Molecular cloning and characterization of 5-enolpyruvylshikimate-3-phosphate synthase gene from Convolvulus arvensis L. |
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Authors: | Zhao-Feng Huang Chao-Xian Zhang Hong-Juan Huang Shou-Hui Wei Yan Liu Hai-Lan Cui Jing-Chao Chen Long Yang Jin-Yi Chen |
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Affiliation: | 1. Institute of Plant Protection, Key Laboratory of Weed and Rodent Biology and Management, Chinese Academy of Agricultural Sciences, Beijing, 100193, China 2. Institute of Plant Protection, Heilongjiang Academy of Land Reclamation Sciences, Harbin, 150038, China
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Abstract: | 5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS), the target enzyme for glyphosate inhibition, catalyzes an essential step in the shikimate pathway for aromatic amino acid biosynthesis. The full-length cDNA of 1,751 nucleotides (CaEPSPS, Genbank accession number: EU698030) from Convolvulus arvensis was cloned and characterized. The CaEPSPS encodes a polypeptide of 520 amino acids with a calculated molecular weight of 55.5 kDa and an isoelectric point of 7.05. The results of homology analysis revealed that CaEPSPS showed highly homologous with EPSPS proteins from other plant species. Tissue expression pattern analysis indicated that CaEPSPS was constitutively expressed in stems, leaves and roots, with lower expression in roots. CaEPSPS expression level could increase significantly with glyphosate treatment, and reached its maximum at 24 h after glyphosate application. We fused CaEPSPS to the CaMV 35S promoter and introduced the chimeric gene into Arabidopsis. The resultant expression of CaEPSPS in transgenic Arabidopsis plants exhibited enhanced tolerance to glyphosate in comparison with control. |
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