双孢蘑菇颜色基因分子标记的初步筛选

摘要

Random amplified polymorphic DNA (RAPD)methodology was used to tentatively identify DNA markers linked to cap color in four white and four brown strains of Agaricus bisporus .Three random primers (S33,S438 and S485)that amplified four specific bands (300 ~2000 bp in size)from the four brown test strains but not the four white test strains were screened out from 500 random primers.After cloning and sequencing these specific bands,four SCAR (Sequence Characterized Amplified Regions)primer pairs were designed accordingly and used to detect SCAR markers specifically associated with brown cap color among 48 (24 brown,24 white)A .bisporus strains.Distribution of the amplified fragments generated by the SCAR4 primer pair revealed a band approximately 1200 bp in size (designated SCAR41200 )in 22 of the 24 brown strains that was absent from all the 24 white strains,indicating that SCAR41200 was closely linked to alleles responsible for the brown cap color gene in A .bisporus .%以4个白色和4个棕色双孢蘑菇(Agaricus bisporus )菌株,对其500个随机扩增多态性 DNA (random amplified polymorphic DNA,RAPD)随机引物进行筛选,筛选出的 S33、S438、S4853个引物在棕色菌株组能扩增出4个特异条带(大小300~2000 bp),将特异片段回收,克隆测序,将合成的4对特征序列扩增区域(sequence characterized amplified regions,SCAR)标记引物对48 个 菌株 进 行 验 证,结果表明：仅SCAR41200在24个棕色菌株的22个中能扩增出特异条带,而所有白色菌株中都没有此条带,表明该标记与棕色菌株的棕色基因相关。