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Journal of Virology, February 2004, p. 1256-1262, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1256-1262.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of Host-Specificity Determinants in Betanodaviruses by Using Reassortants between Striped Jack Nervous Necrosis Virus and Sevenband Grouper Nervous Necrosis Virus

Tokinori Iwamoto,1,{dagger} Yasushi Okinaka,2* Kazuyuki Mise,3 Koh-Ichiro Mori,1 Misao Arimoto,1 Tetsuro Okuno,3 and Toshihiro Nakai2

Kamiura Station, Japan Sea-Farming Association, Oita 879-2602,1 Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528,2 Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan3

Received 27 June 2003/ Accepted 4 October 2003

Betanodaviruses, the causal agents of viral nervous necrosis in marine fish, have bipartite positive-sense RNAs as genomes. The larger genomic segment, RNA1 (3.1 kb), encodes an RNA-dependent RNA polymerase, and the smaller genomic segment, RNA2 (1.4 kb), codes for the coat protein. Betanodaviruses have marked host specificity, although the primary structures of the viral RNAs and encoded proteins are similar among betanodaviruses. However, no mechanism underlying the host specificity has yet been reported. To evaluate viral factors that control host specificity, we first constructed a cDNA-mediated infectious RNA transcription system for sevenband grouper nervous necrosis virus (SGNNV) in addition to that for striped jack nervous necrosis virus (SJNNV), which was previously established by us. We then tested two reassortants between SJNNV and SGNNV for infectivity in the host fish from which they originated. When striped jack and sevenband grouper larvae were bath challenged with the reassortant virus comprising SJNNV RNA1 and SGNNV RNA2, sevenband groupers were killed exclusively, similar to inoculation with SGNNV. Conversely, inoculations with the reassortant virus comprising SGNNV RNA1 and SJNNV RNA2 killed striped jacks but did not affect sevenband groupers. Immunofluorescence microscopic studies using anti-SJNNV polyclonal antibodies revealed that both of the reassortants multiplied in the brains, spinal cords, and retinas of infected fish, similar to infections with parental virus inoculations. These results indicate that viral RNA2 and/or encoded coat protein controls host specificity in SJNNV and SGNNV.

* Corresponding author. Mailing address: Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan. Phone: 81-824-24-7978. Fax: 81-824-22-7059. E-mail: okinaka{at}hiroshima-u.ac.jp.

{dagger} Present address: University of Alabama at Birmingham, Birmingham, AL 35294.

Journal of Virology, February 2004, p. 1256-1262, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1256-1262.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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