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A simple procedure to determine the infectivity and host range of viruses infecting anaerobic and hyperthermophilic microorganisms

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Abstract

Plaque assay is the method traditionally used to isolate and purify lytic viruses, to determine the viral titer and host range. Whereas most bacterioviruses are either temperate or lytic, the majority of known archeoviruses are not lytic (i.e. they are temperate or chronic). In view of the widespread occurrence of such viruses in extreme environments, we designed an original method, called the inverted spot test, to determine the host range and infectivity of viruses isolated from anaerobic hyperthermophilic and sulfur-reducing microorganisms. Here, we used this approach to prove for the first time the infectivity of Pyrococcus abyssi virus 1 (PAV1) and to confirm the host range of Thermococcus prieurii virus 1 (TPV1), the only two viruses isolated so far from any of the described marine hyperthermophilic archaea (Euryarchaeota phylum, Thermococcales order).

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Acknowledgments

This work was financially supported by a MENRT Grant to A. Gorlas, by the CNRS “PID Origines des Planètes et de la Vie” programs (2007–2009) and the Foundation for Research on Biodiversity (FRB) (2009–2012). We thank Anne-Claire Baudoux and Christelle Desnues for their critical reading of the manuscript.

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  1. Laboratory of Microbiology of Extreme Environments (LMEE), UMR 6197/CNRS/UBO IUEM, Place Nicolas Copernic, Technopôle Brest Iroise, 29280, Plouzane, France

    Aurore Gorlas & Claire Geslin

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  1. Aurore Gorlas
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  2. Claire Geslin
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Correspondence to Aurore Gorlas.

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Communicated by F. Robb.

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792_2013_513_MOESM1_ESM.tif

Figure S1 Spot tests results with 10 μl TPV1 virions purified by iodixanol and several controls (10 μl TPV1 virions purified by CsCl, 10 μl iodixanol solution, and 10 μl T. prieurii culture supernatant) on the lawn of T. barophilus after 12 h of incubation (TIFF 203 kb)

792_2013_513_MOESM2_ESM.tif

Table S2a Characteristics distinguishing Thermococcus species tested in this study.1, T. barophilus (Marteinsson et al., 1999); 2, T. onnurineus (Bae et al., 2006); 3, T. kodakaraensis (Atomi et al., 2004); 4, T. celer (Zillig et al., 1983); 5, T. gorgonarius (Miroshnichenko et al., 1998); 6, T. siculi (Grote et al., 1999); 7, T. pacificus (Miroshnichenko et al., 1998); 8, T. fumicolans (Godfroy et al., 1996); 9, T. sibiricus (Miroshnichenko et al., 2001); 10, T. gammatolerans (Jolivet et al., 2003). +, Positive; –, Negative; S, Stimulatory; R, Required. ND: Not done. (TIFF 145 kb)

792_2013_513_MOESM3_ESM.tif

Table S2b Characteristics distinguishing Pyrococcus species tested in this study.1, P. furiosus (Fiala and Stetter, 1986); 2, P. horikoshii (González et al. 1998); 3, P. glycovorans (Barbier et al., 1999).+, Positive; –, Negative; S, Stimulatory; R, Required (TIFF 181 kb)

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Gorlas, A., Geslin, C. A simple procedure to determine the infectivity and host range of viruses infecting anaerobic and hyperthermophilic microorganisms. Extremophiles 17, 349–355 (2013). https://doi.org/10.1007/s00792-013-0513-0

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