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花青素合成酶(ANS,anthocyanidin synthase)是植物花青素合成的关键酶,催化无色花色素转变成有色花色素。本研究从心里美萝卜HX12Q-49中克隆获得了花青素合成酶基因Rs ANS(Gen Bank登录号:KR262954)。该基因全长1137 bp,包含2个外显子和1个内含子;开放阅读框1071 bp,编码356个氨基酸。同源性分析显示Rs ANS蛋白与大白菜、甘蓝和芥菜ANS蛋白同源性较高。qRT-PCR表明Rs ANS在心里美萝卜5个不同发育时期均有表达,且在破肚期表达量最高。通过对花青素合成途径其他8个结构基因和3个调控基因表达进行分析,结果显示心里美萝卜中b HLH转录因子在不同发育时期的表达模式与Rs ANS、CHI、DFR和UFGT基本一致,即在破肚期的表达量最高;WD40转录因子的表达与CHS类似,其表达量在芽期、破肚期、膨大前期和膨大盛期逐渐上升,随后在成熟期下降。
Anthocyanidin synthase (ANS) is a key enzyme in the anthocyanidin biosynthesis of plants, catalyzing the conversion of colorless anthocyanidins into colored anthocyanidins. In this study, anthocyanin synthase gene Rs ANS (Gen Bank accession number: KR262954) was cloned from Meirene radish HX12Q-49. The gene is 1137 bp in length and contains two exons and one intron. The open reading frame is 1071 bp, encoding 356 amino acids. Homology analysis showed that Rs ANS protein has high homology with ANS protein of Chinese cabbage, Brassica juncea and Brassica juncea. qRT-PCR showed that Rs ANS was expressed in 5 different developmental stages of the heart radish, and had the highest expression level during the breakage phase. The results showed that the expression pattern of b HLH transcription factor at different developmental stages was basically consistent with that of Rs ANS, CHI, DFR and UFGT through the analysis of the other eight structural genes and three regulatory genes in anthocyanin synthesis pathway. The expression level of WD40 was similar to that of CHS. The expression level of WD40 gradually increased at budding stage, ruffling stage, pre-swollen stage and sweltering stage, then decreased at maturity stage.