High-Throughput
Single-Cell Technology Reveals the
Contribution of Horizontal Gene Transfer to Typical Antibiotic Resistance
Gene Dissemination in Wastewater Treatment Plants
posted on 2021-08-20, 13:36authored byZiyan Wei, Kai Feng, Zhujun Wang, Yu Zhang, Min Yang, Yong-Guan Zhu, Marko P. J. Virta, Ye Deng
The
spread of antibiotic resistance genes (ARGs) has gained much
attention worldwide, while the contribution of vertical gene transfer
(VGT) and horizontal gene transfer (HGT) is still elusive. Here, we
improved an emerging high-throughput single-cell-based technology,
emulsion, paired isolation, and concatenation polymerase chain reaction
(epicPCR), by lengthening the sequence of ARG in the fused ARG-16S
rRNA fragments to cover the variance of both ARG and its hosts. The
improved epicPCR was applied to track the hosts of a widely detected
ARG, sul1 gene, in five urban wastewater treatment
plants (UWTPs) during two seasons. The sul1 host
bacteria were highly diverse and mostly classified as Proteobacteria
and Bacteroidetes. Clear seasonal divergence of α-diversity
and interaction networks were present in the host community. The consensus
phylogenetic trees of the sul1 gene and their host
demonstrated incorrespondence on the whole and regularity on abundant
groups, suggesting the important role of both HGT and VGT, respectively.
The relative importance of these two ways was further measured; HGT
(54%) generally played an equal or even more important role as VGT
(46%) in UWTPs. The application of the improved epicPCR technology
provides a feasible approach to quantify the relative contributions
of VGT and HGT in environmental dissemination of ARGs.