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Construction and analysis of an intergeneric fusion from Pigmentiphaga sp. strain AAP-1 and Pseudomonas sp. CTN-4 for degrading acetamiprid and chlorothalonil

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Abstract

Pseudomonas sp. CTN-4 degrades chlorothalonil (CTN) but not acetamiprid (AAP), and Pigmentiphaga sp. strain AAP-1 degrades AAP but not CTN. A functional strain, AC, was constructed through protoplast fusion of two parental strains (Pseudomonas sp. CTN-4 and Pigmentiphaga sp. strain AAP-1) in order to simultaneously improve the degradation efficiency of AAP and CTN. Fusant-AC with eight transfers on plates containing two antibiotics and CTN was obtained. For the purpose of identifying and confirming the genetic relationship between fusant-AC and its parents, randomly amplified polymorphic DNA (RAPD), scanning electron microscopy (SEM), and 16S ribosomal DNA (rDNA) analysis were performed. In toto, RAPD fingerprint analysis produced 194 clear bands with 9 primers, which not only had bands in common with strains CTN-4 and AAP-1, but also had its own novel fusant-specific bands. The genetic similarity indices between fusant-AC and parental strains CTN-4 and AAP-1 were 0.40 and 0.69, respectively. The result of SEM indicated that the cell morphology of fusant-AC differed from both its parents. The fusant strain AC possesses a strong capability for AAP and CTN degradation. At AAP concentration (50–300 mg L−1), the degradation was achieved within 5 h. At the initial dose of 50 and 100 mg L−1 CTN, the percentages reached 96 and 91 % over a 36-h incubation period. The present study indicates that the protoplast-fusion technique may have possible applications in environmental pollution control.

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Acknowledgments

This work was supported by grants from the Chinese National Natural Science Foundation (31100083, 31300422, and 41501304), the Foundation for Young Talents in College of Anhui Province, the Natural Science Foundation from Educational Commission of Anhui Province (KJ2015A049), Provincial Natural Science Foundation of Anhui (1508085MC49 and 1408085QC47), Scholar Backbone Supporting Plan of Huaibei Normal University, and Construction Project from College Scientific Research Innovation Team of Anhui Province—Ecological Restoration and Utilization of Coal Mining Subsidence Area.

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  1. College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China

    Guangli Wang, Danfeng Zhu, Minghua Xiong, Hui Zhang & Yuan Liu

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  1. Guangli Wang
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  2. Danfeng Zhu
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  3. Minghua Xiong
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Correspondence to Minghua Xiong or Yuan Liu.

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Wang, G., Zhu, D., Xiong, M. et al. Construction and analysis of an intergeneric fusion from Pigmentiphaga sp. strain AAP-1 and Pseudomonas sp. CTN-4 for degrading acetamiprid and chlorothalonil. Environ Sci Pollut Res 23, 13235–13244 (2016). https://doi.org/10.1007/s11356-016-6482-y

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