玉米蜕皮激素合成酶基因ZmCYP92A1的克隆和表达分析

采用RT-PCR结合RACE技术,从玉米中克隆ZmCYP92A1,GenBank登录号为KY270496.1。该基因全长1 713 bp,开放阅读框为1 551 bp,编码517个氨基酸。预测ZmCYP92A1蛋白分子量为58.58 kD,理论等电点为6.44,存在2个跨膜结构域,1个信号肽,有1个P450保守结构域。多序列比对表明,ZmCYP92A1蛋白与其他物种的CYP92A1蛋白具有极高的相似性。系统发生分析揭示,CYP92A1蛋白与属于CYP75亚家族的类黄酮3-单加氧酶的亲缘关系较近。实时荧光定量PCR结果发现,非生物胁迫(干旱、高盐、低温和脱落酸)均诱导ZmCYP92A1基因的表达,表明该基因参与植物抵御非生物胁迫。

Cloning and Expression of Phytoecdysone Biosynthesis Enzyme Gene ZmCYP92A1 in Maize

Phytoecdysone biosynthesis enzyme gene, ZmCYP92A1 (KY270496.1), was cloned from maize (Zea mays) via real-time quantitative PCR (RT-qPCR) integrated with rapid-amplification of cDNA ends (RACE) technology. The gene is 1 713 bp in length with an open reading frame (ORF) of 1 551 bp, and encodes a protein with 517 amino acid (aa), molecular weight of 58.58 kD, and isoelectric point of 6.44. And ZmCYP92A1 protein possesses two transmembrane domains, one signal peptide and one conserved domain of p450. Multiple sequence alignment analysis indicated that ZmCYP92A1 protein shared high sequence similarities with CYP90B1 in other plants. Meanwhile, phylogenetic analysis revealed that ZmCYP92A1 protein had close relationship with flavonoids 3-monooxygenase of CYP75 subfamily. The expression of ZmCYP92A1 could be induced by some abiotic stresses, such as drought, salt stress, low temperature, and ABA, which indicated that this gene was involved in abiotic stresses defense.