Nischwitz Claudia, Skantar Andrea, Handoo Zafar A, Hult Maria N, Schmitt Mark E, McClure Michael A
Department of Biology, Utah State University, Logan 84322.
United States Department of Agriculture-Agricultural Research Service, Nematology Laboratory, Beltsville, MD 20705.
Plant Dis. 2013 Nov;97(11):1424-1430. doi: 10.1094/PDIS-03-13-0263-RE.
Several species of root-knot nematodes (Meloidogyne spp.) are known to have significant presence on turfgrass in golf course greens, particularly in the western United States. Nematodes isolated from a golf course in King County, WA were identified as Meloidogyne minor based on analysis of the large ribosomal subunit (LSU 28S D2-D3 expansion segment), the internal transcribed spacers 1 and 2 (ITS rDNA), the intergenic spacer region 2 (IGS2), and the nuclear protein-coding gene Hsp90. Sequence-characterized amplified region (SCAR) primers that were originally designed to be specific for M. fallax were found to cross-react with M. minor. A population from California was determined to be M. fallax based on juvenile tail morphology and analysis of the ribosomal markers and Hsp90, comprising the first report of this species in North America. Using trees based on Hsp90 genomic alignments, the phylogenetic relationships of these populations and known root-knot nematode species were congruent with previous trees based on ribosomal genes. Resolution of M. fallax and M. chitwoodi using Hsp90 was equivalent to species separation obtained with 28S or 18S rDNA alignments. The strengths and weaknesses of ribosomal and Hsp90 markers, and the use of SCAR polymerase chain reaction as diagnostic tools are discussed.
已知几种根结线虫(Meloidogyne spp.)在高尔夫球场果岭的草坪草上大量存在,尤其是在美国西部。基于对大核糖体亚基(LSU 28S D2-D3扩展片段)、内部转录间隔区1和2(ITS rDNA)、基因间隔区2(IGS2)以及核蛋白编码基因Hsp90的分析,从华盛顿州金县的一个高尔夫球场分离出的线虫被鉴定为微小根结线虫。最初设计用于特异性鉴定伪根结线虫的序列特征扩增区域(SCAR)引物被发现与微小根结线虫存在交叉反应。基于幼虫尾部形态以及核糖体标记和Hsp90的分析,确定来自加利福尼亚的一个种群为伪根结线虫,这是该物种在北美的首次报道。利用基于Hsp90基因组比对构建的树状图,这些种群与已知根结线虫物种的系统发育关系与之前基于核糖体基因构建的树状图一致。使用Hsp90区分伪根结线虫和奇氏根结线虫的效果与使用28S或18S rDNA比对进行物种区分的效果相当。本文讨论了核糖体和Hsp90标记的优缺点,以及将SCAR聚合酶链反应用作诊断工具的情况。