摘要
从莒县鸡腿菇种植区采集到表现鸡爪症状的样品,分离得到病原菌分离物RZ,采用改良的CTAB法提取其基因组总DNA,利用真菌通用引物ITS1/ITS4扩增其rDNA ITS区域序列,扩增产物纯化克隆后转化大肠杆菌DH5α,挑选阳性克隆进行测序。将获得的序列进行同源性比对和构建系统进化树,发现RZ分离物与GenBank中Bionectria ochroleuca 2个分离物聚为一簇,ITS的核苷酸一致率均为99.17%,从分子水平上证明该分离物为淡色生赤壳菌(B.ochroleuca)。
The Coprinus comatus samples with the symptom of chicken claw were collected from Juxian areas, and a fungi strain named RZ was isolated. Its genome total DNA was obtained by improved CTAB meth- od. The internal transcribed spacers （ITS） were amplified by PCR using fungi universal primers ITS1 and ITS4, and the amplified products were transformed into Escherichia coli DI-ISct after purified. Then the positive clones were screened and sequenced. The homology analysis showed that RZ was clustered with two Bionectria ochroleuca isolates in GenBank and shared 99.17% of nucleotide identity in ITS, which confirmed it to be Bionectria ochroleuca.
出处
《山东农业科学》
2012年第3期84-87,共4页
Shandong Agricultural Sciences
基金
国家现代农业(食用菌)产业体系建设专项(CARS-24)
山东省中青年科学家科研专项基金(BS2011NY021)
泰安市科技发展计划项目(20113008)
关键词
鸡腿菇
病害
转录间隔区
分子鉴定
Coprinus comatus
Pathogen
Internal transcribed spacers （ITS）
Molecular identification
