基于rDNA的PCR-RFLP及序列分析对四种螺旋线虫的分子鉴定  

Molecular Identification of Four Helicotylenchus Species Based on PCR-RFLP and Sequencing of rDNA

作　　者：赵雷[1] 郑炜[1] 梅圆圆[1] 郑经武[1] 

机构地区：[1]浙江大学生物技术研究所,杭州310029

出　　处：《农业生物技术学报》2010年第5期981-984,共4页Journal of Agricultural Biotechnology

基　　金：教育部留学回国基金[教外司留(2007)1108号];国家自然科学基金(No.30870274);浙江省自然科学基金杰出青年团队项目(No.R307396)共同资助

摘　　要：利用rDNAITS-PCR-RFLP及序列分析等技术对来自国内浙江、山西、山东以及德国等地6个不同螺旋线虫地理群体进行了分类和鉴定。结合形态学特征分别鉴定为小头螺旋线虫(Helicotylenchus microcephalus)、双角螺旋线虫(H.digonicus)、双宫螺旋线虫(H.dihystera)和翅尾铗螺旋线虫(H.pteracerus)。其中3个不同地理群体的双角螺旋线虫PCR产物片段长度和限制性内切酶酶切图谱相同,其余3个种的PCR产物长度稍大,4个螺旋线虫种的酶切图谱各不相同。选用的6种限制性内切酶(AluⅠ、AvaⅠ、HaeⅢ、HinfⅠ、MspⅠ和RsaⅠ)组合能有效地鉴别四种螺旋线虫。这是我国有关该属线虫分子特征的首次报道。PCR-RFLP and seq uencing of rDNA ITS regions of six geographic populations in genus Helicotylenchus were made.Four species,Helicotylenchus microcephalus,H.digonicus,H.dihystera,and H.pteracerus,were identified.Both the sequence length and digestion patterns were the same in the three populations of H.digonicus.The digestion patterns varied in different fourspecices,which was identified as H.microcephalus,H.digonicus,H.dihystera and H.pteracerus,respectively based on the morphological method.The combination of six enzymes,AluⅠ,AvaⅠ,HaeⅢ,HinfⅠ,MspⅠ and RsaⅠ,could be used effectively to identify and classify this four Helicotylenchus species.This is the first report about the molecular data of the nematode genus in China.

关 键 词：ITS-PCR-RFLP 小头螺旋线虫 双角螺旋线虫 双宫螺旋线 翅尾铗螺旋线虫 

分 类 号：S432.45[农业科学—植物病理学]