茶树CsMAPK3的全长克隆及其逆境表达分析

以‘福鼎大白’茶树为材料,克隆了CsMAPK3的全长cDNA序列(GenBank登录号:MF034662)、基因组序列及其启动子序列。CsMAPK3的cDNA序列全长1700 bp,含有1119 bp的ORF序列,编码373个氨基酸;CsMAPK3蛋白预测为亲水性蛋白,含有多个磷酸化位点;多序列比对和进化树分析表明CsMAPK3 C–末端含有保守的CD结构域,属于TEY类型的A亚家族MAPK;亚细胞定位预测CsMAPK3主要定位于细胞质和细胞核中。CsMAPK3基因组全长4930 bp,包含5个内含子和6个外显子,第1个内含子和第2个内含子较大,分别为1608和1318 bp,外显子长度在130~350 bp之间。克隆获得起始密码子上游1125 bp的启动子区序列,该启动子上含有干旱、低温、高温以及ABA等相关的顺式作用元件。荧光定量表达分析显示,ABA、低温和盐胁迫均能显著上调茶树叶片中CsMAPK3的表达。蛋白互作预测表明CsMAPK3可能与MYBR1互作来响应ABA依赖途径的非生物胁迫过程。综上表明,CsMAPK3可能与茶树抗逆响应密切相关。

Cloning and Abiotic Stress Expression Analysis of CsMAPK3 Gene in Tea#br# Plant

MAPK（mitogen-activated protein kinase）genes play a crucial role in the plant response to stress. In this study，the full-length of cDNA and genome sequence，and the promoter sequence of CsMAPK3 were isolated from the tea plant（Camellia sinensis）cultivar‘Fudingdabai’. Moreover，the characteristic of bioinformatics and the expression patterns of CsMAPK3 under different stress treatments were investigated. The full-length cDNA of CsMAPK3 was 1 700 bp，with a 1 119 bp ORF，encoding 373 amino acids（Accession No. MF034662）. It was predicted that CsMAPK3 was a hydrophilic protein， containing multiple phosphorylation sites. The homologous alignment and phylogenetic tree analysis showed that CsMAPK3 had a CD domain in the C-terminal region and conserved in TEY motif，which belongs to the group A of MAPKs. Subcellular localization prediction suggested that CsMAPK3 could be located in both the cytoplasm and the nucleus. The genome sequence of CsMAPK3 was 4 930 bp in length and constituted by five introns and six exons；thereinto，the first and the second intron were 1 608 bp and 1 318 bp in length，respectively，which were longer than other introns，whereas the length of the exons was ranged from 130 bp to 350 bp. Moreover，we cloned a promoter sequence of CsMAPK3 which was 1 125 bp in length，and contained several stress-responsive elements involved in drought，cold，high temperature and ABA-signaling. Expression analysis showed that ABA，cold and salt treatments could significant up-regulate the expression of CsMAPK3. Protein interaction network prediction showed that CsMAPK3 could interact with MYBR1 resulted in responsing to abiotic stress in ABA-dependent pathway. In conclusion，CsMAPK3 might be correlated to the abiotic stress-responsive in tea plant.