摘要
The activated sludge process is characterized by high microbial density and diversity,both of which facilitate antibiotic resistance gene transfer.Many studies have suggested that antibiotic and non-antibiotic drugs at sub-inhibitory concentrations are major inducers of conjugative gene transfer.The self-transmissible plasmid pND6-2 is one of the endogenous plasmids harbored in Pseudomonas putida ND6,which can trigger the transfer of another co-occurring naphthalene-degrading plasmid pND6-1.Therefore,to illustrate the potential influence of stimulants on conjugative transfer of pND6-2,we evaluated the effects of four antibiotics (ampicillin,gentamycin,kanamycin,and tetracycline) and naphthalene,on the conjugal transfer efficiency of pND6-2 by filter-mating experiment.Our findings demonstrated that all stimulants within an optimal dose promoted conjugative transfer of pND6-2from Pseudomonas putida GKND6 to P.putida KT2440,with tetracycline being the most effective (100μg/L and 10μg/L),as it enhanced pND6-2-mediated intra-genera transfer by approximately one hundred-fold.Subsequently,seven AS reactors were constructed with the addition of donors and different stimulants to further elucidate the conjugative behavior of pND6-2 in natural environment.The stimulants positively affected the conjugal process of pND6-2,while donors reshaped the host abundance in the sludge.This was likely because stimulant addition enhanced the expression levels of conjugation transfer-related genes.Furthermore,Blastocatella and Chitinimonas were identified as the potential receptors of plasmid pND6-2,which was not affected by donor types.These findings demonstrate the positive role of sub-inhibitory stimulant treatment on pND6-2 conjugal transfer and the function of donors in re-shaping the host spectrum of pND6-2.
基金
supported by the National Natural Science Foundation of China(No. 31670512)
Natural Science Basic Research Plan in Shaanxi Province of China(No. 2018JM3039)。
