平榛MAPKK基因克隆及在非生物胁迫下的表达分析  

Cloning and Expression Profile Analysis of MAPKKK From Corylus Heterophylla Fisch

作　　者：梁丽松[1] 赵天田[1] 马庆华[1] 王贵禧[1] 

机构地区：[1]中国林业科学研究院林业研究所/国家林业局林木培育重点实验室/林木遗传育种国家重点实验室,北京100091

出　　处：《核农学报》2015年第1期49-55,共7页Journal of Nuclear Agricultural Sciences

基　　金：中央级公益性科研院所基本科研业务费专项资金(RIF2013-10);北京市自然科学基金资助项目(6144030);国家林业局林业公益性物业科研专项(201304710)

摘　　要：通过研究外界刺激信号对平榛中促分裂原活化蛋白酶途径的影响,探索该途径中MAPKK类基因对各种非生物胁迫的响应机制。根据平榛花芽转录本高通量测序的结果,采用RACE-PCR方法克隆到一个平榛MAPKK基因,全长1 065bp,推断其编码354个氨基酸,命名为Ch MAPKK2。序列分析表明,该基因含有MAPKK类型基因典型的磷酸一致化序列SLADIDS。构建的系统发育树结果表明,它与葡萄Vv MAPKK2以及苹果Md MAPKK的亲缘关系较近,相似性分别为81.69%和78.53%。采用qRTPCR,以ACTIN为内参对Ch MAPKK2基因在自然条件下花芽部位的表达模式进行分析,结果表明:在自然条件下从11月份到次年4月份Ch MAPKK2的表达量呈现先上升后下降的趋势;对平榛根蘖苗进行4℃、干旱及盐胁迫处理,发现Ch MAPKK2基因均受上述3种非生物胁迫的诱导而上调表达;Ch MAPKK2在不同器官中的表达具有组织表达特异性,雌花芽中表达量最高,其次是树皮,雄花序中表达量最低。上述研究结果表明Ch MAPKK2基因在平榛适应非生物胁迫的过程中发挥着重要作用。Elucidation of the effects of various stimuli on MAPK signal pathway,the response mechanism of MAPKK gene in Corylus heterophylla Fisch was analyzed under various abiotic stress conditions. According to the high-throughput sequencing results of hazelnut buds,a 1 065 bp fragment including the whole coding region was obtained through RACEPCR. It encoded a polypeptide of 354 amino acids,designated as Ch MAPKK2。The sequence showed that this gene had SLADIDS sequence which is the typical phosphorylation consensus sequence of MAPKK family. Phylogeny tree results showed Ch MAPKK2 was close to Vv MAPKK2 from grape and Md MAPKK from apple,with 81. 69% and 78. 53% amino acids similarity,respectively. The Ch MAPKK2 expression in hazelnut buds was analyzed with ACTIN as an internal control gene under natural conditions and the result displayed that the expression level first increased and then decreased during November to next April. Enhancsed expression of Ch MAPKK2 in leaves was induced when suckers were exposed to low temperatures of 4℃,PEG and salt stress treatment. Spatial expression analysis revealed that the transcription level of Ch MAPKK2 was higher in bud than those in bark and male inflorescence indicating tissue-specific expression.The results suggested that the Ch MAPKK2 gene played an important role in C. heterophylla Fisch under various abiotic stresses.

关 键 词：平榛 MAPKK 基因克隆 非生物胁迫 表达模式 

分 类 号：S664.4[农业科学—果树学]