Journal of Ecology and Rural Environment ›› 2014, Vol. 30 ›› Issue (6): 761-767.doi:

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Remediation of 3,5,6-Trichloro-2-Pyridinol Polluted Soil With Ralstonia sp. T6 and Construction of a Chlorpyrifos-Mineralizing Engineered Strain

ZHANG  Yang, WANG  Fei, HUANG  Yan, CUI  Zhong-Li   

  1. College of Life Science,Nanjing Agricultural University
  • Received:2014-04-15 Revised:2014-06-13 Online:2014-11-25 Published:2014-12-05
  • Contact: CUI Zhong-Li College of Life Science,Nanjing Agricultural University E-mail:czl@njau.edu.cn

Abstract: TCP is a kind of metabolic product of degradation of chlorpyrifos. Ralstonia sp. T6, a TCP (3,5,6-trichloro-2-pyridinol) degrading strain of bacteria, was used in an experiment to explore its TCP degrading characteristics in soil. Results show that T6 degraded 80% of 50 mg·kg-1 TCP in 6 days. Temperature, inoculation rate and initial concentration of the substrate were factors affecting its degrading efficiency. The optimum temperature for T6 degrading TCP was 30℃; in soils lower than 10×108 CFU/kg in bacteria content, TCP biodegradation rate rose with rising bacteria content, while in soils higher than that, the rate did not rise, but fall instead with rising bacteria content. A methyl parathion hydrolase gene (mpd) from Stenotrophomonas sp. DSP-1 was inserted into 16S rDNA of Ralstonia sp. T6 by conjugation and a genetically engineered T6-mpd that is supposed to be able to completely mineralize chlorpyrifos was obtained. Culturing tests show that T6-mpd and Ralstonia sp. T6 displayed similar growth characteristics. Chlorpyrifos degrading tests show that in LB medium, the chlorpyrifos hydrolysis efficiency of T6-mpd was basically the same as that of Stenotrophomonas sp. DSP-1, but in inorganic salt medium, T6-mpd‘s 50 mg·L-1 chlorpyrifos hydrolysis rate in 60 h reached only 36%, significantly lower than DSP-1’s. Simulated in-situ soil remediation shows that T6-mpd, 10×108 CFU·kg-1 in content, could degrade 64% of 50 mg·kg-1 chlorpyrifos. It is, therefore, believed that Strain T6-mpd is a promising tool for bioremediation of chlorpyrifos-contaminated environment.

Key words: chlorpyrifos, 3,5,6-trichloro-2-pyridinol, remediation, genetically engineered strain T6-mpd

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