Cloning SmERF B3-45 from Salix matsudana and Functional Analysis on Its Tolerance to Salt
[Objective]To validate whether SmERF B3-45 plays a positive regulatory role in plant response to salt stress may lay the foundation for revealing the role of AP2/ERF transcription factors in regulating the salt tolerance of Salix matsudana.[Method]The cis-acting elements in the promoter region of SmERF B3-45 from the AP2/ERF superfamily of Populus euphratica were analyzed.The full-length CDS sequence of SmERF B3-45 was cloned using specific primers,and bioinformatics and subcellular localization analysis were conducted.The function was elucidated by constructing overexpression vectors and transforming Arabidopsis thaliana mutants ERF-OE1 and ERF-OE2,and by using virus-induced gene silencing(VIGS).[Result]Cis-acting element analysis suggested that SmERF B3-45 may be involved in the expression regulation pathways responding to stress.RT-qPCR results showed that the expression of SmERF B3-45 was induced by NaCl treatment and was widely expressed in different tissues of Populus euphratica.Subcellular localization indicated that the SmERF B3-45 protein was localized in the nucleus.In transgenic Arabidopsis,the expression of SmERF B3-45 significantly increased.Under salt stress,compared with the wild type,the root length of overexpressing SmERF B3-45 Arabidopsis significantly increased,while the total protein content,Na+content,MDA content,and Na+/K+ratio significantly decreased,and the CAT content and K+content significantly increased.Gene-silenced plants showed a significant downregulation of SmERF B3-45 expressions.Compared with the control,the total protein content of the gene-silenced plants significantly reduced,while MDA and proline content were significantly higher than that of the negative control plants.Additionally,the silenced plants demonstrated wilting leaves,indicating that the silencing of SmERF B3-45 reduced the tolerance of Salix matsudana to salt.[Conclusion]SmERF B3-45 is confirmed as a positive regulatory transcription factor in the plant response to salt stress.
万方数据
维普
