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Characterization of a glutamine synthetase gene DvGS1 from Dunaliella viridis and investigation of the impact on expression of DvGS1 in transgenic Arabidopsis thaliana.
- Source :
-
Molecular biology reports [Mol Biol Rep] 2014 Jan; Vol. 41 (1), pp. 477-87. Date of Electronic Publication: 2013 Dec 04. - Publication Year :
- 2014
-
Abstract
- A novel glutamine synthetase (GS) gene DvGS1 showing highest amino acid sequence identity of 78 % with the other homologous GS proteins from green algae, was isolated and characterized from Dunaliella viridis. Phylogenetic analysis revealed that DvGS1 occupied an independent phylogenetic position which was different with the GSs from higher plants, animals and microbes. Functional complement in E. coli mutant confirmed that the DvGS1 encoded functional GS enzyme. Real-time PCR analysis of DvGS1 in D. viridis cells under nitrogen starvation revealed that the mRNA level of DvGS1 was positively up-regulated in 12 h. The DvGS1 levels at the points of 12 and 24 h were separately twofold and fourfold of the level before nitrogen starvation. In order to investigate the potential application of DvGS1 in higher plants, the transgenic study of DvGS1 in Arabidopsis thaliana was carried out. Phenotype identification demonstrated that all three transgenic lines of T3 generation showed obviously enhanced root length (26 %), fresh weight (22-46 % at two concentrations of nitrate supplies), stem length (26 %), leaf size (29 %) and silique number (30 %) compared with the wild-type Arabidopsis. Biochemical analysis confirmed that all three transgenic lines had higher total nitrogen content, soluble protein concentration, total amino acid content and the leaf GS activity than the wild type plants. The free NH4 (+) and NO3 (-) concentration in fresh leaves of three transgenic lines were reduced by 17-26 % and 14-15 % separately (at two concentrations of nitrate supplies) compared with those of the wild types. All the results indicated that over-expression of DvGS1 in Arabidopsis significantly results in the improvement of growth phenotype and the host's nitrogen use efficiency.
- Subjects :
- Amino Acid Sequence
Arabidopsis growth & development
Arabidopsis metabolism
Cloning, Molecular
Glutamate-Ammonia Ligase biosynthesis
Molecular Sequence Data
Nitrogen metabolism
Phenotype
Phylogeny
Plant Proteins biosynthesis
Plant Roots genetics
Plant Roots growth & development
Plant Roots metabolism
Plants, Genetically Modified genetics
Plants, Genetically Modified growth & development
Plants, Genetically Modified metabolism
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Arabidopsis genetics
Chlorophyta enzymology
Glutamate-Ammonia Ligase genetics
Plant Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1573-4978
- Volume :
- 41
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Molecular biology reports
- Publication Type :
- Academic Journal
- Accession number :
- 24307252
- Full Text :
- https://doi.org/10.1007/s11033-013-2882-y