《植物生理学报》 2021, 57(6): 1189-1198

梭梭HaFT-3基因启动子的克隆及功能分析

满玲娟，任燕萍，王波，何仲鑫，游露，马占龙，张桦^*

新疆农业大学农学院, 乌鲁木齐830052

通信作者：张桦；E-mail: hazelzhang@163.com

摘　要：

14-3-3蛋白是一种可调节性蛋白, 在调控植物生长发育和响应非生物胁迫中发挥了重要作用。使用 染色体步移法克隆梭梭14-3-3蛋白基因HaFT-3上游长度1 340 bp的启动子序列。PlantCARE和PLACE在 线工具预测分析HaFT-3基因启动子, 发现大量CAAT-box、TATA-box等基本顺式作用元件及激素响应元件、 脱水响应元件等多个与非生物胁迫相关的顺式作用元件。构建融合表达载体pCAMBIA3301-HaFT-3::Intron-GUS, 使用浸花法转化拟南芥, 并筛选至T3代。不同生长阶段转基因拟南芥GUS组织化学染色显示,  HaFT-3基因启动子仅在成苗期的叶片和根部诱导下游基因表达。GUS染色和GUS定量分析显示HaFT-3 基因启动子受高温、低温、ABA、IAA诱导。梭梭HaFT-3基因启动子可以响应非生物胁迫, 为进一步研 究HaFT-3基因功能提供参考。

关键词：梭梭; 14-3-3蛋白; 启动子; GUS组织化学染色

资助：国家自然科学基金(32060070)和新疆农业大学作物学重点学科发展基金(XNCDKY2018001)

Cloning and functional analysis of HaFT-3 gene promoter from Haloxylon  ammodendron

MAN Lingjuan, REN Yanping, WANG Bo, HE Zhongxin, YOU Lu, MA Zhanlong, ZHANG Hua* College of Agriculture, Xinjiang Agricultural Univ, HE Zhongxin, YOU Lu, MA Zhanlong, ZHANG Hua^*

College of Agriculture, Xinjiang Agricultural University, Urumqi 830052, China

Corresponding author: ZHANG Hua; E-mail: hazelzhang@163.com

Abstract:

14-3-3 protein is a kind of regulatory protein, which plays an important role in regulating plant  growth and development and responding to abiotic stress. The 1 340 bp promoter sequence of Haloxylon  ammodendron 14-3-3 protein gene HaFT-3 was cloned by chromosome walking method. According to the  prediction analysis of PlantCARE and PLACE, HaFT-3 promoter contains a large number of basic cis-acting  elements such as CAAT-box and TATA-box, furthermore, hormone response elements, dehydration response elements and other cis-acting elements related to abiotic stress are also found. The fusion expression vector pCAMBIA3301-HaFT-3::Intron-GUS was constructed and transformed into Arabidopsis thaliana by  flower soaking method, and then screened to T3 generation. GUS histochemical staining showed that  HaFT-3 promoter only induced downstream gene expression in leaves and roots at seedling stage. GUS  staining and GUS quantitative analysis showed that HaFT-3 promoter was induced by high temperature,  low temperature, ABA and IAA. HaFT-3 promoter of H. ammodendron could respond to abiotic stress,  which provides a reference for further study of HaFT-3 gene function.

Key words: Haloxylon ammodendron; 14-3-3 protein; promoter; GUS histochemical staining