思茅松HMGS基因的克隆与表达分析  

Identification and expression of the 3-hydroxy-3-methylglutaryl-CoA synthase gene from Pinus kesiya var.langbianensis

作　　者：李云琴 原晓龙[1,2] 李江 王毅[1,2] 陈伟 LI Yunqin;YUAN Xiaolong;LI Jiang;WANG Yi;CHEN Wei(The Key Laboratory of Forestry Plant Cultivation and Utilization,Kunming,Yunnan 650201,China;The Key Laboratory of Rare and Endangered Forest Plants of National Forestry and Grassland Administration,Yunnan Academy of Forestry and Grassland,Kunming,Yunnan 650201,China)

机构地区：[1]云南省森林植物培育与开发利用重点实验室,云南昆明650201 [2]国家林草局云南珍稀濒特森林植物保护和繁育重点实验室,云南昆明650201

出　　处：《森林与环境学报》2020年第4期428-432,共5页Journal of Forest and Environment

基　　金：云南省林业科学院创新基金资助项目(MS2019-02,QN2018-01);国家林业公益性行业科研专项资助项目(201304105)。

摘　　要：为研究3-羟基-3-甲基戊二酰辅酶A合酶基因(HMGS)在思茅松二萜类生物合成中的作用机制,采用RT-PCR技术从思茅松中获得3-羟基-3-甲基戊二酰辅酶A合酶基因的cDNA序列。序列分析显示,该基因全长1 425 bp,编码474个氨基酸,属于HMGS基因家族。序列比对分析显示PkHMGS基因所编码的氨基酸序列拥有HMGS家族蛋白特有的保守序列GNTEIEGVDSTNACYGGTA,与樟子松HMGS的蛋白序列同源性为99.79%。系统进化分析显示,思茅松与裸子植物的HMGS聚为1支,与樟子松的亲缘关系最近。荧光定量PCR分析表明,在思茅松的树皮中HMGS基因的表达明显受到割脂物理伤害的诱导,在高产脂思茅松松针表达量最高。结果为揭示思茅松高产脂的分子机理提供理论依据。This study investigated the effect of the 3-hydroxy-3-methylglutaryl-Coenzyme A synthase gene(HMGS)on diterpenoids biosynthesis in Pinus kesiya var.langbianensis.For the first time,the complete open reading frame(ORF)of the HMGS gene was cloned from P.kesiya var.langbianensis by RT-PCR.Sequence analysis showed the ORF was 1425 bp,encoded 474 amino acids,and belonged to the HMGS family.Proteinic sequence alignment revealed 99.79%similarity between P.kesiya var.langbianensis and Pinus sylvestris var.mongolica,with a conserved site GNTEIEGVDSTNACYGGTA in the PkHMGS gene.The genetic similarity between the two species was further clarified via a dendrogram.Fluorescent quantitative reverse PCR showed increased PkHMGS gene expression when the bark of P.kesiya var.langbianensis was cut,and PkHMGS gene expression was highest in the pine needles of the high-resin-yield individuals.These results provide a theoretical basis for the molecular mechanism of high-resin-yield P.kesiya var.langbianensis.

关 键 词：思茅松 3-羟基-3-甲基戊二酰辅酶A 合酶 基因克隆 生物信息学分析 

分 类 号：Q753[生物学—分子生物学]