated plants. Maximum activities of POX and PAL were observed at the 5?days after inoculation, respectively. Results suggested that the destruction of eggs and plant defence mechanisms leading to systemic resistance are two main suppression mechanisms used by P. fluorescens against nematode.     

Management of root-knot nematode (Meloidogyne spp.) on sweet pepper (Capsicum annuum L.) with moringa (Moringa oleifera Lam.) leaf powder

A major constraint facing sweet pepper production is infestation by nematodes leading to reduced yields. Field studies were conducted during the 2012 cropping season at the Experimental Farms of the University for Development Studies, Nyankpala, Northern region, Ghana, to determine efficacy of various levels of moringa leaf powder for the control of root-knot nematodes in sweet pepper (Capsicum annuum L.) in the savanna ecology of Ghana. Treatments consisted of three levels of moringa leaf powder (40, 60 and 80?g/L) per plot and 0?g/L (control). The experiment was laid out in a randomised complete block design with each treatment replicated four times. The infestations of root-knot nematodes were significantly lower in the moringa leaf powder-treated plots than the control. Although significant differences were not observed in all the parameters evaluated among the moringa leaf powder treatments, sweet pepper plants treated with 80?g/L of moringa leaf powder per plot recorded the highest mean value of plant height, number of leaves, number of fruits per plant, fruit weight per plant total yield per plot and the thickest plant girth. Similarly, the sweet pepper plants treated with 80?g/L of moringa leaf powder had the lowest infection index (root gall) and nematode population. Application of moringa leaf powder at 40, 60 and 80?g/L increased sweet pepper yield and decreased nematode population confirming their potential in management of root-knot nematodes.     

Evaluation of pomegranate (Punica granatum L. var. Gabsi) peel extract for control of root-knot nematode Meloidogyne javanica on tomato

The present study was carried out to assess the nematicidal potential of Punica granatum L. against the root-knot nematode Meloidogyne javanica responsible for yield losses in tomato. Varied concentrations of methanolic, ethanolic and aqueous extracts from pomegranate peels were investigated for activity against eggs and juveniles of M. javanica in in vitro assays. All extracts used significantly inhibited egg hatch by over than 75%, but viability of second-stage juveniles (J2) was not significantly inhibited by ethanolic extract. Aqueous extract was assessed at the concentration of 10, 25 and 50% against M. javanica on tomato in greenhouse trials; pomegranate peels powder was also tested at the rate of 3, 6 and 9 g as a soil amendment. Both extracts significantly reduced nematode infestations; aqueous extract enhanced plant growth but powder amendment exhibited a phytotoxicity compared to the untreated plants. The reduction in number of galls, egg masses and nematode reproduction rate was recorded.     

Efficacy of Saccharomyces cerevisiae on controlling the root-knot nematode (Meloidogyne javanica) infection and promoting cucumber growth and yield under laboratory and field conditions

Saccharomyces cerevisiae is a promising plant growth-promoting yeast for different crops. Applicability of S. cerevisiae as a biocontrol agent of the root-knot nematode (Meloidogyne javanica) was investigated on cucumber under growth-room and field conditions. The yeast S. cerevisiae similar to the nematicide, Ethoprophos, when applied as a rhizospheric soil drench treatment led to an obvious reduction of root galling caused by M. javanica and resulted in reducing the nematode reproduction ability on cucumber under growth room and field conditions. The yeast was more effective at 10 than at 5?g/l. Furthermore, the application of S. cerevisiae resulted in improving cucumber plant growth and increasing its fruit yield. High content of total phenolics in cucumber roots of S. cerevisiae-treated plants and hydrogen peroxide-treated plants gives a clue on the ability of the yeast to induced plant resistance in a similar way to exogenous hydrogen peroxide.     

Role of cyanide production by Pseudomonas fluorescens CHA0 in the suppression of root-knot nematode, Meloidogyne javanica in tomato

Summary Pseudomonas fluorescens strain CHA0 produces hydrogen cyanide (HCN), a secondary metabolite that accounts largely for the biocontrol ability of this strain. In this study, we examined the role of HCN production by CHA0 as an antagonistic factor that contributes to biocontrol of Meloidogyne javanica, the root-knot nematode, in situ. Culture filtrate of CHA0, resulting from 1/10-strength nutrient broth yeast extract medium amended with glycine, inhibited egg hatch and caused mortality of M. javanica juveniles in vitro. The bacterium cultured under high oxygen-tension conditions exhibited better inhibitory effects towards nematodes, compared to its cultivation under excess oxygen situation. Growth medium amended with 0.50 or 1.0 mM FeEDDHA further improved hatch inhibition and nematicidal activity of the strain CHA0. Strain CHA77, an HCN-negative mutant, failed to exert such toxic effects, and in this strain, antinematode activity was not influenced by culture conditions. Exogenous cyanide also inhibited egg hatch and caused mortality of M. javanica juveniles in vitro. Strains CHA0 or CHA77 applied in unsterilized sandy-loam soil as drench, caused marked suppression of root-knot disease development incited by M. javanica in tomato seedlings. However, efficacy of CHA77 was noticeably lower compared to its wild type counterpart CHA0. An increased bioavailability of iron following EDTA application in soil substantially improved nematode biocontrol potential of CHA0 but not that of CHA77. Soil infestation with M. javanica eggs resulted in significantly lower nematode population densities and root-knot disease compared to the juveniles used as root-knot disease-inducing agents. Strain CHA0 significantly suppressed nematode populations and inhibited galling in tomato roots grown in soil inoculated with eggs or juveniles and treated with or without EDTA. Strain CHA0 exhibited greater biocontrol potential in soil inoculated with eggs and treated with EDTA. To demonstrate that HCN synthesis by the strain CHA0 acts as the inducing agent of systemic resistance in tomato, efficacy of the strain CHA0 was compared with CHA77 in a split root trial. The split-root experiment, guaranteeing a spatial separation of the inducing agent and the challenging pathogen, showed that HCN production by CHA0 is not crucial in the induction of systemic resistance in tomato against M. javanica, because the HCN-negative-mutant CHA77 induced the same level of resistance as the wild type but exogenous cyanide in the form of KCN failed to trigger the resistance reaction. In the root section where both nematode and the bacterium were present, strain CHA0 reduced nematode penetration to a greater extent than CHA77, suggesting that for effective control of M. javanica, a direct contact between HCN-producing CHA0 and the nematode is essential.     

The effects of <Emphasis Type="Italic">β</Emphasis>-amino-butyric acid on resistance of cucumber against root-knot nematode, <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

In this study, the effects of β-amino-butyric acid (BABA) on root-knot nematode(Meloidogyne javanica) infection of cucumber and accumulation of total phenolic compounds, hydrogen peroxide and activity of some enzymes related to plant defense mechanisms, i.e., guaiacol peroxidase (GPOX), polyphenol oxidase (PPO), catalase (CAT) in cucumber roots infected with nematode were investigated. Results of this study show that treating the cucumber seedlings with the above elicitor significantly reduces the nematode infection level (the nematode galls, number of egg masses per plant and number of eggs per individual egg mass) compared to control. Additionally, treatment of cucumber roots by BABA and BABA + nematode, significantly increased peroxidase, polyphenol oxidase and catalase activities in root tissues, 1 day after nematode inoculation in comparison to nematode inoculated plants as control and sterile water-treated plants. Enzyme activities reached to a maximum level at 4, 4 and 3 days after nematode inoculation, respectively. Additionally, the amount of H₂O₂, a product of oxidative stress, was significantly increased in the BABA and BABA + nematode treatments in comparison to control. Such increases have occurred in two phases and maximum levels of it were observed at 5 days after inoculation. Inoculation of cucumber plants by BABA also significantly increased accumulation of total phenol in comparison to control and maximum level of it was observed at 7 days after nematode inoculation. The results suggest that the inhibitory effect of BABA on the root-knot nematode (M. javanica) may be related to its ability to enhance defense responses in the cucumber roots.     

Effects of Pseudomonas fluorescens strain UTPF5 on the mobility,mortality and hatching of root-knot nematode Meloidogyne javanica

The root-knot nematode Meloidogyne spp. includes important plant pathogens worldwide. This study has considered nematode Meloidogyne javanica second stage larvae activity in the extracts of Pseudomonas fluorescens strains UTPF5 and cytotoxic effect of the strain on the nematode. The movement of second stage larvae of nematodes in water agar medium at four concentrations of bacterial extracts and second stage larvae mortality rate of hatching nematode and bacterial strains in vitro were affected. Different concentrates of the strain UTPF5 effect nematode larvae movement and disposal of the same. Bacterial extraction kills almost 100% of the larvae hatching after 24?h and a complete ban on egg hatch of biocontrol nematodes and nematode indicated that root-knot nematode larvae movement on the right attract the bacteria P. fluorescens to extract in the first place.     

Effects on plant growth and root-knot nematode infection of an endophytic GFP transformant of the nematophagous fungus Pochonia chlamydosporia

Pochonia chlamydosporia (Pc123) is a fungal parasite of nematode eggs which can colonize endophytically barley and tomato roots. In this paper we use culturing as well as quantitative PCR (qPCR) methods and a stable GFP transformant (Pc123gfp) to analyze the endophytic behavior of the fungus in tomato roots. We found no differences between virulence/root colonization of Pc123 and Pc123gfp on root-knot nematode Meloidogyne javanica eggs and tomato seedlings respectively. Confocal microscopy of Pc123gfp infecting M. javanica eggs revealed details of the process such as penetration hyphae in the egg shell or appressoria and associated post infection hyphae previously unseen. Pc123gfp colonization of tomato roots was low close to the root cap, but increased with the distance to form a patchy hyphal network. Pc123gfp colonized epidermal and cortex tomato root cells and induced plant defenses (papillae). qPCR unlike culturing revealed reduction in fungus root colonization (total and endophytic) with plant development. Pc123gfp was found by qPCR less rhizosphere competent than Pc123. Endophytic colonization by Pc123gfp promoted growth of both roots and shoots of tomato plants vs. uninoculated (control) plants. Tomato roots endophytically colonized by Pc123gfp and inoculated with M. javanica juveniles developed galls and egg masses which were colonized by the fungus. Our results suggest that endophytic colonization of tomato roots by P. chlamydosporia may be relevant for promoting plant growth and perhaps affect managing of root-knot nematode infestations.     

Interaction between Meloidogyne incognita and Pochonia chlamydosporia and their effects on the growth of Phaseolus vulgaris

An experiment was conducted to test the effect of different doses of 2, 4 and 8?g/2?kg of soil of Pochonia chlamydosporia against the root-knot nematode (Meloidogyne incognita) on Phaseolus vulgaris. It was observed that inoculation of plant with the nematode alone, and 15?days prior to fungal inoculation, reduced the plant growth when compared with the plant with fungal application followed by the nematode. Plant length, fresh and dry weight, chlorophyll, carotenoid, protein contents and nitrate reductase activity decreased in nematode-infested plants. Application of higher dose of 8?g/2?kg of soil of P. chlamydosporia increased all the plant growth parameters as well as biochemical parameters. Highest number of galls per root system was recorded on the plants infested with nematode but not treated with the fungus. However, application of fungus prior to nematode inoculation improved the plant growth and reduced the number of galls and the number of egg masses per root system.     

Nematicidal activity of culture filtrate of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> against <Emphasis Type="Italic">Meloidogyne hapla</Emphasis>

Culture filtrates of Beauveria bassiana at different concentrations were evaluated for nematicidal activity against the northern root knot nematode (Meloidogyne hapla); bioassays included egg hatching, mortality and infectivity on tomato plants in pots under glasshouse conditions. The percentage mortality and inhibition of hatching of root-knot nematode were directly proportional to the concentration of culture filtrates of B. bassiana. Soil drenching with culture filtrate of B. bassiana significantly reduced nematode population densities in soil and in the roots and subsequent gall formation and egg-mass production by M. hapla under glasshouse conditions.     

Influence of Glomus intraradices and Soil Phosphorus on Meloidogyne incognita Infecting Cucumis melo

The interaction among Glomus intraradices, Meloidogyne incognita, and cantaloupe was studied at three soil phosphorus (P) levels in a greenhouse. All plants grew poorly in soil not amended with P, regardless of mycorrhizal or nematode status. In soil amended with 50 μg P /g soil, M. incognita suppressed the growth of nonmycorrhizal plants by 84%. In contrast, growth of mycorrhizal plants inoculated with M. incognita was retarded by only 21%. A similar trend occurred in plants grown in soil with 100 μg P /g soil. Mycorrhizal infection had no effect on the degree of root-knot gall formation and did not affect the number of nematode eggs per egg mass. Mineral levels in plant shoots generally declined as soil P levels increased and were not significantly influenced by G. intraradices or M. incognita.     

Bacillus halotolerans strain LYSX1-induced systemic resistance against the root-knot nematode Meloidogyne javanica in tomato

Purpose

Determination of the nematicidal potential and mode of action of bacteria isolated from tobacco rhizosphere soil against the root-knot nematode Meloidogyne javanica in tomato plants.

Methods

Antagonistic bacteria were isolated from rhizosphere soil of tobacco infested with root-knot nematodes. Culture filtrate was used to examine nematicidal activity and ovicidal action of bacterial strains. Biocontrol of M. javanica and growth of treated tomato plants were assessed in pot experiments. To clarify whether secondary metabolites of bacteria in tomato roots induced systemic resistance to M. javanica, bacterial culture supernatants and second-stage juvenile nematodes were applied to spatially separated tomato roots using a split-root system. Bacterial strains were identified by 16S rDNA and gyrB gene sequencing and phylogenetic analysis.

Results

Of the 15 bacterial strains isolated, four (LYSX1, LYSX2, LYSX3, and LYSX4) demonstrated nematicidal activity against second-stage juveniles of M. javanica, and strain LYSX1 showed the greatest antagonistic activity; there was dose-dependent variability in nematicidal activity and inhibition of egg mass hatching by strain LYSX1. In vivo application of LYSX1 to tomato seedlings decreased the number of egg masses and galls and increased the root and shoot fresh weight. Treatment of half of the split-root system with LYSX1 reduced nematode penetration to the other half by 41.64%. Strain LYSX1 was identified as Bacillus halotolerans.

Conclusion

Bacillus halotolerans LYSX1 is a potential microbe for the sustainable biocontrol of root-knot nematodes through induced systemic resistance in tomato.

     

Effect of plant extracts on the mortality of root-knot nematodes’ J2, Meloidogyne javanica

Root-knot nematode, Meloidogyne javanica, is a major problem confronting greenhouse’s productions, field crops, vegetables, grapevines and almond rootstocks in Kermanshah province, Iran. Nematicides are not affordable to control this nematode. In the search for alternatives to chemicals control of nematodes, this research has dealt with nematicidal effects of crude herbal extracts on the root-knot nematodes. This study aimed to evaluate the effect of 21 endemic and exotic herbal extracts belong to 12 families of flowering plants in comparison with chicken manure and chemical nematicide (Temik) to control root-knot nematodes in in vitro conditions. The nematodes were pured and mass multiplied on tomato in the soil at greenhouse conditions. In order to study the effect of herbal extracts on mortality of second-stage juveniles (J₂), a 6?mL of each extract was poured in sterilised Petri dish and 54?±?4 juveniles were added. Distilled water was used as control and treatments replicated four times and incubated at ambient temperature. The LC₅₀ value of each extract was determined by assessing the mortality of juveniles (in the range of 5–95%) after 24, 48 and 72 h. Comparison between LC₅₀ value of the extracts indicated that Cinnamomum zeylanicum and Eugenia caryophillata are the most effective crude extracts on the mortality of juveniles and they were 15.4 and 17.9?mg?mL^?1, respectively. Meanwhile, the extract of tobacco, ferulago, garlic, eucalyptus, persan lilac, rattle, oliveria, licorice, russian knapweed, turnsole, sicilian sumac and chicken manure did not have any antinematode activity against fresh second-stage juveniles of the root-knot nematode.     

Population Development and Pathogenicity of Meloidogyne javanica on Flue-cured Tobacco as Influenced by Ethoprop and DD

Growth of flue-cured tobacco as influenced by Meloidogyne javanica and the effectiveness of DD and ethoprop to manage this nematode were evaluated over two growing seasons. Populations of M. javanica, root galling, plant height, steam crown diameter, whole plant weight, and yield were monitored at approximately 2-week intervals beginning 28 days after transplanting. Treatment influence on nematode population development, root galling, and plant growth generally followed a pattern in descending order of efficacy: DD (187 liters/ha), ethoprop (27, 18, or 9 kg a.i./ha), and control. In all treatments, nearly season-long increases in M. javanica populations and root galling were observed. Correlation coefficients relating nematode populations or root galling to final tobacco yield suggested either method may be used successfully to evaluate nematicide efficacy in research plots. Plant growth parameters most affected by M. javanica in order of decreasing severity were cured leaf yield, whole plant weight, plant height, and stem diameter.     

The nematicidal potential of local Bacillus species against the root-knot nematode infecting greenhouse tomatoes

Bacillus spp. were isolated from Iranian tomato fields and evaluated for their efficacy against root-knot nematodes, Meloidogyne javanica. The 52 spore-forming bacterium isolates were obtained from tomato rhizosphere following heat treatment. Eight isolates were chosen based on their potency in prevention of M. javanica egg hatch and juvenile mortality and production of proteolytic enzymes in Petri plates. Their ability to form biofilms were also determined in pot experiments. According to phenotypic traits and 16s rDNA sequencing, all selected isolates belonged to Bacillus spp. including B. cereus and B. pumilus. Treatment with bacterial culture filtrates in vitro caused juvenile mortality of 72 to 99% after 48 h. After four days, the percentage of egg hatch ranged from 1.6 to 59% depending on the isolate. Bacillus pumilus (ToIr-MA) and Bacillus sp. (ToIr-10) were found to have significant ability to produce extracellular proteases and to form maximum biofilm, considerably reducing the number of egg masses and root gall index (P = 0.05) in comparison to untreated plants. Application of ToIr-MA and ToIr-10 enhanced the fresh and dry weights of shoot and root systems. There was significant enhancement in dry root weight (45 and 50%) and shoot weight (67 and 75%). Results suggest that these two Bacillus spp. have potential as biocontrol agents against root-knot disease in tomato production.