铁皮石斛转化酶抑制子家族基因的克隆和表达分析  

Cloning and Expression Analysis of Invertase Inhibitor Gene Families from Dendrobium officinale

作　　者：苗小荣 牛俊奇 莫昭展 王爱勤[1] 何龙飞[1] 

机构地区：[1]广西大学农学院,南宁530005 [2]玉林师范学院生物与制药学院,玉林537000

出　　处：《生物技术通报》2018年第1期129-136,共8页Biotechnology Bulletin

基　　金：广西科学研究与技术开发计划项目(桂科重1298001-3-1);广西省南宁市重点研发项目(20162094)

摘　　要：转化酶抑制子(Invertase Inhibitor,Inv Inh)能调控转化酶的活性,在植物的蔗糖代谢过程中具有重要作用。本研究旨在从珍稀濒危兰科药用植物铁皮石斛中克隆Inv Inh家族基因,并对其进行定量表达以及生物信息学分析。结果显示:从铁皮石斛叶中克隆到3个Inv Inh基因,分别命名为Do Inv Inh1、Do Inv Inh2和Do Inv Inh3,分别编码176、177和188个氨基酸。生物信息学分析表明,Do Inv Inh蛋白都具有1个信号肽、2个跨膜结构以及PMEI蛋白的保守结构域。Do Inv Inh蛋白的氨基酸序列同源性较低,但都具有4个保守的Cys位点和SP位点。荧光定量PCR分析发现,在根、茎、叶和花中都能检测到3个Do Inv Inh基因表达。Do Inv Inh2和Do Inv Inh3基因在根中表达量最低,而在花中表达量最高。Do Inv Inh1和Do Inv Inh3基因在铁皮石斛不同生长年限茎中的表达模式相似,都是在1年生茎中表达量最高,2年生茎中次之,在3年生茎中表达量最低。Do Inv Inh2基因在2年生茎中基因表达量最高。Do Inv Inh1和Do Inv Inh3基因表达量与CIN酶活性都呈显著负相关,推测这2个基因可能在调节茎中CIN酶活性中起作用。Invertase inhibitor （ Invlnh ） controls the activity of invertase and plays a significant role in the sucrose metabolic processes. This study aims to clone member of Invlnh gene in a rare and endangered medicinal plant Dendrobium officinale, and to quanlitatively detect the expression in different organs and to conduct bioinformatics analysis. As results, three Invlnh genes were cloned, named as Dolnvlnhl, Dolnvlnh2, and Dolnvlnh3, and they encoded 176, 177, and 177 amino acids, respectively. Bioinformatics analysis of 3 Dolnvlnh proteins showed that there were 1 signal peptide, 2 transmembrane structures, and the conservative domain structure of PMEI protein, Though the homology of Invlnh protein amino acid sequence was very low among different species, there were 4 conservative Cys and SP sites in all of them. The results of quantitative real-time PCR demonstrated that the expressions of all 3 Dolnvlnh genes were detected in the roots, stems, leaves, and flowers. The Dolnvlnh2 and Dolnvlnh3 genes expressed the lowest in roots, while the highest in flowers. The Dolnvlnhl and Dolnvlnh3 presented a similar gene expression pattern in stems at diflerent growth years of 1）. officinale, i.e., the highest expression in the stems of one year, the medium in the stems of two years, and the lowest in stems of three years. The Dolnvlnh2 gene was at the highest expression in the stems of two years. The expressions of Dolnvlnhl and Dolnvlnh3 genes were significantly in negative correlation with CIN activities, speculated that two genes probably played regulatory roles in CIN activities.

关 键 词：铁皮石斛 转化酶抑制子 基因克隆 基因表达 

分 类 号：S567.239[农业科学—中草药栽培]