Cloning,Subcellular Localization and Expression Analysis of TaWRKY70 in Wheat

doi:10.7668/hbnxb.20195906

Abstract

Abstract:

To elucidate the role of WRKY transcription factor family members in the dynamic regulation of wheat growth and development,as well as in responses to abiotic stresses,this study investigated the expression patterns of TaWRKY gene under drought,high salinity,and low-temperature stress conditions.Using the common wheat cultivar Chinese Spring as the experimental material,we obtained the TaWRKY70 gene through molecular cloning.The coding sequence of TaWRKY70 was 885 bp in length,encoding a 294-amino-acid hydrophilic and unstable protein.Bioinformatics analysis revealed that the protein possessed a typical WRKYGQK conserved domain and a C₂HC-type zinc finger structure,classifying it as a Group Ⅲ WRKY transcription factor.Cis-regulatory element analysis of the TaWRKY70 promoter region identified regulatory elements involved in responses to methyl jasmonate,abscisic acid,and ethylene.Co-expression gene analysis suggested that TaWRKY70 was associated with multiple stress response processes in wheat,including hormone signaling,defense against microbial pathogens,and responses to cold stress.Phylogenetic analysis indicated that TaWRKY70 shared a close evolutionary relationship with WRKY70 proteins from other Poaceae species,such as barley,maize,sorghum,and foxtail millet.Subcellular localization experiment further confirmed that TaWRKY70 was localized in the nucleus,consistent with the characteristics of a transcription factor.Expression pattern analysis showed that TaWRKY70 was expressed in wheat roots,stems,leaves,young spikes,and grains,with higher expression levels observed in roots and leaves.Under abiotic stress conditions,TaWRKY70 expression was downregulated in response to abscisic acid and low-temperature treatments but upregulated under salicylic acid,NaCl,PEG6000,and high-temperature treatments.In conclusion,the cloning of TaWRKY70 gene and analysis of its expression pattern provide a basis for the next step to analyze the molecular mechanism of TaWRKY70 involved in wheat stress resistance.

Key words: Wheat, TaWRKY70, Gene cloning, Subcellular localization, Expression analysis

CLC Number:

LU Zhenhua, LIANG Chen, ZHANG Li, TONG Kexin, CHEN Xiaoqiang, LI Ming, DING Bo, QIU Lina, XIE Xiaodong, WANG Junbin. Cloning,Subcellular Localization and Expression Analysis of TaWRKY70 in Wheat[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 12-19. doi: 10.7668/hbnxb.20195906.

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Figures/Tables 9

Fig.1 PCR amplification of TaWRKY70 in wheat M.2K PlusⅡ DNA Marker;1,2,3.Fragments of TaWRKY70; 4.Negative control(H₂O).

Fig.2 Analysis of amino acid sequences and conserved domains encoded by TaWRKY70 The shaded area represents the conserved WRKY domain.The sequence with underline means WRKYGQK core sequence and the C₂HC zinc-finger structure are boxed.

Tab.1 Cis-acting elements and functional annotations of TaWRKY70 promoter

顺式作用元件 Cis-acting elements	数量 Number	功能 Function
TGA-element	1	生长素响应元件
TGACG-motif	8	茉莉酸甲酯响应元件
CGTCA-motif	8	茉莉酸甲酯响应元件
TCA-element	1	水杨酸响应元件
CCGTCC-box	8	乙烯响应元件
TATC-box	1	赤霉素响应元件
ABRE	1	ABA响应元件
TCCC-motif	3	光响应元件
Sp1	2	光响应元件
Box 4	2	光响应元件
MBS	1	与干旱诱导相关的MYB结合位点
GCN4_motif	1	参与胚乳表达的顺式调控元件
GC-motif	3	参与缺氧特异性诱导的增强子样元件

Fig.3 GO enrichment analysis of TaWRKY70 co-expression genes Color represents the enrichment level;size represents the number of enriched genes.

Fig.4 Phylogenetic tree of TaWRKY70 and its homologous WRKY70 proteins

Fig.5 Subcellular localization of TaWRKY70

Fig.6 Tissue-specific expression analysis of the TaWRKY70 gene Different lowercase letters indicate significant differences(P<0.05).The same as Fig.8.

Fig.7 Relative expression heatmap of TaWRKY70 gene under different abiotic stresses

Fig.8 Analysis of expression patterns of TaWRKY70 under different stress treatment conditions

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