Effects of tomato spotted wilt virus isolates, host plants, and temperature on survival, size, and development time of Frankliniella occidentalis

Tomato spotted wilt virus (TSWV) replicates in both its plant hosts and its thrips vectors. Replication of TSWV within thrips suggests the potential for pathological effects that could affect the fitness of its vectors directly, whereas infection of the plant may alter its suitability as a host for thrips development. This study was undertaken to examine the influence of TSWV isolate, host plant, and temperature on potential direct and host-mediated effects of virus infection of the thrips and the plant on Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), an important vector of TSWV. Neonate F. occidentalis were reared to adult eclosion on excised foliage of Datura stramonium (L.) (Solanaceae) or Emilia sonchifolia (L.) (Compositae) infected with either the CFL or RG2 isolate of TSWV, or not infected. Effects of the TSWV isolates and host plants on thrips were measured at 18.3, 23.9, and 29.4 °C. Results demonstrate significantly improved survival and a small but significant decrease in development time of F. occidentalis on TSWV-infected plants. These effects resulted from the combined influence of the direct effects of the virus on infected thrips and plant-mediated effects resulting from virus infection of the thrips’ host plant. Our results extend previous findings and help to explain inconsistencies among previously published reports by demonstrating that the manifestation and magnitude of effects of TSWV on F. occidentalis are dependent on host plant, virus isolate, and temperature.     

Summer weeds as hosts for Frankliniella occidentalis and Frankliniella fusca (Thysanoptera: Thripidae) and as reservoirs for tomato spotted wilt Tospovirus in North Carolina

In North Carolina, Tomato spotted wilt tospovirus (family Bunyaviridae, genus Tospovirus, TSWV) is vectored primarily by the tobacco thrips, Frankliniella fusca (Hinds), and the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). TSWV overwinters in winter annual weeds from which it is spread to susceptible crops in spring. Because most susceptible crops are destroyed after harvest before winter weeds emerge in the fall, infected summer weeds are thought to be the principal source for spread of TSWV to winter annual weeds in fall. A survey of summer weeds associated with TSWV-susceptible crops in the coastal plain of North Carolina conducted between May and October revealed that relatively few species were commonly infected with TSWV and supported populations of F. fusca or F. occidentalis. F. occidentalis made up > 75% of vector species collected from 15 summer weed species during 2002. The number of F. occidentalis and F. fusca immatures collected from plant samples varied significantly among plant species. Ipomoea purpurea (L.) Roth, Mollugo verticillata L., Cassia obtusifolia L., and Amaranthus palmeri S. Wats supported the largest numbers of immature F. occidentalis. Richardia scabra L., M. verticillata, and Ipomoea hederacea (L.) supported the largest numbers of F. fusca immatures. TSWV was present at 16 of 17 locations, and naturally occurring infections were found in 14 of 29 weed species tested. Five of the TSWV-infected species have not previously been reported as hosts of TSWV (A. palmeri, Solidago altissima L., Ipomoea lacunosa L., I. purpurea, and Phytolacca americana L.). Estimated rates of infection were highest in I. purpurea (6.8%), M. verticillata (5.3%), and I. hederacea (1.9%). When both the incidence of infection by TSWV and the populations of F. occidentalis and F. fusca associated with each weed species are considered, the following summer weed species have the potential to act as significant sources for spread of TSWV to winter annual weeds in fall: I. purpurea, I. hederacea, M. verticillata, A. palmeri, C. obtusifolia, R. scabra, Ambrosia artemisiifolia L., Polygonum pensylvanicum L., and Chenopodium album L.     

Preference of the vector thrips Frankliniella occidentalis for plants infected with thrips‐non‐transmissible Tomato spotted wilt virus

The effect of a thrips‐non‐transmissible Tomato spotted wilt virus (TSWV) on insect–host interactions between thrips and Arabidopsis thaliana was analysed. A wild‐type TSWV virulent isolate and a TSWV isolate that induces mild symptoms on inoculated plants (TSWV‐Mo) were used in this study, and TSWV‐Mo isolate was obtained by single local lesion isolation using Petunia x hybrid after several passages on Nicotiana rustica plants. In transmission test, although wild‐type TSWV (TSWV‐wt) was transmitted by two thrips species (transmission ratio; Frankliniella occidentalis, 25%; Thrips tabaci, 10%; and T. palmi, 0%), none of the thrips transmitted TSWV‐Mo. Feeding damage by F. occidentalis in A. thaliana plants was more extensive on TSWV‐wt‐infected plants than on TSWV‐Mo‐infected plants, despite comparable preference. Among the markers of plant defences, salicylic acid‐regulated genes were upregulated threefold to sixfold by TSWV‐wt or TSWV‐Mo infection. In contrast, jasmonate‐regulated genes and jasmonate/ethylene‐regulated genes were not affected by the infections. Pull assays showed that adjacent TSWV‐Mo‐infected plants were preferred over uninfected plants. In conclusion, our results showed that the transmissibility by thrips of TSWV is not related to preference of vector thrips and suggested that TSWV‐Mo‐infected plants may be used as attractants for behaviour control of thrips.     

Does natural deposition of pine pollen affect the ovipositional behavior of Frankliniella occidentalis and Frankliniella fusca?

Tree pollen, especially Pinus spp. (Pinaceae), is shed in large quantities every spring in North America. Pine pollen deposition onto leaves was found to significantly influence the ovipositional behaviors of certain thrips species (Thysanoptera: Thripidae) in peanut and tomato leaf choice and no‐choice tests. Pine pollen (Pinus elliottii Engelm.) increased the oviposition rate 2.9‐fold for Frankliniella occidentalis (Pergande) (western flower thrips) and 1.6‐fold for Frankliniella fusca (Hinds) (tobacco thrips) in choice tests averaged over both plant species. These results support the idea that pollen has a greater impact on F. occidentalis behavior than on F. fusca behavior. The most dramatic increase was in peanut, where F. occidentalis only oviposited on leaves dusted with pollen, suggesting that the addition of pollen stimulated this flower thrips to lay eggs on a poor host‐plant part. The impact of pollen on the rate of oviposition by thrips is important because it is the early‐instar nymphs that acquire tomato spotted wilt virus (TSWV), which these two thrips species vector. In a laboratory bioassay, the addition of pine pollen to TSWV‐infected peanut foliage increased the percentage of infected F. fusca after one generation.     

Obstructor,a Frankliniella occidentalis protein,promotes transmission of tomato spotted wilt orthotospovirus

Tomato spotted wilt orthotospovirus (TSWV) causes substantial economic losses to vegetables and other crops. TSWV is mainly transmitted by thrips in a persistent and proliferative manner, and its most efficient vector is the western flower thrips, Frankliniella occidentalis (Pergande). In moving from the thrips midgut to the salivary glands in preparation for transmission, the virions must overcome multiple barriers. Although several proteins that interact with TSWV in thrips have been characterized, we hypothesized that additional thrips proteins interact with TSWV and facilitate its transmission. In the current study, 67 F. occidentalis proteins that interact with G_N (a structural glycoprotein) were identified using a split-ubiquitin membrane-based yeast 2-hybrid (MbY2H) system. Three proteins, apolipoprotein-D (ApoD), orai-2-like (Orai), and obstructor-E-like isoform X2 (Obst), were selected for further study based on their high abundance and interaction strength; their interactions with G_N were confirmed by MbY2H, yeast β-galactosidase and luciferase complementation assays. The relative expressions of ApoD and Orai were significantly down-regulated but that of Obst was significantly up-regulated in viruliferous thrips. When interfering with Obst in larval stage, the TSWV acquisition rate in 3 independent experiments was significantly decreased by 26%, 40%, and 35%, respectively. In addition, when Obst was silenced in adults, the virus titer was significantly decreased, and the TSWV transmission rate decreased from 66.7% to 31.9% using the leaf disk method and from 86.67% to 43.33% using the living plant method. However, the TSWV acquisition and transmission rates were not affected by interference with the ApoD or Orai gene. The results indicate that Obst may play an important role in TSWV acquisition and transmission in Frankliniella occidentalis.     

Transmission of Tomato spotted wilt virus isolates able and unable to overcome tomato or pepper resistance by its vector Frankliniella occidentalis

Tomato spotted wilt virus (TSWV) causes serious diseases of many economically important crops. Disease control has been achieved by breeding tomato and pepper cultivars with the resistance genes Sw‐5 and Tsw, respectively. However, TSWV isolates overcoming these genetic resistances have appeared in several countries. To evaluate the risk of spread of these resistance‐breaking isolates, we tested their ability of transmission by the main vector of TSWV, the thrips Frankliniella occidentalis. We compared the transmission rate by thrips of six TSWV isolates of different biotype (able or unable to overcome this resistance in pepper and tomato), and with divergent genotype (A and B). Our results indicate that the transmission rate was related to the amount of virus accumulated in thrips but not to virus accumulation in the source plants on which thrips acquired the virus. No correlation was found between transmission efficiency by thrips and the genotype or between transmission efficiency and the ability of overcoming both resistances. This result suggests that resistance‐breaking isolates have the same potential to be transmitted as the isolates unable to infect resistant tomato and pepper cultivars.     

Variation in thrips species composition in field crops and implications for tomato spotted wilt epidemiology in North Carolina

Thrips were surveyed in tomato spotted wilt-susceptible crops in five areas across North Carolina. Tomato, pepper, and tobacco plants in commercial fields were sampled and 30 species of thrips were collected over a 3-year period. The most common species overall was Frankliniella tritici (Fitch). The most common thrips species that are known to vector Tomato Spotted Wilt Virus (TSWV) were F. fusca (Hinds), and F. occidentalis (Pergande). Relatively low numbers of Thrips tabaci Lindeman, another reported vector, were collected. The spatial and temporal occurrence of vectors varied with sampling method, crop species, region of North Carolina, and localized areas within each region. In a laboratory experiment, no difference was detected between the ability of F. fusca and F. occidentalis to acquire and transmit a local isolate of TSWV. Based on vector efficiency and occurrence, F. fusca is considered the most important vector of TSWV in tobacco, whereas both F. fusca and F. occidentalis are important vectors of TSWV in tomato and pepper.     

Relationships between Tospovirus Incidence and Thrips Populations on Tomato in Mendoza, Argentina

The objectives of this work were to estimate the capability of local populations of thrips as vectors of groundnut ringspot virus (GRSV) and tomato spotted wilt virus (TSWV), and to determine the species composition of vectors in tomato crops. Transmission assays were performed. Incidence of tospoviruses was estimated in commercial crops. Random samples of flowers were taken from tomato for identification of thrips. Of the five species of thrips tested, Frankliniella gemina (first record), F. occidentalis and F. schultzei transmitted GRSV and TSWV. F. schultzei was a significantly more efficient vector of GRSV than F. occidentalis under controlled assay conditions. The thrips were identified on flowers from six surveyed tomato crops. F. occidentalis was the most frequently identified species (43.0%), followed by F. schultzei (35.6%) and Thrips tabaci (10.1%). The incidence of tospoviruses was low (1.1–2.8%) in crops planted during August–September and greater (9.5–61.1%) in crops planted in December. GRSV was prevalent (85%) over TSWV (11%).     

Transmission of tomato spotted wilt virus by Frankliniella occidentalism median acquisition and inoculation access period

To quantify the transmission of tomato spotted wilt virus (TSWV) by Frankliniella occidentalis, the median acquisition access period (AAP₅₀) and median inoculation access period (IAP₅₀) were determined. These parameters were established using transmission rates obtained after AAPs and in IAPs which both ranged from 5 to 2560 min. An AAP₅₀ of 106 min was found when larvae acquired virus from TSWV-infected Impatiens plants. IAP₅₀s of 58 or 137 min, respectively, were calculated when petunia or Datura stramonium leaf disks were used to test the inoculation efficiency of viruliferous thrips. The virus could successfully be acquired or inoculated in periods of 5 min. Transmission reached an optimum after an AAP of 21.3 h (AAP_opt) and in an IAP of 42.7 h (IAP_opt). These results show that TSWV can be acquired and transmitted efficiently by F. occidentalis in short feeding periods.     

Virus infection alters the predatory behavior of an omnivorous vector

The interactions between parasites and their hosts can cause profound changes in host behavior, including changes that can alter other trophic interactions. The western flower thrips Frankliniella occidentalis is an important omnivorous insect vector of Tomato spotted wilt virus (TSWV), which infects crops worldwide and also infects its thrips vector. Here, we show that tospovirus‐infected female thrips become more predaceous, illustrating how the functional role of omnivores may change in response to pathogen infection. Our findings support the hypothesis that increased predation among virus‐infected female thrips compensates for the detrimental effects of virus infection. Because predatory behavior is unlikely to increase virus transmission to plants, it is doubtful that this shift in feeding behavior is due to an adaptive parasite manipulation of vector behavior. In this study, increases in predatory behavior were observed in female thrips, but not in male thrips. This sexually dimorphic compensatory response indicates that male and female thrips utilize different feeding strategies to compensate for parasite infection, the expression of which is constrained by resource availability. Our findings demonstrate a novel, but potentially common pathway by which viruses can influence the structure of trophic interactions in food webs.     

Propagation of tomato spotted wilt virus in Frankliniella occidentalis does neither result in pathological effects nor in transovarial passage of the virus

The effect of tomato spotted wilt virus (TSWV) on Frankliniella occidentalis Pergande (Thysanoptera; Thripidae) following a 6-hour acquisition access period on infected plants was investigated. No statistically significant differences were observed among viruliferous, non-viruliferous and control thrips with respect to developmental time, reproduction rate and survival. Thrips larvae, exposed or non-exposed to TSWV, developed from egg to adult in 13.1 and 13.2 days, respectively. Exposed females produced an average of 28.3 larvae whereas control thrips produced 22.3 larvae and longevity was 13.4 and 12.5 days, respectively. None of these values were significantly different. Population reproductive statistics, net reproductive rate (R ₀), mean generation time (T) and intrinsic rate of increase (r _m) were calculated from the life fertility tables. R ₀ and r _m were higher for viruliferous thrips as compared to non-viruliferous and non-exposed thrips. Virus transmission studies revealed that viruliferous thrips were able to transmit virus until death and that TSWV was not transovarially transmitted.     

Temporal and spatial dynamics of Tomato spotted wilt virus and its vector in a potato crop in Argentina

The nature of spatial and temporal dynamics of Tomato spotted wilt virus (TSWV) and its vector in a potato crop cv. Innovator without insecticide application is analysed. Seed tuber was analysed for the presence of TSWV as a source of initial inoculum. The presence of plants with symptoms of TSWV was evaluated by visual observation and DAS‐ELISA analysis to confirm the virus infection. Thrips species were collected from leaves and inflorescences and identified under stereomicroscope. The distribution of symptomatic plants and thrips species was recorded five times at 14 days intervals. The initial seed tuber infection was of 1.1%. Disease incidence was 0% at 29 days after planting (DAP), 0.2% at 43 DAP, 2.2% at 56 DAP, 11.6% at 70 DAP and 14.6% at 84 DAP. The progress of the disease was adequately described by a Logistic model [y = 0.15/(1 + 1205372.93 × exp (?0.22 × DAP))]. Thrips vector species identified as resident in the crop during the whole cycle were Thrips tabaci (n = 423), Frankliniella occidentalis (n = 141) and as occasional species, F. schultzei (n = 34) and F. gemina (n = 5) were found. At 43 and 56 DAP a random distribution pattern was observed and the thrips species found were T. tabaci (n = 188) and F. occidentalis (n = 105). An aggregated pattern was determined at 70 and 84 DAP. Spatial patterns of the disease spread suggest a polycyclic epidemic with TSWV secondary spread in the potato crop. Multiple control measures were deduced from these epidemiological results like virus testing in tubers, removal of external virus infection sources and thrips control.     

Preliminary evidence of recovery from Tomato spotted wilt virus infection in Frankliniella occidentalis individuals

In this study we analysed the ability of individual thrips to transmit Tomato spotted wilt virus (TSWV) in a population of Frankliniella occidentalis over their lifespan as adults (about 10 days). In three experiments a total of 636 thrips were individually tested for their transmission capacity through leaf disc assays using four inoculation access periods (IAPs). Almost half of the transmitting thrips maintained the capacity to infect leaf discs in each of the four IAPs, confirming the persistent propagative nature of the transmission modality. Nevertheless, a relevant number of thrips (9.25% of transmitter thrips) was able to transmit in the early phases of their adult life (for the first two IAPs), but did not transmit the virus for the remainder of their lifetime. We compared the virus titer of these individuals at the end of the fourth IAP with that of individuals that maintained transmission ability in the four IAPs and showed a statistically significant difference. This difference could be evidence for recovery from TSWV infection in individual thrips.     

Direct and indirect effects of a thrips‐transmitted Tospovirus on the preference and fitness of its vector,Frankliniella fusca

Phytoviruses including tospoviruses are known to affect the behavior and fitness of their vectors both positively and negatively. In this study, we investigated the effects of Tomato spotted wilt virus (TSWV) (family Bunyaviridae, genus Tospovirus) infection on the fitness and feeding ability of tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae) using peanut, Arachis hypogaea L. (Fabaceae), as a host. Potentially viruliferous F. fusca laid more eggs than non‐viruliferous F. fusca. In contrast, fewer potentially viruliferous F. fusca developed into adults and required a longer developmental time than non‐viruliferous F. fusca, indicating a direct negative effect of the virus on thrips fitness. In addition, no‐choice feeding tests indicated that non‐viruliferous F. fusca fed more rapidly than potentially viruliferous F. fusca. Typically, phytovirus infections are known to enhance the availability of vital nutrients such as free amino acids in infected host plants and to affect other important physiological processes negatively. Free amino acids are known to play a vital role in egg production and development. Further investigations in this study revealed that leaflets of infected plants had ca. 15 times more free amino acids than non‐infected leaflets. TSWV‐infected leaflets were used to rear potentially viruliferous thrips. Higher amino acid levels in TSWV‐infected leaflets than in non‐infected leaflets could have contributed to increased oviposition by potentially viruliferous F. fusca compared to non‐viruliferous F. fusca. Taken together, these results suggest that increased concentrations of free amino acids in TSWV‐infected plants might serve as an incentive for thrips feeding on otherwise unsuitable hosts, thereby facilitating TSWV acquisition and transmission.     

Vector and virus induce plant responses that benefit a non-vector herbivore

The negative cross-talk between induced plant defences against pathogens and arthropod herbivores is exploited by vectors of plant pathogens: a plant challenged by pathogens reduces investment in defences that would otherwise be elicited by herbivores. This negative cross-talk may also be exploited by non-vector herbivores which elicit similar anti-herbivore defences in the plant. We studied how damage by the thrips Frankliniella occidentalis and/or infection with Tomato spotted wilt virus (TSWV) affect the performance of a non-vector arthropod: the two-spotted spider mite Tetranychus urticae, a parenchym feeder just like F. occidentalis. Juvenile survival of spider mites on plants inoculated with TSWV by thrips was higher than on control and on thrips-damaged plants. However, thrips damage did not reduce spider-mite survival as compared to the control, suggesting that the positive effect of TSWV on spider-mite survival is independent of anti-thrips defence. Developmental and oviposition rates were enhanced on plants inoculated with TSWV by thrips and on plants with thrips damage. Therefore, spider mites benefit from TSWV-infection of pepper plants, but also from the response of plants to thrips damage. We suggest that the positive effects of TSWV on this non-vector species cannot be explained exclusively by cross-talk between anti-herbivore and anti-pathogen plant defences.     

Occurrence and localization of spherical viruslike particles in tissues of apparently healthy tobacco thrips,Frankliniella fusca,a vector of tomato spotted wilt virus

Spherical viruslike particles (VLP) were found in the tissues of apparently healthy tobacco thrips, Frankliniella fusca. The particles occurred in abundance in thrips from Ontario but were absent in thrips from Oklahoma reared under identical conditions. The VLP were not transmissible to any of the seven plant hosts (in four families) of F. fusca suggesting that they may be an insect virus. Transmission of tomato spotted wilt virus (TSWV) by F. fusca, a known vector, was not affected by the presence or absence of the VLP. No TSWV particles were detected in tissues of F. fusca that transmitted TSWV to test plants. The VLP occurred in several internal organs and hemocoele of the thrips and were isolated in vitro by preparing homogenates of gut tissues. Infection of oocytes and presence of VLP in young nymphs suggested transovarial transmission of the particles. The VLP measured 62 ± 4 nm in diameter and usually occurred in dense viroplasms in the cell cytoplasm. Development of the particles within the viroplasms is discussed.     

Life-history parameters of western flower thrips on susceptible and resistant cucumber genotypes

The life-history parameters reproduction rate, developmental time and age specific survival of the western flower thrips,Frankliniella occidentalis (Pergande) [Thysanoptera: Thripidae], were determined on susceptible and resistant cucumber (Cucumis sativus L.) genotypes. Both newly emerged andF. occidentalis females of mixed ages showed a substantial reduction (36 to 50%) of the reproduction rate on all resistant genotypes, in particular after the second day. On the resistant genotypes 9127 and 9140,F. occidentalis had a prolonged developmental period. This was primarily due to a prolongation of the second larval stage. On all resistant genotypes,F. occidentalis suffered from high (82 to 97%) preadult mortality, predominantly at the second larval stage. It is conclude that the resistant genotypes do not cause an immediate intoxication of adult nor preadult thrips stages.     

Seasonal abundance of hemipteran predators in relation to western flower thrips Frankliniella occidentalis (Thysanoptera: Thripidae) on weeds in the eastern Mediterranean region of Turkey

Generalist predators and the prey species Frankliniella occidentalis (Pergande) were periodically sampled from 64 weed species to determine their numerical interrelationships in three ecologically different locations in the eastern Mediterranean region of Turkey in 2002–2003. Adult and nymph stages of seven hemipteran predator species were recorded: Deraeocoris pallens Reuter, Geocoris arenarius (Jakovkev), Nabis punctatus Costa, Orius laevigatus Fieber, Orius majusculus (Reuter), Orius niger (Wolff) and Piocoris erythrocephalus (Peletie and Serville). The most common and abundant predator species was O. niger, which was found on 53 weed species in the winter–spring and summer–fall periods; the highest total numbers of O. niger adults per weed species were 139, 275, 266 and 325 on Urtica urens, Sinapis arvensis, Lamium amplexicaule and Mentha aquatica, respectively. Relatively higher numbers of Orius spp. nymphs were detected on the weed species Echium plantagineum, Cichorium intybus, Heliotropium europeum, Mentha aquatica and Polygonum aviculare. Orius spp. were attracted to flowers in significantly higher numbers than to leaves of the investigated weeds. No significant correlation was found between the numbers of Orius spp. and F. occidentalis on the major weed species, except E. plantagineum. These findings suggest that weeds may provide Orius spp. with resources other than prey, such as nectar and pollen, shelter, and breeding sites. The weed species on which relatively higher numbers of Orius adults and nymphs were found may be considered as candidates for studies aiming to include weeds to enhance integrated pest management of F. occidentalis on crops.     

Internal anatomy and morphology of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) with special reference to interactions between thrips and tomato spotted wilt virus

The internal morphology of the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) (WFT), a primary vector of tomato spotted wilt virus (TSWV), was shown to bear certain similarities to other thrips species, particularly in the composition of the piercing-sucking feeding structures. Striking differences were observed in number, position and ducting of the salivary glands, the morphology of the alimentary canal and the number and arrangement of the malpighian tubules between the WFT and other studied Thysanopterans. These differences provide support for the conclusion that internal morphologies may vary widely in this order, and perhaps among species in the same genus. The results of our investigation support the need for more detailed studies of other thrips species, particularly in light of the potential importance morphological characteristics may play in governing the capacity of thrips species to serve as vectors of TSWV.