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Construction of Transgenetic Strains of Pseudomonas Stutzeri and the Interaction between the Strains and Rice with Highly-efficient Photosynthetic Rate

Author: ZuoChunLing
Tutor: ZhangBaoMing；LinMin
School: Chinese Academy of Agricultural Sciences
Course: Crop Cultivation and Farming System
Keywords: Nitrogen-fixing Pseudomonas stutzeri A1501 Four carbon dicarboxylic acid transport system To turn pepc gene photosynthesis in rice
CLC: S144
Type: Master's thesis
Year: 2003
Downloads: 87
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Abstract

Rice Bacterium Associated nitrogen fixation in rice growth and photosynthesis. Nitrogen Fixation strains build carbon utilization ability, the application of transgenic rice with high photosynthetic rate it can study the interaction between the two. Nitrogen-fixing Pseudomonas stutzeri (Pseudomonas stutzeri) A1501 is present in the rice rhizosphere combined with nitrogen-fixing bacteria. Four carbon dicarboxylic acid transport system (Dicarboxylate acids transport system, Dct-system) is the the diazotrophs energy transportation system. In this study, using a high-frequency conjugative transfer characteristics Tn5-mob suicide plasmid, constructed Tn5: dctPQM recombinant plasmid named pSZY6 to. In helper plasmid pRK2013 assist, three conjugative test methods, carry the import of wild type strain A1501 A1501 four carbon dicarboxylic acid transport structure, gene dctPQM recombinant plasmid pSZY6 of chromosomes containing extra copy dctPQM gene screening on selective plates recombinant strains. Three strains showed higher nitrogenase activity, the growth of the use of four-carbon dicarboxylic acid, tentatively named the A142, A115, A136,. The different nitrogenase activity in different four-carbon dicarboxylic acid as the sole carbon source, recombinant strains using three carbon source: which succinic acid> fumaric acid> malic acid. Carbon source sufficient or restrictions (four carbon dicarboxylic acid in the medium concentration of 20 mM, 10 mM, 5 mM), nitrogenase activity in A142 with dicarboxylic acid concentration to improve and enhance, and were higher than the wild-type strain A1501. The recombinant strain A115 and the wild-type strain A1501 application turn pepc gene \The interaction between the research Rice photosynthesis and nitrogen-fixing bacteria nitrogen fixation. The results show that: the application of nitrogen-fixing bacteria, the net photosynthetic rate turn pepc Gene Rice, the fluorescent kinetic parameters (Fv / Fm) and PEPC enzyme activity was significantly increased, biological and economic yield increased by 38.08% and 26.86% respectively. Comparison with the original species, the the pepc transgenic rice significantly improve the nitrogen-fixing bacteria especially recombinant strains of nitrogen-fixing efficiency and nitrogen fixation total. Recombinant strain A115, A142 and the wild-type strain A1501 administration of low soil nitrogen and organic matter content of Panjin City, Liaoning Province, saline land rice area, found that the application of nitrogen-fixing bacteria accelerated rice tillering increase the crude protein content of rice plants to enhance root vigor, and to promote the rice growth, particularly those containing an additional copy dctPQM gene recombinant strain most significantly. And nitrogen-fixing bacteria significantly better than with chemical fertilizer and organic manure effect. Cloned promoter region A1501 four carbon dicarboxylic acid binding protein encoding gene dCTP by specific primers were designed using the PCR method, and with independent dctPp cloned into the transfer characteristic, the stable presence of a plasmid vector in the Gram-negative bacteria pLA2917 introduced into the lacZ gene, and then carry the coding region in dctPp constructed dctP-lacZ fusion gene expression vector. The use of two conjugative test methods, dctP-lacZ conjugal transfer into P.stutzeri wild-type strains A1501 RpoN mutant, DctB ^{- mutant Filter on selective plates containing X-gal adapter zygote. : Inducible dctP gene expression; dctP for σ ^{54 -dependent promoters; Chinese Academy of Agricultural Sciences, master's degree the papers DctB mutation, dctP not expression, Description dCtP gene expression requires additional activator of transcription ; glucose and lactate induced dctP gene expression, the glucose to suppress dctP gene expression levels, four carbon the two acid rock acid is higher than the level of gene expression induced dCtP fumaric acid.}}

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