Genome-wide Analysis of Dof Transcription Factors in Elephant Grass and Their Response to Cold Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-1105

Abstract

Abstract:

Objective To analyze the characteristics of the Dof transcription factor family in elephant grass (Cenchrus purpureus), and its response mechanism to cold stress, and conduct genome-wide identification of CpDof genes, and providing support for mining cold-tolerant candidate genes in elephant grass. Method Based on the elephant grass genome data, CpDof genes were identified through conserved domain search. Bioinformatics methods were used to analyze their physicochemical properties, chromosome localization, phylogeny, and cis-acting element characteristics. Taking ‘Chuanxu 4’ elephant grass as the material, after the treatment by 4 ℃ cold stress, transcriptome sequencing and quantitative real-time PCR were used to analyze gene expression characteristics. Key genes were cloned to construct yeast expression vectors, and heterologous expression experiments were conducted to verify cold tolerance function. Result A total of 68 CpDof genes were identified in the elephant grass genome, unevenly distributed on 14 chromosomes. Collinearity analysis revealed 51 pairs of segmental duplications and 2 pairs of tandem duplications among CpDof genes. Subcellular localization prediction showed that almost all CpDof family members were localized in the nucleus. Under cold stress induction, 28 CpDof genes were significantly upregulated, among which CpDof17 and CpDof28 had relatively prominent expressions. They were highly expressed in the roots and stems, respectively, and cold stress enhanced their expression in the roots and leaves. Yeast heterologous expression experiments showed that the overexpression of CpDof28 significantly improved the cold tolerance of Saccharomyces cerevisiae, while CpDof17 had no obvious effect. Conclusion CpDof28 may act as a key regulatory factor playing a core role in the response of elephant grass to cold stress. This study clarifies the core characteristics of the Dof gene family in elephant grass and screenes the cold-tolerant candidate gene CpDof28, providing important genetic resources and theoretical basis for cold-resistant molecular breeding and variety improvement of elephant grass.

Key words: Cenchrus purpureus, Dof, cold stress, transcription factor, expression analysis, CpDof28, heterologous expression in yeast, cold tolerance

ZHANG Ya-ning, MAO Chun-li, HU Zhi-yong, YANG Dan, WU Jia-hai, WANG Bi-xian, HUANG Lin-kai. Genome-wide Analysis of Dof Transcription Factors in Elephant Grass and Their Response to Cold Stress[J]. Biotechnology Bulletin, 2026, 42(6): 267-278.

Figures/Tables 10

Fig. 1 Chromosomal distribution of CpDof genesVertical bars indicate chromosome density, and the scale on the left indicates chromosome length

Fig. 2 Distribution and collinearity analysis of Dof in the genome of elephant grass (Cenchrus purpureus)

Fig. 3 Synteny analysis of Dof genes between elephant grass and A. thaliana and O. sativa

Fig. 4 Phylogenetic tree of Dof members in elephant grass, rice (O. sativa) and A. thaliana

Fig. 5 Analysis of cis-acting elements (CREs) of CpDof

Fig. 6 Phylogenetic tree, conserved motifs, conserved domains, and gene structures of CpDof family members

Fig. 7 Expressions of Dof genes in the ‘Chuanxchu No. 4’ elephant grass genome under low-temperature stress conditions

Fig. 8 Expressions of 10 CpDof genes under low temperature (4 ℃) treatmentAll experiments were independently repeated three times. Error bars indicate the standard deviation of three replicates. * indicate significant differences in transcript levels compared to the blank control (0 h) (* P0.05, ** P0.01, *** P0.001, **** P0.000 1)

Fig. 9 Expressions of 10 CpDof genes in different tissuesAll experiments were independently repeated three times. Different letters indicate statistically significant differences (P0.05) according to Tukey’s test

Fig. 10 The tolerance of yeast cells overexpressing CpDof17 and CpDof28 to low-temperature stressA, B: Serially diluted drops of yeast transformants without or with cold treatment. C, D: Growth curves of cold-treated transgenic yeast and blank control group yeast strains in liquid medium

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