黑果枸杞和宁夏枸杞中黄酮醇合酶基因的克隆及表达分析

Molecular Cloning and Expression Analysis of Flavonol Synthase in Lycium ruthenicum Murr. and Lycium barbarum L.

黑果枸杞和宁夏枸杞为枸杞属中两种重要的药用植物,具有极大的药用价值。本研究利用转录组数据克隆了黑果枸杞和宁夏枸杞黄酮醇合酶基因(Flavonol synthase,FLS),分别命名为LrFLS及LbFLS,并通过实时定量PCR(real-time quantitative PCR)分析其在两种枸杞果实四个发育时期中的表达模式。研究结果表明:LrFLS与LbFLS各编码一条由350个氨基酸残基组成的蛋白序列。蛋白序列分析表明:LrFLS、LbFLS相似度高达99%,各含有两个保守的功能域。进化分析表明,LrFLS和LbFLS同属茄科的烟草、马铃薯的FLS蛋白序列相似度最高。实时定量PCR结果表明,LrFLS在S2时期的表达量最高,在果实发育过程中呈现先升高后降低的趋势;而LbFLS在S2时期表达量最低,呈现先降低后升高的趋势。本研究为揭示黄酮醇在黑果枸杞和宁夏枸杞果实中的累积具有一定的启示作用。

Lycium ruthenicum Murr. and Lycium barbarum L. are two important medicinal plants in solanaceae with great medicinal values. In this study, two Flavonol synthases（LrFLS and LbFLS） were retrieved from L. ruthenicum and L. barbarum transcriptome data, respectively. The PCR results showed that both LrFLS and LbFLS encoded a protein composed of 350 amino aicds. Protein sequences analysis indicated that LrFLS and LbFLS shared 99% sequence identity and had two conserved domains, respectively. Phylogenetic analysis displayed that LrFLS and LbFLS had high similarity with other Solanaceae plants such as tobacco and potato. Real-time quantitative PCR results suggested that the expression of LrFLS first increased then decreased during fruit ripening with the highest level in S2 stage, while the expression of LbFLS decreased in S2 stage and followed by an increase. This study will have an inspiration for facilitating the elucidation of biosynthesis flavonol in L.ruthenicum and L. barbarum fruits.