Karanastasi E, Decraemer W, Kyriakopoulou P E, Neilson R
Laboratory of Nematology, Benaki Phytopathological Institute, 8 Stefanou Delta str, 145 61 Kifissia, Attica, Greece.
Department of Invertebrates, Royal Belgian Institute of Natural Sciences, Vautierstraat 29, B-1000 Brussels, Belgium.
Plant Dis. 2005 Jun;89(6):685. doi: 10.1094/PD-89-0685A.
The polyphagous stubby-root nematode species, Paratrichodorus teres (Hooper) Siddiqi, was first described from soil under lettuce near Norwich, UK and subsequently reported from South Africa and the United States, but predominantly from temperate regions within Europe (4). P. teres is one of 13 economically important trichodorid species known to be vectors of Tobacco rattle virus (TRV) (4). Artichokes planted during 2000 in a field located in the Kandia area of the Argolis Region, Greece (37°32'N, 22°56'E) exhibited symptoms of a virus infection. Sampling was done to ascertain the presence of Longidorus fasciatus, a vector of artichoke Italian latent nepovirus known to occur in the area (1,4). In addition to L. fasciatus, an unknown trichodorid species and Tylenchorhyncus sp. were recovered from the root zone of artichoke at a number of sites within the field. Measurements and morphological examination of the female (n = 13, body length = 741.7 ± 25.5 μm, onchiostyle = 43.7 ± 0.8 μm, and position of vulva from anterior region relative to total body length V% = 53.8 ± 0.4 μm) and male (n =1, body length = 720.5 μm, onchiostyle = 43.5 μm, spicule length = 51.7 μm, and number of ventromedian precloacal supplements = 3) trichodorids isolated from soil samples conformed to the original description of P. teres and the generic polytomous key (2). Furthermore, morphological identification was supported by molecular data. DNA was extracted from seven individual trichodorids, each of which were placed into separate 0.5-ml micro-centrifuge tubes containing 20 μl of 0.25 M NaOH and incubated at 25°C overnight. Thereafter, samples were incubated at 99°C for 3 min and 10 μl of 0.25 M HCl, 5 μl of 0.5 M Tris-HCl, (pH 8.0) and 5 μl of 2% Triton X-100 were added to each tube. Samples were incubated at 99°C for a further 3 min and stored at -20°C. Template DNA was amplified using polymerase chain reaction with primers specific for 18S rDNA and sequenced (3). The resultant consensus sequence had 99.8% homology to P. teres populations isolated from Portugal and good homology (95 to 98%) with five other Paratrichodorus spp. listed on public sequence databases, e.g., NCBI GenBank. This constitutes a new geographic record and a possible association of P. teres on artichoke. References: (1) D. J. F. Brown et al. Eur. J. Plant Pathol. 103:501, 1997. (2) W. Decraemer. Fundam. Appl. Nematol. 21:37, 1998. (3) C. M. G. Oliveira et al. J. Nematol. 36:153, 2004. (4) C. E. Taylor and D. J. F. Brown. Nematode Vectors of Plant Viruses, CAB International Mycological Institute, Wallingford, UK, 1997.
多食性短体线虫物种,细尾拟毛刺线虫(Paratrichodorus teres)(胡珀)西迪基,最初是从英国诺维奇附近生菜地的土壤中描述的,随后在南非和美国有报道,但主要来自欧洲的温带地区(4)。细尾拟毛刺线虫是已知的13种经济上重要的毛刺线虫物种之一,是烟草脆裂病毒(TRV)的传播媒介(4)。2000年在希腊阿尔戈利斯地区坎迪亚地区(北纬37°32′,东经22°56′)的一块田地里种植的洋蓟出现了病毒感染症状。进行采样以确定已知存在于该地区的洋蓟意大利潜隐线虫的传播媒介——带纹长针线虫(Longidorus fasciatus)的存在情况(1,4)。除了带纹长针线虫外,还在该田地内的多个地点从洋蓟根区分离出一种未知的毛刺线虫物种和矮化线虫属(Tylenchorhyncus)的一个种。对从土壤样本中分离出的雌虫(n = 13,体长 = 741.7 ± 25.5 μm,口针 = 43.7 ± 0.8 μm,阴门相对于身体前部的位置V% = 53.8 ± 0.4 μm)和雄虫(n = 1,体长 = 720.5 μm,口针 = 43.5 μm,交合刺长度 = 51.7 μm,泄殖腔前腹面补充器数量 = 3)毛刺线虫进行的测量和形态学检查符合细尾拟毛刺线虫的原始描述和通用多歧检索表(2)。此外,分子数据支持形态学鉴定。从7条单独的毛刺线虫中提取DNA,每条线虫分别置于含有20 μl 0.25 M氢氧化钠的单独0.5 ml微量离心管中,并在25°C下孵育过夜。此后,将样品在99°C下孵育3分钟,然后向每个管中加入10 μl 0.25 M盐酸、5 μl 0.5 M Tris - HCl(pH 8.0)和5 μl 2% Triton X - 100。样品再在99°C下孵育3分钟,然后储存在 - 20°C。使用针对18S rDNA的引物通过聚合酶链反应扩增模板DNA并进行测序(3)。所得的共有序列与从葡萄牙分离出的细尾拟毛刺线虫种群具有99.8%的同源性,与公共序列数据库(如NCBI GenBank)中列出的其他5种拟毛刺线虫属物种具有良好的同源性(95%至98%)。这构成了一个新的地理记录以及细尾拟毛刺线虫与洋蓟可能的关联。参考文献:(1)D. J. F. Brown等人,《欧洲植物病理学杂志》103:501,1997年。(2)W. Decraemer,《基础与应用线虫学》21:37,1998年。(3)C. M. G. Oliveira等人,《线虫学杂志》36:153,2004年。(4)C. E. Taylor和D. J. F. Brown,《植物病毒的线虫传播媒介》,CAB国际真菌研究所,英国沃灵福德,1997年。