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

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

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

2016

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

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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