花生MAPK13基因的克隆及表达分析研究

Cloning and Expression Analysis of a Mitogen-activated Protein Kinase 13 Gene in Peanut

为了挖掘花生抗果腐病相关基因,本研究以花生品种花育20号为试验材料,从花生果腐病转录组文库中筛选出促丝裂原活化蛋白激酶13(MAPK13)基因编码区序列设计引物,通过RACE-PCR克隆得到AhMAPK 13基因°结果表明AhMAPK 13基因序列全长1818 bp,含有/非编码区593 bp,5非编码区122 bp,开放阅读框全长1104 bp,编码1条含有368个氨基酸的蛋白序列。预测其分子量为42. 5kDa,含有MAPK家族典型的11个结构域,属于MAPK基因家族B组成员。亚细胞定位显示AhMAPK13定位于细胞质和细胞核中° RT-qPCR分析发现AhMAPK 13基因在茎中表达量高于其他组织,说明该基因具有组织表达特异性;AhMAPK 13基因受干旱、低温、NaCl(ABA诱导时表达量上升,说明该基因可能广泛参与植物非生物胁迫响应过程;AhMAPK 13基因受JA、GA、ET、SA诱导时表达量上调,说明该基因可能参与到这些激素介导的抗逆信号转导途径。本研究结果为花生抗果腐病育种研究提供了新的基因资源。

In this article, our aim is to detect peanut genes associated with peanut pod rot resistance. A novel mitogen-activated protein kinase gene, named AhMAPK13, was isolated from leaf of peanut ( Arachis hypogaea L. cultivar Huayu20) using the previous peanut pod rot transcriptomic library coding area sequence as template by PACE-PCR. The full-length cDNA of AhMAPK13 is 1818 bp, including a 1104 bp ORF, a 122 bp 5′UTR and a 593 bp 3′ UTR. The number of deduced amino acid was 368 and its predicted molecular weight 42.5kDa. Sequence analysis revealed that the sequence contain the 11 MAPK motifs, belongs to the group B MAPKs in plants. Further, subcellular localization analysis showed that AhMAPK13 was located in the nucleus and cytoplasm in plant cells. The tissue specific expression analysis by RT-qPCR showed that AhMAPK13 is a tissue specific expression gene with the most abundant level in stems roots. The expressions of AhMAPK13 were induced by PEG6000,NaCl,4℃ and ABA, suggesting that AhMAPK13 may be involved in ABA mediated abiotic stress processes. The increased expression of the AhMAPK13 gene induced by JA, GA, ET, and SA indicates that the gene may participate in these hormones mediated stress response signal transduction pathways. The results of this study provide a new gene resource for peanut resistance breeding.