烟草DELLA基因家族的克隆与表达模式分析  

Cloning and expression pattern analyses of tobacco DELLA gene family

作　　者：蔡健宇 余婧 张景云[3] 李翔宇[4] 贾蒙骜 尹国英 叶定勇 薛晓兵 张盼 邹颉 郭玉双 CAI Jianyu;YU Jing;ZHANG Jingyun;LI Xiangyu;JIA Mengao;YIN Guoying;YE Dingyong;XUE Xiaobing;ZHANG Pan;ZOU Jie;GUO Yushuang(Key Laboratory of Molecular Genetics of CNTC,Guizhou Academy of Tobacco Science,Guiyang 550081,China;Ninghai County Agriculture&Forestry Bureau of Ningbo City of Zhejiang Province,Ningbo 315600,Zhejiang,China;Institute of Vegetables and Flowers,Jiangxi Academy of Agriculture Sciences,Nanchang 330200,China;Institute of Crop Resources Research,Heilongjiang Academy of Agricultural Sciences,Harbin 150030,China;School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550030,China)

机构地区：[1]贵州省烟草科学研究院烟草行业分子遗传重点实验室,贵阳市550081 [2]浙江省宁波市宁海县农业农村局,浙江省宁波市315600 [3]江西省农业科学院蔬菜花卉研究所,南昌市330200 [4]黑龙江省农业科学院作物资源研究所,哈尔滨市150030 [5]贵州中医药大学药学院,贵阳市550030

出　　处：《烟草科技》2022年第10期10-18,共9页Tobacco Science & Technology

基　　金：国家烟草专卖局基因组计划重大专项“以uORF为靶标的基因超量表达编辑体系的建立及应用”[110202001030(JY-13)];贵州省烟草科学研究院科技项目“贵州省烤烟种子休眠萌发体系的构建与应用”(GZYKY2020-02);贵州省科技厅自然科学项目“黔产金钗石斛DELLA基因的克隆与功能鉴定”(黔科合基础[2019]1035);中国烟草总公司贵州省公司科技项目“贵州烤烟品种区域化布局及配套技术集成应用”(2020XM07)、“烟草白粉病和PVY抗性基因聚合及育种材料创制”(201903)。

摘　　要：为明确烟草受逆境胁迫时DELLA基因家族的转录调节功能,通过烟草EST序列比对分析克隆了3个DELLA基因家族基因(NtDELLA1,NtDELLA2,NtDELLA3),并对3个基因的结构和表达模式进行了分析。生物信息学分析结果表明,烟草DELLA基因全长1700 bp左右,其编码蛋白包含DELLA蛋白的特征结构域DELLA基序和VHYNP基序,与拟南芥DELLA蛋白家族的GAI蛋白高度同源,推测烟草DELLA蛋白与拟南芥DELLA蛋白具有类似的功能;荧光定量PCR分析结果表明,这3个基因在烟草不同组织和关键生长发育时期表达模式类似,尤以茎中表达量最高;胁迫应答分析结果表明,烟草DELLA基因的转录对干旱、低温、烟草花叶病毒(Tobacco mosaic virus,TMV)侵染和外源激素均表现出不同程度的响应,其中干旱、赤霉素(GA)和脱落酸(ABA)可抑制烟草DELLA基因的表达,而低温胁迫可诱导NtDELLA1上调表达,TMV侵染可诱导NtDELLA1和NtDELLA2的大量表达。In order to clarify the transcription regulation function of DELLA gene family in tobacco under adversity stress,three members(NtDELLA1,NtDELLA2,NtDELLA3)of DELLA gene family were cloned by tobacco EST sequence alignment,and their gene structures and expression patterns were analyzed.The results of bioinformatics analysis showed that the full length of tobacco DELLA genes were around 1700 bp,and the proteins encoded by those genes contained DELLA protein feature structures,DELLA motif and VHYNP motif,and were highly homologous to GAI protein of Arabidopsis DELLA protein family.Thus,it was assumed that the functions of tobacco DELLA proteins were similar to those of Arabidopsis.qRT-PCR analysis further confirmed that the three genes had similar expression patterns in different tissues at key development stages,and their expression levels in tobacco stalk were the highest.The results of analysis on stress responses indicated that tobacco responded to drought,low temperature,TMV infection and exogenous hormones at varying degrees,and drought,GA and ABA inhibited the expression of tobacco DELLA gene family,while low temperature induced up-regulated expression of NtDELLA1,TMV induced the abundant expression of NtDELLA1 and NtDELLA2.

关 键 词：烟草 DELLA蛋白 生物信息学 胁迫应答 

分 类 号：TS413[农业科学—烟草工业] Q943.2[生物学—植物学]