生长素反应因子ZmARF1的表达分析及互作网络探究

Expression Analysis and Interaction Network of Auxin Reactive Factor ZmARF1

以抗旱型玉米自交系郑36和弱抗旱型玉米自交系B73为试验材料,利用克隆、qRT-PCR、亚细胞定位、互作蛋白预测等手段验证ZmARF1(GRMZM2G702026)对干旱胁迫的响应模式及初步探究ZmARF1基因的互作网络。克隆及测序分析表明,该基因含有1个2 034 bp的开放阅读框,翻译677个氨基酸。蛋白分析表明,该蛋白无跨膜域,属于高亲水性蛋白,亚细胞定位于细胞核。潜在磷酸化位点分析显示,ZmARF1蛋白含有潜在的丝氨酸、苏氨酸和酪氨酸磷酸化位点数目分别为39、16、6个。复合进化树、功能域和保守motif分析表明,该基因蛋白与其他物种同源性较高,说明不同物种该基因在功能上较保守。ZmARF1属于组成型表达基因,受干旱胁迫的正向诱导,且郑36基因表达量上升幅度大于B73。预测的互作功能蛋白主要通过调节激素应答基因的表达、参与激素介导的信号等途径,来调控植物对逆境的胁迫应答和生长发育等过程。

Drought-resistant Zheng 36 and weak drought-resistant B73 were used as experimental materials.Cloning and sequencing analysis indicated that the gene contained a 2034 bp open reading frame and translated 677 amino acids. Protein analysis showed that the protein has no transmembrane domain and belongs to a highly hydrophilic protein. The subcellular localization is located in the nucleus. Analysis of potential phosphorylation sites revealed that the ZmARF1 protein contains 39, 16, and 6 potential serine, threonine, and tyrosine phosphorylation sites, respectively. The composite phylogenetic tree, functional domain and conserved motif analysis indicated that the homologous gene was highly homologous to other species, indicating that the gene is functionally conserved in different species. ZmARF1 is a constitutively expressed gene, which is positively induced by drought stress, and the expression level of Zheng 36 gene is increased more than B73. The predicted interaction function proteins mainly regulate the stress response and growth and development of plants under stress by regulating hormone response gene expression and participating in hormone-mediated signaling.