Please wait a minute...
浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2012, Vol. 38 Issue (5): 551-558    DOI: 10.3785/j.issn.1008-9209.2011.10.141
生物科学与技术     
番茄SlMAPK12基因的分离及表达特征分析
孔福苓， 王洁， 程琳， 关小燕， 何艳军， 卢钢*
浙江大学 农业与生物技术学院 园艺系，浙江 杭州310058
Isolation of mitogenactivated protein kinase SlMAPK12 gene and characterization of its expression pattern in Solanum lycopersicum
KONG Fuling, WANG Jie, CHENG Lin, GUAN Xiaoyan, HE Yanjun, LU Gang*
Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
 全文: PDF(2576 KB)   HTML ( ) 
摘要： 利用PCR克隆技术，在番茄MicroTom的cDNA中分离出一个促分裂原活化蛋白激酶(mitogenactivated protein kinase, MAPK)，命名为SlMAPK12 (GenBank 登录号为JF795447)。该基因编码621个氨基酸，含有促分裂原活化蛋白激酶11个区域，磷酸化位点为TDY(苏氨酸-天冬氨酸-酪氨酸，属于D组)。氨基酸序列比对发现，它与拟南芥、杨树、蓖麻等D组MAPK具有高度一致性。利用RTPCR及qRT-PCR技术分析SlMAPK12的时空表达特性，发现SlMAPK12在Micro-Tom植株的多个器官中均有表达，在雄蕊中的表达量最高。对Micro-Tom幼苗进行(38±1) ℃高温处理，发现SlMAPK12在雄蕊中的相对表达量增高，于12 h时点达到最高，而后下降。推测SlMAPK12可能参与番茄花粉发育过程中的高温胁迫反应。
Abstract: In nature, plants are often exposed to everchanging external stimuli during their growth and development, including pathogen infection, drought, salinity and various other biotic and abiotic stresses. These extracellular stimuli are perceived by the plant through a highly elaborate signaling network, which leads to an adaptive stress response at cellular and organismal levels. A specific class of serine/threonine
 
protein kinases, mitogenactivated protein kinases (MAPKs) play an important role in the transduction of various extra and intracellular signals and are conserved throughout eukaryotes. Recently, increasing evidences
 indicate that MAPK (mitogenactivated protein kinase) has very important effects on plant growth and development, as well as stress resistance. As we know, current research on tomato (Solanum lycopersicum L.) MAPK gene is relatively limited. While the recently completed Genome Sequencing Project for tomato genome   provides an opportunity to deduce the tomato MAPK genes and infer its evolutionary history. So more detailed biochemical and functional characterization is likely to be carried out to understand the function of tomato MAPKs. The cDNA sequence of novel SlMAPK12 (GenBank No. JF795447) was amplified by PCR using primers designed based on the predicted sequences of tomato unigene.Then  many bioinformatics tools were used to analyze and predict SlMAPK12, such as ProtParam. To analyze the expression pattern of SlMAPK12 in various tissues and during  flower developmental stages, the leaves, stems, petals, stamens, pistils, fruits and 5 stages of floral development from flowering tomato plants were characterized by RT-PCR and furthermore proved by qRT-PCR. For high temperature treatment, the stamen were collected at 0 h, 12 h, 24 h, 48 h and 72 h after (38±1) ℃ heat stress treatment. Then the qRT-PCR experiments were employed to characterize gene expression profiles. SlMAPK12  encoding a protein of 621 amino acids  contained 11 subdomains of the MAPK. Subdomain Ⅷ contained the threonine and tyrosine residues, named T-D-Y (Thr-Asp-Tyr) motif, that was phosphorylated for the activation of  SlMAPK12.  SlMAPK12 protein sequences were found to contain four types of special domains: active site, ATP binding site, substrate binding site, and activation loop (A-loop) and KIM docking site. Amino acid sequence alignment revealed that SlMAPK12 shared high identity with groupD MAPKs from Arabidopsis, Populus, and Ricinus, which indicated that the groupD MAPKs was  conserved in different plant species. The temporal and spatial expression patterns of SlMAPK12 analysed by RT-PCR and qRT-PCR in different tomato tissue indicated that the relative gene expression level of SlMARK12 in stamen was much higher than other organs. Also, the levels of SlMAPK12 expressed in different developmental stages of tomato flower were almost the same. These results indicated that SlMAPK12 might  have very important effects on flower development. The expression pattern of SlMAPK12  gene under hightemperature (38±1) ℃ was analysed by qRTPCR. We found that the transcripts of SlMAPK12 in stamen accumulated markedly when the tomato seedlings were subjected to heat stress. These results above indicated that SlMAPK12 probably involved in the adaptive responses to heat stress during tomato pollen development.  The preferential expression of SlMAPK12 gene provides a new avenue for functional analyses in tomato. The major challenge for future research is to focus on identifying more MAPKs and understanding the specific role of MAPK genes during tomato growth and development.
出版日期: 2012-09-20
CLC:  Q 78  
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　

引用本文:

链接本文:

http://www.zjujournals.com/agr/CN/Y2012/V38/I5/551

[1] 吕琳慧1, 徐幼平2, 任至玄1, 康冬1, 王继鹏1, 蔡新忠1*. Ca2+信号通路对本氏烟叶位介导的核盘菌抗性的影响[J]. 浙江大学学报(农业与生命科学版), 2014, 40(6): 605-610.
[2] 张永亮， 武安泉， 赵锦慧. 野桑蚕丝氨酸蛋白酶样蛋白基因克隆及特征（英文）[J]. 浙江大学学报(农业与生命科学版), 2014, 40(2): 119-124.
[3] 李阳1， 赵媛1， 徐幼平2， 王继鹏1， 蔡新忠1*. 草酸对植物体内Ca2+浓度及信号传导途径的影响[J]. 浙江大学学报(农业与生命科学版), 2014, 40(2): 141-145.
[4] 张健慧， 王首锋*. 膜荚黄芪苯丙氨酸解氨酶基因在毕赤酵母中的分泌表达[J]. 浙江大学学报(农业与生命科学版), 2014, 40(1): 1-8.
[5] 王家庆1*， 边佳2， 李代宗3， 马爽1， 王亮1， 那广宁1. 虹鳟Ndufb2基因全长cDNA序列的克隆与分析[J]. 浙江大学学报(农业与生命科学版), 2013, 39(6): 600-606.
[6] 王亚1,2， 刘建平1， 徐梦云1， 季为捷1， 凃巨民1， 张晓波1*. 水稻泛素缀合酶样蛋白基因OsCROC-1A的克隆与表达分析[J]. 浙江大学学报(农业与生命科学版), 2013, 39(4): 360-368.
[7] 王安可， 何秋伶， 潘晶晶， 孙英超， 祝水金， 陈进红*. 农杆菌介导的VgDGAT1a基因棉花茎尖转化体系优化研究[J]. 浙江大学学报(农业与生命科学版), 2013, 39(3): 253-260.
[8] . 人辅脂酶样3基因hCLPSL3及其同源基因cDNA克隆及鉴定[J]. 浙江大学学报(农业与生命科学版), 2012, 38(5): 575-584.