Mutator转座子介导的PPR插入位点分离与遗传分析

Mutator Transposon Mediated PPR Insertion Site Isolation and Genetic Analysis

利用Mutator(Mu)诱变群体,进行Mu因子插入PPR(Pentatricopeptide Repeats,PPR)位点的分离,及其插入真实性验证。以含活性Mu转座子为父本和玉米自交系综31(Z31)杂交,经与Z31多代回交和自交获得BC3F2为材料,利用Mu-AFLP方法分离Mu插入侧翼序列的PPR位点,且验证插入真实性。采用Mu-AFLP方法获得Mu因子插入侧翼序列14条,去除冗余序列2条和重复序列8条,其余4条为Mu因子插入的基因序列,经基因型遗传分析验证了2条侧翼序列为真实插入。经功能分析,其中1个为PPR突变位点,且Mu因子插入于该基因5’UTR区第121bp和第122bp碱基之间。经基因组定位和功能分析,显示Mu插入位点定位在第6染色体上,为PPR基因家族,能够编码PPR蛋白,推测可能与玉米叶色变化有关。

Using Mutator（Mu） mutation groups,separates and validates the Mu factor mediated PPR（Pentatricopeptide Repeats,PPR） mutation site.Containing active Mu transposons as male parent hybrid with maize inbred lines Z31,after repeated backcross with Z31 and selfing,obtain BC3F2 as material.Using Mu-AFLP method to separate Mu insertion flanking sequences of the PPR sites,and verify the authenticity of the insertion sites.Using Mu-AFLP method,14 Mu factor insertion flanking sequences have been cloned.Removing 2 redundant sequences and 8 repeat sequences,the remained 4 target sequences were the object for further study.Gene genetic analysis verified 2 flanking sequences insertion for real.The function analysis,in which 1 is PPR mutation site and Mu factor inserted into the gene 5＇ UTR 121st bp and 122nd bp between base pairs.The genomic localization and functional analysis shows that Mu insertion site of target sequence located at chromosome 6,and belonged to PPR gene family.This gene could be encoded PPR protein,so we hypothesized that this Mu insertion site influences corn leaf color.