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Arabidopsis E3 ligase KEG associates with and ubiquitinates mkk4 and mkk5 to regulate plant immunity

Journal of Integrative Plant Biology 2021年第2期63卷 327-339页

作者： Chenyang Gao Pengwei Sun Wei Wang Dingzhong Tang State Key Laboratory of Ecological Control of Fujian-Taiwan Crop Pests Key Laboratory of Ministry of Education for GeneticsBreeding and Multiple Utilization of CropsPlant Immunity CenterFujian Agriculture and Forestry UniversityFuzhou 350002China Institute of Genetics and Developmental Biology the Chinese Academy of SciencesBeijing 100101China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China

Mitogen-activated protein kinase(MAPK) cascades are highly conserved signaling modules that regulate plant immune responses. The Arabidopsis thaliana Raf-like MAPK kinase kinase ENHANCED DISEASE RESISTANCE1(EDR1) is a key negative regulator of plant immunity that affects the protein levels of mkk4 and mkk5, two important MAPK cascade members, but the underlying mechanism is poorly understood. Here, genome-wide phosphorylation analysis demonstrated that the E3 ligase KEEP ON GOING(KEG) is phosphorylated in the edr1 mutant but not the wild type, suggesting that EDR1 negatively affects KEG phosphorylation. The identified phosphorylation sites in KEG appear to be important for its accumulation. The keg-4 mutant, a previously identified edr1 suppressor, enhances susceptibility to the powdery mildew pathogen Golovinomyces cichoracearum. In addition, mkk4 and mkk5 protein levels are reduced in the keg-4 mutant. Furthermore,we demonstrate that mkk4 and mkk5 associate with full-length KEG, but not with truncated KEG-RK or KEG-RKA, and that KEG ubiquitinates and mediates the degradation of mkk4 and mkk5. Taken together, these results indicate that mkk4 and mkk5 protein levels are regulated by KEG via ubiquitination, uncovering a mechanism by which plants finetune immune responses by regulating the homeostasis of key MAPK cascade members via ubiquitination and degradation.

关键词： MAPK cascade plant immunity KEG EDR1 mkk4 mkk5 Arabidopsis powdery mildew

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SERK Receptor-like Kinases Control Division Patterns of Vascular Precursors and Ground Tissue Stem Cells during Embryo Development in Arabidopsis

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Molecular Plant 2019年第7期12卷 984-1002页

作者： Huiqiang Li Zeping Cai Xiaojuan Wang Meizhen Li Yanwei Cui Nan Cui Fei Yang Mingsong Zhu Junxiang Zhao Wenbin Du Kai He Jing Yi Frans E.Tax Suiwen Hou Jia Li Xiaoping Gou Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations School of Life Sciences Lanzhou University Lanzhou Gansu 730000 China College of Forestry Hainan University Danzhou Hainan 571737 China Department of Molecular and Cellular Biology University of Arizona Tucson AZ 85721 USA

During embryo development, the vascular precursors and ground tissue stem cells divide to renew them-selves and produce the vascular tissue, endodermal cells, and cortical cells. However, the molecular mech-anisms regulating division of these stem cells have remained largely elusive. In this study, we show that loss of function of SOMATIC EMBRYOGENESIS RECEPTOR-UKE KINASE (SERK) genes results in aberrant em-bryo development. Fewer cortical, endodermal, and vascular cells are generated in the embryos of serk1 serk2bak1 triple mutants. WUSCHEL-RELATED HOMBOBOX5 (WOXS) is ectopically expressed in vascular cells of serkl serk2 bak1 embryos. The first transverse division of vascular precursors in mid-globular em-bryos and second asymmetric division of ground tissue stem cells in early-heart embryos are abnormally altered to a longitudinal division. The embryo defects can be partially rescued by constitutively activated mitogen-activated protein kinase (MAPK) kinase kinase YODA (YDA) and MAPK kinase mkk5. Taken together, our results reveal that SERK-mediated signals regulate division patterns of vascular precursors and ground tissue stem cells, likely via the YDA-mkk4/5 cascade, during embryo development.

关键词： embryo ground tissue stem cell mkk4 mkk5 SERKs vascular precursors

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枣mkk5基因的克隆及表达分析

枣MKK5基因的克隆及表达分析

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中国园艺学会2019年学术年会暨成立90周年纪念大会

作者：张玉冯春芳王玖瑞刘孟军河北农业大学林学院河北农业大学中国枣研究中心

植物中促分裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPK)是一类丝氨酸/苏氨酸蛋白激酶,在细胞中对各种生长因子进行应答。它广泛存在于各种真核生物中,与其他一些信号分子组成MAPK级联(MAPKcascades)。越来越多的试验证... 详细信息

植物中促分裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPK)是一类丝氨酸/苏氨酸蛋白激酶,在细胞中对各种生长因子进行应答。它广泛存在于各种真核生物中,与其他一些信号分子组成MAPK级联(MAPKcascades)。越来越多的试验证据表明,MAPK在植物生长发育调节和抗逆等生理过程中起重要作用。在拟南芥中,MPK6基因不仅参与脱落酸、茉莉酸、乙烯、一氧化氮、臭氧和过氧化氢的信号转导,而且在表皮细胞的决定方式、花药和胚胎发育的过程中起重要作用。MPK6和mkk5参与MAPK级联系统,MPK6能与mkk家族中的mkk2、mkk3、mkk4、mkk5和mkk9相互作用,通过磷酸化信号将外界的刺激信号传递到胞内,活化下游底物调节植物生长发育。本研究基于已经发表的同源基因和枣全基因组数据,筛选和克隆枣mkk5,并进行该基因的表达分析,从而探讨枣树mkk5在花发育中的作用。以花粉活力较低的品种‘冬枣’和花粉活力高的品种‘月光’为试材,在枣基因组数据库中筛选mkk5基因并设计引物。提取花蕾RNA,经反转录获得cDNA,以cDNA为模板进行PCR扩增获取目的基因,并以不同发育时期花蕾以及花不同组成部分为试材,通过qRT-PCR扩增分析该基因的表达差异。依据在枣基因组数据库中筛选获得的mkk5基因设计引物,使用‘冬枣’花蕾cDNA模板进行PCR扩增,获得目的基因。克隆该基因并测序分析,发现该基因cDNA全长为1002bp,编码333个氨基酸,其蛋白等电点为6.88,是一个不稳定的疏水性蛋白。利用Blastp搜索枣mkk5氨基酸序列保守结构域,发现该基因含有MNPKK家族的典型结构。与‘月光’中的拷贝数相比,mkk5在‘冬枣’有扩张,基因拷贝数增加。克隆分析‘月光’和‘冬枣’中的mkk5发现,两品种间该基因序列无差异。经该基因不同器官表达分析发现,mkk5在嫩枝、老枝、叶中均有微量的表达,而且在花蕾中表达量较高,推测其与花发育相关。‘冬枣’和‘月光’比较,前者花蕾中mkk5基因表达高于后者,mkk5的高表达可能影响了‘冬枣’花发育,与‘冬枣’花粉活力较低有关。

关键词：枣 mkk5 基因克隆表达分析花

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