人工栽培蛹虫草退化现象的分子分析
MOLECULAR ANALYSIS OF DEGENERATION OF ARTIFICIAL PLANTED CORDYCEPS MILITARIS
李美娜 吴谢军 李春燕 冯伯森 李庆伟
摘 要：利用PCR-RFLP和RAPD方法对野生驯化蛹虫草及其退化菌种进行了基因水平的分析.PCR-RFLP实验,采用真菌通用引物ITS1和ITS4,扩增出5.8S和其两端的两个转录间隔区(ITS),选用5种识别四个碱基的内切酶(HaeIII、AfaI、TagI、AluI和XspI),其中只XspI酶切结果在两个菌种中存在差异,进一步测序表明,整个片段全长534bp,共有13个位点发生碱基突变,且都为c转换为t.RAPD实验,选用4组计80个随机引物进行PCR扩增,共筛选出9个对所有供试个体均存在明显差异的引物.结果表明了退化菌株较野生驯化菌株在DNA水平上发生了频率较高的突变,这些突变可能直接与导致菌种退化的基因相关联.
关键词：驯化菌株, 退化菌株, 5.8SrRNA, RFLP, RAPD
分类号：Q939.53 文献标识码：A
文章编号：1007-3515(2003)02-0277-0282
基金项目：大连市科技局重点推广项目:(2002-05)
作者简介：李庆伟,通讯作者: email; liqw@yahoo.com.cn
作者单位：李美娜（辽宁师范大学生命科学学院,大连,116029）
吴谢军（大连双天新药研究开发中心,大连,116031）
李春燕（大连双天新药研究开发中心,大连,116031）
冯伯森（辽宁师范大学生命科学学院,大连,116029）
李庆伟（辽宁师范大学生命科学学院,大连,116029）
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