赵烨;国静;王甲威;徐丽;谭钺;陈新;魏海蓉;朱东姿;刘庆忠;宗晓娟;

• 1:山东省果树研究所·山东省果树生物技术育种重点实验室
• 2:山东农业大学林学院

摘要(Abstract)：

【目的】探明PcMPK3基因的表达调控规律。【方法】利用染色体步移技术从甜樱桃矮化砧木Gisela 6中克隆PcMPK3基因的启动子序列PcMPK3pro。利用Neural Network Promotor Prediction、softberry、PLACE和PlantCARE网站在线预测PcMPK3基因的基础启动子、转录起始位点和顺式作用元件。将PcMPK3pro定向替换植物表达载体pBI121-SN1的CaMV35S组成型启动子,构建重组表达载体pBI-PcMPK3pro:GUS,瞬时转化烟草叶片。【结果】结果表明,PcMPK3pro含有启动子核心元件TATA-box和CAAT-box等多种响应胁迫的顺式作用元件。受病原菌丁香假单胞菌番茄致病变种(Pseudomonas syringae pv. tomato DC3000,Pst DC3000)侵染,PcMPK3pro能驱动GUS报告基因表达且GUS酶活性显著提高。【结论】推测PcMPK3基因参与植物响应病原菌感染的胁迫过程。

关键词(KeyWords)： 甜樱桃;砧木;PcMPK3;启动子;GUS基因

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年基金项目(31601732);国家自然科学基金面上项目(31770706);; “十二五”国家科技支撑计划(2013BAD02B03-3-2);; 山东省现代农业产业技术体系果品创新团队(SDAIT-06-04)

作者(Author): 赵烨;国静;王甲威;徐丽;谭钺;陈新;魏海蓉;朱东姿;刘庆忠;宗晓娟;

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参考文献(References)：

• [1] ASAI T,TENA G,PLOTNIKOVA J,WILLMANN M R,CHIU W L,GOMEZ-GOMEZ L,BOLLER T,AUSUBEL F M,SHEEN J. MAP kinase signalling cascade in Arabidopsis innate immunity[J]. Nature,2002,415(6875):977-983.
• [2] MENG X,ZHANG S. MAPK cascades in plant disease resistance signaling[J]. Annual Review of Phytopathology,2013,51(1):245-266.
• [3] ZHANG S,KLESSIG D F. MAPK cascades in plant defense signaling[J]. Trends in Plant Science,2001,6(11):520-527.
• [4] BETHKE G,UNTHAN T,UHRIG J F,POSCHL Y,GUST A A,SCHEEL D,LEE J. Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling[J]. Proceedings of the National Academy of Science USA,2009,106(7):8067-8072.
• [5] LIU Y,SCHIFF M,DINESH-KUMAR S P. Involvement of MEK1 MAPKK,NTF6 MAPK,WRKY/MYB transcription factors,COI1 and CTR1 in N-mediated resistance to tobacco mosaic virus[J]. The Plant Journal,2004,38(5):800-809.
• [6] SEO S,KATOU S,SETO H,GOMI K,OHASHI Y. The mitogen-activated protein kinases WIPK and SIPK regulate the levels of jasmonic and salicylic acids in wounded tobacco plants[J].The Plant Journal,2010,49(5):899-909.
• [7] MOUSTAFA K. Improving plant stress tolerance:potential applications of engineered MAPK cascades[J]. Trends in Biotechnology,2014,32(8):389-390.
• [8] MOUSTAFA K. MAPK transgenic circuit to improve plant stress-tolerance[J]. Plant Signal&Behavior,2014,9(11):e970101.
• [9] SANSAVINI S A L,LUGLI S. New rootstocks for intensive sweet cherry plantations[J]. Acta Horticulturae,2014,1020:411-434.
• [10] KLEMENT Z R D,BáLóE,PáNCZéL M,PRILESZKY M.The effect of cold on development of bacterial canker in apricot trees infected with Pseudomonas syringae pv. syringae[J]. Physiological Plant Pathology,1984,24(2):237-246.
• [11] LITTLE E L,BOSTOCK R M,KIRKPATRICK B C. Genetic Characterization of Pseudomonas syringae pv. syringae Strains from Stone Fruits in California[J]. Applied&Enviromental Microbiology,1998,64(64):3818-3823.
• [12] ZONG X,WANG J,XU L,WEI H,CHEN X,ZHU D,TAN Y,LIU Q. Identification and characterization of 12 Mitogen-activated Protein Kinase genes implicated in stress responses in cherry rootstocks[J]. Journal of the American Society for Horticultural Science,2016,141(5):490-497.
• [13] BLáZQUEZ M A,GREEN R,NILSSON O,SUSSMAN M R,WEIGEL D. Gibberellins promote flowering of arabidopsis by activating the LEAFY promoter[J]. The Plant Cell,1998,10(5):791-800.
• [14] ZHANG S,XU R,LUO X,JIANG Z,SHU H. Genome-wide identification and expression analysis of MAPK and MAPKK gene family in Malus domestica[J]. Gene,2013,531(2):377-387.
• [15] CHEN L,HU W,TAN S,WANG M,MA Z,ZHOU S,DENG X,ZHANG Y,HUANG C,YANG G,HE G. Genome-wide identification and analysis of MAPK and MAPKK gene families in Brachypodium distachyon[J]. PLoS One,2012,7(10):e46744.
• [16] KONG F,WANG J,CHENG L,LIU S,WU J,PENG Z,LU G.Genome-wide analysis of the mitogen-activated protein kinase gene family in Solanum lycopersicum[J]. Gene,2012,499:108-120.
• [17] KOVTUN Y,CHIU W L,TENA G,SHEEN J. Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants[J]. Proceedings of the National Academy of Science USA,2000,97(6):2940-2945.
• [18] YUASA T,ICHIMURA K,MIZOGUCHI T,SHINOZAKI K.Oxidative stress activates ATMPK6,an Arabidopsis homologue of MAP kinase[J]. Plant&Cell Physiology,2001,42(9):1012-1016.
• [19] LU C,HAN M H,GUEVARA-GARCIA A,FEDOROFF N V.Mitogen-activated protein kinase signaling in postgermination arrest of development by abscisic acid[J]. Proceedings of the National Academy of Science USA,2002,99(24):15812-15817.
• [20]杨明星,申磊,文嘉瑜,唐倩,石兰平,刘艳艳,杨晟,胡炯,刘彩玲,吴杨,何水林.辣椒CaMAPK7启动子顺式作用元件及其表达分析[J].热带作物学报,2015,36(4):673-679.YANG Mingxing,SHEN Lei,WEN Jiayu,TANG Qian,SHI Lanping,LIU Yanyan,YANG Sheng,HU Jiong,LIU Cailing,WU Yang,HE Shuilin. Cis-acting elements and expression analysis of the Promoter of CaMAPK7[J]. Chinese Journal of Tropical Crops,2015,36(4):673-679.
• [21]朱斌,陆俊杏,彭茜,翁昌梅,王淑文,余浩,李加纳,卢坤,梁颖.甘蓝型油菜MAPK7基因家族及其启动子的克隆与表达分析[J].作物学报,2013,39(5):789-805.ZHU Bin,LU Junxing,PENG Qian,WENG Changmei,WANG Shuwen,YU Hao,LI Jiana,LU Kun,LIANG Ying. Cloning and analysis of MAPK7 gene family and their promoters from Brassica napus[J]. Acta Agronomica Sinica,2013,39(5):789-805.
• [22] ZHANG D,JIANG S,PAN J,KONG X,ZHOU Y,LIU Y,LI D. The overexpression of a maize mitogen-activated protein kinase gene(ZmMPK5)confers salt stress tolerance and induces defence responses in tobacco[J]. Plant Biology(Stuttg),2014,16(3):558-570.
• [23] ZHANG L,LI Y,LU W,MENG F,WU CA,GUO X. Cotton GhMKK5 affects disease resistance,induces HR-like cell death,and reduces the tolerance to salt and drought stress in transgenic Nicotiana benthamiana[J]. Journal of Experimental Botany,2012,63(10):3935-3951.
• [24] LU W,CHU X,LI Y,WANG C,GUO X. Cotton GhMKK1 induces the tolerance of salt and drought stress,and mediates defence responses to pathogen infection in transgenic Nicotiana benthamiana[J]. PLoS One,2013,8(7):e68503.