一个鉴别低温刺激型秀珍菇的ISSR-SCAR标记建立与应用

为了快速、准确地鉴别低温刺激型秀珍菇菌株,在对18个供试菌株进行拮抗试验、现蕾出菇试验、ISSR图谱分析的基础上获得了秀珍菇菌株的ISSR特异性标记,将其克隆、测序。根据测序结果,应用Primer 6.0软件设计SCAR引物进行引物筛选,获得1对引物并成功转化为低温刺激型秀珍菇的稳定SCAR标记。采用该引物对供试秀珍菇菌株的基因组DNA 进行PCR扩增,低温刺激型秀珍菇菌株均能扩增出大小为205 bp的特异性DNA片段。该ISSR-SCAR标记能快速、有效地鉴别出低温刺激型秀珍菇菌株,准确率达100%,为秀珍菇种质资源的鉴别和分类提供了技术支撑。

Establishing and Applying of a ISSR-SCAR Marker to Identify Low-temperature Stimulating Type of Pleurotus geesteranus

A specific ISSR marker was obtained based on the antagonism test,fruiting test and ISSR spectrum analysis of 18 strains to identify the low-temperature stimulating type of Pleurotus geesteranus rapidly and accurately.The specific ISSR marker was cloned and sequenced.SCAR primers(designed by Primer 6.0)for the specific ISSR marker were screened,than a pair specific primers were obtained,which could convert the specific ISSR marker to stable SCAR mark.Verification was conducted by PCR using the screened specific primers to amplify the genomic DNA.Results showed that 205 bp specific fragment was amplified only in the low-temperature stimulating type of P.geesteranus strains.This ISSR-SCAR marker could distinguish the low-temperature stimulating type of P.geesteranus strains rapidly with accurasy of 100%.This study would provide a technological support for the identification and classification of P.geesteranus germplasm resources.