大豆滞绿突变体的T-DNA插入位点分析

The T-DNA Insertion Sites Analysis of Soybean Stay-green Mutant

通过农杆菌介导的子叶节转化法获得的大豆T-DNA插入突变体ls-1是一个滞绿突变体,该突变体同时表现出花器官异常和种子败育的表型.通过染色体步移的方法分析发现T-DNA插到大豆基因组的7号染色体上,对测序获得的插入位点序列进行了PCR验证.进一步通过半定量RT-PCR和生物信息学分析,推测突变体表型的产生可能由于T-DNA插入编码S1型核酸内切酶的基因Glyma.07G191100和编码线粒体中交替型NAD(P)H脱氢酶的基因Glyma.07G191200之间,使二者表达受到影响造成的.该研究为后续的基因功能研究奠定了基础.

The soybean T-DNA insertion mutant ls-1 obtained by Agrobacterium-mediated cotyledonary node transformation is a stagnant green mutant.In the while,it displayed the phenotype of floral organ abnormalities and abortion.The result of chromosome walking showed that T-DNA was inserted into chromosome 7 of soybean genome,the sequence of insert site was confirmed by PCR.Further analysis of semi-quantitative RT-PCR and bioinformatics suggested that the phenotype of the mutant may be due to the insertion of T-DNA between Glyma.07G191100,which encodes S1 type endonuclease,and Glyma.07G191200,which encodes NAD(P)H-Quinone oxidoreductase,the expression of both genes is influenced.This study laid a foundation for the subsequent study of gene function.