受棉铃虫诱导的棉花糖基转移酶UGT基因家族的生物信息学及表达分析

doi:10.6048/j.issn.1001-4330.2023.10.007

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (10): 2396-2403.DOI: 10.6048/j.issn.1001-4330.2023.10.007

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

受棉铃虫诱导的棉花糖基转移酶UGT基因家族的生物信息学及表达分析

玛依拉·玉素音(), 阳妮, 徐海江, 杨延龙, 张大伟, 李春平, 石必显, 赖成霞()

新疆农业科学院经济作物研究所,乌鲁木齐 830091

收稿日期:2023-02-10 出版日期:2023-10-20 发布日期:2023-11-01
通信作者: 赖成霞(1972-),女,新疆乌鲁木齐人,副研究员,研究方向为棉花分子育种,(E-mail)lchxia2001@163.com
作者简介:玛依拉·玉素音(1987-),女,新疆乌鲁木齐人,助理研究员,研究方向为棉花分子育种,(E-mail)119327132@qq.com
基金资助:
棉花等经济作物分子设计育种及多层组学技术联合创新平台建设”(2020026528-1);新疆农业科学院青年科技骨干创新能力培养项目(xjnkq-2019006)

Bioinformatics and expression analysis of UGT gene family of cotton glycosyltransferase damaged by Helicoverpa armigera

Mayila Yusuyin(), YANG Ni, XU Haijiang, YANG Yanlong, ZHANG Dawei, LI Chunping, SHI Bixian, LAI Chengxia()

Research Institute of Economic Crops, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China

Received:2023-02-10 Online:2023-10-20 Published:2023-11-01
Correspondence author: LAI Chengxia (1972-), female, native place: Urumqi, associate researcher, research direction: cotton molecular breeding, (E-mail)lchxia2001@163.com
Supported by:
The innovation platform construction based onmolecular design breeding combined with multi-level omics technology in xinjiang rape,safflower,and cotton(2020026528-1);Innovation Ability Training Project for Young Sci-Tech Backbone Talents Sponsored by Xinjiang Academy of Agricultural Sciences(xjnkq-2019006)

摘要/Abstract

摘要：

【目的】 筛选分析受棉铃虫诱导的棉花糖基转移酶(Glycosyltransferase)UGT基因家族,为UGTG01、UGTG02、UGTGo4、UGTGo6、UGTGo7基因功能深入研究奠定基础。【方法】 以受棉铃虫诱导筛选出的棉花糖基转移酶UGT基因组数据库为基础,利用生物信息学方法分析棉花糖基转移酶UGT基因家族的理化性质、系统进化树、蛋白质的二级结构预测以及保守基序与结构域。【结果】 从棉花基因组中搜索到5个推定的UTG基因家族成员,其氨基酸数目介于468~534个,理论等电点分布在4.97~5.58,相对分子质量区域为52 743.1~60 208.62,亚细胞定位在细胞质外,均为亲水性蛋白质。基因结构均含有1个外显子,二级结构主要由α-螺旋、无规则卷曲为主,均含3个Motif基序和有Glycosyltransferase_GTB-type superfamily结构域。5个基因在根、茎、子叶、真叶、花、棉桃中具有明显的组织表达差异性,5个基因与UGT73C家族基因,水稻的UGT707A3、大豆的GmUGT在进化树的同一分支上。【结论】 糖基转移酶基因在不同组织中的表达具有显著差异,5个糖基转移酶与UGT73C、UGT707A3、GmUGT都处于进化树的同一个分支上,其可能具有相似的抗虫功能,能通过调控皂苷元、类黄酮、柚皮素等物质响应棉花的抗虫性。

关键词: 糖基转移酶; 亚细胞定位; 系统进化; 生物信息学; 基因家族

Abstract:

【Objective】 This project aims to screen and analyze the UGT gene family of cotton glycosyltransferase damaged by Helicoverpa armigerawere in the hope of providing a basis for in-depth researches into the roles of UGTG01, UGTG02, UGTGo4, UGTGo6, and UGTGo7 genes.【Methods】 Based on the genome database of cotton glycosyltransferase UGT damaged by Helicoverpa armigera, the bioinformatics method was used to systematically analyze the physical and chemical properties, phylogenetic tree, secondary structure prediction, conserved motif, and conserved domain of cotton glycosyltransferase UGT gene family.【Results】 Five putative members of the UTG gene family with amino acid numbers ranged from 468 to 534, whose theoretical isoelectric point distributions of was from 4.97 to 5.58, relative molecular mass ranged between 52,743.1 and 60,208.62, subcellular localization outside the cytoplasm, and hydrophilicity as proteins were discovered through a genome search of cotton. The gene structure contained one exon, and the secondary structure was mainly composed of α-helix, irregularly coiled, with three motifs and a Glycosyltransferase_GTB-type superfamily domain. Expression analysis showed that the five genes displayed notable variations in the root, stem, cotyledon, true leaf, flower, and cotton boll; molecular evolutionary tree exhibited that the five UGTs were found to be on the same branch of the UGT73C family genes, rice UGT707A3 and soybean GmUGT.【Conclusion】 These glycosyltransferase genes have markedly varied levels of expression in various tissues and are found to be on the same evolutionary branch as UGT73C, UGT707A3, and GmUGT in evolutionary tree analysis, which indicates that they may have similar insect resistance functions, and also respond to insect resistance in cotton by regulating saponin elements, flavonoids, and naringenin.

Key words: glycosyltransferase; subcellular localization; systematic evolution; bioinformatics; gene family

中图分类号:

S562

玛依拉·玉素音, 阳妮, 徐海江, 杨延龙, 张大伟, 李春平, 石必显, 赖成霞. 受棉铃虫诱导的棉花糖基转移酶UGT基因家族的生物信息学及表达分析[J]. 新疆农业科学, 2023, 60(10): 2396-2403.

Mayila Yusuyin, YANG Ni, XU Haijiang, YANG Yanlong, ZHANG Dawei, LI Chunping, SHI Bixian, LAI Chengxia. Bioinformatics and expression analysis of UGT gene family of cotton glycosyltransferase damaged by Helicoverpa armigera[J]. Xinjiang Agricultural Sciences, 2023, 60(10): 2396-2403.

图/表 7

表1 定量PCR 引物序列

Tab.1 Primer sequences for RT-qPCR

基因Gene	正向引物Forward primer(5'-3')	反向引物Reversr primer(5'-3')
histone 3	GGTGGTGTGAAGAAGCCTCAT	AATTTCACGAACAAGCCTCTGGAA
UGTG01	TTTCGCTCTGCAATAGGACA	AGCGATCTCCTTTAACTGACC
UGTG02	ATCAAGGGACTAATGCGAAG	CAAATCCAAGCACCTAACGA
UGTGo4	CTCAACCGTTTAACACCTCC	GCCTCTTCCTTTTACCCTT
UGTGo6	ACGCTAAAATCCTTATTAACACC	AGTTAAAGAATGAAGATGGGACT
UGTGo7	GTCATCTAAACTCTTTCCCACT	AGATTCAATAAAGGTCCCACT

表2 棉花糖基转移酶UGT蛋白基本理化参数

Tab.2 Basic physical and chemical parameters of UGT protein of cotton glycosyltransferase

基因 Gene	蛋白长度 Length(aa)	分子质量 Molecular weight (kDa)	等电点 Theoretical pI	不稳定系数 Instability index	脂溶性指数 Aliphatic index	亲水性 Hydrophilic
UGTG01	534	60 208.62	5.22	49.66	85.77	-0.183
UGTG02	469	52 972.26	5.01	48.29	86.84	-0.202
UGTGo4	493	54 852.32	5.58	40.18	93.75	-0.143
UGTGo6	469	52 660.09	5.31	40.46	84.58	-0.233
UGTGo7	468	52 743.1	4.97	49.85	89.12	-0.166

图1 棉花糖基转移酶UGT家族基因结构和二级结构预测注:A:UGT家族基因结构分析;B: UGT家族基因蛋白质二结构预测

Fig.1 Gene structure analysis and the secondary structures prediction of cotton glycosyltransferase UGT family Note: A:the gene structure analysis of UGT gene family; B: the secondary structures prediction of UGT gene family

图2 棉花糖基转移酶的氨基酸序列比对

Fig.2 Comparision analysis of amino acid sequences of cotton glycosyltranserses

图3 棉花糖基转移酶UGT基因家族蛋白质保守基序和保守结构域注:A:UGT家族蛋白质保守基序;B:UGT家族基因保守结构域分析

Fig.3 The analysis of protein conserved motifs and domain of cotton glycosyltransferase UGT gene family Note: A:protein conserved motif of UGT gene family;B.Conserved domain analysis of UGT gene family

图4 5个UGT基因在棉花不同组织的表达

Fig.4 Expression analysis of five UGTs in cotton different tissues

图5 不同植物UGTs分子进化树注:不同物种UGT不同颜色标记,拟南芥、山芥、水稻、大豆及棉花分别由绿色三角形、蓝色菱形、橙色方形、黄色倒三角形及红色圆形标记。

Fig.5 Phylogenetic tree of different plants UGTs Note:UGTs from the different species were labeled with different colored dots Arabidopsis thaliana, sorrel, rice, soybean, and cotton, represented as green triangles, blue diamonds, orange squares, yellow inverted triangles, and red circles, respectively.

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受棉铃虫诱导的棉花糖基转移酶UGT基因家族的生物信息学及表达分析

Bioinformatics and expression analysis of UGT gene family of cotton glycosyltransferase damaged by Helicoverpa armigera

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