甜椒胞质雄性不育恢复系SRAP及SCAR标记

以甜椒胞质雄性不育恢复系5-2R1和相应的保持系5-2为试材,利用SRAP分子标记技术筛选与甜椒恢复基因相关的分子标记。128对SRAP引物共扩增获得了3796条从100bp至800bp大小不同的条带,平均每对引物组合可扩增出30条清晰的条带,检测到4个多态性位点,其中恢复系材料仅有1个。对该特异片段进行回收、克隆和测序,结果表明该片段全长为259bp。经数据库比对分析表明,该片段与线粒体中的NADH脱氢酶第5亚基(GenBank:EF151914.1)具有81%的同源性;同时,该片段与辣椒BAC克隆PEPBAC 158K24(GenBank:FJ597540.1)的部分序列高度同源。根据序列特点设计特异SCAR引物,对恢复系和保持系进行扩增验证,仅在恢复系中扩增出目的条带,表明已成功地将此SRAP标记转化为简单、稳定的SCAR标记。我们推测本研究获得的SCAR标记可能与胞质雄性不育恢复性状连锁。

SRAP and SCAR Marker Linked to Restoring Gene for Cytoplasmic Male Sterility in Sweet Pepper

SRAP(sequence-related amplified polymorphism) was analyzed between restorer line 5-2R1 and its maintainer line 5-2 of Cytoplasmic Male Sterility(CMS) in sweet pepper.Using 128 pairs of SRAP primers,3 796 bands between 100 bp and 800 bp were amplified in these two lines,and averagely 30 bands per primer combination were amplified.There are 4 polymorphic loci in these two lines,but only one polymorphic fragment was found in CMS restoring line.This specific SRAP fragment was retrieved,cloned and sequenced,and the results indicate the length of the sequence is 259 bp.The sequence analysis shows that this fragment has 81% identity with NADH dehydrogenase subunit 5(ND5) gene and part of this fragment is highly homologous with part of BAC clone PEPBAC158K24 in Capsicum frutescens.After cloning and sequencing,specific SCAR primers were designed.By amplification testing the specific fragment was detected in restorer line but not in the maintainer line.This showed that the SRAP marker was successfully transformed to simple and stable SCAR marker.So it could be inferred that the SCAR marker is likely related to CMS restoring gene.