Bunyaviridae : Tospovirus :

Tomato Spotted Wilt Virus

By Connor Thornton
Fall 2013

Connor Thornton

U21655345

Introduction:
The family Bunyaviridae is host to
many vertebrates and invertebrate causing a
wide spread number of zoonotic diseases
but it also includes the genus Tospovirus
which infects plants. Tospovirus is most
notable for the Tomato Spotted Wilt Virus
(TSWV), a cause of plant wilting and death,
and all genre of Tospovirus cause major

Figure 1: Crop injury caused by infection

with Tomato spotted wilt disease.
http://www.plantday12.eu/imgScience/ISHS/i
shs.htm

hindrances to the produce industry as they causes wide spread crop destruction in
tomatoes, tobacco, onions, cucumbers, peppers and a number of other plants with
no known vaccine nor reliable prevention system(1). The crop damage is amplified
as TSWV is spread by a variety of thrip species which are difficult to control due to
their high reproductive rate, high mobility and high resistance to insecticides (15,
16).

Characteristics:
The Tomato Spotted Wilt Virus is a spherical virus about 90 nm in diameter
with an envelope containing many polypeptide spikes 5-10 nm long(2). The genome
is 16.63 kb in length segmented into three gene segments, L, M and S, and encodes
five proteins; envelope glycoprotein (GP), RNA-directed RNA polymerase L (L),
movement protein (NSM), nucleoprotein (N) and non-structural protein NS-S (NSS)
(3, 4, 5). Each of the three segments contains hairpin structures at each end
homologous to those of other Bunyaviridae species (2-5) and are capable of
reassortment to form recombinant viruses (2).

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The L gene segment

contains one large open reading
frame (ORF) which encodes for the
viral RNA-dependant RNA
polymerase as well as viral
encapsulation. It contains
panhandle structures at the
terminal sequences which contain the initiation sequences for replication and aid in
synthesis. Several key motifs in the L gene segment show homology with those of
other Bunyaviridae species as well as the PB1 protein in Thogoto virus of
Orthomyxoviridae which it is believed to share a common ancestor with (3).
The S gene segment is
ambisense, coding two ORF, one of

Figure 2: Representation of gene orginization

and composition.
http://viralzone.expasy.org/all_by_species/25
3.html

which encodes the nucleocapsid protein as well as the nonstructural protein NS1 (5,
6). NS1 has been determined as the causative agent of the virus and is found in
cells infected with TSWV but is not found in assembled virions (6).
The M gene segment, like the S gene segment, is ambisense and has two
ORF. One ORF encodes the precursor to the glycoprotiens G1 and G2 (4). Tospovirus
is the only member of Bunyaviridae with two ambisense RNA segments. TSWV also
contains a gene for a suppressor of RNA silencing (7) moreover has been shown to
work in conjunction with Iris yellow spot Tospovirus to further overcome host
defenses and produce greater infection rates than each alone (25).

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Transmission through the arthropod vector:

Tospovirus species are transmitted solely through thrip species to and from
plants. The western flower thrip (WFT), Frankliniella occidentalis is the most
common thrip to plague agroecosystems (9, 10, 11). Thrips as well as TSWV inhabit
a majority of the world and pose the most threat in western North America as well
as Eastern Europe and South-Eastern Asia. WFT show accelerated population growth
as well as reduced generation time at high temperatures, 30 degree Celsius, with
reduced rates in lower temperatures, 15 degree Celsius (20) but is seen to still
inhabit lower temperature regions if abundany food is present.
Thrips are shown to infect plants with
TSWV when feeding on plant tissue as the
TSWV particles replicate within infected
thrips salivary glands suggesting the golgi
apparatus has a major role in viral
replication or release. Transmission of the
TSWV from thrips to plants is attributed to
viral particles inhabiting the thrips saliva

Figure 3: Thrip lifecycle showing nymph

thrips as soil dwelling.
http://cisr.ucr.edu/avocado_thrips.html

entering the plant during feeding and infecting plant cells with aid of mechanical
damage caused by the thrip (12). The TSWV then replicate and produce nonstructural protein 1 (NS1) which causes cell death, plant wilting and eventual death
of the plant (6). NS1 produces filamentous inclusion bodies within host cells and
suppresses RNA-mediated gene silencing allowing the virus to evade host defenses
in addition to replicating in a wide variety of hosts (21). One infect thrip is able to
infect a multitude of plants and can do so indefinitely.

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Conversely, as thrips feed on infected plants viral particles including N and

NSs proteins are taken into and absorbed by the thrips midgut lumen and epithelial
cells. Here two different courses may occur based on the lifecycle of the thrip. If the
thrip is still a larva then the ingested TSWV particles will circulate through the
endothelial cells and would end up in the salivary glands where they would replicate
for release when the thrip feeds again. On the other hand, if the thrip is an adult
without TSWV in its salivary glands then the TSWV particles will not reach the thrips
head and will remain in the midgut lumen never to infect another host. The adults
that did not ingest TSWV particles as larvae cause the virus to no longer be spread
and it becomes non-virulent. Therefore, to obtain the virus a thrip must be born on
an infected plant as they do not leave it until they are adults (13). In fact, plants
infected with TSWV are better suited to hosting thrips than those without. Female
thrips find infected plants as a better breeding ground as the infected plants appear
to be more attractive. Incidentally, thrip-resistant plants (ones that prevent thrip
feeding) infected with TSWV are visited more often than non-infected thrip-restraint
plants but the females will not reproduce there as there is no food source. This
shows that thrips are drawn to TSWV and will readily spread it (22).

Prevention:
The easiest and most economic way to
remove insect pests is to make use of
insecticides to remove the pests before they
can infect the crop or to reduce the infection
rate. However, Thrips tabaci is susceptible to
diazinon and endosulfan while Frankliniella
occidentalis is resistant to all registered insecticides(8) and European and African

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strains of F. occidentalis also show clear signs of cross-resistance (17). Thrips prove
to be hearty animals that are recorded to avoid insecticide contact by avoiding
protected areas which in conjunction with, detoxification enzymes, esteraces,
glutathione S-transferases, and acetylcholinesterase which prevent any major
damage to be cause by chemical means (16). Another method to remove pests is
the addition of a predator species that wont
endanger the crops. Predatory mites such as

Figure 4: Use of sticky traps to catch thrips.

http://www.omafra.gov.on.ca/english/crops/facts/03075.htm

Amyblyseius cucumeris, Hypoaspis aculeifer and Hypoaspis miles have been

studied to provide significant reduction in the number of thrip larvae that reach
maturity (8, 9, 19). When thrip larvae enter the edaphic phase; propupa and pupa
phase (figure 3), they inhabit the soil and are vulnerable to soil-dwelling predators
(9) once they mature though the thrips become nearly impossible to catch and are
hunted by a few number of animals such as spiders and Hymenoptera [bees].
Nematode parasites, protozoan and fungi also infect thrips but have yet to show
significant research as suitable solutions (11).
A noteworthy reduction in the number of thrips was also shown by the
placement of sticky sheet bug traps where the thrips travel and feed. Traps 2.4m
above the floor with show peak effectiveness along with a color preference of blue
traps to capture females and yellow traps to capture male (18) though no evidence
to suggest gender differences in viral virulence. The removal of the female thrip is
preferred though as the larvae are isolated on the plant they are born on and since
the spread of the virus is dependent on the larva ingesting a plant infected with
TSWV the virus is dependent on female thrips (22). The use of sticky traps is an
effective way to reduce the number of thrips in a region but will not remove them
entirely.

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Fortunately plant species have shown to be able to generate resistance to

different serogroups of TSWV after inoculation of the plant with a nucleocapsid
nucleotide sequence from TSMV. The generated resistance was also shown to
extend to homologous, closely related serogroups and even to several Impatiens
necrotic spot virus (INSV) serogroups (15). This is not true of all plants though as
tobacco inoculated with several N genes
showed resistance to only homologous
strains and not that of less analogous
strains (14). On the other hand, the use of
acibenzolar-S-methyl and Imidacloprid to
prevent not the invasion of TSWV but the
symptoms showed that plants inoculated
before introduction into a TSWV infected
thrip environment are less likely to become

Figure 5: Effect of Tomato Spotted Wilt

Virus on tobacco leaves. Left is a
control leaf while right is a leaf first
innoculated with acibenzolar-S-methyl.
http://www.apsnet.org/publications/imagere
sources/Pages/IW000022.aspx

infected (23, figure 5). Additionally the ambisense nature of the M and S gene
segments causes viral small RNAs (vsiRNAs) to be produced that could be employed
as a means of preventing viral transcription and ceasing NS1 production thus
preventing symptoms in treated plants (24). With more research a vaccine to strains
of TSWV or an elimination or pacification of thrips may be engineered to reduce or
even eliminate the spread and density of occurrence.