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.

A new pathotype characterization of Daxing and Huangyuan populations of cereal cyst nematode(Heterodera avenae) in China

The cereal cyst nematode（CCN, Heteroder aavenae） causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and Huangyuan population in Qinghai Province, China, are two CCN populations. In this study, the CCN pathotypes of Daxing and Huangyuan populations were characterized by tests on 23 standard ＂International Test Assortment＂ with the local species Wenmai 19 as the susceptible control. Tested materials were grouped by three nematode populations＇ virulence on resistant genes（Rha1, Rha2, Rha3, Cre1） and nonresistant genes, varieties and lines. Both Daxing and Huangyuan populations were avirulent to Ortolan（Ha1）. Barley cvs. Ortolan, Siri, Morocco, Bajo Aragon 1-1, and Martin 403-2 were all resistant to both populations. Cultivars Herta, Harlan 43 and wheat Iskamish-K-2-light were all susceptible to Huangyuan population, all of them, however, were resistant to Daxing population. The other five oats were all resistant to the two tested CCN populations. Except Iskamisch K-2-light, all the other wheat cultivars（Capa, Loros×Koga, AUS 10894, and Psathias） were susceptible to Daxing population. Because the pathotypes of the two tested CCN populations in Beijing and Qinghai were not identical to any of the 13 pathotypes previously characterized by the test assortment, we classified Daxing and Huangyuan populations as the new pathotypes, named Ha91.

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.

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.

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.

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%).

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.

The population structure and sex ratios of Bursaphelenchus xylophilus under a-pinene stress

Pine wood nematode(PWN), Bursaphelenchus xylophilus, is a serious pathogen of pines throughout the world. Previous work indicated that different concentrations of a-pinene could affect nematode reproduction,however the mechanism of that influence is not clear. In order to examine the reproductive strategies of PWN in response to the stress of the volatile material a-pinene, we investigated different aspects of population changes of B.xylophilus under two concentrations of a-pinene. The results show that a high concentration(214.5 mg ml-1)promoted population growth while a low concentration(56.33 mg ml-1) decreased the population. Population structure was analyzed and it was found that a high concentration of a-pinene decreased the percentage of adults but increased the percentages of larvae and eggs.Furthermore, from the results of an evaluation of sex ratios(female/male), it was determined that a high concentration could elevate sex ratios but a low concentration decreased ratios sharply. These results suggest that the PWN could regulate its population by changing sex ratios under stress of a-pinene. This study has provided a theoretical basis for the prevention and control of pine wilt disease caused by the pine wood nematode.

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.

Effect of Pleurotus ostreatus on Dynamics of Meloidogyne arenaria Population and Control Effectiveness

The control effects and application methods of Pleurotus ostreatus on the peanut root-knot nematode disease, Meloidogyne arenaria, were systematically studied in the greenhouse. Results of pot experiments showed that P. ostreatus could effectively reduce the 2nd-instar population density in the soil, and the larva reduced by 80% compared with the control. P. ostreatus could markedly lower the infecting number of the peanut root-knot nematodes, the nematode density inside the root knots declined by 40.00% -82.64%, and the peanut root knots also reduced by 86.96% - 94. 03%. The fungus could defer the initial infection of M. arenaria to the roots, the epidemic rate of the peanut root-knot nematode disease was significantly cut down, and the increase of root-knot changed from the way of Logistic curve in the control to the way of linear curve in the treatment. The final control effectiveness was about 80%. The results of pot trial also indicated that the control effectiveness is closely related to the application time and the amount of this fungus. The key factor affecting control effectiveness was the application time of P. ostreatus in the soil and the suitable time was 20 days before sowing.

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.

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.

Chemical mutagenesis and soybean mutants potential for identification of novel genes conferring resistance to soybean cyst nematode

The resistance of soybean(Glycine max(L.) Merr.) to soybean cyst nematode(SCN, Heterodera glycines Ichinohe), which is a devastating pathogen in soybean production and causes a large quantity of annual yield loss worldwide, can shift during the long-term interaction and domestication. It is vital to identify more new resistance genetic sources for identification of novel genes underlying resistance to SCN for management of this pathogen. In the present study, first, two ethane methylsulfonate-mutagenesis soybean M2 populations of PI 437654, which shows a broad resistance to almost all of SCN races, and Zhonghuang 13, which is a soybean cultivar in China conferring strong resistance to lodging, were developed. Many types of morphological phenotypes such as four-and five-leaflet leaves were observed from these two soybean M2 populations. Second, 13 mutants were identified and confirmed to exhibit alteration of resistance to SCN race 4 through the forward genetic screening of 400 mutants of the PI 437654 M2 population, the rate of mutants with alteration of SCNinfection phenotype is 3.25%. Third, these identified mutants were further verified not to show any changes in the genomic sequences of the three known SCN-resistant genes, GmSHMT08, GmSNAP18 and GmSANP11, compared to the wildtype soybean; and all of them were still resistant to SCN race 3 similar to the wild-type soybean. Taken together, we can conclude that the 13 mutants identified in the present study carry the mutations of the new gene(s) which contribute(s) to the resistance to SCN race 4 in PI 437654 and can be potentially used as the genetic soybean sources to further identify the novel SCN-resistant gene(s).

Mating and reproductive characteristics of the pathogenic nematode Bursaphelenchus xylophilus

Pine wood nematode(B ursaphelenchus xylophilus),one of the most destructive invasive species,has caused extremely serious economic,ecological and social losses in many countries throughout the world.Since the high reproductive rate of B.xylophilus PWN is the main cause of rapid death of its pine hosts(Pinus spp.),understanding the reproductive and population biology and the ecology of this nmatode are of great importance.This study mainly focused on analyzing the mating process and population structure under different combinations of sex ratios for mating.Reproductive efficiency of B.xylophilus peaked when the sex ratio(female to male)was 3.4:1.Phases of the mating process for the different sex-ratio combinations indicated that B.xylophilus had evolved alternative reproductive strategies to cope with complex copulating conditions to obtain a suitable population structure for further propagation.This research provides fundamental information on the mechanism that is responsible for the rapid population growth of B.xylophilus.

RNAi of <i>MiASB</i>caused high mortality of <i>Meloidogyne incognita</i>juveniles and inhibited the nematode disease

The southern root-knot nematode, Meloidogyne incognita, is one of the most prevalent and damaging plant-parasitic nematodes in the world and causes serious damages to agricultural production. We cloned a mitochondrial ATP synthase b subunit gene fragment of M. incognita (MiASB) based on the nematode genomics prediction. By soaking in the MiASB dsRNA solution, the hatching of RNAi treated eggs was reduced by 60% compared to negative control and by 64% compared to untreated control. Mortality of RNAi treated second stage juvenile (J2) was 8.6 times higher than that of negative control and 26 times higher than the untreated control. Inoculating the RNAi treated egg masses and J2 to tomato seedlings showed the pathogencity was significantly reduced. For the RNAi treated egg masses, the amount of root galls on silence treated seedlings was reduced by 92% compared to that on the negative control seedlings, and reduced by 93% compared to that on untreated control seedlings. For the treated J2, the amount of root galls on silence treated seedlings was reduced by 83% and 86% compared to negative and untreated control seedlings, respectively. The study revealed the MiASB silence had a positive effect on prevention and control of root-knot nematode disease, and also showed that the MiASB may be involved in the pathogenesis of nematode, which provided new ideas and ways to the research of nematode pathology and nematode disease control.

甘薯茎线虫病抗扩展性遗传特性分析与QTL定位

甘薯茎线虫病由马铃薯腐烂线虫引起,是严重影响甘薯产量和品质的检疫性病害。挖掘抗茎线虫病基因并通过分子设计育种培育抗病品种是防控茎线虫病的有效途径。本研究前期以抗茎线虫病甘薯品种美国红为父本,感病品种徐紫薯8号为母本,通过控制授粉有性杂交方式构建了包含274个F1子代的分离群体。以该F1群体为材料,利用室内人工接种法对F_(1)子代的茎线虫病抗扩展性进行鉴定,结果表明,甘薯茎线虫病抗扩展性呈连续性偏峰态分布,甘薯茎线虫病发病体积比与扩展直径和扩展长度呈极显著正相关,与薯块直径、薯块长度和薯块长宽比无相关性,说明薯块的大小和薯形对抗扩展性鉴定结果无影响。甘薯茎线虫病抗扩展性遗传力为75.7%,表明抗扩展性主要受遗传因素控制。基于前期构建的甘薯SNP遗传图谱对抗扩展性进展QTL定位,获得与抗扩展性紧密连锁的QTL 10个,解释6.6%~10.7%的表型变异。候选基因功能注释表明,苯丙素生物合成、植物激素信号转导、植物病原互作代谢等通路参与抗病胁迫。筛选5个关键候选基因进行荧光定量表达分析,在接种茎线虫后候选基因itf13g19570表达量显著增高。研究结果为甘薯茎线虫病抗病基因挖掘和抗病机理解析提供了重要参考。

不同悉生培养条件下食细菌线虫对细菌种群的影响

1 引言 线虫是地球上物种多样性和功能类群最丰富的生物种之一,土壤自生线虫(Free-living Nematodes)约占线虫总数的52%,而其中大部分属于食细菌线虫[3,4,8,9].来自田间和土培试验的研究表明,食细菌线虫在土壤生态系统有机物分解、养分矿化等生态过程中起着重要作用[1,3-7]但由于土壤的高度异质性和环境的复杂性,迄今对食细菌线虫与细菌在种群水平上的相互作用尚缺乏深入的了解,限制了对该类土壤动物功能及作用机理的认识[6].

松树对松材线虫抗性的研究

通过23种松树、5种松树的105个种源及13种非松属树木对松材线虫Bursaphelenchusxylophilus的抗性研究,结果表明:供试松树的抗性差异很大,火炬松、班克松、刚松均未感病。马尾松1年生幼苗感病,2、4年生幼苗及10年生幼树抗病。湿地松3年生幼苗感病,10年生幼树未感病。美国五针松在不同地方的抗病性差异很大。其它17种均不同程度地感病。华山松21个种源均严重感病,油松21个种源均感病。马尾松、火炬松及湿地松的供试种源均未感病。除雪松外,其它12种非松属树木均感病。环境条件影响松树的抗性。低温不利病害发展。不同来源的松材线虫对同种松树的致病力不同,地理位置不同的线虫致病性差异要大于地理位置相同、寄主不同的松材线虫。接种量和松树枯死率呈正相关。