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从河南省10个地市采集的小麦病根样品中分离得到82株病原分离株,通过形态特征观察、回接致病性测定、以及分子生物学的方法对所分离菌株进行鉴定。其中47个菌株菌丝分支处都形成典型的“∧”状;子囊壳埋生或半埋生,子囊棍棒状,子囊孢子线形,稍弯曲,无色,具有5~10个分隔,大小为68~94μm×2~4μm;对82个分离株rDNA-ITS区进行PCR扩增和序列测定,BLAST序列比对结果表明,47株菌株与小麦全蚀病菌的ITS序列同源性达97%~99%,据此确定47株菌株为小麦全蚀病菌。应用小麦全蚀病菌4个变种的特异性引物进行PCR扩增都得到禾顶囊壳小麦变种(870 bp)的特异性片段,鉴定47株菌株均为禾顶囊壳小麦变种(Gaeumannomyces graminis var.tritici)。
Totally 82 isolates were isolated from diseased wheat samples collected from 10 cities in Henan Province. The pathogenicity of the isolates were determined by morphological observation, pathogenicity test and molecular biology. Among them, 47 mycelium branches formed typical “Λ” shape. The ascospore shell was buried or semi-buried, and the ascochlea was stick-shaped. The ascospore was linear, slightly curved and colorless, with 5 ~ 10 divisions in size (68 ~ 94μm × 2 ~ 4μm). The rDNA-ITS region of 82 isolates were amplified by PCR and sequenced. The results of BLAST alignment showed that the sequences of 47 isolates shared 97% ~ 99%, thus determining 47 strains of wheat eclipses bacteria. The specific fragment of 870 bp was obtained by PCR amplification using four primers of wheat total ectomycorrhizal fungi. All the 47 strains were identified as Gaeumannomyces graminis var. tritici).