MfERF053 enhances drought resistance of Arabidopsis thaliana by improving reactive oxygen species scavenging capacity and promoting expression of stress-resistance-related genes

doi:10.11686/cyxb2025359

Abstract

Abstract:

Drought stress restricts plant growth and agricultural production， and mining drought-tolerannce genes and analyzing their mechanisms are crucial for molecular breeding. Medicago falcataMfERF053 belongs to the AP2/ERF transcription factor family. Previous studies have shown that its expression is upregulated under drought stress， and Arabidopsis thaliana overexpressing this gene exhibits a drought-tolerant phenotype. However， the underlying mechanism remains unclear. In this study， A. thaliana Col-0 wild type （Col） and MfERF053-overexpressing lines （#19， #20， #33） were used as materials. After natural drought stress treatment， plant phenotypic， physiological， and molecular indicators were determined to clarify the drought-tolerance mechanism of MfERF053. The results showed that under drought stress： the overexpressing lines exhibited delayed leaf wilting and better recovery ability after rehydration； their relative water content （RWC） was significantly higher than that of Col （P<0.05）， while their relative electrical conductivity （REC） and malondialdehyde （MDA） content were significantly lower than those of Col （P<0.05）； the function of photosystem Ⅱ （PSⅡ） was more stable， with the maximum photochemical efficiency （F_v/F_m） and electron transport rate （ETR） significantly higher than those of Col （P<0.05）； the degree of nitroblue tetrazolium （NBT）［for superoxide anion （·O??） localization］ and 3，3'-diaminobenzidine （DAB）［for hydrogen peroxide （H?O?） localization］ staining in leaves of the overexpressing lines was significantly lighter than that in Col， indicating reduced accumulation of ·O?? and H?O?， and the activities of catalase （CAT） and peroxidase （POD） were significantly higher than those in Col （P<0.05）. At the molecular level， under drought stress， the expression levels of stress-tolerance-related genes in MfERF053-overexpressing lines， including reactive oxygen species （ROS）-scavenging-related genes （AtCAT1， AtPOD3， AtSOD4）， drought-responsive genes （AtRD22， AtRD29A）， cold/drought cross-responsive genes （AtCOR15A， AtCOR47）， potassium ion transport gene （AtHAK5）， and abscisic acid （ABA） signaling pathway gene （AtABI5）， were all significantly up-regulated （P<0.05）. Most of these genes exhibited significantly higher expression levels in overexpressing lines than in Col （P<0.05）， except that AtRD29A and AtABI5 in individual overexpressing lines showed no significant difference compared with Col. In conclusion， MfERF053 was found to enhance ROS scavenging capacity， upregulate the expression of stress-tolerant-related genes， and synergistically improve the physiological homeostasis and molecular response of A. thaliana under drought， ultimately improving its drought tolerance. This study provides important genetic resources and a theoretical basis for the molecular breeding of drought-tolerant crops/forages.

Key words: MfERF053, Arabidopsis thaliana, drought resistance, reactive oxygen species scavenging, stress-resistant-related genes

Qian LI, Wei DUAN, Xue-li ZHANG, Li-jun LIU, Yu-xiang WANG. MfERF053 enhances drought resistance of Arabidopsis thaliana by improving reactive oxygen species scavenging capacity and promoting expression of stress-resistance-related genes[J]. Acta Prataculturae Sinica, 2026, 35(8): 157-169.

Figures/Tables 7

Table 1 PCR primers

引物 Primer	正向引物/反向引物 Forward primer/reverse primer （5′-3′）
qAtactin-F	CGCCATCCAAGCTGTTCTC
qAtactin-R	TCACGTCCAGCAAGGTCAAG
AtCAT1-F	GTCCTGGGATTCAGACAGGC
AtCAT1-R	GGCTCACGTTAAGACGAGT
AtPOD3-F	CCAATCCAGAAACGGAAGTG
AtPOD3-R	TCTGACGCTTCTGACGACAC
AtSOD4-F	GAAGAACCTTGCTCCTTCCAG
AtSOD4-R	GATTGGCAGTTGTGTCAACAAC
AtRD22-F	ACTTGGTAAATATCACGTCAGGCT
AtRD22-R	CTGAGGTGTTCTTGTGGCATACC
AtRD29A-F	GATATCGACAAGGATGTGCCG
AtRD29A-R	GTATCCAGGTCTTCCCTTCGC
AtCOR47-F	GCTTCGTCGTCGTTTCTCT
AtCOR47-R	CCAATGTATCTGCGGTTTGA
AtHAK5-F	TGCATCACTGGGACGGAGGCA
AtHAK5-R	GTACGCAGCCTGCCCGCAAT
AtABI5-F	AGAGGGATAGCGAACGAGTCTAGTC
AtABI5-R	GTTCGGTTTGGATTAGGTTTAGG

Fig.1 Phenotypes of wild-type and MfERF053-overexpressing A. thaliana under natural drought stress

Fig.2 Comparison of relative water content， relative electrical conductivity， and malondialdehyde content between wild-type and MfERF053-overexpressing A. thaliana under drought stress

Fig.3 Comparison of chlorophyll fluorescence between wild-type and MfERF053-overexpressing A. thaliana under drought stress

Fig.4 Comparison of reactive oxygen species content and antioxidant enzyme activities between wild-type and MfERF053-overexpressing A. thaliana under drought stress

Fig.5 Comparison of antioxidant enzyme-related gene expression between wild-type and MfERF053-overexpressing A. thaliana under drought stress

Fig.6 Relative expression of stress-resistant-related genes in wild-type and MfERF053-overexpressing A. thaliana under drought stress

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