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采用线虫通用引物TW81和AB28对采自陕西的20个小麦禾谷孢囊线虫群体的rDNA-ITS区进行PCR扩增、克隆和测序。利用Mega软件的UPGMA方法分析了陕西省小麦禾谷孢囊线虫群体及其与GenBank公布的近缘种的系统发育关系,结果表明陕西省小麦孢囊线虫群体的rDNA-ITS区片段的长度为1 045 bp,且均为禾谷孢囊线虫(Heterodera ave-nae),ITS区序列同源性为99.56%,只有3处碱基存在差异。其ITS区同中国河南(EU106175)、北京(AY148382)、山东(HM370427)的禾谷孢囊线虫H.avenae,澳大利亚的H.australis(AY148395)及俄罗斯的H.pratensis(AY148351)的亲缘关系最为接近。利用8种限制性内切酶AvaⅠ(Eco88Ⅰ)、AluⅠ、HhaⅠ(CfoⅠ)、HaeⅢ(BsuRⅠ)、HindⅢ、HinfⅠ、RsaⅠ和MvaⅠ(Bst N1)对20个陕西省CCN群体的ITS区PCR扩增产物进行限制性内切酶谱(RFLP)分析,结果表明8种限制性内切酶酶切ITS-PCR产物后共产生22个酶切片段,20个种群的PCR-RFLP谱型一致,表明陕西省小麦禾谷孢囊线虫是同一个群体且与已报道的中国CCN群体的“C”型一致,有别于欧洲的“A”型群体和印度的“B”型群体。这是首次报道陕西省小麦禾谷孢囊线虫种群的分子特征。
The rDNA-ITS regions of twenty wheat cereal cyst nematode populations collected from Shaanxi were amplified by PCR using the nematode common primers TW81 and AB28, and cloned and sequenced. The phylogenies of wheat cereal cyst nematode in Shaanxi Province and its relatives published in GenBank were analyzed by UPGMA method of Mega software. The results showed that the length of rDNA-ITS region of wheat cyst nematode population in Shaanxi Province was 1 045 bp, and all were Heterodera ave-nae. The homology of ITS sequence was 99.56%, only 3 bases were different. The ITS region has the highest genetic relationship with H.pratensis (AY148395) in Australia and H.pratensis (AY148351) in China (EU106175), Beijing (AY148382) and Shandong (HM370427) Close to The ITS region of 20 CCN populations in Shaanxi province were amplified by PCR using eight restriction enzymes Ava Ⅰ (Eco88Ⅰ), AluⅠ, HhaⅠ (CfoⅠ), HaeⅢ (BsuRI), HindⅢ, HinfⅠ, RsaⅠand MvaⅠ (Bst N1) The results of restriction endonuclease analysis (RFLP) analysis showed that the restriction endonucleases (RFLP) analysis showed that 22 restriction enzyme digestion ITS-PCR products produced a total of 22 digested fragments, 20 PCR-RFLP patterns consistent, indicating that Shaanxi Province The wheat cereal cyst nematodes are one and the same as the “C” type of Chinese CCN population reported, which is different from the “A” type in Europe and the “B” type in India. This is the first report on the molecular characteristics of wheat cereal cyst nematode population in Shaanxi Province.