黑麦重复序列在检测小麦品种中外源染色体的应用

本研究根据RAPD引物OPH20在黑麦中扩增出的特异序列pSc20H.2设计一对PCR引物pSc20ht-23/24,以来源于黑麦的小麦抗叶锈近等基因系材料TcLr45及感病对照Thatcher为亲本进行PCR扩增。并对42个小麦抗叶锈近等基因系及103个小麦品种材料进行检测。引物pSc20ht23/24在TcLr45中扩增出一条约750bp的条带,而在Thatcher中无扩增条带。对该特异片段回收、克隆测序为734bp。42个小麦抗叶锈近等基因系检测在TcLr26中扩增出与TcLr45相同的条带,而在同样来源于黑麦的小麦抗叶锈近等基因系TcLr25中未扩增出该条带;中国春-Imperial黑麦附加系1R-7R中除5R外均扩增出该条带;13个1B/1R易位系小麦品种也扩增出该条带;90个地方小麦品种中有16个扩增出该条带,6个品种经系谱分析具有黑麦遗传背景,表明该标记可用于检测小麦中含有的除黑麦5R染色体之外的外源染色质。

The Repetitive Sequence of Secale cereale Applied on Detection of Exogenous Chromosome of Wheat

A pair of PCR primers pSc20ht23/24 was designed based on sequence pSc20H.2 amplified by RAPD primer OPH20 in rye.The primers were used to amplify wheat leaf rust resistance near isogonic lines of TcLr45,derived from rye,and its susceptible background Thatcher.In addition,42 wheat leaf rust resistance near isogonic lines and 103 wheat varieties were detected with the primers.A specific band size about 750 bp was amplified in TcLr45 by the primers pSc20ht23/24,and there were no bands amplified in Thatcher.This specific band was cloned and sequenced,the full length is 734 bp.The result from testing 42 wheat leaf rust resistance near isogonic lines showed that TcLr26 amplified the same size segment as TcLr45,but there was no amplification in TcLr25 derived from rye.The same size specific band was amplified in the Chinese Spring-Imperial addition lines from 1R to 7R except 5R addition line.All of the 13 wheat varieties of 1B/1R rye translocation line were amplified the same band as TcLr45 and TcLr26,and 16 of 90 wheat landraces amplified this band too.The results from pedigree analysis indicated that 6 of 16 varieties had the genetic background of rye,this SCAR marker can be used for detecting exogenous chromosome of rye except 5R in wheat.