【摘要】 测定苹果炭疽菌rDNA全序列,比对苹果炭疽菌和其它炭疽菌ITS序列以及构建系统关系树,发现苹果炭疽菌与胶孢炭疽菌的ITS序列相似性高达99.8%,并与胶孢炭疽菌聚在一起,可以明确苹果炭疽菌应属于胶孢炭疽菌。进一步的序列比对发现,苹果炭疽菌的18S rDNA 3’端比其它胶孢炭疽菌多出一段379 bp的序列,根据这一特有片段设计引物CgF1与通用引物ITS4配对,结果仅能从苹果炭疽菌中扩增出1 232 bp的特异性条带。用苹果炭疽菌接种离体苹果,以接种发病的病组织总DNA为模板,利用引物CgF1/ITS4进行PCR扩增,同样可以扩增出1 232 bp的特异性条带,而健康苹果组织DNA中未能扩增出任何条带,表明该方法可用于苹果炭疽菌的鉴定和快速检测。更多还原

【Abstract】 The rDNA sequence of apple anthracnose pathogen was determined.Sequence comparing of ITS of Colletotrichum was conducted and a phylogenetic tree based on alignment of their ITS nucleotide sequences was constructed.The results showed that Cg-1 and Cg-2 shared the highest sequence similarity(99.8%) with C.gloeosporioides and they could be grouped into one branch with C.gloeosporioides.It was confirmed that apple anthracnose pathogen should belong to C.gloeosporioides.It was found that there is a 379 bp sequence residue in 3 ’ ends of 18S rDNA of apple anthracnose pathogen comparing with the other C.gloeosporioides.Specific primer CgF1 was designed according to the 379 bp sequence, and only a 1 232 bp specific band could be amplified using primer pair CgF1 and ITS4.Apple fruits were inoculated with apple anthracnose pathogen.Total DNA was extracted from the diseased tissue,and an identical 1 232 bp band was amplified.It was suggested that the method could be applied for the detection and rapid identification of apple anthracnose pathogen.更多还原