Cloning and Expression Analysis of Wheat TaCOBL-5 Genes

doi:10.13304/j.nykjdb.2024.0172

Abstract

Abstract:

Wheat yield directly impacts food security in China， but various abiotic stresses such as drought， low temperature， salt and high temperature severely constrain wheat production. In previous analysis， transcriptomic data revealed differential expression of wheat TaCOBL-5D under multiple abiotic stresses. TaCOBL-5D and its homologous genes TaCOBL-5A and TaCOBL-5B were cloned and obtained， and the comprehensive bioinformatics and expression pattern were analyzed. The results showed that TaCOBL-5 genes exhibited significant conservation in gene structure， protein tertiary structure， conserved domains， and promoter regulatory elements with COBL genes from other species. TaCOBL-5 genes exhibited the highest expression level in roots and responded differently to various abiotic stresses， especially significantly down-regulated under drought stress， indicating its importance under drought stress； it also showed different responses to low temperature， high temperature， and salt stress. In addition， the expression level of the TaCOBL-5D gene varied significantly among different drought-resistant and high-temperature-resistant materials， further suggesting its important role in stress resistance. These findings laid the foundation for further understanding of the function of COBL genes in wheat and provided scientific support for wheat breeding efforts aimed at enhancing stress tolerance.

Key words: wheat, COBL genes, bioinformatics analysis, abiotic stress, expression pattern

摘要：

小麦产量关系我国粮食安全，干旱、低温、盐害和高温等非生物胁迫严重制约小麦产量增长。前期转录组分析发现小麦TaCOBL-5D在多种非生物胁迫下差异表达。克隆并获取TaCOBL-5D及其同源基因TaCOBL-5A、TaCOBL-5B，并对其生物信息学特性及表达模式进行了分析。结果显示，TaCOBL-5与其他物种的COBL基因在基因结构、蛋白三级结构、保守结构域以及启动子调控元件方面表现出明显的保守性。TaCOBL-5在根部表达量最高，并对各种非生物胁迫响应不同，尤其是在干旱胁迫下调控显著，表明其在干旱胁迫中的重要性，同时对低温、高温和盐胁迫也有不同响应。此外，TaCOBL-5D基因在不同干旱抗性及高温抗性材料中表达量差异显著，进一步暗示其在逆境胁迫中具有重要作用。这些研究结果有助于理解COBL基因在小麦中的功能，同时为小麦抗逆育种提供科学支持。

关键词: 小麦, COBL基因, 生物信息学分析, 非生物胁迫, 表达模式

CLC Number:

S512

Xinyue BAO, Hongmin CHEN, Weiwei WANG, Yimiao TANG, Zhaofeng FANG, Jinxiu MA, Dezhou WANG, Jinghong ZUO, Zhanjun YAO. Cloning and Expression Analysis of Wheat TaCOBL-5 Genes[J]. Journal of Agricultural Science and Technology, 2024, 26(6): 11-21.

鲍新跃, 陈红敏, 王伟伟, 唐益苗, 房兆峰, 马锦绣, 汪德州, 左静红, 姚占军. 小麦TaCOBL-5基因克隆及表达分析[J]. 中国农业科技导报, 2024, 26(6): 11-21.

Figures/Tables 10

Table 1 Primers used in this study for gene cloning and qPCR

引物名称 Primer name	正向引物序列 Forward primer sequence （3’-5’）	反向引物序列 Reverse primer sequence （3’-5’）
TaCOBL-5A	GCGGCACCCGTGTCTTCTAT	CGTCTCGTCTCGTCGCAGTA
TaCOBL-5B	GCGGCACCCATGTCTTCTAT	CGTCTCGTCTCGTCGCAGTA
TaCOBL-5D	ACGGCACCCGCGTCTTCTAT	TCTCGTCGCTGTAAAAACTG
qTaCOBL-5A	CGTTGGATCTCTCTTGCAGC	TGGGATGGTCATGGGCAAAG
qTaCOBL-5B	GATTACGTGCAGGTTACATTCC	TCTCAAGGCTCCAGGTCAGG
qTaCOBL-5D	CAGCGAATCATAAGCCTCTG	GAGTAGCGGGGCAGGAAATG
TaActin	GGAATCCATGAGACCACCTAC	GACCCAGACAACTCGCAAC

Table. 2 TaCOBL-5 gene information and physicochemical properties analysis

基因

Gene

基因号

Gene ID number

物理位置

Physical position/bp

分子量

Molecular weight/Da

等电点

蛋白长度

Protein length/aa

预测定位

Predicted location

TaCOBL-5A

TraesCS5A02G392000

588 375 577~588 379 139

574 675 118~574 678 654

467 600 940~467 604 428

Fig. 1 Analysis of gene structure， conserved motif and cis-acting regulatory elements of TaCOBL-5A： Gene structure； B： Conserved motif； C： Cis-acting regulatory elements

Fig. 2 Phylogenetic analysis of TaCOBL-5 proteins

Fig. 3 Analysis of conserved COBRA domain and the tertiary structure of COBL proteinsA： COBRA domain； B： Tertiary structure of COBL protein

Fig. 4 Prediction of hydropath and GPI modification sites of TaCOBL-5 proteins

Fig. 5 Prediction putative networks of wheat miRNAs and the interacting proteinsA： Putative network of wheat miRNAs； B： Network of interacting proteins

Fig. 6 Analysis of tissue expression patterns of TaCOBL-5 genes

Fig. 7 Expression profiles of TaCOBL-5 genes under cold， drought， heat and salt stress

Fig. 8 Expression profiles of TaCOBL-5D among different resistant materials under drought and heat stressesA： Drought stress； B： Heat stress

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