梭梭HaFT-1、HaFT-9基因启动子克隆及胁迫响应分析  

Cloning and Stress Response Analysis of HaFT-1 and HaFT-9 Gene Promoters in Haloxylon ammodendron

作　　者：潘绒 任文静 李新平[1] 张桦[1] 任燕萍[1] 马丽[1] 王波[1] Pan Rong;Ren Wenjing;Li Xinping;Zhang Hua;Ren Yanping;Ma Li;Wang Bo(College of Life Sciences,Xinjiang Agricultural University,Urumqi,830052)

机构地区：[1]新疆农业大学生命科学学院,乌鲁木齐830052

出　　处：《分子植物育种》2023年第22期7372-7385,共14页Molecular Plant Breeding

基　　金：国家自然科学基金地区科学基金项目(31760070);自治区研究生科研创新项目(XJ2020G144);新疆农业大学作物学重点学科发展基金项目(XNCDKY2018003)共同资助。

摘　　要：14-3-3蛋白是一类普遍存在于真核生物中的分子伴侣,研究表明梭梭14-3-3蛋白HaFT-1和HaFT-9功能与胁迫响应密切相关。为进一步分析HaFT-1和HaFT-9基因胁迫响应机制,本研究采用染色体步移技术扩增HaFT-1和HaFT-9基因5’端上游调控序列,分别获得961和879 bp启动子序列。序列分析表明二者启动子除含有基本的启动子核心元件外,还包含多种逆境响应元件。分别构建HaFT-1和HaFT-9基因启动子-GUS重组载体瞬时转化烟草、棉花、梭梭幼苗,GUS染色结果显示:二者启动子在正常生长条件或高温、脱水、NaCl胁迫及ABA处理下均能调控GUS表达,并且HaFT-1启动子对以上处理的响应更加明显;在高温胁迫下,梭梭幼苗中二者启动子调控的GUS表达具有组织特异性。RT-qPCR结果显示,在梭梭幼苗中,高温、脱水、NaCl胁迫及ABA处理下HaFT-1和HaFT-9基因表达趋势与以上GUS染色结果基本一致。本研究为进一步阐明HaFT-1和HaFT-9基因在胁迫响应中的作用机制提供了科学依据。14-3-3 proteins are ubiquitous molecular chaperones in eukaryotes.Studies have shown that functions as 14-3-3 proteins HaFT-1 and HaFT-9 are closely related to stress response.In order to further analyse stress response mechanism of HaFT-1 and HaFT-9 genes,the upstream regulatory sequences of 5'ends of HaFT-1 and HaFT-9 genes were amplified with chromosome walking technique,and 961 and 879 bp promoter sequences were obtained,respectively.Sequence analysis showed that both promoters contained not only basic promoter core elements,but also a variety of stress response elements.The recombinant vectors of HaFT-1 or HaFT-9 promoter-GUS were constructed and transiently transformed into tobacco,cotton and Haloxylon ammodendron seedlings.GUS staining results showed:the two promoters could regulate GUS expression under normal growth conditions or heat,dehydration,NaCl and ABA treatments,and the response of HaFT-1 promoter to above treatments was more obvious;under high temperature stress,the two promoters could regulate GUS expression in H.ammodendron seedlings in which the expression of GUS was tissue-specific.RT-qPCR showed that the expression trends of HaFT-1 and HaFT-9 genes in H.ammodendron seedlings under high temperature,dehydration,NaCl stress and ABA treatment were broadly consistent with GUS staining results.This study will provide a scientific basis of further elucidating the mechanism of HaFT-1 and HaFT-9 genes in response to stress.

关 键 词：梭梭 HaFT-1基因 HaFT-9基因 启动子 瞬时转化 

分 类 号：S793.9[农业科学—林木遗传育种]