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Cloning of 9-cis-epoxycarotenoid dioxygenase gene (TaNCED1) from wheat and its heterologous expression in tobacco
Authors:S. J. Zhang  G. Q. Song  Y. L. Li  J. Gao  J. J. Liu  Q. Q. Fan  C. Y. Huang  X. X. Sui  X. S. Chu  D. Guo  G. Y. Li
Affiliation:1. Crop Research Institute, Shandong Academy of Agricultural Sciences; Key Laboratory of Wheat Biology and Genetic Improvement on North Yellow and Huai River Valley, Ministry of Agriculture, National Engineering Laboratory For Wheat and Maize, Jinan, 250100, Shandong, P.R. China
Abstract:Abscisic acid (ABA) regulates plant responses to various environmental stresses. Oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED) is the critical step in the biosynthesis of ABA in higher plants. Using a homologous cloning approach, a NCED-like gene (designated as TaNCED1) was isolated from wheat (Triticum aestivum). It contained an open reading frame of 1 848 bp and encodes a peptide of 615 amino acids. Multiple sequence alignments showed that TaNCED1 shared high identity with NCEDs from other plants. Phylogenetic analysis revealed that TaNCED1 was most closely related to a barley HvNCED1 gene. The predicted 3D structure of TaNCED1 showed high similarity with other homologues. Southern blot analysis indicated that TaNCED1 was a single copy in the genome of wheat. TaNCED1 was differentially expressed in various organs and the expression was up-regulated by low temperature, drought, NaCl, and ABA. Heterologous expression of TaNCED1 in tobacco (Nicotiana tabacum) significantly improved its drought tolerance. Under drought treatment, TaNCED1-overexpressing transgenic tobacco plants exhibited higher germination rate, higher relative water content, content of soluble sugars and of ABA when compared with the wild type plants.
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