【摘要】 蒎烯合成酶(α-pinene synthase, APS)是α-蒎烯生物合成的关键酶。本研究利用同源克隆及基因组步移法从思茅松中克隆得到一个α-蒎烯合成酶基因,命名为PkAPS (GenBank登录号为KX394684),基因全长3 523 bp,含有10个内含子,其c DNA全长1 956 bp,编码651个氨基酸残基。生物信息学分析显示PkAPS属于萜烯合成酶家族蛋白;系统进化树分析显示PkAPS与马尾松α-蒎烯合成酶亲缘关系最近。基因表达分析显示:PkAPS在高产脂思茅松个体各组织的表达量均高于低产产脂思茅松个体;而在高产脂思茅松不同组织中的表达比较中,PkAPS在小枝中的表达量最高。本研究将有利于进一步研究思茅松α-蒎烯合成酶调控机理及培育高产α-蒎烯思茅松新品种的研究。更多还原

【Abstract】 pinene synthase(APS) is a key enzyme during the process of α-pinene biosynthesis. The present study cloned an α-pinene synthase gene from Pinus kesiya var langbianensis with the methods of homologous gene cloning and genomic walking cloning, and named it Pk APS(GenBank ID: KX394684). There is 3 523 bp including 10 introns of Pk APS, its full length of c DNA is 1 956 bp and encodes 651 amino acids residue. The bioinformatics showed that Pk APS is assigned to terpene biosynthetic enzyme families, and the dendrogram revealed that Pk APS should be the closest genetic relationships with the α-pinene synthase gene from P. massoniana. The gene expression clarified the expression levels of gene Pk APS originated from all the individuals in the high-yield oleoresin P. kesiya are higher than that of in the low-yield oleoresin P. kesiya; and the Pk APS gene expression compared with the different tissues of high-yield oleoresin P. kesiya revealed the highest expression levels was in the branchlet. The results of this research has applied some theoretical basis for exploring the mechanism of α-pinene biosynthetic regulation and breeding a new variety of P. kesiya that could produce higher yield α-pinene annually.更多还原