茎瘤芥BjuEAR1-1的克隆与表达分析  

Cloning and Expression Analysis of BjuEAR1-1 in Tuber Mustard

作　　者：程春红[1] 游经番 蔡兆明[1] 叶春宏 陈丽 CHENG Chunhong;YOU Jingfan;CAI Zhaoming;YE Chunhong;CHEN Li(College of Life Science and Technology,Yangtze Normal University,Chongqing 408100,China)

机构地区：[1]长江师范学院生命科学与技术学院,重庆408100

出　　处：《华北农学报》2022年第3期37-43,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31701451);重庆市科技局项目(cstc2020jcyj-msxmX0770);重庆市教育委员会项目(KJQN201901438,KJQN202101432);武陵山区特色植物资源保护与利用实验室开放课题(TEZWKFKT202002);重庆市高校创新研究群体项目(CXQT21029)。

摘　　要：茎瘤芥在生长发育过程中会受到非生物逆境胁迫的影响,导致产量降低。ABA作为植物逆境激素,在调控植株响应非生物逆境胁迫过程中具有重要作用。BjuEAR1-1是ABA信号通路中负调控因子EAR1的同源基因,其突变体ear1表现出对ABA超敏感的表型。因此,探究BjuEAR1-1的生物学功能,可为茎瘤芥抵抗不良环境的调控机制提供方向,也为有效利用基因资源培育耐逆稳产的茎瘤芥新品种提供重要的理论依据。以永安小叶为试验材料,采用PCR方法克隆BjuEAR1-1基因编码序列,采用酶切酶连方法构建植物过表达载体35S∷PTF101-GFP-BjuEAR1-1,采用烟草叶片瞬时转化技术对BjuEAR1-1的亚细胞定位情况进行分析,并对BjuEAR1-1基因启动子顺式元件进行分析,最后采用荧光定量PCR方法,检测BjuEAR1-1在不同非生物逆境胁迫处理下及不同组织器官的基因表达模式。烟草亚细胞定位结果表明,BjuEAR1-1定位于细胞核和细胞质中;BjuEAR1-1在茎瘤芥的根、茎、叶、花、种荚和膨大茎中均有表达,在根中表达量最高;BjuEAR1-1的表达水平显著受低温和ABA诱导,盐胁迫处理下BjuEAR1-1的表达水平与对照组没有显著性差异。BjuEAR1-1在细胞核和细胞质中发挥生物学功能,并且BjuEAR1-1调控茎瘤芥对于低温和ABA的响应。During the growth and development stages,tuber mustard is always affected by abiotic stresses.As a plant stress hormone,ABA plays an important role in regulating plant response to abiotic stress.BjuEAR1-1 was a homologue of negative regulator EAR1 in ABA signaling pathway,and its mutant ear1 exhibited an ABA sensitive phenotype.Therefore,to explore the biological function of BjuEAR1-1 will provide a direction for the regulation mechanism of resistance to adverse environment stresses,and also provide an important theoretical basis for the effective use of gene resources to cultivate new varieties of BjuEAR1-1 with resistance to adverse stresses and stable yield.The BjuEAR1-1 coding sequence was cloned by PCR method and the plant overexpression vector 35S∷PTF101-GFP-BjuEAR1-1 was constructed by enzyme digestion and ligation method.The subcellular localization of BjuEAR1-1 was analyzed by transient transformation technology of tobacco leaves.The cis-element of BjuEAR1-1 gene promoter was analyzed,and the gene expression patterns of BjuEAR1-1 under different abiotic stresses and different tissues and organs were detected by fluorescence quantitative PCR.The result of subcellular localization assay showed that BjuEAR1-1 was localized in the nucleus and cytoplasm of tobacco.BjuEAR1-1 was expressed in root,stem,leaf,flower,seed pod and swollen stem of tuber mustard,with the highest expression in root.The expression level of BjuEAR1-1 was significantly induced under low temperature and ABA stresses,and there was no significant difference under salt stress.BjuEAR1-1 was localized in the nucleus and cytoplasm and regulated the response of tuber mustard to low temperature and ABA stresses.

关 键 词：茎瘤芥 非生物逆境胁迫 BjuEAR1-1 基因克隆 亚细胞定位 基因表达 

分 类 号：S637.3[农业科学—蔬菜学]