草菇有性世代间rDNA序列变异分析及RAPD分析  

Sequence variation of rDNA and RAPD analysis between sexual generations of Volariella volvacea

作　　者：徐学锋[1] 姬宏超[1] 李巧玲[1] 莫美华[1] 

机构地区：[1]华南农业大学食品学院,广东广州510642

出　　处：《食品工业科技》2012年第10期198-201,共4页Science and Technology of Food Industry

基　　金：国家自然科学基金(30800774);华南农业大学校长基金(2007K046)

摘　　要：草菇的遗传与有性生殖过程存在很多的特殊性和不确定性,本研究以草菇V23菌株的成熟子实体弹射孢子,单孢分离后得13个单株,分别摇瓶培养、收集菌丝,提取总DNA,用通用引物ITS4、ITS5进行PCR扩增后连接、转化并测序,将单孢间和单孢与亲本间的ITS序列进行比对,分析草菇ITS序列多样性及变异性;同时以相应单孢菌株DNA为模板,用20条随机引物进行RAPD扩增。结果表明:草菇单孢间和单孢与亲本间的ITS序列相似度均在99%以上,相较于亲本的ITS序列,单孢序列中发现了有5个位点出现碱基转换,但没有两个以上的单孢发生同一转换;20条随机引物分别与14个样本进行RAPD扩增,其中3个引物出现多态性条带,带型结果发现单孢菌株间的带型差异呈现一定规律。草菇ITS序列变异较少,不适于单孢间的变异分析,而RAPD方法对摸索草菇有性世代间的遗传分离规律有重要价值。The life history and sexuality pattern of Volariella volvacea are still uncertain and unclear.13 single sexual spores were isolated from strain V23,and cultivated in liquid medium to obtain mycelia for DNA extraction.Internal Transcribed Spacers（ITS）of these 13 isolates and V23 were amplified and sequenced with primer ITS4 and ITS5.All theses sequences had over 99% similarity to homologous sequence from Genbank with multiple alignments,only five transitions but no transversion had been observed between two sexual Generations of Volariella volvacea.Less variation inter sexual spores revealed that ITS sequence was not the perfect gene for genetic analysis of Volariella volvacea.The random amplified polymorphic DNA（RAPD）analysis was performed with 20 random primers and genomic DNA which isolated from the 13 single spore strains and parent strain V23,PCR with only 3 random primers showed polymorphic bands,and some genetic separation regularity revealed that RAPD analysis had much more important value on research of sexuality pattern of Volariella volvacea.

关 键 词：草菇 ITS RAPD 同宗结合 

分 类 号：Q789[生物学—分子生物学]