胡椒碱合成相关BAHD酰基转移酶基因的克隆与分析  

Cloning and Analysis of BAHD Acyltransferase Genes Related to Piperine Biosynthesis

作　　者：程俊 后思宇 李靖 岑怡 苏岳峰 伍宝朵[2] 范睿[2] 郝朝运[2] 杨建峰[2] 丁元昊 胡丽松[2] Cheng Jun;Hou Siyu;Li Jing;Cen Yi;Su Yuefeng;Wu Baoduo;Fan Rui;Hao Chaoyun;YangJianfeng;Ding Yuanhao;Hu Lisong(College of Tropical Crops,Hainan University,Haikou,570228;Hainan SIM Soonliang Academician Workstation,Key Laboratory of Genetic Resources Utilization of Spice and Beverage Crops(Ministry of Agriculture),Spice and Beverage Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wanning,571533)

机构地区：[1]海南大学热带作物学院,海口570228 [2]中国热带农业科学院香料饮料研究所,农业农村部香辛饮料作物遗传资源利用重点实验室,海南省Sim Soonliang院士工作站,万宁571533

出　　处：《分子植物育种》2023年第2期437-453,共17页Molecular Plant Breeding

基　　金：海南省重点研发计划项目(ZDYF2020187);海南省自然科学基金创新研究团队项目(2019CXTD417);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-11)共同资助。

摘　　要：胡椒碱是胡椒中重要的功能物质和品质指标之一。本研究以中国胡椒的主栽品种‘热引1号’为材料,基于胡椒基因组数据和转录组数据,筛选参与胡椒碱合成相关BAHD酰基转移酶,对候选的酰基转移酶基因进行克隆、分析和诱导表达。结果表明从胡椒基因组中鉴定出BAHD酰基转移酶基因78个。基于多序列比对及motif分析,胡椒BAHD酰基转移酶成员分为7类,特征结构域主要为HXXXD、DFGWG、YFGN。表达模式分析筛选出在胡椒果实中特异表达的BAHD基因7个,并成功克隆到全长序列。其中,Pn1.3303、Pn1.3375编码的氨基酸包含完整且未发生突变的特征结构域,在胡椒种子中特异表达,表达模式与胡椒碱合成紧密相关。将2个目的基因分别构建到原核表达载体pGEX-4T-1上,最终成功诱导表达出N末端与GST标签融合的目的蛋白。本研究为胡椒碱生物合成相关BAHD酰基转移酶的鉴定提供了基因资源。Piperine is one of the most important functional substance and quality trait in Piper nigrum.’Reyin-1’(Piper nigrum L.),the main Piper nigrum variety in China was used as the experimental material in this study.We identified acyltransferases involved in the piperine biosynthesis based on the whole genome sequence and transcriptome data.The main results are as follows:A total of 78 BAHD acyltransferase genes were identified in the piper nigrum genome.According to multiple sequence alignment and motif analysis,78 BAHD genes were divided into 7 subclasses and the highly conserved characteristic domains were found(HXXXD,DFGWG and YFGN).7 piperine biosynthesis-related BAHD genes were identified and cloned.Among them,Pn1.3303 and Pn1.3375 contain conserved and whole characteristic domains and differentially expressed in seeds.They were successfully in duced and expressed proteins in pGEX-4T-1 vector.The results provide available gene resources for further identification of BAHD acyltransferase related to piperine biosynthesis.

关 键 词：胡椒 胡椒碱 BAHD酰基转移酶 

分 类 号：S573.9[农业科学—作物学]