不同分子标记鉴定甜菜雄性不育系与保持系的应用

Application of Different Molecular Markers to Identify Male Sterile Lines and Maintainers in Sugar Beet

准确鉴定甜菜育性对于甜菜育种工作具有重大意义,本试验利用3对甜菜不育系与保持系以及甜菜育性相关分子标记,以探究不同分子标记对于鉴定甜菜不育系与保持系的有效性.本试验使用VNTR分子标记,鉴定甜菜雄性不育细胞质(S)与正常型细胞质(N),使用CAPS标记与DFLP标记鉴定甜菜细胞核Rf1与Rf2位点基因型.还利用根据基因组和转录组序列设计的7对引物,以及2对SSR引物,验证了其鉴定育性的有效性.结果显示,仅TR1、s17、o7、21 E2M14S四对引物产生了多态性,其余引物均未产生多态性.其中,TR1扩增带型与细胞质基因型间具有明显的相关性(P<0.01);s17引物与o7引物基因型生育恢复独立性的无效假设均未被拒绝;引物21 E2M14S扩增条带类型与育性间不存在明显的相关性(P=0.721).总之,TR1引物对于鉴定细胞质育性类型准确性较高,s17引物对于鉴定Rf1位点基因型也较为可靠.而Rf2位点对于育性的恢复可能起到很小的作用.

In this experiment,three pairs of sugar beet sterile and maintainer lines and sugar beet breeding-related molecular markers were used to investigate the effectiveness of different molecular markers for identifying sugar beet sterile and maintainer lines.In this experiment,VNTR molecular markers were used to identify the male sterile cytoplasm(S)and normal cytoplasm(N)of sugar beet.CAPS markers and DFLP markers were used to identify the genotypes of Rfl and Rf2 loci in sugar beet nuclei.The validity of the genotypes was also verified using seven primer pairs designed based on genomic and transcriptomic sequences,as well as two SSR primer pairs to identify fertility.The results showed that only four primer pairs,TR1,s17,o7,and 21 E2M14S,produced polymorphism,while the rest did not produce polymorphism.Among them,there was a significant correlation between TR1 amplified band type and cytoplasmic genotype(P<0.01);the null hypothesis of genotype fertility restoration independence for both s17 primer and o7 primer was not rejected;there was no significant correlation between primer 21 E2M14S amplified band type and fertility(P=0.721).In conclusion,the TRl primer was more accurate for identifying cytoplasmic fertility types,and the s17 primer was more reliable for identifying the genotype of the Rf1 locus.In contrast,the Rf2 locus may play a small role in the recovery of fertility.