Functional analysis of the MaERF058 gene in response to drought stress in Melilotus albus

doi:10.11686/cyxb2023338

Abstract

Abstract:

Drought is one of the most common stresses in agricultural production. It has a deleterious effect on plant growth and many aspects of development， such as photosynthetic function， stomatal aperture， osmotic balance， and hormone regulation， thereby affecting crop yield and quality. Melilotusalbus is a high-quality forage legume that is widely grown in the northern arid regions of China because of its high protein content and tolerance to drought， salinity， and barren environments. Transcription factors （TFs） are important proteins that regulate gene expression. They bind to specific cis-elements in the promoter regions of their target genes to activate or repress their transcription， thereby regulating plant growth and development in response to abiotic stress. Genes in the plant ethylene response factor （ERF） TF family respond to various abiotic stresses， and are important components of plant stress resistance mechanisms. MaERF058 is a key drought tolerance gene in M.albus. In this study， we analyzed the subcellular localization of MaERF058 in tobacco （Nicotiana tabacum）， and found that the expressed protein was located in the nucleus. Overexpression of MaERF058 in Arabidopsisthaliana significantly increased its drought tolerance， compared with that of wild type. Compared with the wild type， the MaERF058-overexpressinglines showed significantly increased proline content （P<0.01） and catalase activity （P<0.05） and significantly decreased malondialdehyde content （P<0.01） under drought conditions. Monoamine oxidase staining using the nitroblue tetrazolium method showed that the reactive oxygen content in the roots was higher in the MaERF058-overexpressinglines than in the control. These results suggest that the TF encoded by MaERF058 in M. albus has a positive regulatory role in the drought stress response.

Key words: Melilotusalbus, ERF transcription factor, drought stress, genetic transformation

Na WEI, Wen-mao JING, Er-wen XU, Rong-xin WANG, Jing-zhong ZHAO, Xue-e MA, Ji-yu ZHANG, Wen-xian LIU. Functional analysis of the MaERF058 gene in response to drought stress in Melilotus albus[J]. Acta Prataculturae Sinica, 2024, 33(8): 159-169.

Figures/Tables 8

Table 1 PCR primers used in the experiment

引物Primer	引物序列Primer sequence （5′-3′）
MsERF058-F	AACCAATGCATTGGATGTACGGAAATAGTAAT
MsERF058-R	GGATCCCGTTAGGAAACCAATAACTGTCC
35S-F	CTATCCTTCGCAAGACCCTTC
MsERF058-R1	GGATCCCGTTAGGAAACCAATAACTGTCC
HPT-F	GGTCGCGGAGGCTATGGATGC
HPT-R	GCTTCTGCGGGCGATTTGTGT

Fig. 1 Sequence and tissue expression pattern analysis of MaERF058

Fig. 2 Subcellular localization of MaERF058 protein in tobacco epidermis

Fig. 3 Identification of the expression level of MaERF058 transgenic Arabidopsis

Fig. 4 Evaluation of plate root length phenotype of Arabidopsis transgenic MaERF058 under drought stress

Fig. 5 PCR detection of MaERF058 transient transformation hairy root and green fluorescent protein detection

Fig.6 Expression identification of MaERF058 transient transformed hairy root

Fig. 7 Physiological indices of WT and transgenic hairy roots under control and drought treatment

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