Bioinformatics Analysis of Peanut Transcription Factor AhHDZ70 and Its Tolerances to Salt and Drought

doi:10.13560/j.cnki.biotech.bull.1985.2025-0901

Abstract

Abstract:

Objective Drought and salinity stress are critical environmental factors that severely affect the yield and quality of peanuts (Arachis hypogaea L.) in China. Investigating the structure and function of AhHDZ70 (Ah12g074600.1) gene will reveal its function in tolerance of peanut to drought and salt, and provide essential gene resources for peanut salt resistance and drought resistance breeding. Method Bioinformatics method were applied to analyze the physical and chemical properties, gene structure, phylogeny, cis-element of promoter, protein interaction of AhHDZ70. Fluorescence quantitative PCR technology was used to identify the expressions of AhHDZ70 gene in different tissues and organs, and under salt, drought, high temperature and low temperature stresses. Subcellular localization analysis was performed using tobacco transient transformation technology. Genetic transformation was carried out in Arabidopsis and biological function of AhHDZ70 was further verified by salt and drought stress treatment. Result AhHDZ70 encoded 326 amino acids, which was an unstable hydrophilic protein with no signal peptide, its secondary and tertiary structures were mainly characterized by irregular curling. Domain analysis showed that AhHDZ70 contained 4 exons and 3 introns. The AhHDZ70 promoter region contained two drought-induced functional elements (MBS motif), one adversity stress response element (TC-richrepeats motif), and one cryogenic response element (LTR motif). The main form of its secondary structure and tertiary structure was irregular curl. Transcriptome combined with RT-qPCR analysis showed that AhHDZ70 had the highest expression in seeds and responded positively to salt, drought, high temperature and low temperature stress. The results of subcellular localization showed that AhHDZ70 was located in the nucleus. AhHDZ70 heterologously overexpressed Arabidopsis to obtain homozygous transgenic plants. Under salt and drought stress, the germination rate of AhHDZ70-overexpressed transgenic lines was significantly higher than that of WT, and the root length was significantly longer than WT. Conclusion AhHDZ70 belongs to the HD-ZIP II transcription factor and is located in the nucleus. AhHDZ70 might play a regulatory role in the growth and development of various tissues in peanuts. Additionally, AhHDZ70 is likely involved in positively regulating peanut's response to salt and drought stress.

Key words: peanut, transcription factor, AhHDZ70, bioinformatics analysis, expression pattern, tolerance to salt, tolerance to drought, gene function

WU Cui-cui, CHEN Deng-ke, LAN Gang, XIA Zhi, LI Peng-bo. Bioinformatics Analysis of Peanut Transcription Factor AhHDZ70 and Its Tolerances to Salt and Drought[J]. Biotechnology Bulletin, 2026, 42(1): 198-207.

Figures/Tables 8

Table 1 Primers used in the study

基因名称 Gene name	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）	实验类型 Type of experiment
AhHDZ70	TTGGAGAGAACACGGGGGACGAATTCATGGAATATGCAACATATTCATCAGC	GCCTGCAGGTCGACTCTAGAGGATCCGGACCAAAAGTCCCACCATTG	过表达实验、亚定位实验、过表达PCR鉴定
β-Actin	CAGGATTTGCCGGTGATGATG	TCTGTTGGCCTTCGGGTTGAG	内参
AhHDZ70	AGATGAGTTTGGGTTTGAGT	GAGGTAGAGGCTTCTGGAT	RT-qPCR

Fig. 1 Phylogenetic tree analysis of homologous proteins between AhHDZ70 and other speciesAh: Arachis hypogaea; Ad: Arachis duranensis; Ca: Cicer arietinum; Cc: Cajanus cajan; Gm: Glycine max; Gs: Glycine soja; La: Lupinus angustifolius; Mt: Medicago truncatula; Ps: Pisum sativum; Pv: Phaseolus vulgaris

Fig. 2 Analysis of AhHDZ70 promoter (A) and prediction of interaction protein (B)

Fig. 3 Analysis of AhHDZ70 expression patterns in tissues, organs, and abiotic stressA: Expression patterns of AhHDZ70 in 22 tissues and organs. B: Expression patterns of AhHDZ70 under abiotic stress. L: Leaf. R: Root

Fig. 4 RT-qPCR identification of AhHDZ70 in different tissues and organs of peanut under abiotic stressA: RT-qPCR analysis of AhHDZ70 in different tissues and organs of peanuts. B-E: RT-qPCR analysis of AhHDZ70 under salt, drought, low temperature and high temperature stress. The difference significance was analyzed by two-tailed t-test. Error bars indicate mean ±SD, *P<0.05, **P<0.01,the same below

Fig. 5 Subcellular localizations of AhHDZ70 proteins in tobacco leaves (bar=50 μm)

Fig. 6 Identification of the germination rate in AhHDZ70 overexpression lines under salt and drought stress

Fig. 7 Root length experiments of salt stress and drought stress in AhHDZ70 overexpressed strain

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