Resistance analysis of the rice variety Huaidao 5 against root-knot nematode Meloidogyne graminicola

Meloidogyne graminicola has emerged as one of the most destructive plant-parasitic nematodes affecting rice(Oryza sativa)production worldwide.Resistance to M.graminicola in rice could be the most effective option for its management.However,sources of germplasm with resistance to M.graminicola in rice remain limited.Here,we describe the root attraction,gall formation and genetic analysis of the resistance to M.graminicola in the rice variety Huidao 5.A nematode attraction assay showed that second-stage juveniles(J2s)of M.graminicola were attracted at the root tip of Huaidao 5 within 8 h without a significant reduction in attraction compared to the susceptible rice variety Nanjing 9108.Microscopic observation of the infection revealed that the J2s invaded root tissues 12 h after inoculation,but their subsequent movement to the root tip was hindered in Huaidao 5,resulting in decreased nematode number compared to Nanjing 9108.Additionally,we used the soil and hydroponic culture systems to simulate upland and flooding conditions in the paddy fields respectively,and found that gall number was significantly reduced,and nematode development was clearly suppressed in Huaidao 5.To investigate the genetic basis of this resistance,cross breeding was performed between the Huaidao 5 and Nanjing 9108 varieties.There was no reduction in the resistance of the F_(1) offspring to M.graminicola in the greenhouse or field trials,suggesting that a dominant gene could control resistance in Huaidao 5.In summary,this study provides a detailed characterization of a novel source of resistance to M.graminicola in rice,which is of great potential for use in crop breeding.

Study on the Activity of Herbal Extract to Kill Root-knot Nematode of Cucumber Seedlings Stage

Present mature plants hydroponic technology was used,combined with some excellent characteristics,such as growth conditions was easy to control and process of root growth was easy to continuously observe,the nematicidal activity of 5 kinds of Chinese herbs extracts and the compound solution of Avermectin,with strong contact toxicity effect indoor,was systematically studied and investigated the affection on the root-knot nematode parasitized on the cucumber seeding stage. It is found that under the premise of no influence on root growth of cucumber,extracts from Picrorhiza scrophulariiflora and Punica granatum showed strong prevention and nematicidal activity,and had the similar efficacy of Avermectin; while the extracts from Cibotium barometz,Aucklandia lappa Decne and Fructus cnidii showed low nematicidal activity and various degrees inhibition effect on plant growth.

Investigation on Pathogenic Factors of Tobacco Root-Knot Nematode Disease in Gengma County,Yunnan Province

Tobacco root-knot nematode disease has caused severe damage in Geng- ma County, Yunnan Province. In order to identify the pathogenic factors of the tobac- co root-knot nematode disease in this county, the pathogenic nematodes, hosts and environment of tobacco fields in Mengsa, Hepai and Sipaishan 3 main tobacco-grow- ing towns in Gengma County were investigated and analyzed based on the local re- lated field survey on tobacco root-knot nematode disease in this county in 2012. The results showed the incidence and severity of the tobacco root-knot nematode disease were all higher than those of previous years. dominant pathogens of the tobacco root-knot The species identification showed the nematode disease were Meloidogyne arenaria and M. javanica in Gengma County. The lacking of disease-resistant culti- vars, poor management and climatic anomaly were the main causes of the tobacco root-knot nematode disease in Gengma. According to the occurrence characteristics of the disease, the agricultural prevention-based control measures were proposed.

Effect of Anti-nematode Preparations on Physiological Traits of Tomato Leaves Affected by Root-knot Nematode

[ Objective ] The paper was to study the effects of anti-nematode preparations with different mechanisms on changes of enzyme systems and membrane permeability of tomato leaves, so as to provide reference basis for effective control of soil root-knot nematode in greenhouse. [ Method] With tomato seedlings af- fected by root-knot nematode as material, changes of superoxide dismutase（SOD）, peroxidase（ POD）, relative conductivity and malondialdehyde （MDA） in toma- toes were tested after the seedling soil was treated by preparations of Wuxianmei, Hailvsu, Duxiandna and Avermectin. [ Result] After treated by different prepara- tions, SOD and POD activity of tomato leaves were higher than control, and that treated by Wuxianmei was the highest. In addition to Duxiandna, the relative con- ductivity and MDA content of other treatments were significantly lower than control. When tomatoes were planted for 70 d, the effect of Avermectin against reot-knot nematode Was the best of 66.3%. [ Conclusion] After tomatoes were infected by root-knot nematode, different preparation treatments all had certain control effect, which made the physical indicators of tomato have obvious change. Integrated control of multiple preparations in greenhouse was beneficial to control soil root-knot nematode.

Study on Controlling Tobacco Root-knot Nematode by Fertilizer and Pesticide Synergy

Synergism between pesticide and fertilizer in controlling root-knot nematode was investigated by field randomized block test. The results showed that the best control efficiency was shared by 0.5% abamectin and 25% mixed pesticide （abamectin and carbosulfan）, 24.1% and 28.0%, respectively. The positive effect was found by synergism between pesticide and fertiUzer in controlling reot-knot nematode. The economic benefit and agricultural benefit were gained by controlling root-knot nematodes. The results suggested that the agricultural and ecological benefit could be gained through the integration of agricultural practice and chemical technology in controlling root-knot nematode.

The synergistic advantage of combining chloropicrin or dazomet with fosthiazate nematicide to control root-knot nematode in cucumber production

The highly-damaging root-knot nematode(Meloidogyne spp.,RKN)cannot be reliably controlled using only a nematicide such as fosthiazate because of increasing pest resistance.In laboratory and greenhouse trials,we showed that chloropicrin(CP)or dazomet(DZ)synergized the efficacy of fosthiazate against RKN.The combination significantly extended the degradation half-life of fosthiazate by an average of about 1.25 times.CP or DZ with fosthiazate reduced the time for fosthiazate to penetrate the RKN cuticle compared to fosthiazate alone.CP or DZ combined with low or medium rate of fosthiazate increased the total cucumber yield,compared to the use of each product alone.A low-dose fosthiazate with DZ improved total yield more than a low dose fosthiazate with CP.Extending the half-life of fosthiazate and reducing the time for fosthiazate or fumigant to penetrate the RKN cuticle were the two features that gave the fumigant-fosthiazate combination its synergistic advantage over these products used singularly.This synergy provides the opportunity for farmers to use a low dose of fosthiazate which lowers the risk of RKN resistance.Farmers could combine DZ at 30 g m^-2 with fosthiazate at a low rate of 0.375 g m^-2 to control RKN and adequately control two major soil-borne diseases in cucumber greenhouses.

Effects of Two Composted Plant Pesticide Residues,Incorporated with Trichoderma viride,on Root-Knot Nematode in Balloonflower

Plant pesticide residues, such as chinaberry （Melia toosendan） residue and sand cypress （Sabina vulgaris） residue, are pesticidal plant materials discarded after the bioactive ingredient has been extracted with organic solvents. The only option for botanical pesticide residue utilization has been as landfill. Chinaberry residue （CBR） and sand cypress residue （SCR） were collected and composted in Yangling, Shaanxi Province, China. We studied the effects of chinaberry residue compost （CBRC）, CBRC incorporated with Trichoderma viride （CBRCT）, sand cypress residue compost （SCRC）, and SCRC incorporated with T. viride （SCRCv） on the root-knot nematode, Meloidogyne incognita, infesting the balloonflower （Platycodon grandiflorum）. Bioassay results indicated that stock solutions of the CBRCT and SCRCT extracts significantly inhibited egg hatching and caused high larval mortality, followed in degree by the CBRC and SCRC extracts. The CBR and SCR extracts caused very low inhibition of eggs and larvae. Supplementing potting mixtures with these four composts reduced the severity of root galling and increased the proportion of marketable roots. The severity of root galling decreased and the average weight of the marketable roots increased with an increase in all the composts when supplemented at rates from 5 to 30%. CBR- and SCR-supplemented pot soils also inhibited the nematodes, but CBR and SCR applied to the soil had a phytotoxic effect and inhibited balloonflower growth. Supplementing field soil with the composts reduced the severity of root galling and the populations of southern root-knot nematodes in the soil. CBRCT and SCRCT clearly enhanced the average weight of the marketable roots by 30.45 and 26.64%, respectively. Continuous supplementation with CBRCT or SCRCT in the same field significantly enhanced the control of the root-knot nematode, and the populations of nematodes continued to decrease with second inoculations. The populations of total Trichoderma spp. were distinctly enhanced and were maintained at high levels for a long time in the supplemented soils.

Preparation and efficacy evaluation of Paenibacillus polymyxa KM2501-1 microbial organic fertilizer against root-knot nematodes

Root-knot nematodes(RKNs) cause huge yield losses to agricultural crops worldwide. Meanwhile, livestock manure is often improperly managed by farmers, which leads to serious environmental pollution. To resolve these two problems, this study developed a procedure for the conversion of chicken manure to organic fertilizer by larvae of Hermetia illucens L. and Bacillus subtilis BSF-CL. Chicken manure organic fertilizer was then mixed thoroughly with Paenibacillus polymyxa KM2501-1 to a final concentration of 1.5×10^(8)CFU g^(-1). The efficacy of KM2501-1 microbial organic fertilizer in controlling root-knot nematodes was evaluated in pot and field experiments. In pot experiments, applying KM2501-1 microbial organic fertilizer either as a base fertilizer or as a fumigant at the dose of 40 g/pot suppressed root-knot disease by 61.76 and 69.05% compared to the corresponding control treatments, respectively. When applied as a fumigant at the dose of 1 kg m;in field experiments, KM2501-1 microbial organic fertilizer enhanced the growth of tomato plants, suppressed root-knot disease by 49.97%, and reduced second stage juveniles of RKN in soil by 88.68%. KM2501-1 microbial organic fertilizer controlled RKNs better than commercial bio-organic fertilizer in both pot and field experiments. These results demonstrate that this co-conversion process efficiently transforms chicken manure into high value-added larvae biomass and KM2501-1 microbial organic fertilizer with potential application as a novel nematode control agent.

Control of Southern Root-knot Nematodes on Tomato and Regulation of Soil Bacterial Community by Biofumigation with Zanthoxylum bungeanum Seed

Biofumigation is an environmentally friendly strategy used to control nematodes and plant diseases.The volatile oil of Zanthoxylum bungeanum has high insecticidal and antibacterial activity.However,it is not known if the seed of Z.bungeanum is a suitable material for biofumigation to control southern root-knot nematodes(SRKN)on tomato,and how it may regulate the soil bacterial community structure.We used pot experiments in the greenhouse to determine the effects of Z.bungeanum seeds on SRKN,plant growth parameters,soil physicochemical and microbial characteristics.A total of 26 volatile components,including nematicidally active substances,were identified from Z.bungeanum seeds.Z.bungeanum seed biofumigation significantly reduced the SRKN population by 88.89%and 81.55%on the 50th and 100th day after transplanting,respectively.Compared to the control,the total soluble sugar,soluble solids,soluble protein,titratable acid,root activity and the fruit yield per tomato plant increased significantly.The content of soil alkali-hydrolyzed nitrogen,available potassium and the soil enzyme activities were also significantly increased.The soil bacterial diversity and the co-occurrence network complexity were increased by Z.bungeanum seed biofumigation.Relativelymore keystone OTUs in biofumigation soil had potential plant growth-promoting capabilities.The function of Z.bungeanum seed increasing tomato production in SRKN-infected soil depends on directly killing SRKN and improving soil properties.These results indicate that Z.bungeanum seed can be used as both a nematicide and a high quality organic fertilizer in tomato production.

Toxicity Test of Different Pesticides on Tobacco Root-knot Nematodes

[Objectives]This study was conducted to quickly screen out effective and safe agents for controlling tobacco root knot nematode disease.[Methods]Six pesticides were tested and screened indoors at five different concentrations.[Results]The six pesticides all had toxic effects on the second-instar larvae of tobacco root-knot nematode,and the corrected mortality was positively correlated with the concentration and time of the pesticides,with the correlation coefficients above 0.8.From the perspective of inhibitory activity,the order was fosthiazate>abamectin>emamectin benzoate>cadusafos>ethoprophos>carbosulfan.Four pesticides that can be used in the field were selected.[Conclusions]This study provides a theoretical basis for field pesticide selection.

Determination of Nematicidal Effects of Some Biopesticides against Root-Knot Nematode （Meloidogyne incognita） on Kiwifruit

The aim of this study was to determine the nematicidal effects of some biopesticides againist root-knot nematodes （RKNs） （Meloidogyne incognita） on Bruno type kiwifruit seedlings. Research was conducted in the Atattirk Central Horticultural Research Institute （ACHRI, Yalova, Turkey） greenhouse during 2013-2014. Biopesticides observed in this study were： sesame oil （Devalone EC）, castor oil （Ricinus communis）, Myrothecium verrucaria （M. verrucaria strain AARC-0255, Inferno DF）, Glomus spp. （Endomycorrhizal fungi, Endo Roots Soluble SP） and Paecilomyces lilacinus （Entomopathogenic fungi, P. lilacinus strain PL1, Bionematon SL）. Nematicide Nemacur EC 400 （fenamiphos） was used as treated control （TC） and trials were set up as randomized parcel design （RPD） with seven characters and 10 replicates. Reproduction ratio was determined according to scale values in trial applications and second stage juvenile （J2） populations of M. incognita in soil. The results showed that the best nematicidal effects against RKNs were determined in castor oil （77.92% in 2013 and 73.61% in 2014） and P. lilacinus （61.03% in 2013 and 55.55% in 2014）.

Efficiency of Some Plant Essential Oils on Root-Knot Nematode Meloidogyne incognita

During the last few decades, researchers have been in the search for environmental friendly ways to cope with agricultural pests, instead of using chemical pesticides. The use of essential oils has a high potential to become an alternative control strategy against plant parasitic nematodes. This study was conducted to determine the toxicity of 10 essential oils （Artemisia absinthium, Citrus bergamia, Eucalyptus citriodora, Hypericum perforatum, Lavandula officinalis, Mentha arvensis, Ocimum basilicum, Piper nigrum, Thymus serpyllum and Zingiber officinale） against the second stage juveniles of the root-knot nematode Meloidogyne incognita. The aqueous solutions of these essential oils have been applied to the second stage juveniles in three different concentrations （1%, 3% and 5%） at four different application time intervals （12, 24, 48 and 72 h）. The interactions between the variables have been examined with repeated measure analysis of variance （ANOVA）. This showed that the interactions of essential oil-time and essential oil-concentration were statistically significant, and it is concluded that L. officinalis, A. absinthium, P. nigrum, C. bergamia and M. arvensis have the most toxic effect in all concentrations and times, respectively.

Morphology and Molecular Identification of Root-knot Nematode in Partial Tobacco-growing Areas in Yunnan Province

[Objectives]The species and distribution of root-knot nematodes in tobacco growing regions of Yunnan were investigated,in order to provide reference for comprehensive prevention of root-knot nematodes disease and tobacco variety layout. [Methods]Fifty-four samples of root-knot nematode host on tobacco crops were collected from Kunming,Qujing and Yuxi cities in Yunnan from August to October in 2015. The species of root-knot nematodes were identified by observing perineal patterns and molecular biological testing（ PCR of ribosomal DNA of ITS region and sequence characterized amplified region,SCAR）. [Results] There were 24 samples belonging to Meloidogyne arenaria,accounting for 44. 44%; seven M. incognita samples,accounting for 12. 96%; one M. javanica sample,accounting for 1. 85%. Eleven samples were compound populations of M. javanica and M. arenaria,accounting for 20. 37%. Five were compound populations of M. incognita and M. arenaria,accounting for 9. 26%. Three were compound populations of M. incognita and M. javanica,accounting for 5. 56%. Three belonged to compound population of M. incognita,M. javanica and M. arenaria,accounting for 5. 56%. [Conclusions] The species of root-knot nematodes in tobacco growing areas of Yunnan mainly include M. incognita,M. javanica and M. arenaria. M. arenaria is the dominant population.

Occurrence of Root-Knot and Reniform Nematodes in Ornamental Plants Grown in Aligarh Muslim University Campus, India

An extensive survey of about 50 ornamental plants growing in A.M.U. campus were carried out to find out the infection of root-knot (Meloidogyne spp.) and reniform nematodes (Rotylenchulus reniformis). Out of 50 species of ornamental plants studied, 29 species of these plants were found to be infected with root-knot nematodes (Meloidogyne spp.) and 15 species were infected with reniform nematode (Rotylenchulus reniformis). The highest percentage of infection of root-knot nematodes was observed in Impatiens balsamina (100%) and reniform nematode was observed in Hibiscus rosa-sinensis (56%). Among the Meloidogyne spp. the highest percentage of infection of M. incognita was observed in Pseuderanthemum atropurpureum (80%) whereas, the highest percentage of infection of M. javanica and M. arenaria was observed in Mirabilis jalapa (72%) and Celosis cristata (28%).

Biological Control of Root-Knot Nematode Meloidogyne incognita in Psoralea corylifolia Plant by Enhancing the Biocontrol Efficacy of Trichoderma harzianum Using Press Mud

Meloidogyne incognita is a plant pathogen causing root-knot disease and loss of crop yield.The present study aimed to use Trichoderma harzianum as a biocontrol agent against plant-parasitic nematodes and used press mud,which is a solid waste by-product of sugarcane,as a biocontrol agent and biofertilizer.Therefore,the combined application of T.harzianum and press mud may enhance nematode control and plant growth.Elemental analysis of press mud using scanning electron microscopy(SEM)integrated with an Energy Dispersive X-ray(EDX)analyzer revealed the presence of different elements such as C,O,Mg,Si,P,K,Ca,Cu and Zn.In addition,a greenhouse study was conducted to investigate the combined effects of press mud and T.harzianum on M.incognita reproduction and growth and the biochemical features of Psoralea corylifolia.The results showed that plant length,dry biomass,leaf area,the number of seeds per plant,chlorophyll a,chl b,carotenoid content,nitrate reductase,carbonic anhydrase,and nitrogen content were significantly increased(P≤0.05)in the T2 plants(plants were treated with 100 g of press mud+50 mL T.harzianum before one week of M.incognita inoculation),over inoculated plants(IC).Antioxidant enzyme activity of ascorbate peroxidase(APX),catalase(CAT),peroxidase(POD),and superoxide dismutase(SOD)in the foliage of P.corylifolia was significantly increased when plants were treated with press mud+T.harzianum.A significant reduction in the number of egg masses,nematode population,and root-knot index(RKI)was found in plants with T2 plants.These results suggest that the combined application of T.harzianum and press mud has the potential to control the M.incognita infection and can be used as an environmentally safe alternative to chemical nematicides and also help in the removal of sugarcane waste that causes environmental pollution.

Preliminary Study on Mixed Use of Agaricus bisporus Residue and Microbial Agent to Control Cucumber Root-knot Nematode

Single or mixed use ofAgaricus bisporus residue and microbial agent had better control effect on cucumber root-knot nematode disease in greenhouse. The results showed that the control effect of single use of A. b/sporus residue was 32.9% - 34.7%, and the yield increasing rate was 2.1% - 8.7% ; the control effect of mixed use ofA. b/sporus residue and microbial agent was 59. 1% - 63. 6%, and yield increasing rate was 2.6% - 3.5%. The experimental dose had no ad- verse effects on soil physical and chemical properties. The mixed use of A. b/sporus residue and microbial agent was one of the effective ways to control root-knot nematode disease.

Genetic Mapping of Root-knot Nematode Resistance Genes in Solanaceous Vegetables

Root-knot nematodes are becoming more and more harmful to solanaceous vegetables. The most economical and effective way to deal with root-knot nematode disease is to breed varieties with disease resistance. This paper reviewed the recent research progress and achievements of tomato, pepper, eggplant and potato on root-knot nematode disease resistance gene characteristics, disease resistance gene mapping and disease resistance gene molecular markers, and prospects for future research directions.

Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus

Pepper （Capsicum annuum. L.） is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode （Meloidogyne spp.） is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chro- mosome library （BAC） via the vector of CopyControFM pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90-120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3.

Pseudomonas oryzihabitans Suppresses Damage Caused by Root-knot Nematode Meloidogyne javanica on Tomato

Agricultural research made in recent years has found that many bacterial organisms act like biological control agents with minimal impact on the environment. These microorganisms and their toxic metabolites should be included in those which are acceptable to qualify a product as organic or integrated and enjoy all the benefits that entails. Hence, interest in the use of microorganisms as biological agents to protect crop plants against plant parasitic nematodes has been increasing. This study investigated the ability of.Pseudomonas oryzihabitans symbiotically associated with the entomopathogenic nematode Steinernema abbasi as a bioagent against plant parasitic nematodes which infected tomato crop. The bacterium is particularly effective against root-knot nematode in vitro affecting the behavior and mobility of root-knot nematode juveniles. Also, studies in planta demonstrated the efficacy of P. oryzihabitans by preventing tomato root system by invasion of juveniles of Meloidogynejavaniea when bacterial cells were applied to the root system before nematodes. This efficacy is dependent on bacterial cell concentration used and the time of the nematode exposure. However, a better nematode control might be achieved with multiple applications of the biocontrol agent. Furthermore, the results of this study provide evidence that the bacterium P. oryzihabitans produces metabolites, which have nematostatic effects.

Efficient control of root-knot nematodes by expressing Bt nematicidal proteins in root leucoplasts

Root-knot nematodes(RKNs)are plant pests that infect the roots of host plants.Bacillus thuringiensis(Bt)nematicidal proteins exhibited toxicity to nematodes.However,the application of nematicidal proteins for plant protection is hampered by the lack of effective delivery systems in transgenic plants.In this study,we discovered the accumulation of leucoplasts(root plastids)in galls and RKN-induced giant cells.RKN infection causes the degradation of leucoplasts into small vesicle-like structures,which are responsible for delivering proteins to RKNs,as observed through confocal microscopy and immunoelectron microscopy.We showed that different-sized proteins from leucoplasts could be taken up by Meloidogyne incognita female.To further explore the potential applications of leucoplasts,we introduced the Bt crystal protein Cry5Ba2 into tobacco and tomato leucoplasts by fusing it with a transit peptide.The transgenic plants showed significant resistance to RKNs.Intriguingly,RKN females preferentially took up Cry5Ba2 protein when delivered through plastids rather than the cytosol.The decrease in progeny was positively correlated with the delivery efficiency of the nematicidal protein.In conclusion,this study offers new insights into the feeding behavior of RKNs and their ability to ingest leucoplast proteins,and demonstrates that root leucoplasts can be used for delivering nematicidal proteins,thereby offering a promising approach for nematode control.