枣E类MADS基因在花和果中的表达及其蛋白互作研究  

Expression of E-type MADS-box Genes in Flower and Fruits and Protein Interaction Analysis in Chinese Jujube

作　　者：高玮林 张力曼 薛超玲 张垚 刘孟军[2] 赵锦[1] GAO Weilin;ZHANG Liman;XUE Chaoling;ZHANG Yao;LIU Mengjun;ZHAO Jin(College of Life Science,Hebei Agricultural University,Baoding,Hebei 071000,China;College of Horticulture,Hebei Agricultural University,Baoding,Hebei 071000,China)

机构地区：[1]河北农业大学生命科学学院,河北保定071000 [2]河北农业大学园艺学院,河北保定071000

出　　处：《园艺学报》2022年第4期739-748,共10页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31772285);国家重点研发计划项目(2018YFD1000607,2019YFD1001605);河北省高校百名优秀创新人才支持计划项目(SLRC2019031)。

摘　　要：以‘金丝小枣’花和果实为试材进行E类MADS-box基因的生物信息、表达、亚细胞定位及蛋白互作研究。3个E-type MADS-box基因分别位于SEP1/2、SEP3和SEP4进化支。与其他物种的同源序列相比,E类蛋白在进化中非常保守。3个E类基因在枣的花和果实中均有表达,ZjSEP1/2在花不同发育时期表达比较稳定,ZjSEP3和ZjSEP4在花发育前期高表达,在感染枣疯病的花变叶中3个基因的表达均显著受抑制。3个蛋白均定位于细胞核。酵母单杂交结果表明ZjSEP1/2和ZjSEP3具有转录激活活性,可以作为转录因子调控下游基因,而ZjSEP4不具有此活性。酵母双杂交结果表明3个E类MADS蛋白都可以和A类、C/D类蛋白发生互作,ZjSEP4与ZjSEP1/2均可与自身互作,但ZjSEP3自身不发生互作。综上,3个E类MADS基因在枣花发育中具有重要作用,但功能和调控模式不同,ZjSEP3不能形成同源二聚体,ZjSEP1/2、ZjSEP4均可形成同源二聚体,3个E类蛋白均可与A、C/D类蛋白形成异源二聚体。In order to study the function of E-type MADS-box genes in flower and fruit of Chinese jujube(Ziziphus jujuba Mill.),their bioinformatics analysis,expression pattern,subcellular location and protein interaction in’Jinsi Xiaozao’were studied systematically. A total of three E-type MADS-box genes were identified in the jujube genome,which are located in the three clades of SEP1/2,SEP3 and SEP4.Compared with homologous sequences of other species,they are very conservative in evolution. Three E-type genes expressed in jujube flowers and fruits. The expression of ZjSEP1/2 was relatively stable at different stages of flower development,ZjSEP3 and ZjSEP4 were highly expressed in the early stage of flower development. After phytoplasma infection,their expressions in phyllody,a typical diseased symptom, were inhibited dramatically. All three proteins were located in the nucleus. According to the results of yeast one-hybrid analysis,ZjSEP1/2 and ZjSEP3 proteins had transcriptional activation activity and could be used as transcription factors to regulate downstream genes,while ZjSEP4 didn’t have this activity.According to the yeast two-hybrid results,all three proteins could interact with A-type and C/D-type proteins. Both ZjSEP4 and ZjSEP1/2 could interact with itself,but ZjSEP3 couldn’t interact with itself. In summary,the three E-type genes play an important role in the development of jujube flowers,but they should have various regulatory modes. ZjSEP3 couldn’t form homodimers,while ZjSEP4 and ZjSEP1/2could form homodimers. But all of three proteins could form heterodimers with A-type and C/D-type proteins.

关 键 词：枣 MADS-BOX E-type 花果发育 表达分析 转录激活活性 蛋白互作 

分 类 号：S665.1[农业科学—果树学]