当归抽薹开花过程中赤霉素代谢水平及其关键酶基因克隆与表达分析  

Analysis on gibberellins metabolic level, key enzyme genes clone and expression pattern during bolting and flowering plants in Angelica sinensis

作　　者：刘迪 崔秀文 黄天苗 李美玲 栗孟飞[1] 魏建和[2] LIU Di;CUI Xiu-wen;HUANG Tian-miao;LI Mei-ling;LI Meng-fei;WEI Jian-he(College of Life Science and Technology/Key Laboratory of Aridland Crop Science,Gansu Agricultural University,Lanzhou 730070,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China)

机构地区：[1]甘肃农业大学生命科学技术学院干旱生境作物学国家重点实验室,甘肃兰州730070 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《中草药》2023年第1期222-234,共13页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(32160083);干旱生境作物学国家重点实验室(甘肃农业大学)基金项目(GSCS-2021-Z03);道地药材生态种植与质量保障项目(202103003);财政部和农业农村部:国家现代农业产业技术体系(CARS-21)。

摘　　要：目的 对当归Angelica sinensis 16个赤霉素(gibberellins,GAs)进行含量测定、GAs代谢酶基因筛选并进行生物信息学分析、关键基因克隆及表达验证,以期为调控当归抽薹开花提供理论依据。方法 利用HPLC-MS/MS对不同材料中GAs含量进行测定与分析,基于当归全长转录组筛选直接参与GAs代谢酶基因,利用在线工具进行生物信息学分析,并对关键酶基因GA2ox6和GA20ox1表达水平进行q RT-PCR检测。结果 在所测定的16个GAs中,8个GAs含量在早薹植株中高于非早薹植株,13个GAs含量随植株发育时期延长逐渐增加。当归全长转录组中含有9个直接参与GAs代谢的酶基因,可分为6个亚家族,共含有6个保守基序,亚细胞定位于细胞质。GA2ox6和GA20ox1基因克隆片段由于插入序列与全长转录组测序片段存在长度差异。GA2ox6基因在抽薹植株、随种苗春化作用和植株发育时期延长呈现低表达,而在冷冻规避春化作用种苗中呈现高表达,GA20ox1基因则相反;GA2ox6和GA20ox1基因在叶和茎中相对于根均呈现高表达。结论 当归含有9个直接参与GAs代谢的酶基因,各成员理化性质和结构存在一定的差异,GA2ox6和GA20ox1基因表达水平与当归抽薹开花生理调控一致。Objective To determine the contents of 16 gibberellins(GAs), screen the genes involved in GAs metabolism and analyze their bioinformatics, as well as clone the key genes and validate their expression level, so as to provide theoretical basis for regulating bolting and flowering of Angelica sinensis. Methods The GAs contents in different materials were determined by HPLC-MS/MS. The genes directly involved in GAs metabolism were screened based on the full-length transcriptomes of A. sinensis,the bioinformatics was analyzed using the online tools, and the expression levels of the GA2ox6 and GA20ox1 genes were validated by qRT-PCR. Results Among of the 16 GAs, the contents of eight GAs were higher in bolted plants than unbolted plants, and the contents of 13 GAs were increased with the extension of plant development. A total of nine genes directly involved in GAs metabolism were identified from A. sinensis, with six subfamily classified, six motifs included, and the sub-cellular located in the cytoplasm. There was a certain difference in the base sequences between the cloned fragment of GA2ox6 and GA20ox1 with the full-length transcriptomes, due to the inserted fragments in the sequence of GA2ox6 and GA20ox1. The expression levels of GA2ox6gene were down-regulated in the bolted plants compared to unbolted plants, and with the extension of seedlings vernalization and plants development, while up-regulated in the seedlings with frozen-avoided vernalization;but the GA20ox1 gene showed opposite results;and the expression levels of GA2ox6 and GA20ox1genes in leaves and stems were higher than roots. Conclusion There are nine genes directly involved in GAs metabolism in A. sinensis, and certain differences in physical and chemical properties and structure among the nine members;the expression levels of the GA2ox6 and GA20ox1 genes are in accordance with the physiological regulation of bolting and flowering of A. sinensis.

关 键 词：当归 赤霉素 赤霉素代谢基因 生物信息学 基因克隆 抽薹开花 

分 类 号：R286.12[医药卫生—中药学]