The Cloning And Functional Analysis Of Transcription Factor GmERF2 And GmNF-YC5 In Soybean

Drought, low temperature, high temperature and salt and other adverse factors seriously endanger the growth and development of plants and its yield and quality, and even lead to death. In the long process of evolution, the plant has formed a complex and effective defense mechanism in order to adapt to these unfavorable factors and adverse environment. Hence, revealing complex signal transduction pathways and gene networks of abiotics stress-responsive gene expression is signicicant for dissecting the mechanisms of plants adaptation to stress, and crop breeding. In addition, the process of gene expression plays a key role in the process of plant response, and transcription factors, as the most important regulation factor, are significant in research.In previous work, overexpression of GmNF-YA20 was proven to enhance stress tolerance of transgenic plants. In order to study regulating mechanism of GmNF-YA20 protein, we cloned some genes encoding proteins interacting with GmNF-YA20 by using yeast two hybrid system. Among them, GmERF2 and GmNF-YC5 were selected, ERF and NF-Y transcription factor family had significant effect on growth and development of plants and plays an important role in regulation of biotic and abiotic stresses. The characteristics and functions of GmERF2 and GmNF-YC5 were showed as following:1. The cloning and function identification of GmERF2 in soybean. In our study, we cloned the GmERF2 gene from soybean. Protein sequences showed that GmERF2 contained an AP2 motif. Subcellular localization analysis showed that GmERF2 gene location in the cell nucleus. The expression pattern of this gene was responsive to under BR, ethylene, drought and salt stress. Through genetic transformation, we obtained GmERF2 transgenic plants in Arabidopsis. Compared to mutant and wild type, we found that transgenic Arabidopsis presented a strong growth phenotype, which suggested that GmERF2 functioned as a positive gene in high concentration salt signaling pathways. In order to study the regulation mechanism of GmERF2 on the protein level, the yeast two hybrid systems was used to screen the library, and the activation of transcription was verified.2. The cloning and function identification of NF-Y transcription factors GmNF-YC5. In our study, the GmNF-YC5 gene was cloned from soybean. Protein sequences showed that GmNF-YC5 contained a HAP5 motif. Subcellular localization analysis showed that GmNF-YC5 gene was located in the cell membrane. The expression pattern of this gene was responsive to under BR and drought st ress. Through genetic transformation, we obtained GmNF-YC5 transgenic plants in Arabidopsis. Compared to mutant and wild type, we found that transgenic Arabidopsis presented a strong growth phenotype, which suggested that GmNF-YC5 functioned as a negative gene in salt signaling pathways.In summary, two important stress-related genes GmERF2 and GmNF-YC5 were cloned from soybean. The results of expression analysis showed that those two genes involved in all kinds of stress responses in plants. It provides important theoretical basis for studying the molecular mechanisms involved in the regulation of plant response to stress and the regulation mechanism of plant stress of those two genes.