title: Construction and analysis of an intergeneric fusion from Pigmentiphaga sp. strain AAP-1 and Pseudomonas sp. CTN-4 for degrading acetamiprid and chlorothalonil.
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.
authors.affiliation: College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China.
authors.forename: Guangli
authors.initials: G
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China.
authors.forename: Danfeng
authors.initials: D
authors.lastname: Zhu
authors.affiliation: College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China. mhx102006@163.com.
authors.forename: Minghua
authors.initials: M
authors.lastname: Xiong
authors.affiliation: College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Life Sciences, Huaibei Normal University, 235000, Huaibei, China. liuyuan0813@aliyun.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
mesh_terms: D001431:Bacteriological Techniques; D001673:Biodegradation, Environmental; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000235:genetics; D004785:Environmental Pollutants / Q000378:metabolism*; D000073943:Neonicotinoids; D009570:Nitriles / Q000378:metabolism*; D011523:Protoplasts; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism*; D011725:Pyridines / Q000378:metabolism*; D019105:Random Amplified Polymorphic DNA Technique
chemical_list: D017931:DNA Primers; D004275:DNA, Ribosomal; D004785:Environmental Pollutants; D000073943:Neonicotinoids; D009570:Nitriles; D011725:Pyridines; C464485:acetamiprid; C005806:tetrachloroisophthalonitrile
keywords: Acetamiprid (AAP); Chlorothalonil (CTN); Fusant strain AC; RAPD · protoplast fusion