《植物生理学报》 2018, 54(2): 247-254

苦荞FtTCPlike1和FtTCPlike2的克隆及表达分析

张嘉丽^1,2, 黄铭坤², 孙伟², 陈庆富³, 刘霞^1,2,*

¹武汉理工大学化学化工与生命科学学院, 武汉430070; ²中国中医科学院中药研究所, 北京100700; ³贵州师范大学荞麦产业技术研究中心, 贵阳550001

通信作者：刘霞；E-mail: lxr1125@126.com

摘　要：

植物TCP转录因子家族是一类调控植物生长、发育的重要转录因子, 主要分为class I和class II两种类型。在本研究中, 我们克隆了苦荞(Fagopyrum tataricum)两个TCP转录因子基因并分别命名为FtTCPlike1和FtTCPlike2。蛋白聚类分析表明FtTCPlike1属于class II而FtTCPlike2属于class I, 并且均含有TCP转录因子典型的保守结构域。亚细胞定位实验也表明FtTCPlike1和FtTCPlike2与报道的其他物种TCP转录因子类似, 都定位在细胞核中。本文同时收集了苦荞芽期的子叶和下胚轴、第1~3片长出的真叶以及5个时期由幼嫩到成熟的种子和花共11种样本进行了实时荧光定量PCR, 发现FtTCPlike1和FtTCPlike2在苦荞的各个不同时期和不同组织中均有不同的表达, 并且在种子的某个特定时期(FtTCPlike1和FtTCPlike2分别在种子发育第3、第4时期和第1、第3时期)有较高的表达量, 表明这两个基因可能参与种子发育。从苦荞芽期来看, FtTCPlike1在子叶中表达量较高但在下胚轴表达较低, 而FtTCPlike2则相反, 这说明FtTCPlike1和FtTCPlike2在苦荞早期形态建成中的调控方式可能有很大区别。

关键词：苦荞; TCP; 基因克隆; 亚细胞定位; 基因表达

收稿：2017-09-18   修定：2018-01-25

资助：国家自然科学基金(81403046和31471562)。

Molecular cloning and analyzing of FtTCPlike1 and FtTCPlike2 in Fagopyrum tataricum

ZHANG Jia-Li^1,2, HUANG Ming-Kun², SUN Wei², CHEN Qing-Fu³, LIU Xia^1,2,*

¹School of Chemistry, Chemical Engineering and Life Sciences, Wuhan University of Technology, Wuhan 430070, China; ²Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; ³Research Center of Buckwheat Industry Technology, Guizhou Normal University, Guiyang 550001, China

Corresponding author: LIU Xia; E-mail: lxr1125@126.com

Abstract:

The TCP transcription factor family is largely involved in regulating plant growth and development. Based on the similarity of protein, TCP family can be divided into two classes, class I and class II. In this study, we isolated two TCP genes from Fagopyrum tataricum, named FtTCPlike1 and FtTCPlike2. Both FtTCPlike1 and FtTCPlike2 contained the reported conserved TCP-domain. Furthermore, phylogenetic tree based on the protein sequences showed that FtTCPlike1 belonged to class II, while FtTCPlike2 clustered into class I. FtTCPlike1 and FtTCPlike2 were found to be sub-located in the nucleolus similar to the previous reported. Next, we collected cotyledons, hypocotyls, three types of true leaves at different developmental stages, flowers and seeds during five stages of development, isolated the RNA and conducted quantitative real-time PCR to analyze the expression of FtTCPlike1 and FtTCPlike2 in these tissues. The results showed that FtTCPlike1 and FtTCPlike2 were expressed in all the tissues analyzed. In addition, FtTCPlike1 and FtTCPlike2 were expressed highly in specific stages (e.g. FtTCPlike1 in stages 3 and 4; FtTCPlike2 in stages 1 and 3) during seed development, indicating that the two genes might play various roles in seed development. Interestingly, FtTCPlike1 was expressed highly in cotyledons but not in hypocotyls, while the expression of FtTCPlike2 was opposite compared to FtTCPlike1, which indicated the distinct role in early F. tataricum morphogenesis.

Key words: Fagopyrum tataricum; TCP; gene cloning; subcellular localization; gene expression