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为进一步探究草莓果实花色苷合成过程中各个酶的作用,明确草莓果实花色苷的代谢机制,以‘阿尔比’草莓(Fragaria×ananassa‘Albion’)为试材,利用RT-PCR从草莓果实中克隆到DFR、ANS和LAR基因的编码序列,对预测的氨基酸序列进行分析,结果表明:各基因与NCBI(美国国立生物技术信息中心)上已有的草莓品种‘Queen Elisa’及‘Korona’(Fragaria×ananassa)所对应基因的同源性高达99%～100%。半定量RT-PCR分析表明,DFR基因依次在幼果期、白熟期和红熟期出现了3个表达峰值,其中红熟期最大;ANS基因的表达量在白熟期后迅速增加,红熟期达最大值;LAR基因的最大表达量出现在幼果期,后呈降低趋势。草莓果实发育过程中,DFR基因在幼果期和红熟期的表达峰值分别与黄烷-3-醇类和花色苷的合成峰值相一致,ANS及LAR分别与花色苷和黄烷-3-醇类的合成密切相关。
In order to further explore the role of each enzyme during anthocyanin synthesis in strawberry fruits, the metabolic mechanism of anthocyanins in strawberry fruits was clarified. The Fragaria × ananassa’Albion ’ The coding sequences of DFR, ANS and LAR genes were cloned and the predicted amino acid sequences were analyzed. The results showed that the genes were closely related to the existing strawberry varieties ’Queen Elisa’ and ’Korona’ (NCBI) on the NCBI (National Center for Biotechnology Information) Fragaria × ananassa) corresponds to the gene homology of up to 99% to 100%. Semi-quantitative RT-PCR analysis showed that DFR gene appeared peak in young fruit, white mature fruit and red ripe fruit, with the highest in red ripening stage. The expression of ANS gene increased rapidly after white ripening, while red The maximum expression of LAR gene appeared in young fruit stage and then decreased. During strawberry fruit development, the peak expression of DFR gene in young fruits and red fruits was consistent with that of flavan-3-alcohols and anthocyanins, respectively. ANS and LAR were correlated with anthocyanin and flavan-3- The synthesis of alcohols is closely related.