太空诱变玉米雄性不育突变体的DNA-AFLP分析

AFLP Analysis of Male Sterile Mutant Induced by Space Flight in Maize

利用AFLP标记及克隆测序技术对太空诱变玉米雄性不育突变体的基因组DNA进行分析,寻找与不育性状紧密连锁的分子标记。在供试的56对引物中,32对引物能够扩增出稳定、清晰的条带,4对引物能够在不育株和可育株之间扩增出多态性条带。通过对多态性片段的回收、克隆测序及序列分析结果表明,不育株特有的条带(Zmd1-302)与玉米基因组的克隆(AC187265)和玉米雄配子cDNA文库中的EST序列(gi33771201)同源性都很高,推测该片段和玉米太空诱变核不育之间的育性差异有关。将具有多态性的AFLP标记转化为SCAR标记,3对SCAR引物在8个不育单株和8个可育单株中均有扩增,通过聚丙烯凝胶电泳未检测到不育和可育之间的长度多态性,序列比对分析发现不育与可育之间存在多个单碱基差异。

In order to get closed molecular marker linked to this male sterile trait, AFLP technology and cloning as well as sequencing were adopted to analyze the difference between male fertile and male sterile which induced by space flight in maize. Total 56 primer combinations were used in this experiment, 32 primer combinations can amplify stable and clear bands. Among them, four differential fragments were eluted, cloned, sequenced. Sequencing analysis through internet blast searching showed that one of the fragments were homology as EST（gi33771201） which was from a maize sperm library and a DNA sequence（AC 187265） which was located at the third chromosome 3.04. According to the sequence obtained by AFLP, three SCAR primers were designed, but the length polymorphism were not detected among the amplified bands between male fertile plants and male sterile plants, but many single different bases were found between the amplified bands from male fertile plants and male sterile plants.