DOI:10.3969/j.issn.1003-5060.2025.02.015
拟南芥 AtCHiC 基因启动子表达载体的构建及鉴定
于鑫,宋慧,胡敏,陈逸凡,叶敏,江力
(合肥工业大学食品与生物工程学院,安徽合肥230601)
摘要
土壤重金属镉污染造成作物产量和品质的下降,严重威胁人类健康。AtCHiC基因编码拟南芥GH-18家族中的一种几丁质酶是否参与植物镉胁迫响应的调控尚不清楚。文章利用聚合酶链式反应(polymerase chain reaction,PCR)技术克隆AtCHiC启动子区域,构建ProAtCHiC-GUS融合表达载体,并通过浸花法将该融合表达载体转化到野生型拟南芥中,鉴定出纯合阳性转基因植株。进一步对转基因植株进行重金属镉胁迫处理,发现其显著诱导植株GUS活性的提高,表明AtCHiC基因可能参与拟南芥镉胁迫响应的调控。该研究为进一步研究非生物胁迫下AtCHiC基因功能提供重要依据。
关键词
拟南芥;AtCHiC基因;镉胁迫;载体构建;GUS染色
中图分类号:Q943.2
文献标志码:A
文章编号:1003-5060(2025)02-0241-04
Construction and identification of promoter expression vectors for Arabidopsis AtCHiC gene
YU Xin, SONG Hui, HU Min, CHEN Yifan, YE Min, JIANG Li
(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China)
Abstract
Heavy metal cadmium pollution in soil leads to the decline of crop yield and quality, which seriously threatens human health. AtCHiC gene encodes a chitinase in GH-18 family of Arabidopsis thaliana, and it is unclear whether it participates in the regulation of cadmium stress response in plants. In this study, the promoter region of AtCHiC was cloned by polymerase chain reaction (PCR), and the ProAtCHiC-GUS fusion expression vector was constructed. The fusion expression vector was transformed into wild-type Arabidopsis thaliana by floral dip method, and homozygous positive transgenic plants were identified. Further, the transgenic plants were treated with heavy metal cadmium stress, and it was found that it significantly induced the increase of GUS activity in plants, which indicated that AtCHiC gene might be involved in the regulation of cadmium stress response in Arabidopsis thaliana. This study provides an important basis for further studying the function of AtCHiC gene under abiotic stress.
Keywords
Arabidopsis thaliana; AtCHiC gene; cadmium stress; vector construction; GUS staining
收稿日期:2022-05-31
修回日期:2022-06-06
基金项目:安徽省自然科学基金资助项目(1508085QC50)