森林草莓‘Ruegen’果胶裂解酶基因的克隆及荧光定量表达分析

The Cloning and Quantitative Expression Analysis of Pectate Lyase Gene in Fragaria vesca

根据森林草莓(Fragaria vesca)果胶裂解酶(Pectate lyase)基因序列信息设计引物,克隆了森林草莓‘Ruegen’的果胶裂解酶基因PLA、PLB和PLC。利用MEGA5.0软件将这3个基因编码的氨基酸序列与其他植物的果胶裂解酶氨基酸序列进行聚类分析,同时采用荧光定量PCR的方法对‘Ruegen’植株不同器官及不同发育时期果实的果胶裂解酶基因表达量进行分析。结果表明:PLA、PLB和PLC在其植株各器官中均有表达,且在发育后期果实中的表达量明显高于其他器官,结合进化树以及果实发育过程中果胶裂解酶活性变化,推测果胶裂解酶基因对草莓果实发育后期的软化具有调节作用。

Primers were designed based on the sequence information of woodland strawberries,and pectate lyase gene(PLA,PLB,PLC)was cloned from‘Ruegen'. Phylogenetic tree is clustered by MEGA 5.0 based on amino acid of pectate lyase gene(Fragaria vesca)and known pectate lyase gene of other plants. At the same time,gene expression levels were analyzed by real-time quantitative PCR method for strawberries on different organs and developmental stages of fruit. The results showed that pectate lyase gene is continuous expression in all organs of strawberry plants. In developing strawberry fruit,pectate lyase gene family was significantly higher than other organs,combined with the results of the phylogenetic tree and the change of pectate lyase activity speculate that pectate lyase gene plays a role in the regulation of strawberry fruit ripening,especially in the late stage of fruit ripening.