白菜类作物开花时间相关基因BraA.FLM.a的CAPS标记开发与利用

The development and utilization of CAPS marker of BraA.FLM.a related to flowering time in Brassica rapa

[目的]本文旨在研究白菜类作物开花时间候选基因BraA.FLM.a的酶切扩增多态性序列(CAPS)标记开发与利用。[方法]PCR扩增2个亲本的BraA.FLM.a基因外显子2到内含子2区域的特异序列,开发CAPS标记BraA.FLM.a-NlaⅢ,用CAPS标记对F2分离群体的基因型进行检测,并进行Pearson相关性分析研究基因型与开花时间表现型的关系。[结果]通过调查亲本早开花油用型品种‘Yellow sarson’材料YS-143和晚开花芜菁品种‘Vegetable turnip’材料VT-044的开花时间和F2群体的开花时间,发现两亲本的开花时间存在显著性差异。通过PCR扩增克隆了两亲本的BraA.FLM.a基因的特异序列,并在突变位点T-A处开发出CAPS标记BraA.FLM.a-NlaⅢ。在136份F_2分离群体上进行此CAPS标记的验证后,发现T基因型开花时间晚于A基因型的开花时间,并且差异显著(P<0.05)。[结论]BraA.FLM.a基因的CAPS标记BraA.FLM.a-NlaⅢ与开花时间表型显著相关,该标记可以用作开花时间的分子选择辅助育种。

[ Objectives ] The aim of this study was to explore the development and utilization of cleaved amplified polymorphic sequences （CAPS） marker for flowering time gene BraA. FLM. a in Brassica rapa. [ Methods ] We cloned the sequence of exon 2 to intron 2 region of the BraA.FLM.a gene from the parents. It was confirmed by sequencing that there was a mutation A-T in LE2＋13 in exon 2, and subsequently a CAPS marker for BraA. FLM. a was developed. We investigated the flowering time of the double haploid segregating populations and tested their genotypes using the CAPS marker. The correlation between genotypes and flowering time were studied by Pearson correlation analysis. [ Results] By investigating the flowering time of early-flowering oil-type＇ Yellow sarson＇ material YS-143 ,the late-flowering＇ Vegetable turnip＇ material VT-044 and the late flowering vegetable turnip and the flowering time of the F2 population,it was found that there was a significant difference in the flowering time between two parents. We cloned the specific sequence of the BraA.FLM.a gene of two parents and developed the CAPS marker BraA.FLM.a-Nla 111 at the mutation site T-A. The validation of this CAPS marker on 136 F2 isolates revealed that the flowering time of T genotype was later than that of A genotype, and there was significant difference （P〈O.05）. [ Conclusions ] This CAPS marker BraA.FLM.a-Nla Ⅱ for the BraA. FLM. a gene was found to be significantly associated with flowering time. Thus,the marker BraA.FLM.a-Nla Ⅲ can be subjected to marker- assisted selection of flowering time in breeding.