棉花GhAPX2基因启动子克隆及其干旱胁迫响应  

Cloning of Cotton GhAPX2 Gene Promoter and Its Response to Drought Stress

作　　者：于月华[1] 祁颢萱 田梦 倪志勇[2] YU Yue-hua;QI Hao-xuan;TIAN Meng;NI Zhi-yong(College of Agronomy,Xinjiang Agricultural University,Urumqi 830052,China;College of Life Sciences,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]新疆农业大学生命科学学院,乌鲁木齐830052

出　　处：《新疆农业大学学报》2022年第6期451-456,共6页Journal of Xinjiang Agricultural University

基　　金：新疆维吾尔自治区重大科技专项项目(2021A02001-3)。

摘　　要：从棉花中克隆了干旱应答基因GhAPX2的3000 bp启动子并将其转化到拟南芥中。采用生物信息学方法分析了GhAPX2启动子的顺式作用元件并对转GhAPX2启动子拟南芥进行干旱胁迫,检测β-葡萄糖苷酸酶(GUS)染色程度,GUS基因表达量和GUS酶活性。Plant CARE在线分析结果表明,3000 bp的GhAPX2启动子中含有2个干旱诱导的MYB转录因子结合位点(MBS)顺式作用元件。为进一步研究GhAPX2启动子在干旱胁迫应答中的功能,用聚乙二醇(PEG)对转GhAPX2启动子拟南芥GhAPX2p∶GUS进行模拟干旱胁迫,随后进行GUS组织化学活性染色,GUS基因表达和GUS酶活性测定。结果表明,相比未处理GhAPX2p∶GUS拟南芥叶片,在PEG胁迫下GhAPX2p∶GUS拟南芥叶片GUS染色程度,GUS基因表达量和GUS酶活性均增加,这些结果表明棉花GhAPX2基因启动子是一个干旱胁迫诱导型启动子。The 3000 bp promoter of the drought response gene GhAPX2 was cloned from cotton and transformed into Arabidopsis and then the cis-acting element of GhAPX2 promoter was analyzed by bioinformatics method and the Arabidopsis transgenic with GhAPX2 promoter was subjected to drought stress,and the degree ofβ-glucosidase(GUS)staining,GUS gene expression and GUS enzyme activity were detected.Plant CARE online analysis showed that the 3000 bp GhAPX2 promoter contained two drought-induced MYB transcription factor binding site(MBS)cis-acting elements.To further study the function of the GhAPX2 promoter in response to drought stress,GhAPX2-promoter-transfected Arabidopsis GhAPX2p∶GUS was subjected to simulated drought stress with polyethylene glycol(PEG),followed by GUS histochemical activity staining,GUS gene expression and GUS enzymatic activity assays.The results showed that compared with untreated GhAPX2p∶GUS Arabidopsis leaves,GhAPX2p∶GUS Arabidopsis leaves under PEG stress increased the degree of GUS staining,GUS gene expression and GUS enzyme activity.These findings indicate that the cotton GhAPX2 gene promoter was a drought stress inducible promoter.

关 键 词：棉花 抗坏血酸过氧化物酶 启动子 GUS报告基因 

分 类 号：Q781[生物学—分子生物学]