小麦TaCOBL-5基因克隆及表达分析

doi:10.13304/j.nykjdb.2024.0172

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (6): 11-21.DOI: 10.13304/j.nykjdb.2024.0172

小麦TaCOBL-5基因克隆及表达分析

鲍新跃¹^,²(), 陈红敏³(), 王伟伟², 唐益苗², 房兆峰², 马锦绣², 汪德州²(), 左静红²(), 姚占军¹()

^1.河北农业大学农学院, 河北保定 071033
^2.北京市农林科学院杂交小麦研究所, 北京 100097
^3.濮阳市现代农业发展中心, 河南濮阳 457177

收稿日期:2024-03-07 接受日期:2024-03-26 出版日期:2024-06-15 发布日期:2024-06-12
通讯作者: 汪德州,左静红,姚占军
作者简介:鲍新跃E-mail：1772145342@qq.com
陈红敏E-mail：hm-chen88@163.com第一联系人：鲍新跃与陈红敏为共同第一作者。
基金资助:
北京市农林科学院协同创新中心资助项目(KJCX201907-2);北京市农林科学院青年科研基金项目(QNJJ202316)

Cloning and Expression Analysis of Wheat TaCOBL-5 Genes

Xinyue BAO¹^,²(), Hongmin CHEN³(), Weiwei WANG², Yimiao TANG², Zhaofeng FANG², Jinxiu MA², Dezhou WANG²(), Jinghong ZUO²(), Zhanjun YAO¹()

^1.College of Agronomy，Hebei Agricultural University，Hebei Baoding 071001，China
^2.Institute of Hybrid Wheat，Beijing Academy of Agriculture and Forestry Sciences，Beijing 100097，China
^3.Puyang Modern Agricultural Development Center，Henan Puyang 457177，China

Received:2024-03-07 Accepted:2024-03-26 Online:2024-06-15 Published:2024-06-12
Contact: Dezhou WANG,Jinghong ZUO,Zhanjun YAO

摘要/Abstract

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

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

中图分类号:

S512

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.

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.

图/表 10

表1 基因克隆和荧光定量的引物序列

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

表2 TaCOBL-5基因信息及其编码蛋白的理化性质分析

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

图1 TaCOBL-5基因结构、保守基序及启动子顺式作用元件分析A：基因结构；B：保守序列；C：启动子顺式作用元件

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

图2 TaCOBL-5蛋白系统进化分析

Fig. 2 Phylogenetic analysis of TaCOBL-5 proteins

图3 COBRA结构域及三级结构预测A： COBRA结构域；B：COBL蛋白三级结构

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

图4 TaCOBL-5蛋白的亲/疏水性和GPI修饰位点预测

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

图5 小麦miRNA调控网络预测及互作蛋白预测A： miRNA调控网络；B：蛋白互作网络

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

图6 TaCOBL-5基因组织表达模式分析

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

图7 低温、干旱、热、盐胁迫下TaCOBL-5基因表达模式

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

图8 不同抗性材料干旱胁迫、高温胁迫处理下TaCOBL-5D表达模式分析A：干旱胁迫；B：高温胁迫

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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小麦TaCOBL-5基因克隆及表达分析

Cloning and Expression Analysis of Wheat TaCOBL-5 Genes

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