| Soybean(Glycine max L.)is one of the most important oil crops in China.Aiotic stresses such as high salt and drought are important environmental factors affecting the growth and development of soybean,leading to the serious reduction of soybean yield and quality,which seriously affects the food security in China.It is of great significance to develop the new varieties and improve the soybean yield and quality.Ethylene response factors(ERFs)are an important class of transcription factors,is one of the most important transcription factors in plants,involved in plant growth and development,signal transduction,biological and abiotic stress response and other biological processes.Currently,there are few reports on the involvement of soybean ERF ethylene response factors in regulating plant stress resistance.The functional study of soybean ERF in stress stress response will provide basic information for the development of new salt-tolerant and drought-tolerant soybean varieties.In this study,the whole-genome identification and bioinformatics analysis of soybean GmERF gene family were carried out,and based on the transcriptomic data of soybean salt and drought stress and q RT-PCR expression pattern analysis,the highest expressed GmERF13 gene was selected as the research objects,and the molecular characteristics of GmERF13 gene and the functional identification of salt and drought resistance were further investigated.The main results are as follows:1.Genome-wide identification of the GmERF gene family.A total of 129 GmERF gene family members(GmERF1-GmERF129)were identified in the whole soybean genome and were unevenly distributed on 20 chromosomes.Phylogenetic analysis revealed that the predicted GmERF proteins can be divided into eight subfamilies.Analysis of the promoter region of GmERF contains cis-acting elements associated with hormonal regulation and abiotic stress.GmERF13 Family genes of the subfamily have relatively conserved gene structure and conserved motifs.2.Analysis of the expression patterns of GmERF13 genes under salt and drought stress.The q RT-PCR results indicated that the GmERF13 gene expression was significantly up-regulated under both salt and drought stress,possibly related to soybean response to salt and drought stress.Further molecular characterization and functional identification of salt tolerance and drought resistance.3.GmERF13 Molecular characterization and analysis.Bioinformatic analysis revealed that the GmERF13 gene has a CDS of 648 bp,encoding 215 amino acids,a GmERF13 molecular weight of 23.84 k Da and an isoelectric point of 5.12,and is a stable hydrophilic protein.Subcellular localization analysis indicated that the GmERF13 protein localized to the nucleus.4.Overexpression of GmERF13 increased salt tolerance and drought resistance of transgenic Arabidopsis plants.Arabidopsis inflorescence was infected with Agrobacterium tumefaciens containing the p CAMBIA1302:GmERF13 recombinant plasmid and selected positive plants with stable inheritance of the GmERF13 gene in T3 generation.The Arabidopsis strain of the GmERF13 gene in T3 was tested under salt and drought stress.The results showed that the chlorophyll and proline contents of GmERF13 transgenic plants were significantly higher than WT and significantly lower than WT Arabidopsis plants under drought and salt stress.Suggesting that the transgenic GmERF13 Arabidopsis lines have strong salt tolerance and drought resistance.5.Overexpression of GmERF13 significantly enhanced salt tolerance and drought tolerance in transgenic soybean hairy root complex.We constructed soybean hairy root complex plants with GmERF13 overexpression and RNAi.The results of stress resistance under salt and drought stress showed that the content of proline,catalase,peroxidase,superoxide dismutase and chlorophyll of the GmERF13 gene,while the malondialdehyde was significantly lower than that of the control group.The results showed that GmERF13 transgenic hairy root complex plants under salt and drought stress. |
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