Journal
GENE
Volume 549, Issue 1, Pages 77-84Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.gene.2014.07.049
Keywords
Ammopiptanthus nanus; Betaine aldehyde dehydrogenase; Heat; Heterologous expression; Salt; Tolerance
Categories
Funding
- National Key Science and Technology Special Project [2013ZX08003-005]
- National Natural Science Foundation of China [31071433]
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Betaine aldehyde dehydrogenase (BADH) catalyzes the synthesis of glycine betaine, a regulator of osmosis, and therefore BADH is considered to play a significant role in response of plants to abiotic stresses. Here, based on the conserved residues of the deduced amino acid sequences of the homologous BADH genes, we cloned the AnBADH gene from the xerophytic leguminous plant Ammopiptanthus nanus by using reverse transcription PCR and rapid amplification of cDNA ends. The full-length cDNA is 1868 bp long without intron, and contains an open reading frame of 1512 bp, and 3'- and 5'-untranslated regions of 294 and 62 bp. It encodes a 54.71 kDa protein of 503 amino adds. The deduced amino add sequence shares high homology, conserved amino acid residues and sequence motifs crucial for the function with the BADHs in other leguminous species. The sequence of the open reading frame was used to construct a prokaryotic expression vector pET32a-AnBADH, and transform Escherichia coli. The transformants expressed the heterologous AnBADH gene under the induction of isopropyl beta-D-thiogalactopyranoside, and demonstrated significant enhancement of salt and heat tolerance under the stress conditions of 700 mmol L-1 NaCl and 55 degrees C high temperature. This result suggests that the AnBADH gene might play a crucial role in adaption of A. nanus to the abiotic stresses, and have the potential to be applied to transgenic operations of commercially important crops for improvement of abiotic tolerance. (C) 2014 Elsevier B.V. All rights reserved.
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