Identification of the Aggregation Pheromone of the Melon Thrips,Thrips palmi

The objective of this study was to identify the aggregation pheromone of the melon thrips Thrips palmi, a major pest of vegetable and ornamental plants around the world. The species causes damage both through feeding activities and as a vector of tospoviruses, and is a threat to world trade and European horticulture. Improved methods of detecting and controlling this species are needed and the identification of an aggregation pheromone will contribute to this requirement. Bioassays with a Y-tube olfactometer showed that virgin female T. palmi were attracted to the odour of live males, but not to that of live females, and that mixed-age adults of both sexes were attracted to the odour of live males, indicating the presence of a male-produced aggregation pheromone. Examination of the headspace volatiles of adult male T. palmi revealed only one compound that was not found in adult females. It was identified by comparison of its mass spectrum and chromatographic details with those of similar compounds. This compound had a structure like that of the previously identified male-produced aggregation pheromone of the western flower thrips Frankliniella occidentalis. The compound was synthesised and tested in eggplant crops infested with T. palmi in Japan. Significantly greater numbers of both males and females were attracted to traps baited with the putative aggregation pheromone compared to unbaited traps. The aggregation pheromone of T. palmi is thus identified as (R)-lavandulyl 3-methyl-3-butenoate by spectroscopic, chromatographic and behavioural analysis.     

An observation on the embryonic development in Thrips palmi (Thysanoptera: Thripidae) eggs obtained by an artificial oviposition setup

Thrips palmi is an important insect pest of vegetables and ornamental crops worldwide. Besides direct damage caused by feeding, it transmits several tospoviruses in a persistent-propagative manner. Eggs of T. palmi are microscopic and embedded within plant tissue by the sharp ovipositor of adult female. In the present study, an artificial oviposition setup has been standardized for T. palmi. Eggs of T. palmi were harvested in sterile water between two thin membranes. The developmental stages of T. palmi embryo were studied starting from oviposition up to hatching by inverted and confocal reflection microscopy. Energids were homogeneously distributed at an early stage of development. The anterior end of the egg curved with a constriction post 38 h. Initiation of tissue organization, mouthparts, appendages, compound eyes were observed at different time points. Appendages were well developed and segmentation was prominent post 70 h. The embryo was completely developed at around 80 h and hatched by 86 h post oviposition at 28 °C temperature. The study first time reports the embryonic development of T. palmi that would be helpful in detailed investigations of thrips developmental biology and evolution.     

Antagonistic plant defense system regulated by phytohormones assists interactions among vector insect, thrips and a tospovirus

The western flower thrips (Frankliniella occidentalis) is a polyphagous herbivore that causes serious damage to many agricultural plants. In addition to causing feeding damage, it is also a vector insect that transmits tospoviruses such as Tomato spotted wilt virus (TSWV). We previously reported that thrips feeding on plants induces a jasmonate (JA)-regulated plant defense, which negatively affects both the performance and preference (i.e. host plant attractiveness) of the thrips. The antagonistic interaction between a JA-regulated plant defense and a salicylic acid (SA)-regulated plant defense is well known. Here we report that TSWV infection allows thrips to feed heavily and multiply on Arabidopsis plants. TSWV infection elevated SA contents and induced SA-regulated gene expression in the plants. On the other hand, TSWV infection decreased the level of JA-regulated gene expression induced by thrips feeding. Importantly, we also demonstrated that thrips significantly preferred TSWV-infected plants to uninfected plants. In JA-insensitive coi1-1 mutants, however, thrips did not show a preference for TSWV-infected plants. In addition, SA application to wild-type plants increased their attractiveness to thrips. Our results suggest the following mechanism: TSWV infection suppresses the anti-herbivore response in plants and attracts its vector, thrips, to virus-infected plants by exploiting the antagonistic SA-JA plant defense systems.     

Comparison of the predation capacities of two soil-dwelling predatory mites,Gaeolaelaps aculeifer and Stratiolaelaps scimitus (Acari: Laelapidae), on three thrips species

Soil-dwelling predatory mites are natural enemies of various soil pest insects and mites. Both Gaeolaelaps aculeifer (Canestrini) and Stratiolaelaps scimitus (Womersley) are commercialized natural enemies of thrips, but there is little information on the predation rate of these predatory mites on different thrips species. We compared their predation capacities on three thrips species, Frankliniella occidentalis, F. intonsa, and Thrips palmi, which are major pests of various horticultural plants. The predatory rate of G. aculeifer was higher than that of S. scimitus. Both predator species fed on more T. palmi thrips than F. occidentalis or F. intonsa thrips, which may be attributable to the smaller body size of T. palmi than the other thrips. Predation rates of female adults were 2.6–2.8 times higher than those of deutonymphs in both species. Predation rates were not separated according to the various developmental stages (i.e., second instar larva, pupa, or adult) of thrips; however, deutonymphs fed on fewer adults than larvae or pupae of F. occidentalis. Our results suggest that both G. aculeifer and S. scimitus are active predators that can prey during any of their developmental stages and on any species of thrips tested.     

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.     

In the presence of red light,cucumber and possibly other host plants lose their attractability to the melon thrips <Emphasis Type="Italic">Thrips palmi</Emphasis> (Thysanoptera: Thripidae)

The melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae), is a serious agricultural pest of many crops. Previous studies have shown that red light decreases the number of Thrips palmi in greenhouses. In order to understand how red light affects T. palmi, we examined the behavioral responses to host plants that were irradiated with a red light-emitting diode panel (660 nm) in an environment with natural or fluorescent (normal-white) light. When T. palmi were allowed to move freely around in the experimental arena, we found that fewer individuals were attracted to plants irradiated by red light than to plants under normal light illumination. We then used a sticky trap of green coloration to exclude olfactory and visual stimuli associated with the host plants in order to test binary choice behavior in T. palmi. The number of thrips attracted to the green sticky trap irradiated with red light was approximately half of that without red light irradiation. This is the first study to show that an addition of red light can change the behavior of insects, leading to an avoidance of green targets in an environment of normal illumination.     

Detection of the Quarantine Species Thrips palmi by Loop-Mediated Isothermal Amplification

Thrips palmi (from the order Thysanoptera) is a serious insect pest of various crops, including vegetables, fruits and ornamental plants, causing significant economic losses. Its presence constitutes a double threat; not only does T. palmi feed on the plants, it is also a vector for several plant viruses. T. palmi originated in Asia, but has spread to North and Central America, Africa, Oceania and the Caribbean in recent decades. This species has been sporadically noted in Europe and is under quarantine regulation in the European Union. For non-specialists its larval stages are indistinguishable morphologically from another widespread and serious insect pest Frankliniella occidentalis (a non-quarantine species in the European Union) as well as other frequently occurring thrips. In this study, we have developed a loop-mediated isothermal amplification protocol to amplify rDNA regions of T. palmi. The results were consistent whether isolated DNA or crushed insects were used as template, indicating that the DNA isolation step could be omitted. The described method is species-specific and sensitive and provides a rapid diagnostic tool to detect T. palmi in the field.     

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.     

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.     

Population ecology of Thrips palmi (Thysanoptera:Thripidae) in orchid farms in Thailand

The population dynamics and in-farm distribution of Thrips palmi Karny were evaluated in two orchid farms, one in the Bangkok metropolitan area and the other in Ratchaburi Province, Thailand. Yellow sticky traps were hung above the orchid canopy (three traps/220 m²), and the trapped adults were counted weekly from November 2009 to October 2010 at the first location and from January to December of 2012 at the second location. T. palmi nymphs and adults were both counted weekly on 300 orchid panicles in an area of 2,200 m² at both locations. We found that the T. palmi population dynamics at the first location was represented by a W-shaped curve, with the distribution clumped throughout the year, whereas the curve was J-shaped at the second location, with the distribution mostly clumped. Qualitative surveys of thrips species were conducted on ten different orchid genera in ten locations and on 20 different Dendrobium hybrids in 20 locations. Five random sampling plots (1 × 4 m²) were examined on each farm. The results indicated that only T. palmi was found on every farm. Moreover, quantitative comparison among five orchid types revealed that Dendrobium flowers with darker color are more attractive to T. palmi. However, this phenomenon does not coincide with Oncidium sp. and Mokara sp. because of their unfavorable morphology for thrips behavior.     

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.     

Herbivore arthropods benefit from vectoring plant viruses

Plants infected with pathogens often attract the pathogens’ vectors, but it is not clear if this is advantageous to the vectors. We therefore quantified the direct and indirect (through the host plant) effects of a pathogen on its vector. A positive direct effect of the plant‐pathogenic Tomato spotted wilt virus on its thrips vector (Frankliniella occidentalis) was found, but the main effect was indirect; juvenile survival and developmental rate of thrips was lower on pepper plants that were damaged by virus‐free thrips than on unattacked plants, but such negative effects were absent on plants that were damaged and inoculated by infected thrips or were mechanically inoculated with the virus. Hence, potential vectors benefit from attacking plants with virus because virus‐infected plants are of higher quality for the vector's offspring. We propose that plant pathogens in general have evolved mechanisms to overcome plant defences against their vectors, thus promoting pathogen spread.     

Economic thresholds of Thrips palmi (Thysanoptera: Thripidae) for eggplants in a greenhouse

Thrips palmi Karny has recently been implicated in damage to eggplant fruits. This thrips causes cosmetic scars on and deforms fruits, thus lowering their market value. To investigate the relationship between the density of thrips and the resulting damage, experimental plots with initial release densities of 0, 20, 40, 70, and 100 adults thrips per plot were established under greenhouse conditions. Thrips density (for flower sampling = x ₁; for sticky trap = x ₂) was monitored using flower sampling and blue sticky trap counts, and was found to be directly related to the proportion of damaged-fruits (y) and the reduction in fruit yield: significant relationships were found for the flower samples (y = 1.2261x ₁ ? 0.6232, r ² = 0.8582) and for the trap catches (y = 11.667 ln(x ₂) ? 9.5, r ² = 0.8896). The proportion of damaged fruits that could be tolerated from an economic perspective, based on the cost of controlling the thrips population chemically and the market value of the fruit, was 6.67–11.76 %; this translated into economic thresholds of 1.05–1.5 adults and/or larvae of T. palmi per flower or 4.91–10.17 adults T. palmi per four-day sticky card count.     

Olfactory response of the anthocorid predatory bug Orius sauteri to thrips-infested eggplants

The predator Orius sauteri (Poppius) (Heteroptera: Anthocoridae) is an effective arthropod natural enemy of thrips, especially Thrips palmi Karny (Thysanoptera: Thripidae), a serious pest of vegetables. First, we studied O. sauteri's response to the odour from thrips-infested eggplant [Solamum melongena L. (Solanaceae)] in a laboratory by two-choice experiments using a Y-tube olfactometer. When detached eggplant leaves were used as odour sources, O. sauteri preferred the volatiles from uninfested leaves to clean air. Concerning preferences among differently infested leaves, O. sauteri preferred the volatiles from plants infested with 10–100 thrips per leaf to uninfested leaves, but showed no significant preference for artificially damaged leaves over uninfested leaves. Similar results were obtained when complete plants were tested as the odour source. Second, release and recapture experiments in a greenhouse, a more realistic set of conditions, were conducted to confirm whether a significant preference for infested plants occurred at similar infestation levels as in the laboratory. Trends favouring infested plants were detected at densities of five and 500 thrips per plant; however, at the five thrips per plant, this trend was due to the large deviation seen in infested plants in only one replicate. In light of the low tolerable thrips density of eggplant, it is necessary to confirm whether artificial treatments with chemicals induce the emission of herbivore-induced plant volatiles that can attract O. sauteri and prolong its residence time on the leaf.     

Cuticular hydrocarbon pattern as a chemotaxonomy marker to assess six species of thrips

Thrips constitute several families of slender insects with fringed wings and unique asymmetrical mouthparts. They have become globally important pests, infesting a variety of agriculturally important crops. Species of thrips are difficult to identify due to their small size and similarities in morphology. Recently, in addition to morphology, both molecular and non-molecular taxonomic tools have been used to identify species differences. Insect cuticular hydrocarbons have been widely used in chemotaxonomy. In this study, a Thermal Separation Probe was used to identify the cuticular hydrocarbons of Frankliniella occidentalis, Frankliniella intonsa, Thrips palmi, Thrips hawaiiensis, Haplothrips chinensis and Gynaikothrips ficorum. We analyzed the hydrocarbon composition of adults in all 6 species, and in the larvae of F. occidentalis, T. hawaiiensis and T. palmi. The results showed that the composition of cuticular hydrocarbons differed between species. All 6 species of adults and 3 species of larvae were easily distinguishable by quantitative analysis of hydrocarbon profiles. These results provide a possible method for the identification of thrips.     

Colour preference and potato cultivar oviposition choice by onion thrips,Thrips tabaci (Thysanoptera: Thripidae)

Thrips tabaci (Lindeman) is an important polyphagous pest, and vector of Tomato spotted wilt virus responsible for sporadic, but devastating epidemics in potato. T. tabaci shows significant preferential differences between potato cultivars that may be important for field resistance. To investigate the role of visual cues in host choice we tested colour preference using two-choice assays. Thrips tabaci showed a significant preference for mid-green over red, blue, and white coloured cards, a preference for both light-green and mid-green over dark-green, and light-green over yellow, but no preferential difference between mid-green and yellow, and between light-green and mid-green. Analysis of the spectral reflectance of potato cultivars differing in thrips preference, revealed significant differences within the 400–700 nm wavelengths. In most cases cultivars preferred by T. tabaci had lighter green foliage and higher reflectance at 552 nm, and thus colour preference may be important for host selection. Oviposition choice of T. tabaci for potato cultivars was determined from counts of larvae and unhatched eggs from leaf disks under choice and no-choice conditions. In contrast to the colour choice experiments, onion thrips showed oviposition preference for cultivars with darker green foliage and lower reflectance at 552 nm.     

Cold tolerance, overwintering and establishment potential of Thrips palmi

The cold tolerance and overwintering survival of the quarantine regulated pest and virus vector, Thrips palmi (Karny) (Thysanoptera: Thripidae), is examined and discussed in terms of its establishment potential in the U.K. Thrips palmi adults and first‐instar larvae have a wide distribution of supercooling points (SCPs) but show ‘pre‐freeze’ mortality as a result of both acute and chronic exposures to temperatures above the SCP range. Thrips palmi did not develop enhanced cold tolerance in response to cues previously shown to cold‐acclimate other thrips species. The acute cold tolerance of T. palmi is higher than that of the recently established and biologically similar species, Frankliniella occidentalis, which is thought to be capable only of very limited winter survival outdoors in the U.K. However, the more ecologically meaningful chronic assays reveal the opposite pattern. If introduced to the U.K., overwintering of T. palmi would thus be largely restricted to protected environments, as its cold tolerance is insufficient to permit outdoor survival for a complete winter. This assertion was demonstrated by caged populations that died out after as little as 25 days in outdoor winter conditions in Yorkshire, U.K. The reversal of relative tolerance of the two species when considering chronic and acute assays suggests that these forms of low temperature mortality have different physiological bases, and emphasizes the need to use both forms of assay in assessments of overwintering capacity.     

Complete Genome Sequence of Mulberry Vein Banding Associated Virus,a New Tospovirus Infecting Mulberry

Mulberry vein banding associated virus (MVBaV) that infects mulberry plants with typical vein banding symptoms had been identified as a tentative species of the genus Tospovirus based on the homology of N gene sequence to those of tospoviruses. In this study, the complete sequence of the tripartite RNA genome of MVBaV was determined and analyzed. The L RNA has 8905 nucleotides (nt) and encodes the putative RNA-dependent RNA polymerase (RdRp) of 2877 aa amino acids (aa) in the viral complementary (vc) strand. The RdRp of MVBaV shares the highest aa sequence identity (85.9%) with that of Watermelon silver mottle virus (WSMoV), and contains conserved motifs shared with those of the species of the genus Tospovirus. The M RNA contains 4731 nt and codes in ambisense arrangement for the NSm protein of 309 aa in the sense strand and the Gn/Gc glycoprotein precursor (GP) of 1,124 aa in the vc strand. The NSm and GP of MVBaV share the highest aa sequence identities with those of Capsicum chlorosis virus (CaCV) and Groundnut bud necrosis virus (GBNV) (83.2% and 84.3%, respectively). The S RNA is 3294 nt in length and contains two open reading frames (ORFs) in an ambisense coding strategy, encoding a 439-aa non-structural protein (NSs) and the 277-aa nucleocapsid protein (N), respectively. The NSs and N also share the highest aa sequence identity (71.1% and 74.4%, respectively) with those of CaCV. Phylogenetic analysis of the RdRp, NSm, GP, NSs, and N proteins showed that MVBaV is most closely related to CaCV and GBNV and that these proteins cluster with those of the WSMoV serogroup, and that MVBaV seems to be a species bridging the two subgroups within the WSMoV serogroup of tospoviruses in evolutionary aspect, suggesting that MVBaV represents a distinct tospovirus. Analysis of S RNA sequence uncovered the highly conserved 5’-/3’-ends and the coding regions, and the variable region of IGR with divergent patterns among MVBaV isolates.     

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.