玉米线虫矮化病病原鉴定

为明确东华北春玉米区暴发的一种危害严重、引起玉米植株矮化的新病害病原,从病区采集大量病株及其根际土壤,采用室内人工接种的方法对分离频率最高的3种镰孢菌,即亚粘团镰孢菌Fusarium subglutinans、拟轮枝镰孢菌F.verticillioides和层出镰孢菌F.proliferatum及线虫(群体)进行致病性测定。结果表明:有线虫(群体)的处理能够引起该病害典型症状,即植株矮化,叶片上有沿叶脉方向的黄色或白色失绿条纹,茎基部组织从内向外呈纵向或横向开裂,推断该病的发生和线虫有关。将回接发病植株病土中分离的矮化线虫属Tylenchorhynchus sp.、短体线虫属Pratylenchus sp.、发垫刃线虫属Trichotylenchus sp.、拟盘旋线虫属Pararotylenchus sp.、小环线虫属Criconemella sp.、垫刃线虫属Tylenchus sp.、丝尾垫刃线虫属Filenchus sp.、螺旋线虫属Helicotylenchus sp.和真滑刃线虫属Aphelenchus sp.进行分属后再回接,只有发垫刃属线虫的处理出现与田间发病症状一致的植株。综合形态特征、致病力和分子序列分析结果,确定该病病原为长岭发垫刃线虫T.changlingensis,并将该病害称为玉米线虫矮化病(maize nematode stunt disease)。     

深圳市观赏植物根际茎线虫的种类鉴定

在对深圳市观赏植物线虫进行调查监测的过程中,采集了长势不良的观赏植物的根和根际土壤样品,用改良贝曼漏斗法分离到植物线虫.其中从万年青(A glaonema modestum)、国王椰子(Ravenea rivularis)和袖珍椰子(Collinia elegans)的根际分离到的2种茎属线虫,经形态鉴定确认分别为食菌茎线虫(Ditylenchus myceliophagus Goodey,1958)和短小茎线虫(Ditylenchus nanus Siddiqi,1963),并对其形态和测量数值进行了详细记述.     

健康与感染青枯病烟株根际土壤与茎秆真菌群落结构与多样性

为研究感染青枯病后烟株根际土壤与茎秆真菌群落结构与多样性的变化,对健康和感染青枯病烟株的根际土壤、病株茎秆发病组织和健株茎秆健康组织等样品中真菌ITS区的rDNA进行了PCR扩增、用Illumina MiSeq测序技术对扩增DNA片段进行高通量测序,并分析不同样品的真菌群落组成与多样性。结果表明,所有烟株根际土壤中优势门为子囊菌门Ascomycota和接合菌门Zygomycota;所有茎秆样品中优势门为担子菌门Basidiomycota和子囊菌门。在属水平,被孢霉属Mortierella、镰刀菌属Fusarium和隐球菌属Cryptococcus为所有土壤中的主要菌属,Boeremia主要存在于发病烟株根际土壤中,而木霉属Trichoderma主要存在于健康烟株根际土壤。发病茎秆病害组织中优势属为小画线壳属Monographella、隐球菌属、鬼伞属Coprinopsis和赤霉属Gibberella;发病茎秆病健交界处组织中优势属为隐球菌属、红酵母属Rhodotorula和小画线壳属。健康烟株茎秆组织中优势属为隐球菌属、链格孢属Alternaria和红酵母属;健康烟株中与发病茎秆病健交界处组织等高茎秆中优势属为镰刀菌属、隐球菌属、链格孢属和Gibellulopsis。青枯菌侵染烟株后根际土壤、发病茎秆病害组织和发病茎秆病健交界处组织的真菌群落中物种丰富度与多样性均显著提高,且发病茎秆病害组织与发病茎秆病健交界处组织真菌群落的变化大于根际土壤。研究结果为烟草青枯病的生物防治提供了参考。     

北方春玉米区长岭发垫刃线虫的分布及遗传多样性分析

 为明确长岭发垫刃线虫(Trichotylenchus changlingensis)的分布,2018年对黑龙江、吉林、辽宁、河北、内蒙古、甘肃6省(自治区)118个地块的玉米根际土壤中长岭发垫刃线虫进行分离及鉴定,结果表明:在6个省(自治区)20个地块的土样中分离到该线虫,分离比例为17.0%。同时采用ISSR分子标记技术对分离到的20个种群进行遗传多样性分析。结果共筛选出16条引物,扩增出93条多态性条带,多态性比率为93.55%。基因分化系数(Gst)为0.4771,遗传变异的47.71%存在于种群间,52.29%存在于种群内。基因流(Nm)值为0.5479,说明所调查地区种群间存在基因流,基因流动较小,但种群间已发生一定程度的遗传分化。UPGMA 聚类分析结果表明,这20个种群具有丰富的遗传多样性,但群体间的遗传关系远近与地理距离无显著相关性。     

菌株CSR-2对玉米茎腐病的生防作用及其全基因组序列分析

本研究从玉米根际土壤中分离获得一株具有良好生防效果的细菌菌株CSR-2,该菌株对多种植物病原真菌均具有拮抗作用,抑制禾谷镰孢菌丝生长、形态及孢子萌发.将菌株CSR-2与玉米膜下滴灌水肥药一体化技术联用田间试验表明,该菌株对玉米茎腐病的防治效果为52.3％,可显著降低玉米茎腐病的田间发病率.利用第三代PacBio测序技术...     

上海盆景植物寄生线虫种类与群体动态

调查上海地区出口盆景，发现15属植物寄生线虫，其中垫刃线虫、螺旋线虫、环线虫、根结线虫、毛刺线虫、短体线虫、剑线虫、矮化线虫、滑刃线虫等是上海口岸盆景植物主要寄生线虫种类。盆景根围线虫群体密度在1年中的变化与气候变化明显相关。从春到秋，种群数量呈增长趋势，群体结构呈多样化发展，秋天是线虫种群数量、种类最为繁盛时期。     

甘肃酒泉甘草根际土壤线虫群落多样性研究

为明确甘肃酒泉地区甘草根际土壤线虫群落特征,于2014年7月和9月,采集甘草根际土壤,利用淘洗-过筛-蔗糖离心法分离线虫,根据线虫形态学特征进行鉴定。结果表明,从甘草根际土壤样品中共分离到植物寄生线虫1目4科5属,非植物寄生线虫4目7科16属。真滑刃属(Aphelenchus)和中杆属(Mesorhabditis)为甘草根际土壤线虫的优势属。与7月份相比,9月份甘草根际土壤线虫个体密度和类群数显著减少,多样性指数(H′)和丰富度指数(SR)降低,且丰富度指数(SR)降幅显著,均匀度指数(J′)显著高于7月份,月份间PPI/MI差异显著,表明外界环境及人类活动对甘草土壤线虫功能类群影响较大;瓦斯乐斯卡指标(WI)为6.55,且7月份显著大于9月份,表明酒泉地区土壤健康指数较高,土壤健康状况良好。     

连作和轮作对大豆胞囊线虫群体数量及土壤线虫群落结构的影响

大豆胞囊线虫Heterodera glycinesIchinohe病害是大豆生产中的毁灭性病害,对世界大豆生产造成重大损失。采用根染色法和常规土壤线虫分类鉴定技术研究了连作和轮作两种耕作方式下大豆胞囊线虫群体数量和土壤线虫群落结构的变化。结果表明,连作使大豆田土壤中胞囊和根系上各龄期胞囊线虫数量显著增加(95.6→335.5),而轮作积累很少(10.3→31.8),感病品种中J2的侵入数量决定了大豆胞囊线虫形成胞囊的数量(22.8→95.6,129.4→335.5)。玉米-玉米-大豆的轮作方式使胞囊积累最少(10.3)。轮作大豆田土壤线虫总数高于连作,主要由于轮作大豆田土壤线虫的优势属Helicotylenchus和优势营养类群植物寄生线虫PP的相对丰度高于连作,但大豆胞囊线虫的相对丰度正相反,连作(33.65%)高于轮作(0.91%),说明Heterodera glycines可以作为土壤线虫的关键属种用以指示土壤发病的严重程度。研究中重要的优势属和关键属种cp值均为3,说明其繁殖和抗干扰能力比较强。研究证明土壤线虫优势营养类群、优势属种和关键属种可以有效地指示农业土壤健康状况和大豆田的大豆胞囊线虫病害的发生程度。     

一株拮抗链霉菌SM3-7的鉴定及其对玉米茎基腐病的防效

为获得对玉米茎基腐病具有良好防效的拮抗菌株,从玉米根际土壤中分离以玉米茎基腐病拟轮枝镰孢菌Fusarium verticillioides为靶标菌的拮抗放线菌菌株。采用平板对峙法测定不同菌株的抑菌活性,用抑菌圈法筛选效果良好的拮抗菌株并进行种类鉴定、抑菌谱和防效测定。结果表明,从土壤中分离并筛选获得1株对拟轮枝镰孢菌具有明显抑制作用的菌株SM3-7。经形态学、生理生化特征和16S rDNA系统发育分析,将菌株SM3-7鉴定为核粒链霉菌Streptomyces sclerogranulatus。该菌株抑菌谱广,对玉米上的8种植物病原菌有良好的抑制作用,对拟轮枝镰孢菌抑菌圈直径可达（28.03±0.45mm）。盆栽防效结果表明,菌株SM3-7发酵液对抗病品种“良玉99”和感病品种“全玉1233”的玉米茎基腐病14d防效分别达77.28%和76.12%,21d防效达72.20%和71.89%。核粒链霉菌菌株SM3-7发酵液可有效防治玉米茎基腐病,为利用微生物防治玉米茎基腐病拟轮枝镰孢菌提供参考。     

长岭发垫刃线虫(Trichotylenchus changlingensis)双重PCR检测方法研究

正长岭发垫刃线虫(Trichotylenchus changlingensis)是一种迁移性植物外寄生线虫,该线虫于2011年首次在我国吉林省长岭县发现,并根据其形态特征及最新分类系统将其归为发垫刃属[1,2]。该线虫能引起玉米叶片黄化,植株矮化,茎基部开裂,茎节缩短等症状,该病发生率普遍在21%～67%,给玉米生产造成了严重影响[3]。     

Prevalence and diversity of plant parasitic nematodes in Northern Ireland grassland and cereals,and the influence of soils and rainfall

The prevalence and diversity of plant parasitic nematodes in Northern Ireland cereal and grassland was determined from 191 agricultural fields. A total of 18 nematode genera were detected, including economically important pests, Meloidogyne spp., Heterodera spp. and Pratylenchus spp., each of which were above economic damage thresholds in a significant proportion of the sites (92.4%, 70% and 28.6%, respectively). The detection of the root knot nematode, Meloidogyne minor (6% prevalence), was significant given its recent emergence across the turf grass sector and the prospect of M. minor becoming a common agricultural pest. Analyses of nematode prevalence and abundance highlighted significant associations with grass and cereals, soil types, soil grade (proxy for soil quality) and rainfall levels. Specifically, nematode populations varied between the two major soils (brown earths and gleys), while significant trends for increased nematode diversity and greater prevalence of both Meloidogyne and Pratylenchus with increasing rainfall were also observed. Multivariate analyses were performed to determine interactive effects and the relative importance of the factors affecting nematode populations. Notably, rainfall, in combination with either crop type or soil grade, had a significant effect on nematode abundance and diversity. The findings suggest significant changes in nematode populations have occurred over the last several decades and the possibility that these are linked to changing climate and cropping practices are discussed, as well as future concerns for plant parasitic nematode management.     

河南省禾谷类作物孢囊线虫的发生分布与种类鉴定

为探究河南省主要禾谷类作物孢囊线虫的发生分布,明确孢囊线虫对不同作物的危害情况,于2017—2020年对河南省18个市50个县（区）的小麦、玉米和水稻作物的孢囊线虫种类和发生分布进行随机取样调查,采用形态学特征、分子生物学鉴定和rDNA-ITS序列进化树分析技术鉴定不同作物孢囊线虫的种类,并根据土壤中孢囊基数和单孢囊卵数明确不同作物孢囊线虫的发病严重度。结果显示,共采集土壤样品308份,其中224份样品检测到孢囊,孢囊检出率为72.7%。小麦孢囊线虫发生分布范围覆盖40个县（区）,其中15个县（区）为禾谷孢囊线虫Heterodera avenae侵染,23个县（区）为菲利普孢囊线虫H. filipjevi侵染,南阳市西峡县西坪镇和开封市尉氏县张市镇为禾谷孢囊线虫和菲利普孢囊线虫混合侵染发生区;玉米孢囊线虫H. zeae在濮阳市清丰县韩村镇、许昌市长葛市董村镇和禹州市范坡镇检测点首次被发现;旱稻孢囊线虫H. elachista在信阳市潢川县魏岗乡、来龙乡和新乡市获嘉县亢村镇检测点首次被发现。孢囊线虫发病严重度数据表明,小麦田平均孢囊含量高达17.3个/100 mL;玉米田平均孢囊量为11.0个/100 mL;水稻田平均孢囊量为4.4个/100 mL。表明河南省孢囊线虫高发地块主要集中在豫北、豫东和豫中平原区。     

Analysis of nematodes and soil-borne fungi fromAmmophila arenaria (Marram grass) in Dutch coastal foredunes by multivariate techniques

A survey was carried out at nine locations in the Dutch coastal foredunes to identify the species of soil borne fungi and nematodes associated withAmmophila arenaria (Marram grass).Ammophila arenaria is a sand binding grass that is very important for the stabilization of coastal foredunes. Degeneration of the plants occurs at stabilized sites and is supposed to be caused by a combination of soil-borne fungi and nematodes. Canonical correspondence analysis (CCA) and two-way indicator species analysis (TWINSPAN) were used to examine which fungal and nematode species usually coexist in the rhizosphere of vigorous and early declining stands ofA. arenaria. In total, 47 species of fungi and 10 genera of plant-parasitic nematodes were found. According to CCA, the community of soil organisms of stands that were more than 10 years old was significantly different from recently established stands of 3 years old. Also, the community of soil organisms isolated from calcareous locations differed significantly from that of lime-poor locations. No relationship between the vigour of the plants (vigorous vs. early declining) and the soil borne species composition was found, although in roots of vigorous stands, the number of nematodes was higher than that of early declining stands. A relatively large group of soil organisms occurred generally. This group possibly contains an ubiquitous pathocomplex that cause the growth reducing effects of biotic origin which generally occur inA. arenaria. Analysis of this group of nematodes and fungi by TWINSPAN resulted in 9 different combinations of concurring soil organisms of which 5 combinations were present at all investigated locations. Two of the latter combinations contained both nematodes and fungi. The first contained three endoparasitic nematodes (Meloidogyne maritima, Heterodera spp. andPratylenchus sp.) that concurred with the fungusMucor hiemalis. The second group containedHeterodera spp.,Telotylenchus ventralis, Filenchus sp. together with the potentially plant-pathogenic fungiMicrodochium bolleyi and Fusarium culmorum, as well as the fungiMortierella sp. andTrichoderma harzianum, all in relatively high numbers.It is concluded that both CCA and TWINSPAN are valuable exploratory techniques, especially when used in combination, to detect possible combinations of soil organisms which may be involved in the degeneration ofA. arenaria. Further identifications of harmful organisms should be obtained from experiments.     

Influence of different fertilization regimes on plant-parasitic nematodes in the black soil region of Northeast China

Plant-parasitic nematodes can cause serious plant diseases and adversely affect crop production. We investigated the temporal and vertical dynamics of plant-parasitic nematodes in northeast China to determine if long-term fertilizer application suppresses the population of plant-parasitic nematodes and influences their vertical distribution in the black soil region. The long-term fertilizer application lasted for 14 years, and included three treatments: pig manure combined with chemical fertilizer (MCF), chemical fertilizer (urea and ammonium phosphate, CF), and no fertilizer (NF). A 3-year corn-wheat-soybean rotation was grown on the field site, soil samples were obtained in the corn phase and nematodes were extracted, identified and counted. Six plant-parasitic nematode genera with relative abundance over 0.1% were found in this study. Heterodera was the dominant genus in all three fertilizer treatments, and its relative abundance was highest in NF (45.4%) and lowest in MCF (32.8%). Fertilizer application had a significant effect on abundance of total soil nematodes but not on either plant-parasitic nematodes or Heterodera. Significant differences in the abundance of total soil nematodes, plant-parasitic nematodes and Heterodera were observed among soil depths and interaction of fertilizer × soil depth. Total soil nematodes were mainly distributed in the 0–20 cm soil layer, and plant-parasitic nematodes and Heterodera were mostly distributed in the 10–30 cm soil layer. Fertilizer application did not show a significant effect on plant-parasitic nematodes or Heterodera at any of the sampling depths from 0 to 80 cm. The results indicated that fertilizer has no effective control on plant-parasitic nematodes when dominant genera and their respective plant hosts exist in relatively high abundance.     

一种准确测定土壤根结线虫种群数量的方法

为探索一种准确测定土壤根结线虫种群数量的方法,采用NaOCl消解附着在土壤病残体上的根结线虫卵囊与线虫常规离心技术相结合的方法,测试了不同NaOCl浓度、离心时间对根结线虫回收效率的影响,筛选出最佳测定条件,并与浅盘法、离心法、Byrd法等线虫常规分离方法进行了比较。结果表明,在9个NaOCl处理浓度中,1.0%~2.5%NaOCl处理对土壤根结线虫的回收效率显著高于3.0%NaOCl处理,达到71.6%~99.4%。在1.0%、1.5%、2.0%和2.5%NaOCl处理30 s后,分别离心2~5、2~3、2~3 min和2 min对根结线虫的回收效率明显高于其它时间处理,在每50 m L土壤平均接种976粒线虫卵的人工病土中,上述处理分离线虫数量分别达到804.3~930.0、810.7~838.7、843.7~867.0和820.7。本研究的改良方法对自然病土中根结线虫的分离数量分别是浅盘法、离心法和Byrd法的29.3、13.4和2.1倍,显著提高了土壤中根结线虫的分离效率,可准确测定土壤根结线虫种群的数量。     

Pratylenchus species of soybean in Croatia1

The numbers and species of phytoparasitic nematodes on soybean were studied over several years in Croatia, especially in the regions of Slavonija and Baranja. 19 phytoparasitic species were found, and of these Pratylenchus spp. predominated. These nematodes were found in mixed populations and their numbers were, at times, enormous. The average number per 100 ml of soil was about 100 individuals, with a maximum of 3348. The relationship between the number of nematodes and yield of soybean was calculated by a single regression equation. The soybean cultivars Sava and OS-88 apparently possess tolerance to relatively high numbers of Pratylenchus spp. in the rhizosphere.     

Root-lesion nematodes of potato: Current status of diagnostics,pathogenicity and management

Root-lesion nematodes of the genus Pratylenchus are migratory endoparasites with worldwide economic impact on several important crops including potato, where certain species like P. penetrans, P. neglectus, and P. scribneri reduce the yield and quality of potato tubers. Morphological identification of Pratylenchus spp. is challenging, and recent advancements in molecular techniques provide robust and rapid diagnostics to differentiate species without the need of specialist skills. However, the fact that molecular diagnostics are not available for all Pratylenchus species means that there are limitations in worldwide application. In general, root-lesion nematodes are difficult to manage once introduced into agricultural land and damage can be related to pathogenicity and population densities. In addition, root-lesion nematodes interact with fungi such as Verticillium dahliae, resulting in disease complexes that enhance the damage inflicted on the potato crop. Management interventions are often focused on limiting nematode reproduction before planting crops and include the application of nematicides, and cultural practices such as crop rotation, cover crops, biofumigation, and biological control. Understanding the limitations of the available crop protection strategies is important and there are many gaps for further study. This review discusses the status of the diagnosis, distribution, pathogenicity, and management of the main species of root-lesion nematodes, reported to infect potatoes worldwide, and highlights areas for potential future research.     

Development of a real‐time PCR method for the detection of the dagger nematodes Xiphinema index,X. diversicaudatum,X. vuittenezi and X. italiae,and for the quantification of X. index numbers

The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.