The Expression BdDREB-30 Gene from Brachypodium distachyon and Functional Analysis of Its Promoter
HUANG Gang1*, TAN Sheng-Long2*, SUN Jing1, CHEN Li-Hong1**
1 Institute for Systems Biology, Jianghan University, Wuhan 430056, China;
2 School of Information and Communication Engineering, Hubei University of Economics, Wuhan 430205, China
Abstract：Dehydration responsive element binding protein (DREB) transcription factors play an important role in plant development and response to various stresses. The study on its DREB transcription factors of Brachypodium distachyon would promote the elucidation of molecular mechanism of the stress resistance of DREB transcription factors in gramineous crops. The early research work on the transcription profiles from the microarray study of DREB transcription factors showed that BdDREB-30 (GenBank No. XM_003561067.4) responds to various stresses. In order to further explore its function, the expression patterns of BdDREB-30 gene in different tissues and under 7 abiotic stresses were studied by qPCR, and the function of its promoter were studied by GUS staining. The results showed that the expression of BdDREB-30 was the highest in stems and its expression under drought and H2O2 stresses was significantly higher than that of the control. The results of multi-sequence alignment with other proteins showed that BdDREB-30 protein contained the typical AP2/ERF domain of DREB/CBF subfamily and the characteristic sequence of A-1 subclass. Evolutionary analysis showed that BdDREB-30 was relatively close to the homologous of wheat crops such as barley (Hordeum vulgare) and rye (Secale cereale). The analysis results of cis-acting elements of promoter showed that the promoter of BdDREB-30 gene contained many light response elements, plant hormone response elements and stress response elements. In order to further study the expression characteristics of the promoter, it was constructed into plant expression vector and transferred into tobacco (Nicotiana tabacum). The plant leaves of transgenic tobacco under low temperature, drought and salt stresses were stained with GUS. GUS staining showed that the promoter of BdDREB-30 gene was strongly induced under drought stress, indicated that the promoter of BdDREB-30 gene was a drought inducible promoter. This study provides a theoretical basis for the study of the molecular mechanism of BdDREB-30 gene in response to stresses.
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