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. 2012 Aug 24:3:295.
doi: 10.3389/fmicb.2012.00295. eCollection 2012.

Differential virus host-ranges of the Fuselloviridae of hyperthermophilic Archaea: implications for evolution in extreme environments

Ruben M Ceballos  1 , Caleb D Marceau, Joshua O Marceau, Steven Morris, Adam J Clore, Kenneth M Stedman
Affiliations

Differential virus host-ranges of the Fuselloviridae of hyperthermophilic Archaea: implications for evolution in extreme environments

Ruben M Ceballos et al. Front Microbiol. .

Abstract

An emerging model for investigating virus-host interactions in hyperthermophilic Archaea is the Fusellovirus-Sulfolobus system. The host, Sulfolobus, is a hyperthermophilic acidophile endemic to sulfuric hot springs worldwide. The Fuselloviruses, also known as Sulfolobus Spindle-shaped Viruses (SSVs), are "lemon" or "spindle"-shaped double-stranded DNA viruses, which are also found worldwide. Although a few studies have addressed the host-range for the type virus, Sulfolobus Spindle-shaped Virus 1 (SSV1), using common Sulfolobus strains, a comprehensive host-range study for SSV-Sulfolobus systems has not been performed. Herein, we examine six bona fide SSV strains (SSV1, SSV2, SSV3, SSVL1, SSVK1, SSVRH) and their respective infection characteristics on multiple hosts from the family Sulfolobaceae. A spot-on-lawn or "halo" assay was employed to determine SSV infectivity (and host susceptibility) in parallel challenges of multiple SSVs on a lawn of a single Sulfolobus strain. Different SSVs have different host-ranges with SSV1 exhibiting the narrowest host-range and SSVRH exhibiting the broadest host range. In contrast to previous reports, SSVs can infect hosts beyond the genus Sulfolobus. Furthermore, geography does not appear to be a reliable predictor of Sulfolobus susceptibility to infection by any given SSV. The ability for SSVs to infect susceptible Sulfolobus host does not appear to change between 65°C and 88°C (physiological range); however, very low pH appears to influence infection. Lastly, for the virus-host pairs tested the Fusellovirus-Sulfolobus system appears to exhibit host-advantage. This work provides a foundation for understanding Fusellovirus biology and virus-host coevolution in extreme ecosystems.

Keywords: Archaea; Crenarchaea; Fusellovirus; Sulfolobus; Sulfolobus spindle-shaped virus; halo assay; host-range; hyperthermophilic.

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Figures

Figure 1
Figure 1
Sulfolobus Spindle-shaped Virus (SSV) infection. Scanning Electron Micrograph (SEM) of Sulfolobus solfataricus strain Gθ either uninfected (A) or infected with SSVRH (B); (C) Transmission Electron Micrograph (TEM) of typical SSVRH virions used to infect Sulfolobus solfataricus strain Gθ; (D) TEM of SSVRH virus particles with infected Sulfolobus solfataricus strain Gθ (lower left of image).
Figure 2
Figure 2
SSV-strain specific halo formation on Sulfolobus solfataricus strain Gθ by all six SSV tested strains. Virus susceptibility is determined by the production of halos of growth inhibition around a 1.0 μL spot of virus concentrate applied to a Gelriteθ softlayer containing S. solfataricus strain Gθ. Locations of spots and identities of SSVs are indicated with arrows. Plate were incubated for 4 days at 78°C. Halo morphologies are typical for each SSV-strain. Triton X-100 and sterile H2O serve as positive and negative controls, respectively.
Figure 3
Figure 3
Representative virus susceptibility of the three host classes. Virus infection was determined as in Figure 2. (A) Completely susceptible host: All 6 SSVs tested inhibit growth of a Sulfolobus strain isolated from Lassen Volcanic National Park. (B) Completely resistant host: No SSVs inhibit growth of S. tokodaii. (C) Selectively susceptible host: 4 of 6 SSV strains inhibit the growth of “S. islandicus” strain HVE 10/4. Spots on plates are (clockwise from arrow): Triton X-100 (positive control), SSV1, SSV2, SSV3, SSVL1, SSVK1, SSVRH, and sterile H2O (negative control). The complete dataset for host-virus interactions is shown in Table 1.
Figure 4
Figure 4
Sulphurisphaera ohwakuensis is susceptible to SSV infection. Virus infectivity was assessed as in Figures 2 and 3. SSV3, SSVRH, and SSVK1 produce halos on Sulphurisphaera ohwakuensis, a Japanese isolate, closely related to Sulfolobus tokodaii. SSV1, SSV2, and SSVL1 did not inhibit growth of this strain.
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