燕山葡萄MAPK基因的克隆、亚细胞定位及表达分析  

Cloning, Subcellular Localization and Expression Analysis of MAPK Genes from Vitis yeshanesis

作　　者：朱自果 张庆田[1] 李秀杰[1] 韩真[1] 李桂荣 李勃[1] ZHU Zi-Guo;ZHANG Qing-Tian;LI Xiu-Jie;HAN Zhen;LI Gui-Rong;LI Bo(Shandong Institute of Pomology,Taian 271000,China;College of Horticulture and Landscape Architecture,Henan Institute of Science and Technology,Xinxiang 453003,China)

机构地区：[1]山东省果树研究所,泰安271000 [2]河南科技学院,园艺园林学院,新乡453003

出　　处：《农业生物技术学报》2020年第3期429-440,共12页Journal of Agricultural Biotechnology

基　　金：山东省林业科技创新项目(LYCX-2018-13);山东省农业科学院创新工程项目(CXGC2016A03);泰安市农业良种工程(2017LZ006);河南省自然科学基金(182300410031)。

摘　　要：促分裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)级联途径在植物的生长发育和应答逆境的信号传导中起着重要作用,但有关MAPK家族成员在中国野生葡萄(Vitis sp.)中的作用研究较少。本研究从中国野生燕山葡萄(Vitis yeshanesis)中获得2条MAPK新基因VyMAPK2和VyMAPK3(GenBank No. MK942078;MK942077),对其进行生物信息学及亚细胞定位分析,并进行生长调节剂及干旱、高盐和低温等逆境处理下的表达分析。VyMAPK2和VyMAPK3蛋白序列中包含MAPK家族所特有的11个保守蛋白激酶亚结构域、TEY基序、活化环(activition-loop)、停靠区域(common docking domains)、磷酸化环(P-loop)和催化位点(C-loop)等结构域,分别归属于MAPK家族的B和A亚家族。亚细胞定位分析表明,VyMAPK2蛋白主要分布在细胞核,VyMAPK3蛋白也主要分布在细胞核,但在细胞质中也有分布基因表达分析显示,VyMAPK2表达量在根中最高,VyMAPK3表达量在叶中最高;生长调节剂处理下,生长素、赤霉素、细胞分裂素、乙烯、脱落酸、水杨酸和茉莉酸甲酯均能够显著诱导VyMAPK3基因的表达(P<0.05),而VyMAPK2的表达变化不明显;在逆境下,干旱、高盐和低温均能够显著诱导VyMAPK3基因的表达(P<0.05),而VyMAPK基因仅受干旱的诱导。以上结果表明,VyMAPK3基因在燕山葡萄生长发育和抵御外界逆境胁迫方面起着重要的作用,此研究有助于了解中国野生葡萄的抗逆机制,为葡萄抗逆育种提供基因资源。Mitogen-activated protein kinase(MAPK) cascade pathway plays an important role in plant growth and development and signal transduction in response to stress. However less the role of MAPK family members were reported in Chinese wild grape(Vitis sp.). In this study, two novel MAPK genes VyMAPK2 and VyMAPK3(GenBank No. MK942078, MK942077) were obtained from Vitis yeshanesis. VyMAPK2 and Vy MAPK3 proteins belonged to the B and A subgroup of the MAPK family, respectively, which all contained11 conserved protein kinase subdomains, TEY motifs, activation-loop, common docking domain, P-loop and C-loop. Subcellular localization analysis showed that VyMAPK2 was mainly distributed in the nucleus,Vy MAPK3 was mainly distributed in the nucleus and cytoplasm. Gene expression analysis showed that VyMAPK2 was mainly expressed in root and VyMAPK3 was mainly expressed in leaves. Under exogenous hormone treatments, auxin, gibberellin, 6-benzyladenine, ethylene, abscisic acid, salicylic acid, and methyl jasmonate could significantly induced the expression of VyMAPK3, but there was no significant change of the expression of VyMAPK2(P<0.05). Under stress, drought, high salinity and low temperature could significantly induced the expression of VyMAPK3 gene(P<0.05), while VyMAPK2 was only induced by drought. The above results showed that VyMAPK3 gene played an important role in the development and resistance to external stresses in V. yeshanesis. This study will be helpful to understand the stress resistance mechanism of wild grape in China and supply the genes for breeding.

关 键 词：燕山葡萄 MAPK基因 生物信息学分析 表达分析 

分 类 号：S663.1[农业科学—果树学]