Archives of Virology
Identication and molecular characterization of a novel circular
single‑stranded DNA virus associated with yerba mate in Argentina
· Soledad de Breuil
· Claudia Nome
Received: 6 March 2018 / Accepted: 27 April 2018
© Springer-Verlag GmbH Austria, part of Springer Nature 2018
A single-stranded DNA (ssDNA) virus was detected in Yerba mate samples showing chlorotic linear patterns, chlorotic
rings and vein yellowing. The full-genome sequences of six diﬀerent isolates of this ssDNA circular virus were obtained,
which share > 99% sequence identity with each other. The newly identiﬁed virus has been tentatively named as yerba
mate-associated circular DNA virus (YMaCV). The 2707 nt-long viral genome has two and three open reading frame on
its complementary and virion-sense strands, respectively. The coat protein is more similar to that of mastreviruses (44%
identity), whereas the replication-associated protein of YMaCV is more similar (49% identity) to that encoded by a recently
described, unclassiﬁed ssDNA virus isolated on trees in Brazil. This is the ﬁrst report of a circular DNA virus associated
with yerba mate. Its unique genome organization and phylogenetic relationships indicates that YMaCV represents a distinct
evolutionary lineage within the ssDNA viruses and therefore this virus should be classiﬁed as a member of a new species
within an unassigned genus or family.
Yerba mate (Ilex paraguariensis St. Hil., Aquifoliaceae)
is one of the most important subtropical trees or shrub
crops in Argentina, Brazil, and Paraguay. Its leaves and
stems are widely used in an infusion popularly known as
“mate”. In Argentina, the cultivated area with this tree is
about 165,200 ha with a total yield of 775 t per year .
In 2010, yerba mate plants showing chlorotic linear pat-
terns, chlorotic rings, and veins yellowing in symptomatic
leaves, were observed in the northeast region of Argentina
. Deep sequencing of small RNAs of symptomatic yerba
mate plants collected in Cerro Azul, Misiones Province,
Argentina, revealed the presence of a new virus: cytorhab-
dovirus yerba mate chlorosis associated virus (YmCaV) .
Two assembled sequences (contigs) showed a low degree of
similarity to the capsid and replication associated proteins
encoded by mastreviruses and the ssDNA virus temperate
fruit decay-associated virus (TFDaV), respectively, based
on BlastX analysis.
In this study we describe the complete genome sequence
of a new circular single strand DNA (ssDNA) virus isolated
from yerba mate in Argentina, which was named yerba mate-
associated circular DNA virus (YMaCV).
Leaf samples with the described symptoms were col-
lected in six diﬀerent locations around Corrientes and Mis-
iones Provinces in Argentina (Supplementary Figure 1).
DNA extractions were performed using 100 mg of infected
tissue with the cetyltrimethylammonium bromide (CTAB)
method . Total DNA from each isolate was subjected
to Phi29 DNA polymerase treatment (TempliPhi™, GE
Healthcare, USA) to obtain a copy of the complete genome
of the ssDNA virus by rolling circle ampliﬁcation (RCA).
A linear DNA fragment of about 2.7 kb was generated with
BamHI digestion, cloned and three clones per isolate were
sequenced bi-directionally with the universal M13 primers.
Additionally, primers were designed, by primer-walking,
to sequence the full genome of each clone. The sequence
fragments obtained for each plasmid DNA were edited and
Handling Editor: Elvira Fiallo-Olivé.
Electronic supplementary material The online version of this
article (https ://doi.org/10.1007/s0070 5-018-3910-y) contains
supplementary material, which is available to authorized users.
* Nicolás Bejerman
Instituto de Patología Vegetal, Centro de Investigaciones
Agropecuarias, Instituto Nacional de Tecnología
Agropecuaria (IPAVE-CIAP-INTA), Camino 60 Cuadras
Km 5,5 (X5020ICA), Córdoba, Argentina
Consejo Nacional de Investigaciones Cientíﬁcas y Técnicas
(CONICET), Godoy Cruz 2290 (1425 FBQ) CABA,
Buenos Aires, Argentina