两个低谷蛋白基因插入缺失标记的设计与验证

低谷蛋白稻米是肾脏病人极有效的食疗辅助品,培育低谷蛋白功能性水稻品种具有重大意义。低谷蛋白基因Lgc1作为培育低谷蛋白功能性水稻品种的优质资源,受到了育种家的青睐。为提高低谷蛋白基因Lgc1分子标记辅助选择的准确性,我们根据其突变体在两个谷蛋白基因GluB4和GluB5间的碱基缺失,设计出了Lgc1基因的插入缺失标记InDel-Lgc1-A和InDel-Lgc1-B。利用InDel标记对W3660(Lgc1低谷蛋白品种)/南粳46(谷蛋白正常品种)的F2分离群体和13份水稻品种进行检测验证。依据其PCR扩增产物的电泳带型,可准确地区分出低谷蛋白纯合基因、谷蛋白正常纯合基因和杂合基因型3种带型,且3种带型与其植株或品种相应的蛋白性状表现完全一致,表明这两对InDel标记可用于Lgc1低谷蛋白基因资源的鉴定以及分子辅助育种。

Design and Validation of Two InDel Markers for Low Glutelin Content Gene (Lgc 1) in Rice

Rice with low glutelin content is quite effective as a kind of functional food for patients with kidney disease.It is of great importance to develop commercial rice varieties of low glutelin content.In recent years,Lgc1 gene,as an excellent genetic resource for rice varieties with low glutelin content,has been given more and more attention by breeders in functional rice breeding.To improve the precision of marker-assisted selection for Lgc1 gene in rice,we designed two pairs of functional markers for Lgc1 gene based on a 3.5 kb deletion between two highly similar glutelin genes GluB4 and GluB5 in two rice mutants.Two InDel markers were designated as InDel-Lgc1-A and InDel-Lgc1-B,respectively.These markers were used to analyze 13 rice varieties and the F2 segregation population derived from the cross of the low glutelin content rice variety W3660 and normal variety Nangjing-46.The PCR results showed that according to the band types the homozygous genotype with low glutelin content,the heterozygous genotype,and the homozygous genotype with normal glutelin content could be clearly distinguished,which was completely consistent with the results of the protein analysis.Thus,these two InDel markers could be used to distinguish varieties with Lgc1 gene,and can also be used in marker-assisted selection.