白木香转录因子AsbZIP59的鉴定和表达分析  

Identification and Expression Analysis of a Transcription Factor AsbZIP59 in Aquilaria sinensis

作　　者：张浩[1,2] 蔡彩虹[2,3] 朱家红 彭世清[2,3] 戴好富 梅文莉[2,3] Zhang Hao;Cai Caihong;Zhu Jiahong;Peng Shiqing;Dai Haofu;Mei Wenli(College of Forestry,Hainan University,Haikou,570228;Hainan Engineering Research Center of Agarwood,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,57110l;Key Laboratory of Conservation and Utilization of Tropical Agro-bioresources of Hainan Province,Hainan Academyof Tropical Agricultural Resources,Haikou,571101)

机构地区：[1]海南大学林学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海南省沉香工程技术研究中心,海口571101 [3]海南热带农业资源研究院,海南省热带农业生物资源保护与利用重点实验室,海口571101

出　　处：《分子植物育种》2022年第23期7751-7758,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31870668,32171824);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-21)共同资助。

摘　　要：转录因子具有调控基因转录的重要作用,是植物生长发育及次生代谢物质合成的关键调控因子。碱性亮氨酸拉链蛋白(bZIP)是一类分布广泛且功能多样的转录因子,因而可能对白木香等药用植物合成具有药效的次生代谢物质及抵抗外界(非)生物压力等方面也具有重要作用。本研究基于结香剂诱导白木香产生沉香的转录组数据进行拼接注释,并以克隆及RT-PCR等方法,首次鉴定了1条白木香bZIP基因的CDS序列,命名为AsbZIP59。AsbZIP59基因全长751 bp,包含1个438 bp的ORF区,共编码145个氨基酸,其蛋白序列与无患子科植物文冠果的XsbZIP相似度达80.42%,且与拟南芥的AtbZIP53也具有较高同源性(69.18%),同属于bZIP-S亚族。生物信息学分析显示,AsbZIP59包含bZIP蛋白的BRLZ特征性结构域,为非分泌型蛋白,且属于非跨膜蛋白,含有20个潜在磷酸化位点,可能主要分布于白木香的细胞核中;其二级结构主要由α-螺旋和无规则卷曲构成;其启动子区域发现与植物生长发育、激素响应和内、外压力响应等相关的多种顺式作用元件,其中与激素、压力响应相关的顺式作用元件数量占总数50%以上。荧光定量PCR检测结果表明随结香时间变化,AsbZIP59基因表达量呈现先快速升高后降低的趋势,于12 h表达量达到最高,为对照组的8.72倍,随后持续下降,在第5天降至对照组的1.81倍。本研究为进一步研究bZIP转录因子在白木香形成沉香过程中所发挥的作用提供理论依据。Transcription factors(TFs) play the crucial roles in biosynthesis of plant secondary metabolites. The basic region/leucine zipper motif(bZIP) transcription factors are ubiquitous in plants and have been reported that contributed into almost all biological processes including signal transduction, environmental stress responses, secondary metabolism and promoting the accumulation of effective ingredients in the medicinal plants, which suggest that might participate into regulating agarwood formation in Aquilaria sinensis. Based on the transcriptome data and genome database from our previous study, a bZIP gene, named AsbZIP59, was cloned from Aquilaria sinensis and verified by RT-PCR. The AsbZIP59 contained 751 nucleotides and its complete open reading frame included 438 base pairs which encoding 145 amino acids. The AsbZIP59 is 80.42% similarity with a bZIP of Xanthoceras sorbifolium on protein sequence level. In addition, AsbZIP59 showing high homologous(69.18%) with AtbZIP53 and both of them are the member of group S from plant bZIP gene family. The bioinformatics analysis indicated that Asb ZIP59 contained a special BRLZ motif and AsbZIP59 not only had no transmembrane domain,but also was a non-secretory protein and it played a major physiological role in the nucleus. This protein included20 phosphorylation sites and its secondary structure mainly consist of α-helix accounted and random coil. Various cis-acting elements, such as growth and development of plant, phytohormone effects and stress responsiveness,were predicted in the promoter region of AsbZIP59 and more than half of them involved in phytohormone or stress responses. According to the real-time fluorescence quantitative PCR, the expression profiles of AsbZIP59 changed during agar-wood formation presented trend of earlier increase and later decrease, specifically, its expression level after treatment 12 hours was almost 8.72 times higher than the untreated control subjects, then gradually decreased to 1.81 times after 5 days treatment higher than in th

关 键 词：白木香(Aquilaria sinensis) 沉香 BZIP 转录因子 表达分析 

分 类 号：S567.19[农业科学—中草药栽培]