PCR—SSCP与测序技术相结合检测小麦耐盐突变体

根据位于小麦第四同源群上与耐盐有关的gf-2.8基因的编码区序列设计1对引物，分别以两个耐盐突变体及其亲本的总DNA为模板进行PCR扩增，在5个供试材料中均扩增出1条约685bp的目的条带，SSCP电泳显示突变体974915与其他供试材料之间存在差异。测序表明冀麦24和其耐盐突变体8901-17的扩增产物序列与gf-2.8基因的发表序列相同，这表明突变体8901-17的突变位点不在该基因上，而另一耐盐突变体974915的序列中则至少存在2个单碱基突变，有一处突变导致了氨基酸的变化，该突变位点位于gf-2.8基因的保守区域内。

Detection of the Wheat Salt-tolerant-mutant Using PCR-SSCP Combining with Direct Sequenceing

gf-2.8 is a gene located on the chromosomal homologous group 4, which was reported to have relationship with salt tolerance of wheat. A pair of primers were designed to amplify the coding region of gf-2.8 in the two salt- tolerant mutants and their parents. Agarose gel electrophoresis showed that the 685bp band was amplified among all the materials. SSCP analysis suggested that 974915 (one of the mutants) was different from the other materials. The sequencing results showed that Jimai 24 and its salt-tolerant mutant 8901-17 bad the same sequence as published, which indicated that the mutation site of 8901- 17 was not on the gf-2.8 gene. However, there were at least two single base mutations in the gf-2.8 of 974915, one of which caused the alternation of amino acid, this mutation occurred in the conservative region of gf-2.8.