ANNOTATED SEQUENCE RECORD
Sequence determination and analysis of the NSs genes of two
Mikhail O. Leastro
Mirtes F. Lima
Alice K. Inoue-Nagata
Renato O. Resende
Received: 20 August 2011 / Accepted: 6 December 2011 / Published online: 21 December 2011
Ó Springer-Verlag 2011
Abstract The tospoviruses groundnut ringspot virus
(GRSV) and zucchini lethal chlorosis virus (ZLCV) cause
severe losses in many crops, especially in solanaceous and
cucurbit species. In this study, the non-structural NSs gene
and the 5
UTRs of these two biologically distinct tospo-
viruses were cloned and sequenced. The NSs sequence
of GRSV and ZLCV were both 1,404 nucleotides long.
Pairwise comparison showed that the NSs amino acid
sequence of GRSV shared 69.6% identity with that of
ZLCV and 75.9% identity with that of TSWV, while the
NSs sequence of ZLCV and TSWV shared 67.9% identity.
Phylogenetic analysis based on NSs sequences conﬁrmed
that these viruses cluster in the American clade.
Among the most detrimental plant viruses in Brazil, tospo-
viruses are especially important due to the devastating
damage they cause on many economically important crops.
Tomato spotted wilt virus (TSWV) is the type species of
the genus Tospovirus, which currently includes at least 23
recognized and tentative species [6, 7, 15, 30]. Based on
the genome structure and organization of its members,
the genus Tospovirus is classiﬁed within the family Bunya-
viridae and it is the only genus in this family whose
members infect plants, while the viruses in all of the other
genera infect vertebrates.
In Brazil, the tospoviruses cause high economic losses in
vegetable crops, such as sweet pepper, chilli pepper, lettuce,
tomato, cucurbits and ornamentals. Within the last decades,
in addition to Tomato spotted wilt virus (TSWV), members
of ﬁve other Tospovirus species or tentative species have
been identiﬁed in Brazil: Tomato chlorotic spot virus
(TCSV), Groundnut ringspot virus (GRSV), Chrysanthe-
mum stem necrosis virus (CSNV), Iris yellow spot virus
(IYSV) and Zucchini lethal chlorosis virus (ZLCV).
TSWV has a wide host range and it is found in different
crops worldwide. Like TSWV, IYSV has been reported on
several continents, but it has a narrow host range, infecting
iris, onion, garlic, lisianthus and alstroemeria [13, 21, 23].
CSNV isolates are found in Asia, Europe and South America
and infect tomato and chrysanthemum plants [14, 16].
GRSV has been reported in Africa and South America,
infecting plants of peanut, soybean, tomato, sweet pepper
and lettuce , and TCSV is found in South America,
infecting a number of distinct hosts . Recently, a natural
M RNA reassortant of GRSV and TCSV (S and L RNA
from GRSV and M RNA from TCSV-L
number NC_015467) was found in the United States in
tomato plants in Florida . ZLCV is restricted to Brazil,
and it has been reported in cucurbit plants in the states of Sa
Paulo, Tocantins and Distrito Federal [1, 3, 18].
The tripartite genome of tospoviruses consists of pseudo-
circular RNA segments, known as L (negative polarity), M
and S (ambisense polarity) RNA. The L RNA encodes the
RNA-dependent RNA polymerase (RdRp) . The M
RNA encodes the movement protein (NS
) and the
glycoproteins precursor . The S RNA encodes the
nucleocapsid protein (N) and the non-structural protein
(NSs), a silencing suppressor gene demonstrated for TSWV
M. Hallwass Á M. O. Leastro Á R. O. Resende (&)
Departamento de Biologia Celular, Universidade de Brası
Brasilia, DF 70910-900, Brazil
M. F. Lima Á A. K. Inoue-Nagata
Embrapa Hortalic¸as, Caixa Postal 218, Brasilia,
DF 70359-970, Brazil
Arch Virol (2012) 157:591–596