Identification of the beet cyst nematode Heterodera schachtii by PCR

Identification of the beet cyst nematode Heterodera schachtii by PCR
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DOI:
10.1023/a:1019974101225
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发表时间:
2002-07-01
影响因子:
1.8
通讯作者:
Moens, M
Moens, M
中科院分区:
农林科学3区
文献类型:
--
作者:
Amiri, S;Subbotin, SA;Moens, M

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利用ITS-rDNA的PCR-RFLP和种特异性引物PCR技术对甜菜胞囊线虫Heterodera schachtii孢囊和幼虫进行了鉴定。MvaI或ScrFI对PCR产物的限制性反应可区分H. schachtii,H. betae,刺蛾H. trifolii和H. medicaginis。这四种线虫的RFLP图谱与8种限制性内切酶。根据Schachtii类群几个种的群体的内转录间隔区序列,设计了一对Schachtii类群的特异性引物。设计了schachtii,允许扩增来自甜菜胞囊线虫幼虫和胞囊的目标序列。描述了用广泛的线虫样品测试的双重PCR协议。
PCR-RFLPs of ITS-rDNA and PCR with species-specific primers were developed for identification of cysts and juveniles of the beet cyst nematode Heterodera schachtii. Restrictions of PCR product by MvaI or ScrFI distinguish H. schachtii, H. betae, H. trifolii and H. medicaginis. RFLP profiles with eight restriction enzymes for these four nematode species are presented. Based on Internal Transcribed Spacer sequences of populations from several Schachtii group species, a specific primer for H. schachtii was designed, permitting amplification of the target sequence from juveniles and cysts of the beet cyst nematode. A duplex PCR protocol tested with a wide range of nematode samples is described.