马铃薯细胞壁蔗糖转化酶基因StCWIN1启动子克隆与表达及其在干旱胁迫下的作用分析
Cloning and Expression Analysis of Potato StCWIN1 Gene Promoter and Its Role under Drought Stress
张玉 1杨文静 1刘璇 1聂峰杰 1张丽 1石磊 1张国辉 2郭志乾 2巩檑1
作者信息
- 1. 宁夏农林科学院农业生物技术研究中心/宁夏农业生物技术重点实验室,750002,宁夏银川
- 2. 宁夏农林科学院固原分院,756000,宁夏固原
- 折叠
摘要
植物细胞壁蔗糖转化酶(cell wall invertase,CWIN)是源、库组织蔗糖代谢及胁迫应答的关键酶.本研究利用基因步移法克隆马铃薯StCWIN1启动子片段,应用PlantCARE在线软件对启动子区域的作用元件进行分析,将融合StCWIN1启动子与GUS报告基因的表达载体转化拟南芥野生型,并利用组织化学染色和GUS实时定量PCR技术探究启动子表达活性、组织表达特性和响应干旱胁迫的表达规律.结果表明,克隆获得StCWIN1基因上游1956 bp启动子序列,其中包含核心调控、植物激素、防御及胁迫、光响应等关键元件;StCWIN1启动子在根、柱头和果英组织中的表达活性高于其他组织;转StCWIN1启动子拟南芥株系叶片中GUS表达量高于野生型,且干旱胁迫显著抑制了 GUS相对表达量.本研究克隆得到具有活性的StCWIN1启动子,基于研究结果推测目的基因可能参与根、花和果实等器官发育,对干旱胁迫也发挥应答调节作用.
Abstract
Cell wall invertase(CWIN)is a key enzyme for stress responses and sucrose metabolism in source and sink tissues.In this study,the promoter sequence of StCWIN1 was cloned by genomic walking,and its cis-regulatory elements were analysed by using PlantCARE.A GUS-fused StCWIN1 promoter expression vector was constructed and subsequently transformed into Arabidopsis thaliana.Promoter activity,tissue expression characteristics and responsiveness to drought stress were investigated by histochemical staining and real-time quantitative PCR.The results showed that the StCWIN1 promoter was 1956 bp long and contained enhancer-regulated,phytohormone-responsive,protective/stress-responsive and light-responsive elements.The expression activity of StCWIN1 promoter were higher in roots,stigmas,and pods than other tissues.The expression of GUS in leaves of Arbidopsis Thaliana with StCWIN1 promoter was higher than that of wild type,and the value was significantly reduced under drought stress.The identification of the active StCWIN1 promoter in the present study suggests that StCWIN1 may be involved in the development of organs such as roots,flowers and fruit pods,and may also function in response to drought stress.
关键词
马铃薯/细胞壁蔗糖转化酶/启动子/GUS活性/干旱胁迫/表达分析Key words
Solanum tuberosum L./Cell wall invertase/Promoter/GUS activity/Drought stress/Expression analysis引用本文复制引用
基金项目
国家自然科学基金(31860419)
国家自然科学基金(32260504)
宁夏回族自治区农业育种专项(2019NYYZ01-2)
宁夏回族自治区自然科学基金(2021AAC05015)
出版年
2024
国家科技期刊平台
NSTL
万方数据