大葱胞质雄性不育位点RAPD标记转化为SCAR标记的研究

Study on Conversion of RAPD Markers Linked to Cytoplasmic Male Sterile loci to SCAR markers in Welsh Onion(Allium fistulosum L.)

S132800、S2002400是与大葱胞质不育位点相连锁的RAPD标记,回收、纯化特异性片段S132800、S2002400,连接至pGEM-T-easy载体,转化受体菌后筛选出阳性克隆,并对插入片段进行测序。根据测序结果,设计了3对SCAR引物:SCS13L/SCS13R(S132800的SCAR标记引物)、SCS200L/SCS200R1、SCS200L/SCS200R2(S2002400的SCAR标记引物)。其中SCS13L/SCS13R可成功地区分N、S两类胞质,适宜退火温度为59℃。而S2002400转化为SCAR后多态性消失,讨论了转化SCAR标记失败的可能原因。

Specific bands S13_2800 and S200_2400, which linked to cytoplasmic male sterile loci in welsh onion, were recovered, purified and inserted into pGEM-T-easy vector, and then transferred into E. coli. DH5α. Positive colony was identified for sequencing. According to the sequences, three pairs of 22-mer specific primers, SCS13L/SCS13R, SCS200L/SCS200R1 and SCS200L/ SCS200R2 were designed. SCS13 L/SCS13R (SCAR marker of S13_2800) could be used to identify N and S cytoplasm, and its optimal annealing temperature was 59℃. Unfortunately the conversed SCAR marker from S200_2400 could not reproduce the polymorphic S200_2400. The Reasons for failing polymorphism when RAPD marker was converted to SCAR marker were also discussed.