独行菜UGT74B1基因的序列结构分析及时空表达特性  

Sequence Structure Analysis and Spatiotemporal Expression Characteristics of UGT74B1 Gene in Lepidium apetalum

作　　者：王敏[1] 王熙林 宋钢钢 郁国良 刘家勤 李佳佳 葛风伟 赵惠新[1,2] Wang Min;Wang Xilin;Song Ganggang;Yu Guoliang;Liu Jiaqin;Li Jiajia;Ge Fengwei;Zhao Huixin(Laboratory of Plant Stress Biology,Arid Regions,College of Life Science,Xinjiang Normal University,Urumqi,830054;Beijing Key Laboratory of Resistance Gene Resources and Molecular Development,School of Life Science,Beijing Normal University,Beijing,100087)

机构地区：[1]新疆师范大学生命科学学院,干旱区植物逆境生物学实验室,乌鲁木齐830054 [2]北京师范大学生命科学学院,抗性基因资源与分子发育北京市重点实验室,北京100087

出　　处：《分子植物育种》2021年第10期3243-3250,共8页Molecular Plant Breeding

基　　金：新疆师范大学新疆特殊环境物种保护与调控生物学实验室开放课题基金(XJDX1414-2016-02);国家自然科学基金(31660079)共同资助。

摘　　要：芥子油苷是独行菜种子的重要药用活性物质,也是重要的植物代谢调节物。但独行菜芥子油苷的合成途径及其调节还不清楚。为了探讨独行菜芥子油苷合成与生长阶段、组织器官的关系,本研究基于独行菜种子转录组数据库,以UGT(尿苷二磷酸-糖基转移酶)为关键词筛选在芥子油苷核心结构起修饰作用的关键酶基因序列,并对其结构及表达特性进行分析。结果表明,独行菜种子萌发中有98条Unigenes,其中一条c19350_g1与拟南芥UGT74B1相似性最高。进一步采用RT-PCR技术克隆到独行菜UGT74B1基因,命名为LaUGT74B1,结构分析显示,该基因的c DNA序列编码区为1386 bp,编码产物为461个氨基酸的蛋白,蛋白的理论等电点为5.38,其不稳定系数为34.70,为稳定蛋白质,不具有跨膜结构。其二级结构主要由α螺旋和无规则卷曲构成,与独行菜与琴叶拟南芥、亚麻荠、荠菜的UGT74B1的序列相似性较高。另外,LaUGT74B1表达具有时空特异性,萌发阶段表达量呈现先下降后上升的变化趋势,苗期主要在叶中表达,花期主要在根中,结实期主要在老茎中。本研究结果可为独行菜芥子油苷生物合成途径阐明提供初步的试验基础。Glucosinolates is an important medicinal active substance in Lepidium apetalum seeds and an important plant metabolism regulator.However,the synthetic pathway and regulation of glucosinolates from Lepidium apetalum are not clear.In order to explore the relationship between glucosinolate synthesis,plant growth stage,tissues and organs,this study based on the Lepidium apetalum seed transcriptome database,using UGT(UDP-glycosyltransferases)as the key to screen the core structure of glucosinolate The key enzyme gene sequence that plays a role in modification,and its structure and expression characteristics were analyzed.The results showed that there were 98 Unigenes in the germination of Lepidium apetalum seeds,one of which c19350_g1 was most similar to Arabidopsis lyrata UGT74 B1.Furthermore,the UGT74 B1 gene was cloned by RT-PCR and named LaUGT74 B1.Structural analysis revealed that the gene’s c DNA sequence coding region is 1386 bp and the coding product is a protein of 461 amino acids.The stability coefficient is 34.70,which is a stable protein and does not have a transmembrane structure.Its secondary structure is mainly composed ofα-helix and random coils,and has high sequence similarity with UGT74 B1 of Arabidopsis lyrata subsp.lyrata,Camelina sativa and Capsella rubella.In addition,LaUGT74 B1 expression is spatio-temporal specific,and the expression level at the germination stage first decreases and then rises.It is mainly expressed in leaves at the seedling stage,mainly in the root at flowering stage,and mainly in the old stem at seedling stage.The results of this study can provide a preliminary experimental basis for elucidating the biosynthetic pathways of glucosinolates.

关 键 词：独行菜 芥子油苷 LaUGT74B1 结构分析 时空表达特性 

分 类 号：S567.2[农业科学—中草药栽培]