Pseudomonasnitroreducens with paraquat resistance

A Pseudomonas and paraquat technology, applied in the field of a paraquat-resistant nitrifying Pseudomonas, can solve the problems of paraquat-resistant crops that have not yet been seen

Inactive Publication Date: 2012-10-24
JIANGSU ACAD OF AGRI SCI
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Problems solved by technology

Paraquat is second only to glyphosate in the international market, but paraquat-resistant crops have not yet appeared

Method used

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  • Pseudomonasnitroreducens with paraquat resistance
  • Pseudomonasnitroreducens with paraquat resistance
  • Pseudomonasnitroreducens with paraquat resistance

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Embodiment Construction

[0026] Soil samples (sandy loam, 7% water content) were mainly collected from Nanjing Red Sun Group Co., Ltd. and Shenzhen Nuopuxin Agrochemical Co., Ltd. The collection site was about 200m away from the production workshop, and the collection depth was 0-30cm.

[0027] 2) Isolation and screening of strains

[0028] Using the dilution plate separation method (Li Shunpeng. Microbial Experiment Guide [M]. 2003), the soil suspension was spread on the beef extract peptone medium containing different concentrations of paraquat (0, 1, 10, 20, 50mmol / L) Place the Petri dish at 30°C for 48 hours to observe the growth. Four monoclones resistant to 20mmol / L PQ (paraquat standard samples were purchased from Dr. Ehrenstorfer-Schafers's laboratory in Germany) were successfully isolated from the soil suspension, and after further screening, one clone resistant to 50mmol / L PQ was selected Single clones were purified for further research work. This single clone was named S3.

[0029] The p...

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Abstract

The invention relates to a pseudomonasnitroreducens with resistance to paraquat (PQ). The strain number is S3. Pseudomonasnitroreducens is separated from PQ contaminated soil, belongs to Gram-negative brevibacterium, grows on a culture medium containing strong oxidant PQ or sodium nitroprusside (SNP), has nitrification reduction property and denitrification reduction property, shows positive in a catalase experiment and shows negative in an indole experiment. Pseudomonasnitroreducens is beneficial to the research of oxidation resistance, nitrification reduction and denitrification mechanism of bacteria and also plays an important role in the control of weeds and the research of other agricultural chemical industries. In the invention, the PnPQR gene is cloned from S3, and the gene codes a member of an atypical major facilitator superfamily (MFS). PnPQR is expressed in the escherichia coli BL21 strain and can increase resistance to paraquat. A transgenic tobacco plant with PnPQR excessively expressed has paraquat resistance.

Description

technical field [0001] The invention relates to a strain of Pseudomonas nitroreducens resistant to paraquat (Paraquat, PQ), which belongs to the field of biotechnology, specifically a strain that can contain strong oxidant PQ (50mmol / L) or Gram-negative brevibacterium strains grown on the medium of sodium nitroprusside (SNP, 200 μmol / L). [0002] A gene PnPQR is isolated from the strain, and the gene is transformed into Escherichia coli to obtain a strain resistant to Escherichia coli, and transforming tobacco through the Agrobacterium-mediated method can improve the resistance of tobacco to paraquat. Background technique [0003] The herbicide paraquat (paraquat, PQ) is one of the most widely used herbicides in the world. The application area is second only to glyphosate. Glyphosate's near-monopoly position in the global market is, to some extent, also benefited from the successful breeding of transgenic glyphosate-resistant crops. Single herbicide pressure will inevitab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/82C12N15/66C12R1/38
Inventor 郭书巧倪万潮束红梅巩元勇沈新莲张香桂徐鹏
Owner JIANGSU ACAD OF AGRI SCI
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