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Tian Y.-S.,Nanjing Agricultural University | Tian Y.-S.,Shanghai Key Laboratory of Agricultural Genetics and Breeding | Tian Y.-S.,Shanghai Ruifeng Agricultural Science and Technology Co. | Yao Q.-H.,Shanghai Key Laboratory of Agricultural Genetics and Breeding | And 4 more authors.
Applied Biochemistry and Microbiology | Year: 2015

To date, only aroA variants derived from Agrobacterium tumefaciens CP4 have been used to generate the commercial glyphosate-resistant crops currently available in the market. Therefore, it is of interest to increase aroA gene diversity and seek new glyphosate-tolerant genes for developing glyphosate-tolerant crops. In this study, a novel aroA gene encoding 5-enolpyruvylshikimate-3-phosphate synthase from Paracoccus denitrificans was isolated using the genomic library construction and complementary screening. Furthermore, the transgenic Arabidopsis plants with the aroA gene from P. denitrificans were obtained to confirm the potential of the novel aroA gene in developing glyphosate-resistant crops. © 2015, Pleiades Publishing, Inc.


Tian Y.-S.,Biotechnology Research Institute of Shanghai Academy of Agricultural science | Tian Y.-S.,Shanghai Ruifeng Agricultural Science and Technology Co. | Tian Y.-S.,Nanjing Agricultural University | Xu J.,Biotechnology Research Institute of Shanghai Academy of Agricultural science | And 5 more authors.
Molecular Breeding | Year: 2015

To date, only AroA variant derived from Agrobacterium tumefaciens CP4 has been used to generate the commercial glyphosate-resistant crops currently available in the market. This single source of the EPSPS gene may have caused the decrease in herbicide tolerance, which has become a major concern of those involved in field management programs. Therefore, it is of interest to increase aroA/EPSPS gene diversity and seek new glyphosate-tolerant genes for developing glyphosate-tolerant crops. In the current study, EPSPS gene from Vitis vinifera (VvEPSPS) was cloned using reverse transcription polymerase chain reaction. However, wild type VvEPSPS cannot be directly used for developing transgenic crops because of its extreme glyphosate sensitivity. Recent studies have demonstrated that DNA shuffling is an effective strategy in producing multi-mutated EPSPS resourced from plants (EPSPSplant) with improved glyphosate resistance in bacteria and plants. After performing DNA shuffling on VvEPSPS gene, one highly glyphosate-resistant mutant with seven amino acid variations was isolated after five rounds of shuffling and screening. The mutant showed seven amino acid changes in the EPSPS gene, namely, Q93R, T113A, P117L, G126A, C160Y, N239H, and V343A. The assay of glyphosate resistance further confirmed the potential of the VvEPSPS mutant in developing glyphosate-resistant crops. © 2015, Springer Science+Business Media Dordrecht.


Tian Y.-S.,Shanghai Key Laboratory of Agricultural Genetics and Breeding | Tian Y.-S.,Shanghai Ruifeng Agricultural Science and Technology Co. | Jin X.-F.,Shanghai Key Laboratory of Agricultural Genetics and Breeding | Fu X.-Y.,Shanghai Key Laboratory of Agricultural Genetics and Breeding | And 5 more authors.
Journal of Toxicological Sciences | Year: 2014

Environmental levels of bisphenol A (BPA) are a global concern because the compound can cause damage to reproductive organs, the thyroid gland, and brain tissues at developmental stages. Plants are important in removing BPA from the atmosphere, soil, and water. However, knowledge on the mechanism by which plants respond to this compound is limited. To determine the response mechanism of plants to BPA, we used a microarray system to analyze the gene expression patterns of Arabidopsis thaliana after irrigation with 3.0 mM BPA. We identified 651 genes that were differentially expressed upregulated and 470 genes that were downregulated by BPA. These genes may specifically contribute to BPA uptake, transformation, conjugation, and compartmentation in plants. The potential function of upregulated genes in plant defense against BPA was also determined.


Lin H.,Shanghai JiaoTong University | Lin H.,University of Nottingham | Rao J.,Shanghai JiaoTong University | Shi J.,Shanghai JiaoTong University | And 7 more authors.
Journal of Integrative Plant Biology | Year: 2014

Soybean [Glycine max (L.) Merr.] is one of the world's major crops, and soybean seeds are a rich and important resource for proteins and oils. While "omics" studies, such as genomics, transcriptomics, and proteomics, have been widely applied in soybean molecular research, fewer metabolomic studies have been conducted for largescale detection of low molecular weight metabolites, especially in soybean seeds. In this study, we investigated the seed metabolomes of 29 common soybean cultivars through combined gas chromatography-mass spectrometry and ultra-performance liquid chromatography-tandem mass spectrometry. One hundred sixty-nine named metabolites were identified and subsequently used to construct a metabolic network of mature soybean seed. Among the 169 detected metabolites, 104 were found to be significantly variable in their levels across tested cultivars. Metabolite markers that could be used to distinguish genetically related soybean cultivars were also identified, and metabolite-metabolite correlation analysis revealed some significant associations within the same or among different metabolite groups. Findings from this work may potentially provide the basis for further studies on both soybean seed metabolism and metabolic engineering to improve soybean seed quality and yield. © 2014 Institute of Botany, Chinese Academy of Sciences.


Tian Y.-S.,Shanghai Academy of Agricultural science | Xu H.,Shanghai Academy of Agricultural science | Peng R.-H.,Shanghai Academy of Agricultural science | Yao Q.-H.,Shanghai Academy of Agricultural science | Wang R.-T.,Shanghai Ruifeng Agricultural Science and Technology Co.
Biotechnology and Biotechnological Equipment | Year: 2014

The gene (CcLcc2) encoding laccase from the basidiomycete Coprinopsis cinerea Okayama-7 #130 was synthesized by polymerase chain reaction-based two-step DNA synthesis, and heterologously expressed in Pichia pastoris. The recombinant protein was purified by ammonium sulphate precipitation and nickel nitrilotriacetic acid chromatography. The molecular mass of CcLcc2 was estimated to be 54 kDa by denaturing polyacrylamide gel electrophoresis. The optimum pH and temperature for laccase catalysis for the oxidation of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) were 2.6 and 45 °C, respectively. The Km values of the enzyme towards the substrates ABTS, 2,6-dimethoxyphenol (2,6-DMP) and guaiacol were 0.93, 1.02 and 28.07 mmol·L-1, respectively. The decolourization of methyl orange, crystal violet and malachite green, commonly used in the textile industry, was assessed. The decolourization percentage of crystal violet and malachite green was 80% after 4 h of reaction, and that of methyl orange was 50% at 4 h. These results show that the CcLcc2 has enormous potential for the decolourization of highly stable triphenylmethane dyes. © 2014 The Author(s). Published by Taylor & Francis.

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