三七香叶基香叶基焦磷酸合酶基因的克隆及表达分析  

Cloning and expression analysis of GGPPS gene from Panax notoginseng

作　　者：闵丹丹[1] 唐美琼[2] 李刚[2] 屈啸声[2] 缪剑华[2] 

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西壮族自治区药用植物园广西药用资源保护与遗传改良重点实验室,广西南宁530001

出　　处：《中国中药杂志》2015年第11期2090-2095,共6页China Journal of Chinese Materia Medica

基　　金：广西科学研究与技术开发项目(桂科重1298001-1-4;桂科重14124002-1);广西自然科学基金项目(2013GXNSFBA019087);广西药用植物园青年基金项目(桂药基20)

摘　　要：基于三七转录组测序结果,选择萜类代谢骨架上香叶基香叶基焦磷酸合酶基因(GGPPS),结合RACE技术,克隆得到全长1 203 bp的c DNA序列,最长开放阅读框为1 035 bp,命名为Pn GGPPS,Gene Bank登录号为KM486564,Pn GGPPS基因全长2 370 bp,包含1个内含子和2个外显子,该基因编码344个氨基酸,与蓖麻Ricinus communis、丹参Salvia miltiorrhiza等植物GGPPS的同源性达到73%以上,具有富天冬氨酸区等多个结构域,属于类异戊二烯合成酶超家族。实时荧光定量PCR结果显示,Pn GGPPS在一至三年生三七的根、茎、叶和三年生的花等不同器官组织中均有表达,但以三年生叶中具有显著水平的高表达量,暗示其既具有调节三七生长发育的基本功能,也与三七叶绿素和类胡萝卜素的合成、叶绿体的发育、阴生适应性,以及品质的形成有关。According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene （GGPPS） in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE （rapid amplification of cDNA ends） technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.

关 键 词：三七 PnGGPPS基因 实时荧光定量PCR 阴生习性 

分 类 号：S567.236[农业科学—中草药栽培]