论文部分内容阅读
脱落酸(ABA)在植物生长发育和逆境胁迫响应等过程中具有重要作用。脱落酸8’-羟化酶是ABA分解及其代谢的关键酶,可降解内源ABA水平。本研究利用Reverse Transcription PCR(RT-PCR)的方法从花生品种闽花6号叶片中克隆了脱落酸8’-羟化酶基因AhCYP707A4的开放阅读框(Open Reading Frame,ORF)。AhCYP707A4基因ORF长1 446 base pairs(bp),编码481个氨基酸。生物信息学分析表明,该蛋白N端有一个长32个氨基酸残基的信号肽序列,其后为P450蛋白保守区,符合典型的P450蛋白的结构特征。氨基酸多序列比对显示AhCYP707A4具有较高的保守序列。实时荧光定量PCR分析表明,该基因在不同发育时期的果皮、种皮和胚中均有表达。在果皮和种皮中,该基因在花生果针入土后40 d(days after pegging,DAP)表达量最高,而胚中在60 d中表达量达到最高。AhCYP707A4基因可能在花生发育过程起重要的调控作用,是调控花生种子发育特别是胚发育的重要候选基因。
Abscisic acid (ABA) plays an important role in the process of plant growth and stress response. Abscisic acid 8’-hydroxylase is the key enzyme in ABA decomposition and its metabolism, which can degrade the level of endogenous ABA. In this study, the open reading frame (ORF) of abscisic acid 8’-hydroxylase gene AhCYP707A4 was cloned from Peanut Variety Minhua 6 using Reverse Transcription PCR (RT-PCR). The AhCYP707A4 gene ORF has 1 446 base pairs (bp) encoding 481 amino acids. Bioinformatics analysis showed that the N-terminal of the protein has a signal peptide sequence with 32 amino acid residues, followed by the conserved region of P450 protein, which is consistent with the structural characteristics of a typical P450 protein. Amino acid multiple sequence alignment showed that AhCYP707A4 has a higher conserved sequence. Real-time PCR analysis showed that the gene was expressed in pericarp, seed coat and embryo at different developmental stages. In peel and seed coat, the expression of DAP was the highest at 40 days after peanut needles inoculation, while it reached the highest at 60 days in embryo. The AhCYP707A4 gene may play an important regulatory role in peanut development and is an important candidate gene for the regulation of peanut seed development, especially embryo development.