本文主要研究内容
作者任亚娟（2019）在《晚疫病菌RxLR效应子PITG22926靶定StMAP3Kβ2调控马铃薯免疫应答的机制研究》一文中研究指出：马铃薯在生产中遭受各种病害的侵袭,其中由卵菌致病疫霉（Phytophthora infestans）引起的晚疫病是最具毁灭性的病害。为了抵御病原菌的侵袭,植物进化出一套复杂天然的免疫系统来激活自身的防卫反应,包括病原相关分子模式激发的免疫反应（PTI）和效应蛋白激发的免疫反应（ETI）。晚疫病菌分泌效应子进入寄主细胞并通过与靶标互作来抑制植物的免疫反应,促进自身的侵染。前期,英国研究团队通过酵母双杂筛选出70多个晚疫病菌RxLR效应子的潜在靶蛋白。其中效应子Pi22926在马铃薯中筛选到的互作蛋白为丝裂原活化蛋白激酶MAP3Kβ2。本研究利用农杆菌介导的遗传转化、瞬时表达、酵母双杂交、免疫共沉淀、蛋白亚细胞定位及病毒介导的基因沉默技术系统研究了Pi22926靶向StMAP3Kβ2调控马铃薯免疫应答的机制。主要研究结果如下:1.效应子Pi22926基因在晚疫病菌侵染马铃薯早期持续上调表达;在本氏烟上瞬时表达Pi22926或在本氏烟和马铃薯中稳定表达Pi22926能促进晚疫病菌侵染;瞬时表达Pi22926特异抑制抗病基因Cf4与其相应无毒基因Avr4及抗性基因Pto与其无毒基因Avrpto介导的细胞死亡信号转导途径。表明该效应子具有明显的毒性功能。2.酵母点对点杂交及免疫共沉淀证明Pi22926与马铃薯丝裂原活化蛋白激酶StMAP3Kβ2及其激酶结构域StMAP3Kβ2（KD）互作,但不能与其激酶结构域ATP结合位点突变体StMAP3Kβ2（KD）Lys430Arg互作,表明StMAP3Kβ2完整的激酶结构域是互作所必需的,预示Pi22926特异靶向StMAP3Kβ2激酶结构域关键位点来抑制其激酶活性。另外,Pi22926不能与另外一个马铃薯丝裂原活化蛋白激酶StMAP3Kε及其激酶结构域互作,表明Pi22926与StMAP3Kβ2互作具有特异性。3.蛋白亚细胞定位表明GFP-Pi22926定位在细胞核中;StMAP3Kβ2-GFP主要定位在细胞质中,少量在细胞核质中;双分子荧光互补实验表明YN-Pi22926与YC-StMAP3Kβ2互作主要位于核质中,表明Pi22926对StMAP3Kβ2的作用主要发生在细胞核中。4.本氏烟中瞬时超量表达StMAP3Kβ2能够诱发叶片注射点细胞死亡;沉默MAP3Kβ2促进晚疫病菌侵染,然而瞬时超量表达MAP3Kβ2抑制病原菌侵染,提高晚疫病抗性。表明MAP3Kβ2正调控植物免疫应答反应。5.Pi22926能够特异抑制MAP3Kβ2在本氏烟上激发的细胞死亡,但不能抑制StMAP3Kε诱导的细胞死亡,而PexRD2能够特异抑制StMAP3Kε在本氏烟上激发的细胞死亡,但不能抑制MAP3Kβ2诱导的细胞死亡,表明Pi22926特异抑制MAP3Kβ2介导的信号途径。6.本氏烟中沉默MAP3Kβ2或MAP3Kε能够部分抑制Avr4/Cf4及Avrpto/Pto介导的细胞死亡,表明MAP3Kβ2和MAP3Kε共同参与Avr4/Cf4及Avrpto/Pto介导的细胞死亡信号途径;沉默MAP3Kβ2不影响StMAP3Kε诱发的细胞死亡,同时沉默MAP3Kε也不能抑制StMAP3Kβ2诱发的细胞死亡,表明StMAP3Kβ2和StMAP3Kε是共同位于Avr4/Cf4信号传导下游平行起作用的两个激酶;沉默丝裂原蛋白激酶信号通路中MEK2和SIPK后,StMAP3Kβ2介导的细胞死亡明显受到抑制,表明StMAP3Kβ2位于MEK2及SIPK上游。丝裂原蛋白激酶信号通路在调控植物免疫应答中发挥关键作用。本研究和前人研究结果表明,晚疫病菌效应子Pi22926和PexRD2分别特异靶向Avr4/Cf4下游平行起作用的两个激酶来抑制免疫应答。本研究揭示了两个效应子如何通过协同作用来有效抑制Avr4/Cf4（及Avrpto/Pto）介导的丝裂原蛋白激酶信号通路、进而操控寄主抗性免疫应答的一种机制。
Abstract
ma ling shu zai sheng chan zhong zao shou ge chong bing hai de qin xi ,ji zhong you luan jun zhi bing yi mei （Phytophthora infestans）yin qi de wan yi bing shi zui ju hui mie xing de bing hai 。wei le di yu bing yuan jun de qin xi ,zhi wu jin hua chu yi tao fu za tian ran de mian yi ji tong lai ji huo zi shen de fang wei fan ying ,bao gua bing yuan xiang guan fen zi mo shi ji fa de mian yi fan ying （PTI）he xiao ying dan bai ji fa de mian yi fan ying （ETI）。wan yi bing jun fen bi xiao ying zi jin ru ji zhu xi bao bing tong guo yu ba biao hu zuo lai yi zhi zhi wu de mian yi fan ying ,cu jin zi shen de qin ran 。qian ji ,ying guo yan jiu tuan dui tong guo jiao mu shuang za shai shua chu 70duo ge wan yi bing jun RxLRxiao ying zi de qian zai ba dan bai 。ji zhong xiao ying zi Pi22926zai ma ling shu zhong shai shua dao de hu zuo dan bai wei si lie yuan huo hua dan bai ji mei MAP3Kβ2。ben yan jiu li yong nong gan jun jie dao de wei chuan zhuai hua 、shun shi biao da 、jiao mu shuang za jiao 、mian yi gong chen dian 、dan bai ya xi bao ding wei ji bing du jie dao de ji yin chen mo ji shu ji tong yan jiu le Pi22926ba xiang StMAP3Kβ2diao kong ma ling shu mian yi ying da de ji zhi 。zhu yao yan jiu jie guo ru xia :1.xiao ying zi Pi22926ji yin zai wan yi bing jun qin ran ma ling shu zao ji chi xu shang diao biao da ;zai ben shi yan shang shun shi biao da Pi22926huo zai ben shi yan he ma ling shu zhong wen ding biao da Pi22926neng cu jin wan yi bing jun qin ran ;shun shi biao da Pi22926te yi yi zhi kang bing ji yin Cf4yu ji xiang ying mo du ji yin Avr4ji kang xing ji yin Ptoyu ji mo du ji yin Avrptojie dao de xi bao si wang xin hao zhuai dao tu jing 。biao ming gai xiao ying zi ju you ming xian de du xing gong neng 。2.jiao mu dian dui dian za jiao ji mian yi gong chen dian zheng ming Pi22926yu ma ling shu si lie yuan huo hua dan bai ji mei StMAP3Kβ2ji ji ji mei jie gou yu StMAP3Kβ2（KD）hu zuo ,dan bu neng yu ji ji mei jie gou yu ATPjie ge wei dian tu bian ti StMAP3Kβ2（KD）Lys430Arghu zuo ,biao ming StMAP3Kβ2wan zheng de ji mei jie gou yu shi hu zuo suo bi xu de ,yu shi Pi22926te yi ba xiang StMAP3Kβ2ji mei jie gou yu guan jian wei dian lai yi zhi ji ji mei huo xing 。ling wai ,Pi22926bu neng yu ling wai yi ge ma ling shu si lie yuan huo hua dan bai ji mei StMAP3Kεji ji ji mei jie gou yu hu zuo ,biao ming Pi22926yu StMAP3Kβ2hu zuo ju you te yi xing 。3.dan bai ya xi bao ding wei biao ming GFP-Pi22926ding wei zai xi bao he zhong ;StMAP3Kβ2-GFPzhu yao ding wei zai xi bao zhi zhong ,shao liang zai xi bao he zhi zhong ;shuang fen zi ying guang hu bu shi yan biao ming YN-Pi22926yu YC-StMAP3Kβ2hu zuo zhu yao wei yu he zhi zhong ,biao ming Pi22926dui StMAP3Kβ2de zuo yong zhu yao fa sheng zai xi bao he zhong 。4.ben shi yan zhong shun shi chao liang biao da StMAP3Kβ2neng gou you fa xie pian zhu she dian xi bao si wang ;chen mo MAP3Kβ2cu jin wan yi bing jun qin ran ,ran er shun shi chao liang biao da MAP3Kβ2yi zhi bing yuan jun qin ran ,di gao wan yi bing kang xing 。biao ming MAP3Kβ2zheng diao kong zhi wu mian yi ying da fan ying 。5.Pi22926neng gou te yi yi zhi MAP3Kβ2zai ben shi yan shang ji fa de xi bao si wang ,dan bu neng yi zhi StMAP3Kεyou dao de xi bao si wang ,er PexRD2neng gou te yi yi zhi StMAP3Kεzai ben shi yan shang ji fa de xi bao si wang ,dan bu neng yi zhi MAP3Kβ2you dao de xi bao si wang ,biao ming Pi22926te yi yi zhi MAP3Kβ2jie dao de xin hao tu jing 。6.ben shi yan zhong chen mo MAP3Kβ2huo MAP3Kεneng gou bu fen yi zhi Avr4/Cf4ji Avrpto/Ptojie dao de xi bao si wang ,biao ming MAP3Kβ2he MAP3Kεgong tong can yu Avr4/Cf4ji Avrpto/Ptojie dao de xi bao si wang xin hao tu jing ;chen mo MAP3Kβ2bu ying xiang StMAP3Kεyou fa de xi bao si wang ,tong shi chen mo MAP3Kεye bu neng yi zhi StMAP3Kβ2you fa de xi bao si wang ,biao ming StMAP3Kβ2he StMAP3Kεshi gong tong wei yu Avr4/Cf4xin hao chuan dao xia you ping hang qi zuo yong de liang ge ji mei ;chen mo si lie yuan dan bai ji mei xin hao tong lu zhong MEK2he SIPKhou ,StMAP3Kβ2jie dao de xi bao si wang ming xian shou dao yi zhi ,biao ming StMAP3Kβ2wei yu MEK2ji SIPKshang you 。si lie yuan dan bai ji mei xin hao tong lu zai diao kong zhi wu mian yi ying da zhong fa hui guan jian zuo yong 。ben yan jiu he qian ren yan jiu jie guo biao ming ,wan yi bing jun xiao ying zi Pi22926he PexRD2fen bie te yi ba xiang Avr4/Cf4xia you ping hang qi zuo yong de liang ge ji mei lai yi zhi mian yi ying da 。ben yan jiu jie shi le liang ge xiao ying zi ru he tong guo xie tong zuo yong lai you xiao yi zhi Avr4/Cf4（ji Avrpto/Pto）jie dao de si lie yuan dan bai ji mei xin hao tong lu 、jin er cao kong ji zhu kang xing mian yi ying da de yi chong ji zhi 。
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论文作者分别是来自华中农业大学的任亚娟，发表于刊物华中农业大学2019-07-15论文，是一篇关于马铃薯论文,晚疫病论文,效应子论文,细胞死亡论文,丝裂原活化蛋白激酶论文,抗病免疫应答论文,华中农业大学2019-07-15论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自华中农业大学2019-07-15论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：马铃薯论文; 晚疫病论文; 效应子论文; 细胞死亡论文; 丝裂原活化蛋白激酶论文; 抗病免疫应答论文; 华中农业大学2019-07-15论文;