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

doi:10.6048/j.issn.1001-4330.2023.10.007

Abstract

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

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

CLC Number:

S562

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.

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.

Figures/Tables 7

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

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

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

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

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

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

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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