ANNOTATED SEQUENCE RECORD
Molecular characterization of a novel amalgavirus
from the entomopathogenic fungus Beauveria bassiana
Received: 3 November 2014 / Accepted: 30 March 2015 / Published online: 10 April 2015
Ó Springer-Verlag Wien 2015
Abstract Beauveria bassiana is a ubiquitous ento-
mopathogen infecting hundreds of insect species. We have
determined the genomic organization and the complete
nucleotide sequence of a novel virus isolated from the
isolate A24 of B. bassiana. Phylogenetic analysis of the
polymerase gene reveals that the virus, tentatively named
Beauveria bassiana virus 1, belongs to the family Amal-
gaviridae and represents a distinct lineage of amal-
gaviruses infecting fungi.
A growing number of viruses have been detected recently
in ascomycetous and, less frequently, basidiomycetous
(pathogenic as well as industrially cultivated micro- and
macrofungi) fungal species. The taxonomic system for
these viruses is continuously growing and being reﬁned.
Since 2009, a few new viruses with an undivided, relatively
small dsRNA genome separately infecting fungal and plant
hosts have been described. Due to similarities in their
RdRp sequences with those of members of the families
Totiviridae and Partitiviridae but signiﬁcant differences in
genome organization, a family named Amalgaviridae has
been newly established .
Here, we describe a new RNA virus infecting the as-
comycete fungus Beauveria bassiana (Balls.-Criv.) Vuill.
The virus has been provisionally named Beauveria bassi-
ana virus 1 (BbV1-A24). B. bassiana is a ubiquitous en-
tomopathogen infecting hundreds of insect species. It is
commercially used for microbial control as a mycoinsec-
ticide, and its isolates are used for producing a large
number of biologically active secondary metabolites .
To date, two viruses have been reported to infect B.
bassiana cultures. Both of these are members of the family
Totiviridae [3, 4].
A series of fungal isolates belonging to Beauveria
bassiana sensu lato obtained from the collection of the
Faculty of Agriculture at the University of South Bohemia
were screened for the presence of dsRNA elements.
Selective extraction of dsRNA was carried out as described
by Castillo et al. . The isolate A24 was found to harbor a
dsRNA element with an approximate molecular size of
3 kbp and was further investigated. A random cDNA li-
brary was prepared from the excised band using a Maxima
First Strand cDNA Synthesis Kit for RT-qPCR (Fermentas,
Lithuania) with 0.5 pmol of dN6 random primer 5
by Generi Biotech, Czech Republic) according to a pro-
tocol described by Darissa et al. . PCR reactions were
carried out using 2x PPP Master Mix (Top-Bio, Czech
Republic) in total volume 30 lL with 0.5 lMof5
SPA primer (Generi Bio-
tech, Czech Republic). The products obtained were re-
solved on a 2 % agarose gel and stained with SYBR Green
I dye. The PCR products were visualized as a smear with
sizes of 100 bp to *1 kbp. Fragments larger than 500 bp
were excised from the gel, puriﬁed using a GeneJET Gel
Electronic supplementary material The online version of this
article (doi:10.1007/s00705-015-2416-0) contains supplementary
material, which is available to authorized users.
& Igor Koloniuk
Department of Plant Virology, Institute of Plant Molecular
Biology, Biology Centre of the Czech Academy of Sciences,
31, 370 05 C
Department of Genetics, Faculty of Science, University of
South Bohemia in C
370 05 C
jovice, Czech Republic
Arch Virol (2015) 160:1585–1588