Pyrosequencing investigation into the bacterial community in permafrost soils along the China-Russia Crude Oil Pipeline (CRCOP).

Abstract

The China-Russia Crude Oil Pipeline (CRCOP) goes through 441 km permafrost soils in northeastern China. The bioremediation in case of oil spills is a major concern. So far, little is known about the indigenous bacteria inhabiting in the permafrost soils along the pipeline. A pilot 454 pyrosequencing analysis on the communities from four selected sites which possess high environment risk along the CRCOP is herein presented. The results reveal an immense bacterial diversity than previously anticipated. A total of 14448 OTUs with 84834 reads are identified, which could be assigned into 39 different phyla, and 223 families or 386 genera. Only five phyla sustain a mean OTU abundance more than 5% in all the samples, but they altogether account for 85.08% of total reads. Proteobacteria accounts for 41.65% of the total OTUs or 45% of the reads across all samples, and its proportion generally increases with soil depth, but OTUs numerically decline. Among Proteobacteria, the abundance of Beta-, Alpha-, Delta- and Gamma- subdivisions average to 38.7% (2331 OTUs), 37.5% (2257 OTUs), 10.35% (616 OTUs), and 6.21% (374 OTUs), respectively. Acidobacteria (esp. Acidobacteriaceae), Actinobacteria (esp. Intrasporangiaceae), Bacteroidetes (esp. Sphingobacteria and Flavobacteria) and Chloroflexi (esp. Anaerolineaceae) are also very common, accounting for 8.56% (1237 OTUs), 7.86% (1136 OTUs); 7.35% (1063 OTUs) and 8.27% (1195 OTUs) of total libraries, respectively. The ordination analysis indicates that bacteria communities in the upper active layer cluster together (similar), while bacterial consortia from the lower active layer and permafrost table scatter (less similar). The abundance of Rhodococcus (12 OTUs), Pseudomonas (71 OTUs) and Sphingomonas (87 OTUs) is even less (<0.01%). This effort to profile the background diversity may set the first stage for better evaluating the bacterial dynamics in response to accidental oil spills.