福建省直播稻根结线虫病的病原种类鉴定

[目的] 福建省福清市江镜镇与福安市潭溪镇水稻区直播稻苗期分别发生严重的根结线虫病,本研究对其病原物进行形态和分子鉴定,明确病原物种类,以期为福建省直播稻根结线虫病害防控提供理论依据。[方法] 通过根结线虫各虫态形态学特征进行观测;同时对其rDNA-ITS区进行测序,通过贝叶斯法与最大似然法构建了系统发育树来确定种类;利用拟禾本科根结线虫特异性引物Mg-F/Mg-R检测种群。[结果] 根结线虫的雌虫、雄虫和2龄幼虫的形态学特征与拟禾本科根结线虫原始描述种一致;rDNA-ITS区序列长度为576 bp,与GenBank拟禾本科根结线虫种群相似度均达99%以上;系统发育树明确了该根结线虫与拟禾本科根结线虫处于同一分支;特异性引物Mg-F/Mg-R检测进一步明确病原为拟禾本科根结线虫单一种群。[结论] 本研究通过形态与分子特征,明确了在福建直播稻上发现的根结线虫为拟禾本科根结线虫。拟禾本科根结线虫在福建省最早于2011年在政和县小范围水稻田发现,此后未在其他水稻种植区发现。本次在福建直播稻上首次发现大面积的根结线虫为害。随着直播稻的种植面积扩大,拟禾本根结线虫引起的水稻病害可能会成为制约其发展的新问题,应引起足够重视。     

Host response of rice genotypes to the rice root-knot nematode (Meloidogyne graminicola) under aerobic soil conditions

Aerobic rice is a production system where adapted rice varieties are established via direct seeding in non-puddled, non-flooded, non-saturated fields and grown under conditions similar to upland conditions. On land cultivated continuously with aerobic rice, a yield reduction has been observed. The rice root-knot nematode Meloidogyne graminicola is considered one of the possible causes of these yield reductions. Resistance to and tolerance for M. graminicola are essential traits for aerobic rice cultivars in alleviating this problem. In our study, the host response of nineteen aerobic, seven upland and four lowland rice genotypes which are either being used in the International Rice Research Institute’s aerobic rice breeding programme or already cultivated by farmers in Asia was evaluated under aerobic soil conditions in an outdoor raised-bed experiment. Our study showed a large variation in susceptibility and sensitivity to M. graminicola infection among the rice genotypes examined. Resistance comparable to the resistant reference genotypes included in the experiment (CG14, TOG5674, TOG7235) was not found but in terms of susceptibility, the upland genotype Morobereken may be an interesting less-susceptible genotype. Tolerance was found and in terms of sensitivity, the high yielding aerobic genotype IR78877-208-B-1-2 may be an interesting tolerant genotype. Our study also allowed the identification of rice genotypes that are either highly susceptible or sensitive to M. graminicola infection and of which the cultivation should be discouraged. On average, M. graminicola caused an almost 30% reduction in yield. Excluding the two susceptible and three resistant reference genotypes included in the experiment, most affected was dry-shoot biomass (23.6% reduction) followed by root length, which was more affected than fresh-root weight (19.8 vs. 8%) and grain filling (17.3%), while plant height and the number of spikelets/panicle were less affected (10.2 and 8.1%, respectively). Neither tillering nor the number of panicles/plant were affected.     

Comparison of Reproduction by Meloidogyne graminicola and M. incognita on Trifolium Species

The reproductive potential of Meloidogyne graminicola was compared with that of M. incognita on Trifolium species in greenhouse studies. Twenty-five Trifolium plant introductions, cultivars, or populations representing 23 species were evaluated for nematode reproduction and root galling 45 days after inoculation with 3,000 eggs of M. graminicola or M. incognita. Root galling and egg production by the two root-knot nematode species was similar on most of the Trifolium species. In a separate study, the effect of initial population densities (Pi) of M. graminicola and M. incognita on the growth of white clover (T. repens) was determined. Reproductive and pathogenic capabilities of M. graminicola and M. incognita on Trifolium spp. were similar. Pi levels of both root-knot nematode species as low as 125 eggs per 10-cm-d pots severely galled white clover plants after 90 days. Meloidogyne graminicola has the potential to be a major pest of Trifolium species in the southeastern United States.     

Occurrence of root-knot disease in green onion in Allahabad

Onion is an important indispensable item in every kitchen as condiment and vegetable, hence commands an extensive internal market. The green leaves and immature and mature bulbs are eaten raw or used in preparation of vegetables. Among the various pests and diseases associated with onion, the root knot nematode (Meloidogyne incognita) had proved itself as an important limiting factor for successful cultivation of this crop. Recently root-knot nematode was observed to cause serious losses to green onion crop grown mostly in the field located near the Jamuna & Ganga river belt of Allahabad and the surrounding area in U.P. Differential host test was done to confirm the root knot disease in onion. The specific identity of the nematode was determined by cutting perineal pattern of the females and was confirmed as Meloidogyne incognita.     

Expression of Tolerance for Meloidogyne graminicola in Rice Cultivars as Affected by Soil Type and Flooding

The effects of different water regimes on the pathogenicity of Meloidogyne graminicola on six rice cultivars were determined in two soil types in three greenhouse experiments. Two water regimes, simulating continuous flooding and intermittent flooding, were used with five of the cultivars. All cultivars were susceptible to the nematode, but IR72 and IR74 were more tolerant than IR20 and IR29 under intermittent flooding. All were tolerant under continuous flooding. UPLRi-5 was grown under multiple water regimes: no flooding; continuous flooding; flooding starting at maximum tillering, panicle initiation, or booting stage; and flooding from sowing until maximum tillering or booting. In sandy loam soil, M. graminicola reduced stem and leaf dry weight, root dry weight, and grain weight under all water regimes. In clay loam soil, the nematode reduced root weight when the soil was not flooded or flooded only for a short time, from panicle initiation, or booting to maturity, and from sowing to maximum tillering. In clay loam soil, stem and leaf dry weight, as well as grain weight, were reduced by the nematode under all water regimes except continuous flooding or when the soil was flooded from sowing to booting stage. These results indicate that rice cultivar tolerance of M. graminicola varies with water regime and that yield losses due to M. graminicola may be prevented or minimized when the rice crop is flooded early and kept flooded until a late stage of development.     

Evaluation of 31 Potential Biofumigant Brassicaceous Plants as Hosts for Three Meloiodogyne Species

Brassicaceous cover crops can be used for biofumigation after soil incorporation of the mowed crop. This strategy can be used to manage root-knot nematodes (Meloidogyne spp.), but the fact that many of these crops are host to root-knot nematodes can result in an undesired nematode population increase during the cultivation of the cover crop. To avoid this, cover crop cultivars that are poor or nonhosts should be selected. In this study, the host status of 31 plants in the family Brassicaceae for the three root-knot nematode species M. incognita, M. javanica, and M. hapla were evaluated, and compared with a susceptible tomato host in repeated greenhouse pot trials. The results showed that M. incognita and M. javanica responded in a similar fashion to the different cover cultivars. Indian mustard (Brassica juncea) and turnip (B. rapa) were generally good hosts, whereas most oil radish cultivars (Raphanus. sativus ssp. oleiferus) were poor hosts. However, some oil radish cultivars were among the best hosts for M. hapla. The arugula (Eruca sativa) cultivar Nemat was a poor host for all three nematode species tested. This study provides important information for chosing a cover crop with the purpose of managing root-knot nematodes.     

植物罹病组织中南方、爪哇、花生根结线虫的LAMP快速检测

【目的】建立一种基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,从植物罹病组织中直接检测3种常见的根结线虫,为根结线虫的监测和防治提供技术支持。【方法】分别采用3种根结线虫的种类特异性引物对所选择的根结线虫的DNA片段进行PCR扩增,扩增产物纯化、回收并测序。根据3种根结线虫的测序结果,针对种类特异区段,采用PrimerExplorerV4软件,分别设计3种根结线虫的LAMP引物。设计的引物组人工合成后,以提取的纯化种群线虫DNA为模板,分别进行引物组的特异性测试,筛选出分别针对3种根结线虫的最佳引物组。【结果】研究设计的3种根结线虫的LAMP特异性引物能够直接从植物根结中检测出南方、花生、爪哇3种常见根结线虫,LAMP快速检测体系为:dNTPS浓度为1 mmol·L~(-1),Mg~(2+)的浓度为5 mmol·L~(-1),不添加甜菜碱,反应时间为45 min。【结论】本实验建立的南方、花生、爪哇根结线虫LAMP快速分子检测方法,具有特异性强、灵敏度高、简单、快速、经济等特征,能够从罹病植物组织中快速准确地检测出南方、花生和爪哇根结线虫,具有极高的实践应用价值。     

Host response of Oryza glaberrima and O. sativa rice genotypes to the rice root-knot nematode Meloidogyne graminicola in a hydroponic system under growth chamber

The host response of 25 rice genotypes belonging to Oryza glaberrima and Oryza sativa to Meloidogyne graminicola infection was examined in a hydroponic system. The M. graminicola can build up high population densities in a hydroponic system. Resistance to this nematode species was found in O. glaberrima genotypes which supported significantly lower nematode numbers per plant and per unit root than O. sativa genotypes. The M. graminicola-infected O. sativa genotypes showed a higher root galling index than the O. glaberrima genotypes. The hydroponic system is efficient and reliable method to examine the host response of rice genotypes to M. graminicola infection, and can be useful for the fast screening of high numbers of rice genotypes for the selection of M. graminicola-resistant rice germplasm for breeding purposes.     

Biological control of rice root-knot nematode,Meloidogyne graminicola through mixture of Pseudomonas fluorescens strains

The plant growth promoting rhizobacterium, Pseudomonas fluorescens strains PF1, TDK1, and PY15 were evaluated individually and in combinations for their efficacy against root-knot nematode, Meloidogyne graminicola, in rice plants under in vitro, glass house and field conditions. Culture filtrates of these strains either individually or as mixture inhibited egg hatching and caused mortality of juveniles of M. graminicola in vitro. The efficacy was more pronounced when filtrates of the strain were used as mixtures than as individual strains. Mixtures of P. fluorescens strains signficantly reduced M. graminicola infestation when applied as bacterial suspensions through seed treatment. The higher activity of peroxidase and chitinase enzymes was observed in plants treated with P. fluorescens mixtures than the plants treated with individual strains, two strain mixtures and untreated control. In field trials on rice, talc formulations of the P. fluorescens strains individually as well as mixtures were evaluated as seed treatment, soil treatment and combination of both. A mixture of the three strains was the most effective when applied either as seed + soil treatment or as seed treatment alone. The introduced P. fluorescens strains survived endophytically on rice roots. The application of the P. fluorescens mixture PF1 + TDK1 + PY15 in seed + soil treatment resulted in higher grain yield which provided a 27.3% increase over the control followed by P. fluorescens mixture PF1 + TDK1 + PY15 in seed treatment alone, which increased the grain yield of rice by 24.7% compared to the control.     

Mustard biofumigation disrupts biological control by Steinernema spp. nematodes in the soil

Mustard green manures or seed meal high in glucosinolates, which produce a natural biofumigant upon incorporation into the soil, form an alternative to synthetic fumigants. However, the non-target impacts of these biofumigants in the field are unclear. We examined the effectiveness of soil incorporation of Brassica carinata seed meal both in controlling the plant-parasitic Columbia root-knot nematode (Meloidogyne chitwoodi), and on the biological control exerted by the entomopathogenic nematodes Steinernema feltiae and Steinernema riobrave on root-knot nematodes and the Colorado potato beetle (Leptinotarsa decemlineata). Singly, both the seed meal and Steinernema spp. reduced root-knot nematode damage to potato tubers and increased marketable tuber yields. However, there was a negative interaction between the two bioagents such that their combination did not further improve suppression of plant-parasitic nematodes. Thus, mustard seed meal applications harmful to the target root-knot nematode also disrupted the ability of Steinernema spp. to act as biocontrol agents. Further, we observed modest disruption of the biological control of potato beetles following biofumigation. But, the potato beetles were less likely to lay eggs on potato plants grown in mustard-amended soil, suggesting a counteracting benefit of mustard application. Multiple, complementary controls must be integrated to replace the very effective pest suppression typical of synthetic soil fumigants. Our study suggests significant interference between biofumigation and biocontrol agents in the soil, presenting challenges in combining these two environmentally friendly approaches to managing plant-parasitic nematodes and other pests.     

Utilization of noxious weeds for the management of plant-parasitic nematodes infesting some vegetable crops: Einsatz von schädlichen unkräutern zur bekämpfung von pflanzenparasitären nematoden auf verschiedenen gëmusekulturen

Significant reduction was observed in the population of plant-parasitic nematodes, Meloidogyne incognita, Rotylenchulus reniformis and Tylenchorhynchus brassicae infesting eggplant and cauliflower when given root-dip treatment in the leaf extracts of Argemone maxicana and Solanum xanthocarpum at different concentrations and dip durations. The root-knot development and larval penetration of second stage juveniles of M. incognita were also inhibited, may be due to bare-root dip treatment in leaf extracts of both the weed plants. Leaf extracts of S. xanthocarpum caused more inhibition in root-knot development, nematode multiplication of reniform and stunt nematodes than that of A. maxicana. Plant growth improvement was noted which seems to be due to dip treatment and reduction in the population of parasitic nematodes. The efficacy of root-dip treatment with respect to improvement in plant growth of eggplant and cauliflower and reduction in root-knot development and nematode population, increased with increasing the concentration of leaf extracts and dip durations.     

Characterization of Root-Knot Nematode Resistance in Medicago truncatula

Root knot (Meloidogyne spp.) and cyst (Heterodera and Globodera spp.) nematodes infect all important crop species, and the annual economic loss due to these pathogens exceeds $90 billion. We screened the worldwide accession collection with the root-knot nematodes Meloidogyne incognita, M. arenaria and M. hapla, soybean cyst nematode (SCN-Heterodera glycines), sugar beet cyst nematode (SBCN-Heterodera schachtii) and clover cyst nematode (CLCN-Heterodera trifolii), revealing resistant and susceptible accessions. In the over 100 accessions evaluated, we observed a range of responses to the root-knot nematode species, and a non-host response was observed for SCN and SBCN infection. However, variation was observed with respect to infection by CLCN. While many cultivars including Jemalong A17 were resistant to H. trifolii, cultivar Paraggio was highly susceptible. Identification of M. truncatula as a host for root-knot nematodes and H. trifolii and the differential host response to both RKN and CLCN provide the opportunity to genetically and molecularly characterize genes involved in plant-nematode interaction. Accession DZA045, obtained from an Algerian population, was resistant to all three root-knot nematode species and was used for further studies. The mechanism of resistance in DZA045 appears different from Mi-mediated root-knot nematode resistance in tomato. Temporal analysis of nematode infection showed that there is no difference in nematode penetration between the resistant and susceptible accessions, and no hypersensitive response was observed in the resistant accession even several days after infection. However, less than 5% of the nematode population completed the life cycle as females in the resistant accession. The remainder emigrated from the roots, developed as males, or died inside the roots as undeveloped larvae. Genetic analyses carried out by crossing DZA045 with a susceptible French accession, {"type":"entrez-protein","attrs":{"text":"F83005","term_id":"11352626","term_text":"pir||F83005"}}F83005, suggest that one gene controls resistance in DZA045.     

Techniques for image analysis of movement of juveniles of root-knot nematodes encumbered with Pasteuria penetrans spores

Root-knot nematodes (Meloidogyne spp.) are the most significant plant-parasitic nematodes that damage many crops all over the world. The free-living second stage juvenile (J2) is the infective stage that enters plants. The J2s move in the soil water films to reach the root zone. The bacterium Pasteuria penetrans is an obligate parasite of root-knot nematodes, is cosmopolitan, frequently encountered in many climates and environmental conditions and is considered promising for the control of Meloidogyne spp. The infection potential of P. penetrans to nematodes is well studied but not the attachment effects on the movement of root-knot nematode juveniles, image analysis techniques were used to characterize movement of individual juveniles with or without P. penetrans spores attached to their cuticles. Methods include the study of nematode locomotion based on (a) the centroid body point, (b) shape analysis and (c) image stack analysis. All methods proved that individual J2s without P. penetrans spores attached have a sinusoidal forward movement compared with those encumbered with spores. From these separate analytical studies of encumbered and unencumbered nematodes, it was possible to demonstrate how the presence of P. penetrans spores on a nematode body disrupted the normal movement of the nematode.     

The role of thionins in rice defence against root pathogens

Thionins are antimicrobial peptides that are involved in plant defence. Here, we present an in‐depth analysis of the role of rice thionin genes in defence responses against two root pathogens: the root‐knot nematode Meloidogyne graminicola and the oomycete Pythium graminicola. The expression of rice thionin genes was observed to be differentially regulated by defence‐related hormones, whereas all analysed genes were consistently down‐regulated in M. graminicola‐induced galls, at least until 7 days post‐inoculation (dpi). Transgenic lines of Oryza sativa cv. Nipponbare overproducing OsTHI7 revealed decreased susceptibility to M. graminicola infection and P. graminicola colonization. Taken together, these results demonstrate the role of rice thionin genes in defence against two of the most damaging root pathogens attacking rice.     

Host response of rice genotypes from crosses of high-yielding and drought-tolerant Oryza sativa advanced breeding lines to the rice root-knot nematode Meloidogyne graminicola

The host response to Meloidogyne graminicola infection of 30 advanced breeding lines developed from crosses between high-yielding and drought-tolerant Oryza sativa genotypes was evaluated in outdoor raised beds. None of the advanced breeding lines showed resistance to M. graminicola comparable to the M. graminicola-resistant African rice (O. glaberrima) genotype TOG5674. However, rice genotype IR86815-23-4-1-2 was identified tolerant which showed no yield reduction even when infected with M. graminicola. IR82635-B-B-143-1, IR85733-19-4-1-1, IR85733-19-4-2-4, IR85,735-42-1-4-1 and IR 85735-42-1-4-4 were less sensitive to M. graminicola infection with less than 20% yield reduction compared to other rice genotypes. These rice genotypes could alleviate yield losses in M. graminicola-infested rice fields in regions with limited resources to control M. graminicola and when resistant cultivars are not available.     

Distribution and Identification of Root-knot Nematodes from Turkey

Root knot nematodes are causing serious losses in protected cultivation fields in the West Mediterranean region of Turkey. Correct and confident identification of the plant parasitic nematodes is important for vegetable growing and breeding. Therefore, ninety-five populations of plant parasitic nematodes were collected from regional greenhouses. Previously described species-specific primers were used to identify Meloidogyne populations. The present study indicated that SEC-1F/SEC-1R and INCK14F-INCK14R primers for identifying of M. incognita, Fjav/Rjav and DJF/DJR primers for M. javanica and Far/Rar for M. arenaria primers can be effective tools to identify the Turkish root-knot nematode species. Dissemination ratios of the population were 64.2%, 28.4% and 7.3% for Meloidogyne incognita, M. javanica and M. arenaria, respectively. The results showed that M. incognita was the prominent root-knot nematode species in the West Mediterranean coastal areas of Turkey.     

Characterization of Isolates of Meloidogyne from Rice-Wheat Production Fields in Nepal

Thirty-three isolates of root-knot nematode were recovered from soil samples from rice-wheat fields in Nepal and maintained on rice cv. BR 11. The isolates were characterized using morphology, host range and DNA sequence analyses in order to ascertain their identity. Results indicated phenotypic similarity (juvenile measurements, perennial pattern, host range and gall shape) of the Nepalese isolates with Meloidogyne graminicola, with minor variations. The rice varieties LA 110 and Labelle were susceptible to all of the Nepalese isolates, but differences in the aggressiveness of the isolates were observed. Phylogenetic analyses based on the sequences of partial internal transcribed spacer (ITS) of the rRNA genes indicated that all Nepalese isolates formed a distinct clade with known isolates of M. graminicola with high bootstrap support. Furthermore, two groups were identified within the M. graminicola clade. No correlation between ITS haplotype and aggressiveness or host range was found among the tested isolates.     

Survey report on occurrence of root knot disease in winged bean

Winged bean (Psophocarpus tetragonolobus) is of rapidly increasing interest as a high-protein multipurpose crop; in the future the winged bean could become as important as the soybean in world agriculture, with the added bonus of yielding substantial quantities of edible, high-protein root tubers. During the survey of root-knot nematodes (Meloidogyne species) on vegetable crops in Uttar Pradesh (India), it was observed that winged bean was restricted to home gardens and backyard cultivation where it was found severe galling of roots and reduces tuber production and may affect pod and seed yield. Sometimes leaf anthracnose disease was also observed but it does not cause severe loss to the crop.