利用广亲和基因S5-n的功能标记鉴定水稻资源和检测杂交种纯度

Germplasm Screening and Hybrid Rice Purity Testing with Functional Marker of Wide Compatibility Gene S5-n

为开发利用水稻广亲和基因S5-n的功能标记筛选了部分资源以及鉴定了以培矮64S不育系为母本的杂交种种子纯度。应用广亲和基因S5-n的功能标记S5136从不育系中筛选出携带广亲和基因S5-n的不育系粤泰A,并利用测序证实,同时发现在缺失下游有1个单碱基的突变,与来源于印度的广亲和材料相同;对以培矮64S为母本的杂交稻两优986的种子中不育系自交结实情况进行了判定,从233棵苗中发现4株培矮64S,自交结实率为1.72%。同时比较了SDS方法和简易DNA快速提取方法的PCR结果,发现利用简易方法提取的DNA当天扩增也可以获得较好的PCR结果,但常温保存2周后,以简易方法提取的DNA为模板难以获得PCR产物。因此,S5-n基因的功能标记S5136可以用于广亲和基因的资源鉴定以及鉴定以培矮64S为母本的杂交种自交结实率。

In order to improve the precision of marker-assisted selection for S5-n,a functional marker S5136 based on the 136 bp deletion in the DNA sequence of S5-n was developed.The sterile line Yuetai A with S5-n has been identified with marker S5136,and conformed by sequencing.There was a SNP in the downstream of the 136 bp DNA deletion region of Yuetai A,which has been discovered in the wide compatible germplasm resources from India.The polymorphism was identified with marker S5136 between the photo-thermo sensitive sterile line Peiai 64S and the parent R986.Therefore S5136 was a perfect marker to test the purity of commercial hybrid seed of Liangyou 986（Peiai64s/R986）.Four Peiai 64S plants were identified from the 233 sampled plants with S5136,with self seed setting rate of Peiai64S being 1.72%.The PCR efficiency was compared between the SDS DNA extracting method and the simple-way DNA extracting method.The results showed that the DNA extracted by the simple-way was better,except that the DNA could not be kept for more than 2 weeks at room temperature.Therefore,the functional marker S5136 was a powerful molecular marker in S5-n germplasm screening and seed purity testing for the two line hybrids with Peiai 64S as maternal parent.