Elevated abundance of bacteriophage infecting bacteria in soil

Abstract

Here we report the first direct counts of soil bacteriophage and show that substantial populations of these viruses exist in soil (grand mean = 1.5 x 10(7) g(-1)), at least 350-fold more than the highest numbers estimated from traditional viable plaque counts. Adding pure cultures of a Serratia phage to soil showed that the direct counting methods with electron microscopy developed here underestimated the added phage populations by at least eightfold. So, assuming natural phages were similarly underestimated, virus numbers in soil averaged 1.5 x 10(8) g(-1), which is equivalent to 4% of the total population of bacteria. This high abundance was to some extent confirmed by hybridizing colonies grown on Serratia and Pseudomonas selective media with cocktails of phage infecting these bacteria. This showed that 8.9 and 3.9%, respectively, hybridized with colonies from the two media and confirmed the presence of phage DNA sequences in the cultivable fraction of the natural population. Thus, soil phage, like their aquatic counterparts, are likely to be important in controlling bacterial populations and mediating gene transfer in soil.

FIG. 1.

TEM images of phages negatively stained with 2% methylamine tungstate. (a) Phage CP6-1 suspended in distilled water without homogenization; (b and c) CP6-1-like phage in phage CP6-1-spiked homogenized rhizosphere soil; (d and e) non-CP6-1-like phage in phage CP6-1-spiked homogenized rhizosphere soil; (f to i) tailed phage from soil and rhizosphere samples; (j to m) VLPs from soil and rhizosphere samples. Bars = 50 nm.