番茄灰霉病菌的鉴定及系统发育树分析

Identification and Phylogenetic Tree Analysis of Tomato Gray Mould Pathogen

从感染番茄灰霉病的病果上分离并纯化得到菌株t08016b,其形态学特征与灰葡萄孢属极其相似。通过扩增菌株t08016b的18SrDNA及内转录间隔区(ITS)序列,对其进行分子鉴定和分类。结果表明:该菌的18SrDNA序列与Botryotinia fuckeliana(EF110887.1)的同源性为99.94%;ITS rDNA序列与Botryotinia fuckeliana(AB444949.1)的同源性为100%,无碱基差别。对菌株t08016b进行系统发育树分析,表明t08016b属于灰葡萄孢属。利用该菌对健康番茄进行侵染,番茄灰霉病的发病率可达97%,证明其具有致病力。综合形态学鉴定、分子鉴定及致病力分析,表明菌株t08016b是一株具有强致病力的灰葡萄孢菌。

Strain t08016b, which morphological characteristics was very similar to the Botrytis spp., was isolated and purified from infected tomato fruits with typical symptoms of grey mold rot. The sequences of 18S rDNA and ITS1-5. 8S-ITS2 rDNA of strain t08016b were specifically amplified and sequenced using universal primer pairs NS1/NS8 and ITS4/ITSS. The results showed that the 18S sequence of t08016b had 99.94% similarity with fuckeliana fuckeliana （EFl10887. 1） and the ITS1-5. 8S-ITS2 rDNA sequence had 100% similarity with Botryotinia fuckeliana （AB444949. i）. The results of sequence alignment and neighbor-joining tree of i8S and ITS1-5. 8S-ITS2 rDNA showed that t08016b was one strain of Botryotinia fuckeliana. The incidence of tomato grey mould could up to 97% after infected by strain t08016b which indicated that t08016b possess strong pathogenicity. Comprehensive morphological identification and molecular identification and pathogenicity analysis, indicated that strain t08016b was a Botrytis cinerea strain with strong pathogenicity.