陈雪凤 吴圣进 王灿琴 韦仕岩 王晓国 郞宁 张雯龙

摘  要：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。

关键词：秀珍菇;低温刺激;ISSR特异性标记;SCAR分子标记

Abstract： 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.

Keywords： Pleurotus geesteranus; low-temperature stimulating; specific ISSR marker; SCAR marker

秀珍菇（Pleurotus geesteranus）隸属于真菌门、担子菌纲、伞菌目、侧耳科、侧耳属，商品名或俗称凤尾菇，其味道鲜美、营养丰富，是一种深受广大消费者喜爱的食用菌。近年来随市场的需求，秀珍菇的栽培规模日益增大，且国内栽培的秀菇菌株品种繁多。秀珍菇根据出菇温度可分为高温型品种和低温型品种，根据出菇是否需要低温刺激可分为自然出菇型（俗称假秀珍）和低温刺激型（也称反季节秀珍菇或真秀珍）。自然出菇型秀珍菇特性是菌丝长满袋且生理成熟后不需要低温刺激即可出菇。低温刺激型秀珍菇特性是菌丝长满袋且生理成熟后，需要低温刺激方能整齐出菇。真秀珍菇和假秀珍由于出菇时对低温温差要求程度不一样，因而栽培管理方法差异较大。……