水稻干尖线虫(Aphelenchoides besseyi)人工培养研究

利用真菌培养水稻干尖线虫的结果表明:不同的真菌培养线虫的效果有较大差异.链格孢菌(Alternariasp.)、镰刀菌(Fusarium sp.)的效果明显优于常用的灰葡萄孢菌(Botrytis cinerea),可选用于培养水稻干尖线虫.在真菌菌落上,不同的接虫量以及不同的温度和培养时间对水稻干尖线虫的培养均有一定的影响.     

鸢尾种苗中截获的草莓滑刃线虫形态和分子鉴定

草莓滑刃线虫是一种重要的叶和芽寄生线虫,可侵染多种重要花卉、经济作物,被列入我国检疫性有害生物名录。目前,草莓滑刃线虫的鉴定以形态学鉴定为主,但一线口岸常分离不到雌虫,鉴定困难。本研究采用形态特征和分子特征相结合的方法对草莓滑刃线虫进行鉴定:对鸢尾种苗中截获的滑刃属线虫群体雌虫、雄虫进行形态观察和形态测计并与已报道文献进行比较;PCR扩增该群体的rDNA-ITS区并测序,构建基于ITS序列的系统发育进化树;采用草莓滑刃线虫特异引物对该群体DNA样本进行扩增。结果表明:该群体与草莓滑刃线虫俄罗斯种群的测量值相接近;系统进化关系分析显示,该群体与NCBI公布的草莓滑刃线虫均处于同一进化支;采用草莓滑刃线虫特异引物PCR扩增可产生片段大小为169 bp的目的条带。     

水稻干尖线虫(Aphelenchoides besseyi)人工培养研究

利用真菌培养水稻干尖线虫的结果表明：不同的真菌培养线虫的效果有较大差异。链格孢菌（Alternaria sp.）、镰刀菌（Fusarium sp.）的效果明显优于常用的灰葡萄孢菌（Botrytis cinerea）,可选用于培养水稻干尖线虫。在真菌菌落上,不同的接虫量以及不同的温度和培养时间对水稻干尖线虫的培养均有一定的影响。     

肾形肾状线虫（Rotylenchulus reniformis）浙江杭州 群体形态学和分子特性及寄主范围

 肾形肾状线虫（Rotylenchulus reniformis）是一种重要的蔬菜线虫,寄主范围和分布很广。本文根据形态学特征和形态测量将来自浙江省杭州市蔬菜上的肾状线虫群体鉴定为肾形肾状线虫,为浙江省首次报道。该群体的形态学特征及形态测量值与模式标本基本吻合,个别形态测量值(比值)有变异。通过对该肾形肾状线虫群体的rDNA中ITS区和编码28S RNA基因中的D2D3区的PCR扩增和序列测定,明确二序列长度分别为808 bp和786 bp;序列分析发现该群体ITS序列与肾形肾状线虫种内一些两性生殖群体的亲缘关系较近,同源性高达99.5%～100%,而与一孤雌生殖群体同源性为92.4%;编码28S RNA基因中的D2D3区与已报道群体的同源性达到99.4%,相似度较高;本研究还发现杭州肾形肾状线虫群体的寄主涉及11个科16个属的果蔬。     

禾谷孢囊线虫(Heterodera avenae)的分子鉴定及抗性基因研究进展

禾谷孢囊线虫是温带禾谷类作物上的世界性重要病原线虫。本文回顾关于禾谷孢囊线虫的分子鉴定和亲缘分析等方面的研究,以及对国内外小麦品种抗性基因的筛选和应用等结果。根据上述方面的结果拟定更为合理的防治策略。     

中国4省(区)葡萄根际剑线虫属和针线虫属种类的鉴定

从中国广西、陕西、山东、宁夏4省(区)36个葡萄品种的230份葡萄根际土样中鉴定出4种剑线虫和1种针线虫,分别为Xiphinema americanum,X.insigne,X.hunaniese,X.elongatum和Paratylenchus audriellus. ,其中X.elongatum为葡萄上的新记录种;P.audriellus在中国属首次记录,葡萄为其新寄主植物。     

水稻干尖线虫14-3-3(Ab-14-3-3-a)基因克隆与功能分析

水稻干尖线虫是水稻上重要的寄生线虫,严重危害水稻安全生产。14-3-3蛋白调控生物体的细胞代谢、生长发育和逆境适应等一系列生物过程。本研究通过c DNA末端快速扩增技术(rapid amplification of c DNA ends,RACE)分离获得了一个水稻干尖线虫(Aphelenchoides besseyi)14-3-3基因,命名为Ab-14-3-3-a。Ab-14-3-3-a全长c DNA序列含有59 bp的5'非翻译区(UTR)、编码251个氨基酸756 bp的开放阅读框。Ab-14-3-3-a DNA序列包含4个外显子和3个内含子。Ab-14-3-3-a蛋白与其他植物寄生线虫14-3-3蛋白氨基酸序列高度相似,系统发育树显示与植物寄生线虫位于同一进化分支。原位杂交显示Ab-14-3-3-a特异定位于成虫的背食道腺细胞中,表明其在线虫取食和寄生过程中发挥潜在功能。qRT-PCR显示,Ab-14-3-3-a在水稻干尖线虫各个龄期均表达,其中在成虫表达水平最高,而在发育过程中幼虫表达水平最低。当线虫取食不同寄主时,Ab-14-3-3-a的表达水平具有差异性,线虫取食感病水稻早期,Ab-14-3-3-a表达水平上升1.5倍,而取食灰葡萄孢(Botrytis cinerea)时Ab-14-3-3-a表达可提高数百倍。利用体外RNA干扰技术将Ab-14-3-3-a的表达进行有效抑制,结果显著影响了线虫的产卵、成虫发育以及线虫种群数量。综上,Ab-14-3-3-a参与调控水稻干尖线虫的取食行为和发育进程。本研究相关结果有助于拓展植物寄生线虫14-3-3蛋白功能,为深入研究线虫与植物互作机制提供了理论依据。     

较小拟毛刺线虫不同群体的形态及分子特征比较研究

毛刺类线虫系一类重要的植物外寄生线虫,其内包括一些可传播植物病毒的种类。本研究基于形态学和rDNA分子特征从我国海南儋州、云南呈贡和昆明、福建厦门等植物根围土壤样品中分离鉴定出8个较小拟毛刺线虫(Paratrichodorus minor)群体;通过对种群数量较大的海南儋州群体与福建厦门群体的形态及测量值比较,发现不同地理群体间主要形态测量值存在一定差异;各群体18S RNA区、r DNA-ITS1区、r DNA-ITS2区和28S RNA基因中D2D3区序列与Gen Bank已登录的较小拟毛刺线虫不同群体相应序列(AJ438053、AJ438054及AJ438056;KJ934126;JN123380和JN123381;JN123396和KJ513001)相似度分别介于99.1%~99.9%、95.8%~96.3%、99.5%~99.8%和99.4%~99.9%。对上述较小拟毛刺线虫群体r DNA-ITS1区、r DNA-ITS2区和28S RNA基因中D2D3区序列比较和酶切分析显示,该3段序列在较小拟毛刺线虫种内群体间都较为保守。其中28S RNA基因中D2D3区序列在种内群体稳定,种间群体有明显的差异,系毛刺类线虫鉴定一较好的分子靶标。有关结果对较小拟毛刺线虫的鉴定及鉴别有直接的指导意义。     

Studies on the weed hosts of the rice root nematode, Hirschmanniella spinicaudata Sch. Stek. 1944

Weed hosts of the rice root nematode, Hirschmanniella spinicaudata were surveyed in a swamp rice field at Badeggi, Nigeria. Forty-five weed species, mainly grasses and sedges occurred very frequently and were very good hosts of the nematode. A wild rice, Oryza longistaminata was a preferred host. The importance of the weed hosts is discussed.     

山西短颈剑线虫(Xiphinema brevicollum)形态与rDNA分子特征

采用浅盘分离法在山西太谷松树根际土壤中分离得到一种剑线虫,通过Seinhorst脱水制片后利用显微镜观察其形态特征并进行测量,其主要特征为:雌性虫体经热杀死后呈腹面弯曲的"C"形,体长1.71~2.26mm,两端渐细;齿针全长127.3~140.8μm;a(体长/体宽)为35.9~51.4μm,b(体长/食道外宽)为5.8~8.7μm,c(体长/尾长)为50.7~103μm。前卵巢与后卵巢长短相近,尾形近似圆锥形。未见雄虫。对该线虫的rDNA-ITS1,rDNAITS2和28SRNA序列进行扩增、测序及系统进化分析,结合形态学特征进行鉴定,将分离到的剑线虫鉴定为短颈剑线虫(Xiphinema brevicollum),这是山西首次发现报道剑线虫属的一个种。     

Morphological and molecular characterization of Phytophthora species associated with root and crown rot of pomegranate in Iran

The purpose of this research was to identify the pathogens causing root and crown rot in major pomegranate-growing areas of Iran. Infected tissue samples were collected from trees with symptoms from 49 pomegranate orchards in four provinces of Iran: Fars, Markazi, Isfahan, and Kohgiluyeh va Boyer-Ahmad. In total, 23 Phytophthora spp. isolates were obtained, which were identified as P. cryptogea species complex (12 isolates) and P. cinnamomi (11 isolates) based on morphological characters. Molecular confirmation of identification was performed by inference of phylogeny of ITS-rDNA regions, β-tubulin gene, and the mitochondrial gene cytochrome c oxidase subunit 1. The results of our phylogenetic analysis confirmed the morphological identification of P. cinnamomi isolates and placed them in Clade 7c of Phytophthora. In addition, the P. cryptogea species complex isolates, despite morphological similarities, were in fact four distinct species including P. cryptogea sensu stricto (two isolates), P. pseudocryptogea (one isolate), P. sp. kelmania (one isolate), and P. erythroseptica (six isolates). This is the first report of pomegranate root and crown rot caused by P. cinnamomi and P. cryptogea species complex.     

危害红掌的穿孔线虫形态和分子鉴定

对一批引进后隔离培养中患病的红掌种苗,进行了根系线虫检测,通过病症分析、线虫形态测量、rDNA-ITS区序列分析等鉴定,结果表明:检测的线虫形态和香蕉穿孔线虫基本一致,rDNA-ITS区测序结果序列一致性为98％～100％.本报道再次证明香蕉穿孔线虫在红掌上的分布和危害,应该对种苗的跨区引进和种植过程进行严格的检疫.     

日本鸡爪槭中旱稻胞囊线虫形态和分子鉴定

从宁波口岸进境的日本鸡爪槭(Acer palmatum)景观树根际介质中分离到1种胞囊线虫属的2龄幼虫,通过形态鉴定和分子特征分析,鉴定其为旱稻胞囊线虫(Heterodera elachista Ohshima,1974)。该日本群体的2龄幼虫体长374~421μm;唇区有3个唇环;口针粗壮,长16.5~19.2μm,基部球圆形或前端略凹陷;侧线3条;尾长圆锥形,末端细圆,透明尾长22.9~30.7μm,约占尾长的50%。基于核糖体DNA的28S-D2/D3区和ITS区序列构建的系统发育树以及ITS序列酶切分析证实该鉴定结论,这是我国首次截获旱稻胞囊线虫。     

Population structure and species delimitation of rice white tip nematode,Aphelenchoides besseyi (Nematoda: Aphelenchoididae), in China

Aphelenchoides besseyi is a major nematode pathogen in rice known as the causal agent of white tip disease. In the present study, A. besseyi was recovered from 32 seed samples collected from the major rice-growing regions in China. Nine mitochondrial COI haplotypes and 95 ITS genotypes were identified, suggesting a high genetic diversity and endemism level of A. besseyi in China. Therefore, A. besseyi is likely to have experienced a long history of host–parasite coevolution in China, rather than being recently introduced from other countries. The results of population structure analysis indicated the presence of four clusters within Chinese A. besseyi, but these did not correspond to their geographic distribution. Haplotype H2 was found to be the most widespread. Haplotypes H9 and H13 were most genetically divergent and have been recognized as two cryptic species by molecular species delimitation methods, and with limited support from morphometric measurements. The COI-based phylogeny suggested the ability to parasitize rice has independently evolved at least four times in the genus Aphelenchoides.     

Morphological and molecular characterization of Globodera populations from Oregon and Idaho

An unusual population of cyst nematode was found in soils collected from a Powell Butte, OR field with a cropping history including potato, wheat, other crops, and significant weed presence. These nematodes could not be placed with certainty into any known species and exhibited some unique morphological features in some specimens. Compared with Globodera pallida, the cyst body length was slightly longer and the second-stage juvenile stylet length was slightly shorter. In some individuals, the J2 stylet knob height was greater and the tail annules were more prominent than in G. pallida, and the tail abruptly narrowed, with a slight constriction near the posterior third of the hyaline terminus. Compared with G. rostochiensis, the hyaline tail terminus had a larger number of refractive bodies, and cysts of this population had a smaller Granek's ratio and fewer cuticular ridges between the anus and vulva. In some individuals, the tail termini of second-stage juveniles were more bluntly pointed, and the stylet knobs were more anteriorly directed with greater height. Unlike G. tabacum, the cyst wall often lacked a network-like pattern and, in some individuals, the juvenile tail terminus distinctly narrowed after a constriction. Molecularly, the population was distinct from G. pallida, G. rostochiensis, and G. tabacum. Multiplex polymerase chain reaction of the internal transcribed spacer (ITS) rDNA region gave results similar to G. tabacum; however, ITS restriction fragment length polymorphism patterns were observed to have individual bands in common with G. rostochiensis and G. pallida. Phylogenetic analysis based on ITS1 and -2 rDNA sequences showed greatest similarity to populations from Argentina and Chile; together, they form a moderately supported clade, distinct from G. rostochiensis, G. tabacum, G. "mexicana," European type G. pallida, and several G. pallida populations from South America.