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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。
The full-length cDNA sequence of CsMAPK3 (GenBank accession number: MF034662), genomic sequence and its promoter sequence were cloned from ’Fuding Da Bai’ tea plant. The cDNA sequence of CsMAPK3 is 1700 bp in length and contains an ORF of 1119 bp encoding 373 amino acids. CsMAPK3 protein is predicted to be a hydrophilic protein with multiple phosphorylation sites. Multiple sequence alignment and phylogenetic tree analysis show that CsMAPK3 C- The end contains a conserved CD domain, belonging to the TEY type A subfamily MAPK; subcellular localization predicted CsMAPK3 mainly located in the cytoplasm and nucleus. The complete genome of CsMAPK3 is 4930 bp in length, containing 5 introns and 6 exons. The first intron and the second intron of CsMAPK3 gene are 1608 and 1318 bp in length with exon lengths of 130 ~ Between 350 bp. Cloning of the start codon 1125 bp upstream of the promoter sequence, the promoter contains drought, low temperature, high temperature and ABA-related cis-acting elements. Fluorescence quantitative analysis showed that ABA, low temperature and salt stress could significantly up-regulate the expression of CsMAPK3 in tea leaves. Protein interaction prediction indicates that CsMAPK3 may interact with MYBR1 in response to ABA-dependent abiotic stresses. In summary, CsMAPK3 may be closely related to the resistance of tea plants.