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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。
AFLP markers and cloning sequencing techniques were used to analyze the genomic DNA of space-mutated maize male-sterile mutants to find molecular markers closely linked with the sterile traits. Among 56 pairs of primers tested, 32 pairs of primers amplified stable and clear bands, and 4 pairs of primers amplified polymorphic bands between sterile and fertile plants. Through the recovery of the polymorphic fragments, cloning and sequence analysis showed that the unique bands (Zmd1-302) and maize genome clones (AC187265) and the maize male gamete cDNA library (gi33771201) Homology is high, it is speculated that the fragment and maize space-induced nuclear fertility differences between fertility. AFLP markers with polymorphism were transformed into SCAR markers. Three pairs of SCAR primers were amplified in 8 sterile plants and 8 fertile plants, and infertility was not detected by polypropylene gel electrophoresis The results of sequence alignment showed that there were multiple single base differences between male sterile and fertile.