橡胶草Tk-bZIP11转录因子基因的克隆及其表达模式分析  

Cloning and Expression Pattern Analysis of Tk-bZIP11 Transcription Factor from Taraxacum kok-saghyz Rodin

作　　者：李闯 王肖肖 杨玉双[2] 陈秋惠 聂秋海 王萌[4] 刘实忠[2] 覃碧[2] Li Chuang;Wang Xiaoxiao;Yang Yushuang;Chen Qiuhui;Nie Qiuhai;Wang Meng;Liu Shizhong;Qin Bi(College of Tropical Crops,Hainan University,Haikou,570228;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101;Beijing Linglong Dandelion Technology and Development Ltd.,Beijing,101102;College of Plant Protection,Hainan University,Haikou,570228)

机构地区：[1]海南大学热带作物学院,海口570228 [2]中国热带农业科学院橡胶研究所,海口571101 [3]北京玲珑蒲公英科技发展有限公司,北京101102 [4]海南大学植物保护学院,海口570228

出　　处：《分子植物育种》2024年第5期1444-1453,共10页Molecular Plant Breeding

基　　金：国家自然科学基金面上项目(31970364);北京玲珑蒲公英科技发展有限公司研发课题组(橡胶草种质收集、创新与良种繁育)共同资助。

摘　　要：橡胶草(Taraxacum kok-saghyz)种植过程中经常受干旱等胁迫影响,提高品种的抗逆性具有重要意义。为了研究bZIP转录因子在橡胶草逆境响应中的功能,本研究从橡胶草中克隆了Tk-bZIP11基因,其ORF全长468 bp,编码156个氨基酸。蛋白结构分析结果表明,Tk-bZIP11蛋白具有bZIP转录因子特有的保守结构域,同时具有核定位信号。在进化关系上,Tk-bZIP11与莴苣(Lactuca sativa)Ls-bZIP11亲缘关系最近,并具有相同的保守基序。亚细胞定位结果表明,Tk-bZIP11定位在细胞核中。实时荧光定量PCR(RT-qPCR)分析结果表明,Tk-bZIP11在橡胶草的不同组织均有表达,其在叶片中表达量最高,是花梗中表达量的3.28倍。在经甘露醇诱导的渗透压胁迫下,Tk-bZIP11在处理后呈现显著下调表达。在NaCl和PEG-4000模拟的胁迫处理下,Tk-bZIP11在处理后12 h内均呈现上调表达,24 h后呈现下调的规律。在植物激素茉莉酸甲酯、乙烯利和脱落酸处理下,Tk-bZIP11表达量也是在处理的前12 h显著上调表达,之后下调表达。以上研究结果表明Tk-bZIP11参与橡胶草对逆境胁迫的响应和激素信号传导过程。本研究为解析橡胶草bZIP转录因子逆境应答机制,创制抗逆新种质提供科学依据和理论基础。Taraxacum kok-saghyz is often affected by drought and other stresses during the planting process,and improving the stresses resistance of the variety is of great significance.To investigate the functions of bZIP transcription factors in response to stress conditions,the Tk-bZIP11 gene was cloned from Taraxacum kok-saghyz.The ORF of Tk-bZIP11 is 468 bp in length and encodes 156 amino acids.Protein structural analysis showed that TkbZIP11 protein contained a conserved domain specific to the bZIP transcription factors,and a nuclear localization signal.In terms of evolutionary relationship,Tk-bZIP11 showed the highest similarity to Lactuca sativa Ls-bZIP11,and they contained the same conserved motifs.Subcellular localization indicated that Tk-bZIP11 was localized to the nucleus.The results of RT-qPCR analysis showed that Tk-bZIP11 was expressed in different tissues of Taraxacum kok-saghyz,with the highest expression in the leaves,which was 3.28 times higher than those in the stalks.Under osmotic stress induced by mannitol,Tk-bZIP11 expression showed a significant down-regulated after treatment.Under stress conditions treated by NaCl and PEG-4000,Tk-bZIP11 expression was obviously up-regulated within the 12 h of treatments,but down-regulated after 24 h.Analogously,after application of phytohormones(i.e.,methyl jasmonate,ethephon and abscisic acid),Tk-bZIP11 expression was also significantly up-regulated within the 12 h of treatments,and then down-regulated.The above findings suggestted that Tk-bZIP11 was involved in the response to stress and hormonal signaling processes in Taraxacum kok-saghyz.This study provided a scientific basis and theoretical foundation for studying the mechanism of bZIP transcription factors in response to stress,and creating new germplasm resistant to stress in Taraxacum kok-saghyz.

关 键 词：橡胶草 Tk-bZIP11 逆境胁迫 表达分析 亚细胞定位 

分 类 号：S576[农业科学—作物学]