Isolation And Characteirzation Of A DREB/CBF Transcirption Factor, GmDREB1A, From Soybean (Glycine Max L.)

Soybean(Glycine max L.) is a most important grain and oill crop,it’s the mainsource of vegetable protein and edible oil, and also important industrial rawmaterials. However, the adverse environmental factors, such as drought, soilsalinization and cold, seriously affect the growth and development of soybean whichbring terrible loss to it. Therefore, cultivating and planting cold,drought,saltresistance soybean varieties is one way of achieving continued to increase soybeanyield and production for sustainable development. Molecular biology and geneticanalysis showed that the transcription factor involved in the expression of a numberof stress-related genes. With the rapidly development of molecular biology andplant genetic engineering technology, it’s of great importance for improvingsoybean resistance by genetically reforming transcription factors and coloingresistance-related transcription factors,transcription factors played vital roles inresponse to biotic and abiotic stresses. In this paper,a new DREB/CBF transcriptionfactors, GmDREB1A, were isolated from soybean Jilin32and analyzed it’s function.The experiment results are as following:1. GmDREB1A was cloned by RT-PCR in soybean Jilin32cDNA. The ORF ofGmDREB1A is957bp,encoding a putative protein of318amino acids with apredicted molecular mass of35.57kDa. The predicted GmDREB1A protein containeda conserved AP2/ERF domain,and analysis of online software suggests that theputative protein located in nucleus. The amino acid sequence alignment andphylogenetic tree analysis indicated that GmDREB1A was grouped into ClassⅥ inDREB/CBF transcription factors subfamily.2. Expressional analysis of GmDREB1A in E.coli Rosetta by SDS-PAGE showed that GmDREB1A protein highly expressed in prokaryotic systems.3. The GmDREB1A∷eGFP fusion gene was obtained by overlapping PCR andtransformed into onion epidermal cells by Agrobacterium-mediated transientexpression. The results showed that the GmDREB1A protein was targeted to thenucleus.4. GmDREB1A was expressed in yeast to analyze transcriptional activation. Theresults indicated that GmDREB1A has transcriptional activation ability in yeast whichsuggested it’s a transcription activator.5. The expression patterns of GmDREB1A was analyzed using quantitative real-time PCR. The results showed GmDREB1A had no tissue-specific expressioncharacteristic which expressed in roots, leaves, flowers, stemsand immature embryos.GmDREB1A responded differently to biotic and abiotic stresses such as salt,drought,cold, wounding, ABA and SA,.6. The plant expression vector pCB35SR1R2-GFP-GmDREB1A was constructedusing GATEWAY clone technology. and transformed into ArabidopsisCol-0usingfloral dipping method. T3generation transgenic Arabidopsis was used to analyze thestress characteristics. The results indicated that overexpression of GmDREB1Aenhanced tolerance to high salt and drought in transgenic Arabidopsis.