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Wang Y.,South China University of Technology | Peng D.,South China University of Technology | Peng D.,Central South University of forestry and Technology | Peng D.,Academy of Seed Industry of Hunan Yahua | And 6 more authors.
Plant Molecular Biology Reporter | Year: 2015

Sapium sebiferum (L.) Roxb is one of the most important oil trees in China. Diacylglycerol acyltransferases (DGATs) esterify sn-1, 2-diacylglycerol with a long-chain fatty acyl-CoA, the last step and the rate-limiting step of triacylglycerol (TAG) biosynthesis in prokaryotic and eukaryotic organisms. At least 74 DGAT2 sequences from 61 organisms have been identified, but the SsDGAT2 gene had not been reported to date. To clarify the function of SsDGAT2, we cloned the CDS (rapid amplification of cDNA end) of SsDGAT2 by RACE technology. The full-length CDS of SsDGAT2 contains 1011 bp and encodes a protein of 336 amino acids. Recombinant SsDGAT2 restored TAG biosynthesis to the yeast strain Saccharomyces cerevisiae H1246 TAG-deficient mutant and preferentially incorporated unsaturated C18 fatty acids into lipids. To investigate the biotechnological potential of SsDGAT2, it was expressed under the control of the 35S promoter in Arabidopsis Col-4. The oleic acid content increased by 50 % in transgenic plants relative to the control. The results indicated that most of the oleic acid increase was at the expense of linolenic acid (18:3) content, which suggests that high-oleic-acid-content seeds can be created by the overexpression of SsDGAT2 in S. sebiferum (L.) Roxb. © 2015 Springer Science+Business Media New York Source

Zhou B.,South China University of Technology | Zhou B.,Central South University of forestry and Technology | Peng D.,South China University of Technology | Peng D.,Academy of Seed Industry of Hunan Yahua | And 4 more authors.
Protein Expression and Purification | Year: 2015

This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. © 2015. Source

Zhou B.,Hunan University | Peng D.,Hunan University | Lin J.,Hunan University | Huang X.,Hunan University | And 6 more authors.
Journal of Plant Biology | Year: 2011

Gibberellins (GAs) are endogenous hormones that play an important role in regulating plant stature by increasing cell division and promoting seed germination. The GA2-oxidase gene from Arabidopsis thaliana (AtGA2ox8) was introduced into Brassica napus L. by Agrobacterium-mediated floral-dip transformation with the aim of decreasing the amount of bioactive GA and hence reduced the plant height. As anticipated, the transgenic plant exhibited dwarf phenotype. Importantly, compared with the wild type, the transgenic plants had delayed the seed germination, increased the chlorophyll content (28.7-36.3%) and photosynthesis capacity (14.3-18.7%) in a single leaf. At the same time, the photosynthesis capacity of the whole plants was significantly enhanced (35.7-48.6%) due to the extra leaves and branches. © 2010 The Botanical Society of Korea. Source

Zhou Y.,Hunan University | Liu H.,Fujian Agriculture and forestry University | Zhou X.,Hunan University | Yan Y.,Academy of Seed Industry of Hunan Yahua | And 10 more authors.
Molecular Breeding | Year: 2014

Glutamate dehydrogenase (GDH) tends to have a lower affinity for ammonium than glutamine synthetase (GS) in higher plants. Consequently, nitrogen is mostly assimilated as ammonium by the GS/glutamate synthase pathway which requires 2-oxoglutarate (2-OG) as carbon skeletons. In contrast, the NADP(H)-dependent GDH in fungi has a higher affinity for ammonium than that in higher plants and plays a more significant part in ammonium assimilation. We isolated an NADP(H)-GDH gene (PcGDH) from the fungus Pleurotus cystidiosus and heterologously expressed it in rice (Oryza sativa L.). Alterations in nitrogen assimilation, growth, metabolism, and grain yield were observed in the transgenic plants. An investigation of the kinetic properties of the purified recombinant protein demonstrated that the amination activity (7.05 ± 0.78 μmoL min-1 mg soluble protein-1) of PcGDH was higher than the deamination activity (3.36 ± 0.42 μmoL min-1 mg soluble protein-1) and that the Km value for ammonium (Km = 3.73 ± 0.23 mM) was lower than that for the glutamate (Km = 15.97 ± 0.31 mM), indicating that the PcGDH tends to interconvert 2-OG and glutamate. Examination of the activity of NADP(H)-GDH in control and transgenic lines demonstrated that NADP(H)-GDH activity in the transgenic lines was markedly higher than that in the control lines; in particular, the amination activity was significantly higher than the deamination activity in shoots of the transgenic lines. The results of the hydroponics experiment revealed that shoot and root length, fresh weight, chlorophyll content, nitrogen content, and amino acid levels (glutamate, glutamine, and total amino acids) were elevated in transgenic lines in comparison with those of the control line under different nitrogen conditions at seedling stage. The 1,000-grain weight and the panicle number in transgenic lines were considerably augmented in the field condition, yet the filled grain rate dropped slightly and there was no apparent change in the grain yield. The levels of glutelin and prolamine in the transgenic seeds were considerably higher than those in control seeds. In conclusion, these results demonstrate that heterologous expression of P. cystidiosus GDH (PcGDH) could improve nitrogen assimilation and growth in rice. © 2014 Springer Science+Business Media Dordrecht. Source

Huang X.Q.,Hunan University | He R.Q.,Hunan University | Liao X.Y.,Hunan University | Zhou B.,Hunan University | And 6 more authors.
Genetics and Molecular Research | Year: 2014

Exogenous gibberellins (GAs) are widely applied to increase crop yields, with knowledge about the physiological functioning and biochemistry mechanisms of these phytohormones improving; however, information remains limited about the effect of GAs on seed filling. In this study, the siliques (containing the seeds) of oilseed rape (Brassica napus L.) were treated with GA3 at 3 stages of seed filling. We confirmed that GA3 regulates the deposition of storage reserves in developing seeds. The percentage of crude fat in the seeds increased during the early stage, but remained stable during the middle and late stages. In comparison, the percentage of total protein decreased during the early and middle stages, but significantly increased during the late stage. In addition, Q-PCR was employed to analyze the expression level of related genes in response to GA3. It was found that the expression of WRI and ABI3 transcription factors corresponded to crude fat content and total protein content, respectively. The expression of storage reserve related genes DGAT, MCAT, SUC2, and GPT was consistent with crude fat content, whereas the expression of Napin corresponded to total protein content. The results of this study indicate that exogenous GA3 has a different effect on storage reserve deposition in seed during different stages of seed filling, and the effect might be achieved via changing the expression of related genes. © FUNPEC-RP. Source

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