花椰菜花球发育相关基因BoCAL的KASP标记开发和应用

BoCAL基因是控制花椰菜花球发生的关键基因之一。为了提高对花椰菜BoCAL基因型鉴定效率,根据该基因第5个外显子中第16个碱基（Ex5+16）G-T的功能性单核苷酸多态性（SNP）突变,开发了竞争性等位基因特异性PCR（KASP）标记(BoCAL-KASP1)。利用该标记对甘蓝类7个变种的蔬菜作物与野生种共计86份材料进行了BoCAL等位基因的分型,发现共有36份材料的基因型是T:T,包括28份花椰菜、6份青花菜与2份引进的地方种材料;43份材料的基因型是G:G,包括2份花椰菜、9份青花菜、6份结球甘蓝、5份芥蓝、3份苤蓝、7份羽衣甘蓝、2份抱子甘蓝、4份引进的地方种与5份甘蓝野生种材料;7份材料显示杂合基因型T:G,包括2份青花菜、2份羽衣甘蓝和3份引进的地方种材料。这86份材料的BoCAL基因型KASP鉴定结果和酶切扩增多态性标记(BoCAL-CAPS1)的鉴定结果完全一致。以上结果说明BoCAL-KASP1标记可以高效准确地鉴定BoCAL基因目标位点的基因型,从而为花椰菜花球发育研究和分子育种提供可靠的功能性标记。

Development and application of KASP marker of BoCAL gene related to curd development in cauliflower

BoCAL is one of the key genes controlling the curd formation in cauliflower. In order to improve the efficiency of BoCAL genotype identification of cauliflower, a competitive allele specific PCR (KASP) marker (BoCAL-KASP1) was developed according to the functional single nucleotide polymorphism (SNP) mutation of the 16th base (Pe5 + 16) G-T in the fifth exon of BoCAL. Using this KASP marker, BoCAL alleles were genotyped in 86 core accessions from seven variants of Brassica. oleracea and related wild species. It turned out that a total of thirty-six accessions showed the genotype of T:T, including twenty-eight cauliflower, six broccoli and two imported landraces; forty-three accessions had the genotype of G:G, including two cauliflower, nine broccoli, six cabbage, five Chinese kale, three kohlrabi, seven kale, two Brussels sprouts, four imported landraces and five related wild species; Seven accessions showed heterozygous genotype T:G, including two broccoli, two kale and three imported landraces. The results of KASP identification of BoCAL genotype of these 86 accessions were completely consistent with those of cleaved amplified polymorphic sequences (BoCAL-CAPS1) identification. Thus, the KASP marker developed in this study could efficiently and accurately identify the genotype of BoCAL target locus, so as to provide reliable functional markers for cauliflower curd development research and molecular breeding.