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Whiteflies are sap-sucking insects responsible for high economic losses. They colonize hundreds of plant species and cause direct feeding damage and indirect damage through transmission of devastating viruses. Modern agriculture has seen a history of invasive whitefly species and populations that expand to novel regions, bringing along fierce viruses. Control efforts are hindered by fast virus transmission, insecticide-resistant populations, and a wide host range which permits large natural reservoirs for whiteflies. Augmentative biocontrol by parasitoids while effective in suppressing high population densities in greenhouses falls short when it comes to preventing virus transmission and is ineffective in the open field. A potential source of much needed novel control strategies lays within a diverse community of whitefly endosymbionts. The idea to exploit endosymbionts for whitefly control is as old as identification of these bacteria, yet it still has not come to fruition. We review where our knowledge stands on the aspects of whitefly endosymbiont evolution, biology, metabolism, multitrophic interactions, and population dynamics. We show how these insights are bringing us closer to the goal of better integrated pest management strategies. Combining most up to date understanding of whitefly–endosymbiont interactions and recent technological advances, we discuss possibilities of disrupting and manipulating whitefly endosymbionts, as well as using them for pest control.

Pepper crops in Israel are infected by poleroviruses, Pepper vein yellows virus 2 (PeVYV-2) and Pepper whitefly-borne vein yellows virus (PeWBVYV). Herein we characterize the transmission of PeWBVYV and the aphid-transmitted PeVYV-2, and show that PeWBVYV is specifically transmitted by MEAM1 species of the whitefly Bemisia tabaci, with a minimum latency period of 120 h, and not by the Mediterranean (MED). PeWBVYV and PeVYV-2 were detected in the hemolymph of MED and MEAM1, respectively, however, amounts of PeWBVYV in the hemolymph of MED or PeVYV-2 in MEAM1 were much lower than PeWBVYV in hemolymph of MEAM1. Moreover, we show that PeWBVYV does not interact with the GroEL protein of the symbiont Hamiltonella and thus does not account for the non-transmissibility by MED. An insect glycoprotein, C1QBP, interacting in vitro with the capsid proteins of both PeWBVYV and PeVYV-2 is reported which suggests a putative functional role in polerovirus transmission.

Made available in DSpace on 2022-04-28T19:40:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Pepper cultivation in Israel has been constrained by two sympatric poleroviruses, Pepper vein yellows virus-2 (PeVYV-2) and Pepper whitefly-borne vein yellows virus (PeWBVYV), which are transmitted specifically by aphids and whiteflies, respectively. This study investigated the interaction between PeVYV-2 and PeWBVYV inside the host plant and the insect vectors. Our results show that PeVYV-2 and PeWBVYV compete against each other inside the host plant and also inside aphids. PeWBVYV was the weaker competitor inside the host plant, with diminished transmission rates when inoculated simultaneously or successively after PeVYV-2 and could only be transmitted efficiently when inoculated first and then challenged by PeVYV-2. Successive inoculations of plants with viruliferous whiteflies with PeWBVYV followed by viruliferous aphids with PeVYV-2 led to a coinfection rate of 60%, but with severely reduced titers of PeWBVYV in the coinfected plants compared with singly infected plants. In contrast, PeVYV-2 was the weaker competitor inside the insect vector, with reduced quantities of the acquired virus and a reduced transmission rate by aphids when given prior acquisition on PeWBVYV. However, we also show that the transmission efficiency of PeVYV-2 and PeWBVYV from coinfected plants by whiteflies and aphids remained comparable to that from singly infected plants. This is likely attributable to the reduced titers of PeWBVYV inside coinfected plants causing lesser impact on transmission of PeVYV-2 by aphids and the stronger competitiveness of PeWBVYV inside the whitefly. Competitive interactions between PeVYV-2 and PeWBVYV inside the host plant and insect vector can thus be beneficial for their coexistence. Department of Entomology Volcani Center Faculdade de Ci^encias Agron^omicas São Paulo State University (UNESP) Faculdade de Ci^encias Agron^omicas São Paulo State University (UNESP)

Made available in DSpace on 2021-06-25T11:15:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Since the detection of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) species in Brazil in the early 1990s, this whitefly has been the predominant species and the main viral vector in the country. In this work, whiteflies were collected from commercial soybean fields near and far from greenhouses where Mediterranean (MED) species had previously been detected infesting vegetable crops. Results indicated that MEAM1 was the predominant whitefly species in soybean in most sampling sites, while MED was found colonizing soybean plants in open field conditions alone and/or with MEAM1 in several places. Among the tested insects, MED species was mostly detected harbouring the facultative endosymbiont Hamiltonella. We also detected cowpea mild mottle virus (CPMMV) infecting soybean plants. Transmission assays demonstrated that MED species was more efficient transmitting CPMMV from beans to beans and from soybean to soybean plants than MEAM1, while MEAM1 was more efficient in transmitting the virus from soybean to bean than MED. Performance assays indicated that adult emergence of both species on CPMMV-infected soybean plants was higher when compared with the emergence on healthy plants. Moreover, nonviruliferous MED and MEAM1 adults preferred to settle more often on healthy plants, while viruliferous adults settled more often on CPMMV-infected soybean plants. As MED has already been found in soybean open fields in São Paulo and Paraná States, Brazil, and it is a good vector of CPMMV, we anticipate problems related to whitefly management and to increase in the incidence of the virus in soybean. UNESP – Faculdade de Ciências Agronômicas Corteva Agriscience Instituto Federal Catarinense – Campus Gulf Coast Research and Education Center University of Florida Instituto Agronômico The Volcani Center Institute of Plant Protection UNESP – Faculdade de Ciências Agronômicas

The sweet potato (cotton) whitefly Bemisia tabaci is a major agricultural pest in various fields and vegetable crops worldwide. It causes extensive damage by direct feeding on plants, reducing quality, secreting honeydew and transmitting plant viruses. B. tabaci is known for its genetic diversity and considered a complex of biotypes or, as suggested, a complex of distinct cryptic species. Management of whiteflies relies mainly on the use of insecticides; however, its ability to develop resistance to major insecticide classes creates a serious challenge to farmers and pest control specialists. Among the cryptic species of B. tabaci, MED is considered more resistant than the MEAM1 to insecticides such as pyriproxyfen and neonicotinoids; however, in recent years there are other species of B. tabaci including MEAM1, Asia I and Asia II-1 that have developed high resistance to various groups of insecticides. Advanced methods based on molecular and gene sequence data obtained from resistant and susceptible field-collected B. tabaci populations resulted in a better understanding of resistance mechanisms in this pest. Several components of IPM-IRM (Integrated Pest Management-Insecticide Resistance Management) programs such as selective and biorational insecticides, insecticide rotation with different modes of action and nonchemical control methods are among the countermeasures of insecticide resistance management for this pest. In the current review, we concentrate on insecticide resistance and resistance management of B. tabaci, focusing on reports published mainly over the past 10 years.

Many plant viruses depend on insect vectors for their transmission and dissemination. The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is one of the most important virus vectors, transmitting more than four hundred virus species, the majority belonging to begomoviruses (Geminiviridae), with their ssDNA genomes. Begomoviruses are transmitted by B. tabaci in a persistent, circulative manner, during which the virus breaches barriers in the digestive, hemolymph, and salivary systems, and interacts with insect proteins along the transmission pathway. These interactions and the tissue tropism in the vector body determine the efficiency and specificity of the transmission. This review describes the mechanisms involved in circulative begomovirus transmission by B. tabaci, focusing on the most studied virus in this regard, namely the tomato yellow leaf curl virus (TYLCV) and its closely related isolates. Additionally, the review aims at drawing attention to the recent knowhow of unorthodox virus—B. tabaci interactions. The recent knowledge of whitefly-mediated transmission of two recombinant poleroviruses (Luteoviridae), a virus group with an ssRNA genome and known to be strictly transmitted with aphids, is discussed with its broader context in the emergence of new whitefly-driven virus diseases.

Bacterial endosymbionts play essential roles in the biology of their arthropod hosts by interacting with internal factors in the host. The whitefly Bemisia tabaci is a worldwide agricultural pest and a supervector for more than 100 plant viruses. Like many other arthropods, Be. tabaci harbours a primary endosymbiont, Porteira aleyrodidarum, and an array of secondary endosymbionts that coexist with Portiera inside bacteriocyte cells. Unlike all of the other secondary symbionts that infect Be. tabaci, Rickettsia has been shown to be an exception by infecting insect organs and not colocalizing with Portiera, and has been shown to significantly impact the insect biology and its interactions with the environment. Little is known about the molecular interactions that underlie insect-symbiont interactions in general, and particularly Be. tabaci-Rickettsia interactions. Here we performed transcriptomic analysis and identified vitellogenin as an important protein that influences the levels of Rickettsia in Be. tabaci. Vitellogenin expression levels were lower in whole insects, but higher in midguts of Rickettsia-infected insects. Immunocapture-PCR assay showed interaction between vitellogenin and Rickettsia, whereas silencing of vitellogenin resulted in nearly complete disappearance of Rickettsia from midguts. Altogether, these results suggest that vitellogenin plays an important role in influencing the levels of Rickettsia in Be. tabaci.

The whitefly Bemisia tabaci is a major pest to agriculture. Adults are able to fly for long distances and to colonize staple crops, herbs and ornamentals, and to vector viruses belonging to several important taxonomic groups. During their early development, whiteflies mature from eggs through several nymphal stages (instars I to IV) until adults emerge from pupae. We aim at reducing whitefly populations by inhibiting the emergence of adults from nymphs. Here we targeted dystrophin, a conserved protein essential for the development of the muscle system in humans, other animals and insects. We have exploited the fact that whitefly nymphs developing on tomato leaves feed from the plant phloem via their stylets. Thus, we delivered dystrophin-silencing double-stranded RNA to nymphs developing on leaves of tomato plantlets with their roots bathing in the silencing solution. Downregulation of dystrophin expression occurred mainly in pupae. Dystrophin silencing induced also the downregulation of the dystrophin-associated protein genes actin and tropomyosin, and disrupted F-actin. Most significantly, the treatment inhibited the emergence of adults from pupae, suggesting that targeting dystrophin may help to restrain whitefly populations. This study demonstrates for the first time the important role of dystrophin in the development of a major insect pest to agriculture.

Single aphids can simultaneously or sequentially acquire and transmit multiple potato virus Y (PVY) strains. Multiple PVY strains are often found in the same field and occasionally within the same plant, but little is known about how PVY strains interact in plants or in aphid stylets. Immuno-staining and confocal microscopy were used to examine the spatial and temporal dynamics of PVY strain mixtures (PVYO and PVYNTN or PVYO and PVYN) in epidermal leaf cells of ‘Samsun NN’ tobacco and ‘Goldrush’ potato. Virus binding and localization was also examined in aphid stylets following acquisition. Both strains systemically infected tobacco and co-localized in cells of all leaves examined; however, the relative amounts of each virus changed over time. Early in the tobacco infection, when mosaic symptoms were observed, PVYO dominated the infection although PVYNTN was detected in some cells. As the infection progressed and vein necrosis developed, PVYNTN was prevalent. Co-localization of PVYO and PVYN was also observed in epidermal cells of potato leaves with most cells infected with both viruses. Furthermore, two strains could be detected binding to the distal end of aphid stylets following virus acquisition from a plant infected with a strain mixture. These data are in contrast with the traditional belief of spatial separation of two closely related potyviruses and suggest apparent non-antagonistic interaction between PVY strains that could help explain the multitude of emerging recombinant PVY strains discovered in potato in recent years.

Several vector-borne plant pathogens have evolved mechanisms to exploit and to hijack vector host cellular, molecular, and defense mechanisms for their transmission. In the past few years, Liberibacter species, which are transmitted by several psyllid vectors, have become an economically important group of pathogens that have devastated the citrus industry and caused tremendous losses to many other important crops worldwide. The molecular mechanisms underlying the interactions of Liberibacter species with their psyllid vectors are poorly studied. “Candidatus Liberibacter solanacearum,” which is associated with important vegetable diseases, is transmitted by the carrot psyllid Bactericera trigonica in a persistent manner. Here, we elucidated the role of the B. trigonica Arp2/3 protein complex, which plays a major role in regulation of the actin cytoskeleton, in the transmission of “Ca. Liberibacter solanacearum.” “Ca. Liberibacter solanacearum” colocalized with ArpC2, a key protein in this complex, and this colocalization was strongly associated with actin filaments. Silencing of the psyllid ArpC2 disrupted the colocalization and the dynamics of F-actin. Silencing of RhoGAP21 and Cdc42, which act in the signaling cascade leading to upregulation of Arp2/3 and F-actin bundling, showed similar results. On the other hand, silencing of ArpC5, another component of the complex, did not induce any significant effects on F-actin formation. Finally, ArpC2 silencing caused a 73.4% reduction in “Ca. Liberibacter solanacearum” transmission by psyllids, strongly suggesting that transmission of “Ca. Liberibacter solanacearum” by B. trigonica is cytoskeleton dependent and “Ca. Liberibacter solanacearum” interacts with ArpC2 to exploit the intracellular actin nucleation process for transmission. Targeting this unique interaction could lead to the development of a novel strategy for the management of Liberibacter-associated diseases. IMPORTANCE Plant diseases caused by vector-borne pathogens are responsible for tremendous losses and threaten some of the most important agricultural crops. A good example is the citrus greening disease, which is caused by bacteria of the genus Liberibacter and is transmitted by psyllids; it has devastated the citrus industry in the United States, China, and Brazil. Here, we show that psyllid-transmitted “Candidatus Liberibacter solanacearum” employs the actin cytoskeleton of psyllid gut cells, specifically the ArpC2 protein in the Arp2/3 complex of this system, for movement and transmission in the vector. Silencing of ArpC2 dramatically influenced the interaction of “Ca. Liberibacter solanacearum” with the cytoskeleton and decreased the bacterial transmission to plants. This system could be targeted to develop a novel approach for the control of Liberibacter-associated diseases.

The destiny of tomato yellow leaf curl viruses (TYLCVs), as well as other begomoviruses, ingested by Bemisia tabaci during feeding on infected plants is quite intriguing. During the first 12 to 48 h, the virus load reaches a threshold of approximately 600 million genomes per insect. This amount remains nearly stable, even though the insect continues to feed for days and weeks, raising several questions. How is such a limit maintained? What are the contributions of ingestion, egestion, excretion, replication, and immunity in maintaining this limit? How is the virus distributed along the circulative pathway and where is it stored? What are the constraints regulating the amount of virus in the insect vector, and what may happen to the insect (and/or the virus) if the regulation is impaired? This review presents and discusses the data implying that several different processes are involved in regulating the amount of begomovirus in B. tabaci, from the side of the insect as well as that of the virus, perhaps to prevent the virus to become pathogenic to its whitefly host.

BACKGROUND Intensive application of chemical insecticides is required for aphid pest control. Among the biorational alternatives, entomopathogenic fungi are the most sustainable biocontrol agents; those of the order Hypocreales attack and cause fungal disease in arthropod hosts, with variations in host susceptibility attributed to both fungal and host characteristics. We evaluated inter- and intraspecies variations in Metarhizium spp. virulence and differences in fungal disease progression on adult and nymph stages of the green peach aphid, Myzus persicae (Sulzer), a parthenogenetically reproducing insect species. RESULTS Minor interspecies diversity was detected between the generalist Metarhizium species examined. Interestingly, significant intraspecies diversity was observed between Metarhizium brunneum isolates Mb7 and MbK. Infected adult aphids demonstrated similar disease progression for both isolates, mortality rates of more than 80% and fivefold reduction in fecundity. However, nymph mortality was detected only following MbK infection, with 50% mortality and significant reduction in molting rates. Confocal laser scanning microscopy demonstrated the variation in the disease stages of conidial adhesion and hemocoel colonization on each examined day post inoculation for each isolate. Significantly faster disease progression was observed in MbK-infected versus Mb7-infected nymphs, the latter demonstrating a higher percentage of uninfected nymphs accompanied with aphid molting. CONCLUSIONS The observed intraspecies variation suggests that altered conidial adhesion to the nymph cuticle is a major factor affecting virulence. We prove the role of nymph ecdysis as a defense mechanism disrupting fungal infection. Because significant differences were observed between closely related isolates, this study emphasizes the importance of appropriate isolate selection for biological control. © 2021 Society of Chemical Industry.

Recent reports of transmission of poleroviruses by whiteflies is indicative of evolution of new virus-vector relationships. Pepper whitefly-borne vein yellows virus (PeWBVYV), was the first report of a polerovirus infecting pepper in Israel which was transmitted by whiteflies (MEAM1) and not aphids. This study reports the inability of the Mediterranean species (MED, Q biotype) of B. tabaci to transmit PeWBVYV. However we show that non-transmission of PeWBVYV by MED is not due to the lack of interaction with the GroEL protein of the Hamiltonella symbiont. Although not transmitted by MED, PeWBVYV was detected in its hemolymph, indicating its translocation across the MED midgut barrier. The aphid transmitted Pepper vein yellows virus 2 (PeVYV-2) was also detected in the hemolymph of MEAM1 whiteflies but PeWBVYV could not be detected in the aphid hemolymph. Interestingly, relative amounts of PeWBVYV in the hemolymph of the, MED was much lower than in hemolymph of MEAM1 whiteflies. We also identified a candidate receptor protein, complement component 1Q sub-complement binding protein (C1QBP) which interacts with the capsid proteins of PeWBVYV and PeVYV-2 but not with the whitefly transmitted Tomato yellow leaf curl virus by a yeast two-hybrid approach using the minor capsid protein (RTD) as bait to screen for interacting proteins against the whitefly cDNA library. C1QBP, is a known receptor of bacterial and viral pathogens but this is the first report of its interaction with a plant virus.

Simple Summary Whiteflies are one of the most important and widespread pests in the world. In South America, the currently most important species occurring are Bemisia afer, Trialeurodes vaporariorum, and the cryptic species Middle East-Asia Minor 1, Mediterranean, and New World, from Bemisia tabaci complex. The present review compiles information from several studies conducted in South America regarding these insects, providing data related to the dynamics and distribution of whiteflies, the associated viruses, and the management strategies to keep whiteflies under the economic damage threshold. Abstract By having an extensive territory and suitable climate conditions, South America is one of the most important agricultural regions in the world, providing different kinds of vegetable products to different regions of the world. However, such favorable conditions for plant production also allow the development of several pests, increasing production costs. Among them, whiteflies (Hemiptera: Aleyrodidae) stand out for their potential for infesting several crops and for being resistant to insecticides, having high rates of reproduction and dispersal, besides their efficient activity as virus vectors. Currently, the most important species occurring in South America are Bemisia afer, Trialeurodes vaporariorum, and the cryptic species Middle East-Asia Minor 1, Mediterranean, and New World, from Bemisia tabaci complex. In this review, a series of studies performed in South America were compiled in an attempt to unify the advances that have been developed in whitefly management in this continent. At first, a background of the current whitefly distribution in South American countries as well as factors affecting them are shown, followed by a background of the whitefly transmitted viruses in South America, addressing their location and association with whiteflies in each country. Afterwards, a series of management strategies are proposed to be implemented in South American fields, including cultural practices and biological and chemical control, finalizing with a section containing future perspectives and directions for further research.

BackgroundMany plant viruses are vector-borne and depend on arthropods for transmission between host plants. Begomoviruses, the largest, most damaging and emerging group of plant viruses, infect hundreds of plant species, and new virus species of the group are discovered each year. Begomoviruses are transmitted by members of the whitefly Bemisia tabaci species complex in a persistent-circulative manner. Tomato yellow leaf curl virus (TYLCV) is one of the most devastating begomoviruses worldwide and causes major losses in tomato crops, as well as in many agriculturally important plant species. Different B. tabaci populations vary in their virus transmission abilities; however, the causes for these variations are attributed among others to genetic differences among vector populations, as well as to differences in the bacterial symbionts housed within B. tabaci.ResultsHere, we performed discovery proteomic analyses in 9 whitefly populations from both Middle East Asia Minor I (MEAM1, formerly known as B biotype) and Mediterranean (MED, formerly known as Q biotype) species. We analysed our proteomic results on the basis of the different TYLCV transmission abilities of the various populations included in the study. The results provide the first comprehensive list of candidate insect and bacterial symbiont (mainly Rickettsia) proteins associated with virus transmission.ConclusionsOur data demonstrate that the proteomic signatures of better vector populations differ considerably when compared with less efficient vector populations in the 2 whitefly species tested in this study. While MEAM1 efficient vector populations have a more lenient immune system, the Q efficient vector populations have higher abundance of proteins possibly implicated in virus passage through cells. Both species show a strong link of the facultative symbiont Rickettsia to virus transmission.

Control of the crop pest African cotton leafworm, Spodoptera littoralis (Boisduval), by chemical insecticides has led to serious resistance problems. Ajuga plants contain phytoecdysteroids (arthropod steroid hormone analogs regulating metamorphosis) and clerodanes (diterpenoids exhibiting antifeedant activity). We analyzed these compounds in leaf extracts of the Israeli Ajuga iva L. by liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) and thin-layer chromatography (TLC), and their efficiency at reducing S.littoralis fitness. First and third instars of S. littoralis were fed castor bean leaves (Ricinus communis) smeared with an aqueous suspension of dried methanolic crude extract of A. iva phytoecdysteroids and clerodanes. Mortality, larval weight gain, relative growth rate and survival were compared to feeding on control leaves. We used &lsquo, 4&rsquo, 6-diamidino-2-phenylindole (DAPI, a fluorescent stain) and phalloidin staining to localize A. iva crude leaf extract activity in the insect gut. Ajuga iva crude leaf extract (50, 100 and 250 &micro, g/&micro, L) significantly increased mortality of first-instar S. littoralis (36%, 70%, and 87%, respectively) compared to controls (6%). Third-instar larval weight gain decreased significantly (by 52%, 44% and 30%, respectively), as did relative growth rate (&minus, 0.05 g/g per day compared to the relevant controls), ultimately resulting in few survivors. Crude leaf extract (250 &micro, L) reduced gut size, with relocation of nuclei and abnormal actin-filament organization. Ajug iva extract has potential for alternative, environmentally safe insect-pest control.

The African cotton leafworm Spodoptera littoralis, a major crop pest worldwide, is controlled by chemical insecticides, leading to serious resistance problems. Ajuga plants contain phytoecdysteroids (analogs of arthropod steroid hormones that regulate metamorphoses) and clerodanes (diterpenoids exhibiting antifeedant activity). We analyzed phytoecdysteroids and clerodanes in leaf extracts of the Israeli Ajuga iva by LC-TOF-MS and TLC, and their efficiency at reducing S. littoralis fitness. Castor bean leaves were smeared with an aqueous suspension of dried methanolic crude extract of phytoecdysteroid and clerodanes from A. iva leaves (50, 100 and 250 µg/µl). First and third instars of S. littoralis larvae were fed with 1 treated leaf for 3 and 4 days, respectively. Mortality, larval weight gain, relative growth rate and survival were compared to feeding on control leaves. To evaluate and localize A. iva crude leaf extract activity in the insect gut, we used DAPI and phalloidin staining. Crude extract of A. iva leaves (50, 100 and 250 µg/µl) significantly increased mortality of first instar S. littoralis larvae (36%, 70% and 87%, respectively) compared to controls (6%). Third instar larval weight gain decreased significantly (by 52%, 44% and 30%, respectively), as did relative growth rate (–0.05 g/g day, compared to the relevant controls). S. littoralis larvae were further affected at later stages, with few survivors. Insect-gut staining showed that 250 µg/µl crude leaf extract reduces gut size, with relocation of nuclei and abnormal actin-filament organization. Our results demonstrate the potential of A. iva extract for alternative, environmentally safe insect-pest control. Key Message Insects cause severe damage to numerous crops and their control relies on pesticides. Green control is becoming increasingly popular due to concerns about the negative impacts of pesticides on the environment. Phytoecdysteroids are found in Ajuga plants and affect a wide range of insects at very low concentrations. Here we demonstrate that crude extract from Ajuga iva alters the development of Spodoptera littoralis. Phytoecdysteroids may therefore be beneficial in IPM programs.

Many species of the genus Ajuga (family Lamiaceae) contain phytoecdysteroids and clerodane diterpenes. Phytoecdysteroids are triterpene-derived analogues of steroid hormones that control molting and metamorphosis in arthropods, whereas clerodanes deter phytophagous insects. We identified and quantified phytoecdysteroid and clerodane contents in three Ajuga plant species in Israel. Leaves and roots of Ajuga iva, Ajuga chamaepitys (Ajuga chia), and Ajuga orientalis were collected from three different populations. Using liquid chromatography–time of flight–mass spectrometry analysis, we identified three phytoecdysteroids: 20-hydroxyecdysone (ecdysterone), makisterone A, and cyasterone and two clerodanes: dihydroajugapitin and columbin. Their contents varied significantly among plant species, organs, and populations. The highest concentrations of 20-hydroxyecdysone, makisterone A, and cyasterone were recorded in leaves and roots of A. iva. Cyasterone content tended to be higher in leaves of A. chamaepitys. Cl...

The African cotton leafworm Spodoptera littoralis, a major crop pest worldwide, is controlled by chemical insecticides, leading to serious resistance problems. Ajuga plants contain phytoecdysteroids (analogs of arthropod steroid hormones that regulate metamorphoses) and clerodanes (diterpenoids exhibiting antifeedant activity). We analyzed phytoecdysteroids and clerodanes in leaf extracts of the Israeli Ajuga iva by LC-TOF-MS and TLC, and their efficiency at reducing S. littoralis fitness. Castor bean leaves were smeared with an aqueous suspension of dried methanolic crude extract of phytoecdysteroid and clerodanes from A. iva leaves (50, 100 and 250 µg/µl). First and third instars of S. littoralis larvae were fed with 1 treated leaf for 3 and 4 days, respectively. Mortality, larval weight gain, relative growth rate and survival were compared to feeding on control leaves. To evaluate and localize A. iva crude leaf extract activity in the insect gut, we used DAPI and phalloidin staining. Crude extract of A. iva leaves (50, 100 and 250 µg/µl) significantly increased mortality of first instar S. littoralis larvae (36%, 70% and 87%, respectively) compared to controls (6%). Third instar larval weight gain decreased significantly (by 52%, 44% and 30%, respectively), as did relative growth rate (–0.05 g/g day, compared to the relevant controls). S. littoralis larvae were further affected at later stages, with few survivors. Insect-gut staining showed that 250 µg/µl crude leaf extract reduces gut size, with relocation of nuclei and abnormal actin-filament organization. Our results demonstrate the potential of A. iva extract for alternative, environmentally safe insect-pest control.Key MessageInsects cause severe damage to numerous crops and their control relies on pesticides. Green control is becoming increasingly popular due to concerns about the negative impacts of pesticides on the environment. Phytoecdysteroids are found in Ajuga plants and affect a wide range of insects at very low concentrations. Here we demonstrate that crude extract from Ajuga iva alters the development of Spodoptera littoralis. Phytoecdysteroids may therefore be beneficial in IPM programs.

Made available in DSpace on 2018-12-11T17:33:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) In Brazil, the first major invasion event of Bemisia tabaci was that of Middle East–Asia Minor 1 (MEAM1) species, formerly termed as B biotype, which commenced in the 1990s mainly by ornamental plants in São Paulo State. More than two decades after this invasion, the presence of the Mediterranean (MED) species of B. tabaci, formerly Q biotype, was reported in Rio Grande do Sul, the southernmost State of Brazil, and now in São Paulo and Paraná States, in southeastern Brazil. Specimens of whiteflies collected from commercial begonia, hydrangea, petunia and poinsettia greenhouses in São Paulo, and also from begonias and poinsettias collected in flower shops in Paraná, were all identified as belonging to MED species. Furthermore, the secondary endosymbionts Arsenophonus, Hamiltonella and Rickettsia of MED from São Paulo and Paraná were detected by PCR and their presence confirmed by sequencing and FISH analysis, and those results differed from MED detected in Rio Grande do Sul that harbored only Hamiltonella and Cardinium. Our results suggest a new MED invasion into Brazil and is associated with ornamental plants. The two MED populations are genetically different and suggest that they are separate invasions. São Paulo State University (UNESP) School of Agriculture Instituto Agronômico de Campinas Centro de Fitossanidade Department of Entomology The Volcani Center Institute of Plant Protection Instituto Federal do Sudeste de Minas Gerais, Campus Rio ARC Centre of Excellence in Plant Energy Biology and School of Chemistry and Biochemistry The University of Western Australia, Crawley São Paulo State University (UNESP) School of Agriculture FAPESP: 2014/21773-0

The green peach aphid, Myzus persicae, is a vector of the Potato leafroll virus (PLRV, Luteoviridae), transmitted exclusively by aphids in a circulative manner. PLRV transmission efficiency was significantly reduced when a clonal lineage of M. persicae was reared on turnip as compared with the weed physalis, and this was a transient effect caused by a host-switch response. A trend of higher PLRV titer in physalis-reared aphids as compared with turnip-reared aphids was observed at 24 h and 72 h after virus acquisition. The major difference in the proteomes of these aphids was the up-regulation of predicted lysosomal enzymes, in particular the cysteine protease cathepsin B (cathB), in aphids reared on turnip. The aphid midgut is the site of PLRV acquisition, and cathB and PLRV localization were starkly different in midguts of the aphids reared on the two host plants. In viruliferous aphids that were reared on turnip, there was near complete colocalization of cathB and PLRV at the cell membranes, which was not observed in physalis-reared aphids. Chemical inhibition of cathB restored the ability of aphids reared on turnip to transmit PLRV in a dose-dependent manner, showing that the increased activity of cathB and other cysteine proteases at the cell membrane indirectly decreased virus transmission by aphids. Understanding how the host plant influences virus transmission by aphids is critical for growers to manage the spread of virus among field crops.

Here, we report the genome of a putative densovirus infecting the carrot psyllid Bactericera trigonica, obtained by inverse PCR and named Bactericera trigonica densovirus (BtDNV). The ambisense genome of BtDNV is identical in structure to those of the ambidensoviruses, and its encoded proteins share the highest sequence identity with the Asian citrus psyllid Diaphorina citri densovirus.

Candidatus Liberibacter solanacerum (CLso), transmitted by Bactericera trigonica in a persistent and propagative mode causes carrot yellows disease, inflicting hefty economic losses. Understanding the process of transmission of CLso by psyllids is fundamental to devise sustainable management strategies. Persistent transmission involves critical steps of adhesion, cell invasion, and replication before passage through the midgut barrier. This study uses a transcriptomic approach for the identification of differentially expressed genes with CLso infection in the midguts, adults, and nymphs of B. trigonica and their putative involvement in CLso transmission. Several genes related to focal adhesion and cellular invasion were upregulated after CLso infection. Interestingly, genes involved with proper functionality of the endoplasmic reticulum (ER) were upregulated in CLso infected samples. Notably, genes from the endoplasmic reticulum associated degradation (ERAD) and the unfolded protein response (UPR) pathway were overexpressed after CLso infection. Marker genes of the ERAD and UPR pathways were also upregulated in Diaphorina citri when infected with Candidatus Liberibacter asiaticus (CLas). Upregulation of the ERAD and UPR pathways indicate induction of ER stress by CLso/CLas in their psyllid vector. The role of ER in bacteria&ndash, host interactions is well-documented, however, the ER role following pathogenesis of CLso/CLas is unknown and requires further functional validation.

Many animal and plant viruses depend on arthropods for their transmission. Virus-vector interactions are highly specific, and only one vector or one of a group of vectors from the same family is able to transmit a given virus. Poleroviruses (Luteoviridae) are phloem-restricted RNA plant viruses that are exclusively transmitted by aphids. Multiple aphid-transmitted polerovirus species commonly infect pepper, causing vein yellowing, leaf rolling, and fruit discoloration. Despite low aphid populations, a recent outbreak with such severe symptoms in many bell pepper farms in Israel led to reinvestigation of the disease and its insect vector. Here we report that this outbreak was caused by a new whitefly (Bemisia tabaci)-transmitted polerovirus, which we named Pepper whitefly-borne vein yellows virus (PeWBVYV). PeWBVYV is highly (>95%) homologous to Pepper vein yellows virus (PeVYV) from Israel and Greece on its 5′ end half, while it is homologous to African eggplant yellows virus (AeYV) on its 3′ half. Koch's postulates were proven by constructing a PeWBVYV infectious clone causing the pepper disease, which was in turn transmitted to test pepper plants by B. tabaci but not by aphids. PeWBVYV represents the first report of a whitefly-transmitted polerovirus. IMPORTANCE The high specificity of virus-vector interactions limits the possibility of a given virus changing vectors. Our report describes a new virus from a family of viruses strictly transmitted by aphids which is now transmitted by whiteflies (Bemisia tabaci) and not by aphids. This report presents the first description of polerovirus transmission by whiteflies. Whiteflies are highly resistant to insecticides and disperse over long distances, carrying virus inoculum. Thus, the report of such unusual polerovirus transmission by a supervector has extensive implications for the epidemiology of the virus disease, with ramifications concerning the international trade of agricultural commodities.

The whitefly B. tabaci is a global pest and transmits extremely important plant viruses especially begomoviruses, that cause substantial crop losses. B. tabaci is one of the top invasive species worldwide and have developed resistance to all major pesticide classes. One of the promising alternative ways for controlling this pest is studying its genetic makeup for identifying specific target proteins which are critical for its development and ability to transmit viruses. Tomato yellow leaf curl virus (TYLCV) is the most economically important and well-studied begomovirus transmitted by B. tabaci, in a persistent-circulative manner. Recently, we reported that B. tabaci Cyclophilin B (CypB) and heat shock protein 70 proteins (hsp70) interact and co-localize with TYLCV in the whitefly midgut, on the virus transmission pathway, and that both proteins have a significant role in virus transmission. Here, we extended the previous work and used the Tobacco rattle virus (TRV) plant-mediated RNA silencing system for knocking down both genes and testing the effect of their silencing on whitefly viability and virus transmission. Portions of these two genes were cloned into TRV constructs and tomato plants were infected and used for whitefly feeding and transmission experiments. Following whitefly feeding on TRV-plants, the expression levels of cypB and hsp70 in adult B. tabaci significantly decreased over 72 h feeding period. The knockdown in the expression of both genes was further shown in the first generation of silenced whiteflies, where phenotypic abnormalities in the adult, wing, nymph and bacteriosomes development and structure were observed. Additionally, high mortality rates that reached more than 80% among nymphs and adults were obtained. Finally, silenced whitefly adults with both genes showed decreased ability to transmit TYLCV under lab conditions. Our results suggest that plant-mediated silencing of both cypB and hsp70 have profound effects on whitefly development and its ability to transmit TYLCV.

We have recently shown that Rickettsia, a secondary facultative bacterial symbiont that infects the whitefly B. tabaci is implicated in the transmission of Tomato yellow leaf curl virus (TYLCV). Infection with Rickettsia improved the acquisition and transmission of the virus by B. tabaci adults. Here we performed a transcriptomic analysis with Rickettsia-infected and uninfected B. tabaci adults before and after TYLCV acquisition. The results show a dramatic and specific activation of the immune system in the presence of Rickettsia before TYLCV acquisition. However, when TYLCV was acquired, it induced massive activation of gene expression in the Rickettsia uninfected population, whereas in the Rickettsia-infected population the virus induced massive down-regulation of gene expression. Fitness and choice experiments revealed that while Rickettsia-infected whiteflies are always more attracted to TYLCV-infected plants, this attraction is not always beneficiary for their offspring. These studies further confirm the role of Rickettsia in many aspects of B. tabaci interactions with TYLCV, and possibly serves as an important factor in the dissemination of the virus.

The onion thrip, Thrips tabaci (Thysanoptera: Thripidae) is a major polyphagous pest that attacks a wide range of economically important crops, especially Allium species. The thrip&rsquo, s damage can result in yield loss of up to 60% in onions (Allium cepa). In the past few decades, thrip resistance to insecticides with various modes of actions have been documented. These include resistance to spinosad, a major active compound used against thrips, which was reported from Israel. Little is known about the molecular mechanisms underlying spinosad resistance in T. tabaci. We attempted to characterize the mechanisms involved in resistance to spinosad using quantitative transcriptomics. Susceptible (LC50 = 0.6 ppm) and resistant (LC50 = 23,258 ppm) thrip populations were collected from Israel. An additional resistant population (LC50 = 117 ppm) was selected in the laboratory from the susceptible population. De novo transcriptome analysis on the resistant and susceptible population was conducted to identify differently expressed genes (DGEs) that might be involved in the resistance against spinosad. In this analysis, 25,552 unigenes were sequenced, assembled, and functionally annotated, and more than 1500 DGEs were identified. The expression levels of candidate genes, which included cytochrome P450 and vittelogenin, were validated using quantitative RT-PCR. The cytochrome P450 expression gradually increased with the increase of the resistance. Higher expression levels of vitellogenin in the resistant populations were correlated with higher fecundity, suggesting a positive effect of the resistance on resistant populations. This research provides a novel genetic resource for onion thrips and a comprehensive molecular examination of resistant populations to spinosad. Those resources are important for future studies concerning thrips and resistance in insect pests regarding agriculture.

Endosymbionts harbored inside insects play critical roles in the biology of their insect host and can influence the transmission of pathogens by insect vectors. Bactericera trigonica infests umbelliferous plants and transmits the bacterial plant pathogen Candidatus Liberibacter solanacearum (CLso), causing carrot yellows disease. To characterize the bacterial diversity of B. trigonica, as a first step, we used PCR-restriction fragment length polymorphism (PCR-RFLP) and denaturing gradient gel electrophoresis (DGGE) analyses of 16S rDNA to identify Sodalis and Spiroplasma endosymbionts. The prevalence of both symbionts in field-collected psyllid populations was determined: Sodalis was detected in 100% of field populations, while Spiroplasma was present in 82.5% of individuals. Phylogenetic analysis using 16S rDNA revealed that Sodalis infecting B. trigonica was more closely related to symbionts infecting weevils, stink bugs and tsetse flies than to those from psyllid species. Using fluorescent in situ hybridization and immunostaining, Sodalis was found to be localized inside the nuclei of the midgut cells and bacteriocytes. Spiroplasma was restricted to the cytoplasm of the midgut cells. We further show that a recently reported Bactericera trigonica densovirus (BtDNV), a densovirus infecting B. trigonica was detected in 100% of psyllids and has reduced titers inside CLso-infected psyllids by more than two-fold compared to CLso uninfected psyllids. The findings of this study will help to increase our understanding of psyllid&ndash, endosymbiont interactions.

Made available in DSpace on 2019-10-06T15:57:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 The Bemisia tabaci is a polyphagous insect and a successful vector of plant viruses. B. tabaci is a species complex and in Brazil native species from the New World (NW) group, as well as the invasive species, Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) were reported. For better understanding the distribution of the different species four years after the Mediterranean species invasion in Brazil, whiteflies were collected from 237 locations throughout the country between the years of 2013 and 2017, species were identified and the facultative endosymbionts detected. The survey revealed that MEAM1 was the prevalent species found on major crops across Brazil. It is the only species present in North, Northwestern and Central Brazil and was associated with virus-infected plants. MED was found in five States from Southeast to South regions, infesting mainly ornamental plants and was not associated with virus-infected plants. The prevalent endosymbionts identified in MEAM1 were Hamiltonella and Rickettsia; and the mtCOI analysis revealed low genetic diversity for MEAM1. In contrast, several different endosymbionts were identified in MED including Hamiltonella, Rickettsia, Wolbachia and Arsenophonus; and two distinct genetic groups were found based on the mtCOI analysis. Monitoring the distribution of the whiteflies species in Brazil is essential for proper management of this pest. São Paulo State University UNESP-FCA Department of Plant Protection CEP Corteva Agriscience Instituto Agronômico de Campinas CEP Universidade Federal de Pelotas Department of Plant Protection CEP Santa Catarina State University UDESC Department of Agronomy/Plant Pathology Institute of Plant Protection Department of Entomology The Volcani Center The University of Western Australia ARC Centre of Excellence in Plant Energy Biology and School of Chemistry and Biochemistry Crawley São Paulo State University UNESP-FCA Department of Plant Protection CEP

The sweet potato whitefly Bemisia tabaci Genn (Hemiptera: Aleyrodidae) has been recorded to differentially prefer rosemary (Rosmarinus officinalis) varieties in commercial fields in Israel. As chemical signaling is a significant component in plant-insect interaction, the present study examined the involvement of rosemary essential oil volatiles in this differential colonization to elucidate the rosemary-whitefly ecological interaction. Thirty-two rosemary varieties with different chemical profiles were used. The average whitefly preference was 25.1% with a significant variation of 51.4%, partitioning the sampled varieties into five preference groups, hence suggesting rosemary as a non-preferred host for the insect. All relations between preference and the major volatiles 1,8-cineole, camphor, linalool, verbenone, bornyl acetate and borneol were significantly (P ≤ 0.05) or notably (0.05 < P ≤ 0.09) negative (r < 1). Therefore, revealing that whitefly preference for rosemary is based on a continuum of repellency rather than attraction. 'Choice' bioassays with a range of the major volatile concentrations and gas chromatography-mass spectrometry (GC-MS) volatile blends (fractions) validated this observation. Principle component analysis of the entire chemical profile of two extreme varieties, representing high and low preferences, identified that approximately 43% of the volatiles in the essential oil were directly associated with repellency. Keeping in mind the remaining 57% of the compounds, this myriad of volatiles exhibit the ecological complexity of the rosemary-whitefly eco-system, explaining that whitefly preference to rosemary is repellency based.

Carrot yellows disease has been associated for many years with the Gram-positive, insect-vectored bacteria, ‘Candidatus Phytoplasma’ and Spiroplasma citri. However, reports in the last decade also link carrot yellows symptoms with a different, Gram-negative, insect-vectored bacterium, ‘Ca. Liberibacter solanacearum’. Our study shows that to date ‘Ca. L. solanacearum’ is tightly associated with carrot yellows symptoms across Israel. The genetic variant found in Israel is most similar to haplotype D, found around the Mediterranean Basin. We further show that the psyllid vector of ‘Ca. L. solanacearum’, Bactericera trigonica, is highly abundant in Israel and is an efficient vector for this pathogen. A survey conducted comparing conventional and organic carrot fields showed a marked reduction in psyllid numbers and disease incidence in the field practicing chemical control. Fluorescent in situ hybridization and scanning electron microscopy analyses further support the association of ‘Ca. L. solanacearum’ with disease symptoms and show that the pathogen is located in phloem sieve elements. Seed transmission experiments revealed that while approximately 30% of the tested carrot seed lots are positive for ‘Ca. L. solanacearum’, disease transmission was not observed. Possible scenarios that may have led to the change in association of the disease etiological agent with carrot yellows are discussed. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Begomoviruses comprise an emerging and economically important group of plant viruses exclusively transmitted by the sweetpotato whitefly Bemisia tabaci in many regions of the world. The past twenty years have witnessed significant progress in studying the molecular interactions between members of this virus group and B. tabaci. Mechanisms and proteins encoded by the insect vector and its bacterial symbionts, which have been shown to be important for virus transmission, have been identified and thoroughly studied. Despite the economic importance of this group of viruses and their impact on the global agriculture, progress in investigating the virus-vector interactions is moving slowly when compared with similar virus-vector systems in plants and animals. Major advances in this field and future perspectives will be discussed in this review.

Biological control approaches such as seeding and augmentation releases of populations of natural enemies mostly rely on the indoor production of predator or parasitoid species, often with the use of alternative prey/host species. In this study, we investigated several development parameters of four egg parasitoid species: Ooencyrtus fecundus, O. near fecundus, O. pityocampae and O. telenomicida, and compared their performance on their natural host, the variegated caper bug (CB) Stenozygum coloratum, and on an alternative, factitious host, the silk moth (SM) Bombyx mori. Survival was higher and development duration shorter in CB eggs, making the CB a better host for these congeners. However, adult longevity was generally longer for individuals that developed in SM eggs. Moreover, O. fecundus and O. pityocampae females that had developed in SM eggs displayed higher fecundity than all other female/host combinations. Survival also varied according to the age of the SM eggs: parasitoid survival rates were significantly higher in 9- to 12-month-old (post-diapausing) eggs than in young (about 1 month old) ones. These results were probably influenced by differences among the egg sizes of the studied hosts. The number of non-laying females and self-superparasitism rates were exceptionally high in O. near fecundus. These findings suggest that SM eggs, and especially those which are being utilized after a long storage, could serve for mass rearing of the studied Ooencyrtus spp.

The variegated caper bug Stenozygum coloratum Klug is common in the Middle East, where it is a minor agricultural pest. Recently, it was found that S. coloratum is an alternative host for the egg parasitoid Ooencyrtus pityocampae Mercet, an important natural enemy of the pine processionary moth Thaumetopoea wilkinsoni Tams, a major pest of pines. This discovery, coupled with the general lack of knowledge of S. coloratum, prompted the present study, which focused on seasonal activity of S. coloratum in Israeli pine forests, bidirectional interactions between the bug and its host the caper plant Capparis spp., and parasitism of its eggs. Stenozygum coloratum activity lasted mainly from May to October, and was followed by an adult winter diapause. Individuals formed large aggregations, causing early withering of plants and a shortening their annual growth periods. Reproductive activity peaked in August and ended in September. Egg parasitism rates increased throughout this period. Several Ooencyrtus spp., including known natural enemies of various pests, parasitized the eggs of S. coloratum and other associated pentatomids found on the caper plants. Thus, capers probably serve as a refuge for these potentially important egg parasitoids. The potential of caper plants and their fauna for biological control programmes is discussed.

The way in which species coexist is an important ecological question. Often, several parasitoid species may share a common host species. Five species of Ooencyrtus Ashmead (Hymenoptera: Encyrtidae) co-inhabit the egg clusters of Stenozygum coloratum Klug (Hemiptera: Pentatomidae), in the East Mediterranean region. Their relative abundance and spatial and seasonal occurrence were investigated. Parasitism and parasitoid male ratios gradually increased to about 50 and 26.8 %, respectively, from May to October. O. telenomicida Vassiliev was dominant in Mediterranean climatic regions and absent from arid areas, O. fecundus Ferriere and Voegele dominated semi-arid areas, and O. near nigerrimus was the commonest in hot-arid areas. O. near fecundus occurred everywhere but was most abundant in the more extreme weather conditions. O. pityocampae Mercet was uncommon in most areas and absent from arid ones. O. telenomicida abundance was positively, and that of O. fecundus negatively, correlated with annual rainfall. O. near fecundus and O. near nigerrimus abundance was positively correlated with temperature extremes. The results display a clear picture for climatic based spatial niche partitioning, although there is evidence that interspecific competition also plays a significant role in the frequency of occurrence of the studied Ooencyrtus spp.

The variegated caper bug (CB) Stenozygum coloratum (Klug, 1845) is common in the Eastern Mediterranean region and a minor agricultural pest. CB eggs were recently shown to serve as alternative hosts for Ooencyrtus pityocampae Mercet, 1921, an important natural enemy of the pine processionary moth (PPM) Thaumetopoea wilkinsoni Tams, 1924 (Lepidoptera: Notodontidae). In this study various aspects of the life history of CB were studied, including its distribution, host range, development, reproduction and ability to develop on various cultivated species of plants. CB occurs in almost all areas and habitats in Israel, except in the coldest regions, attacking several caper species (Capparis spp.). When reared on Capparis zoharyi at 25°C, one generation lasted 6 weeks and females laid a single 12-egg cluster every 3 days. Decreases in temperature, but not changes in day length, terminated reproductive activity. CB nymphs survived for up to three months, or even completed their development, on some agricultural plants. However, reproductive activity was initiated only if adults were fed capers. These findings emphasize the importance of capers in the CB life cycle and account for why other plants are rarely attacked. These plants may serve as a temporary refuge for CB when capers are scarce or unsuitable, mainly late in the season. Morphology of developmental stages, egg deposition, cannibalistic behaviour, sex ratio and effects of temperature on egg production were also studied. The possibility of using CB for enhancing the biological control of PPM is discussed.

Background The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is among the 100 worst invasive species in the world. As one of the most important crop pests and virus vectors, B. tabaci causes substantial crop losses and poses a serious threat to global food security. Results We report the 615-Mb high-quality genome sequence of B. tabaci Middle East-Asia Minor 1 (MEAM1), the first genome sequence in the Aleyrodidae family, which contains 15,664 protein-coding genes. The B. tabaci genome is highly divergent from other sequenced hemipteran genomes, sharing no detectable synteny. A number of known detoxification gene families, including cytochrome P450s and UDP-glucuronosyltransferases, are significantly expanded in B. tabaci. Other expanded gene families, including cathepsins, large clusters of tandemly duplicated B. tabaci-specific genes, and phosphatidylethanolamine-binding proteins (PEBPs), were found to be associated with virus acquisition and transmission and/or insecticide resistance, likely contributing to the global invasiveness and efficient virus transmission capacity of B. tabaci. The presence of 142 horizontally transferred genes from bacteria or fungi in the B. tabaci genome, including genes encoding hopanoid/sterol synthesis and xenobiotic detoxification enzymes that are not present in other insects, offers novel insights into the unique biological adaptations of this insect such as polyphagy and insecticide resistance. Interestingly, two adjacent bacterial pantothenate biosynthesis genes, panB and panC, have been co-transferred into B. tabaci and fused into a single gene that has acquired introns during its evolution. Conclusions The B. tabaci genome contains numerous genetic novelties, including expansions in gene families associated with insecticide resistance, detoxification and virus transmission, as well as numerous horizontally transferred genes from bacteria and fungi. We believe these novelties likely have shaped B. tabaci as a highly invasive polyphagous crop pest and efficient vector of plant viruses. The genome serves as a reference for resolving the B. tabaci cryptic species complex, understanding fundamental biological novelties, and providing valuable genetic information to assist the development of novel strategies for controlling whiteflies and the viruses they transmit. Electronic supplementary material The online version of this article (doi:10.1186/s12915-016-0321-y) contains supplementary material, which is available to authorized users.

We investigated the dual effects of bacterial infections and diseased cassava plants on the fitness and biology of the Bemisia tabaci infesting cassava in Africa. Isofemale B. tabaci colonies of sub-Saharan Africa 1-subgroup 3 (SSA1-SG3), infected with two secondary endosymbiotic bacteria Arsenophonus and Rickettsia (AR+) and those free of AR infections (AR−), were compared for fitness parameters on healthy and East African cassava mosaic virus-Uganda variant (EACMV-UG)-infected cassava plants. The whitefly fecundity and nymph development was not affected by bacterial infections or the infection of cassava by the virus. However, emergence of adults from nymphs was 50 and 17% higher by AR− on healthy and virus-infected plants, respectively, than AR+ flies. Development time of adults also was 10 days longer in AR+ than AR−. The whiteflies were further compared for acquisition and retention of EACMV-UG. Higher proportion of AR− acquired (91.8%) and retained (87.6%) the virus than AR+ (71.8, 61.2%, respectively). Similarly, the AR− flies retained higher quantities of virus (~ninefold more) than AR+. These results indicated that bacteria-free whiteflies were superior and better transmitters of EACMV-UG, as they had higher adult emergence, quicker life cycle and better virus retention abilities than those infected with bacteria. Electronic supplementary material The online version of this article (doi:10.1007/s10340-017-0910-8) contains supplementary material, which is available to authorized users.

The whitefly Bemisia tabaci (Gennadius) is a cosmopolitan insect pest and causes great damage to many agricultural crops by direct feeding, and transmitting many plant viruses causing disastrous effects. The global distribution of B.tabaci was previously investigated, but so far, it has not been confirmed in Serbia. Sampling conducted on plants at three localities in Serbia revealed the presence of three whitefly species, one is B.tabaci Mediterranean (MED) species, a quarantine pest. Additionally, Trialeurodes vaporariorum Westwood and an Aleurodes sp. that was not identified to the species level were also confirmed. The last two are common and widespread species in Serbia. Whitefly collections were made from cabbage, kale and oilseed rape plants (Brassica oleracea, B.oleracea var. sabauda, B.napus, respectively) grown in the field, on sunflower (Helianthus annuus) grown in the greenhouse and on potted hibiscus (Hibiscus sp.) and poinsettia (Euphorbia pulcherrima) plants grown indoors. DNA sequence analysis from a portion of the Cytochrome Oxidase I gene (COI) revealed the presence of B.tabaci MED species, which is considered to be a very important pest worldwide and has been reported from neighbouring countries, however, not yet from Serbia. Bemisia tabaci MED is recorded here for the first time in Serbia. This first interception suggests its introduction with ornamentals grown indoors, with no outdoor reports and that it is not yet spread. This report is an important alert for local authorities to take the necessary steps in monitoring and preventing its possible expansion.

Citrus greening disease known also as Huanglongbing (HLB) caused by the phloem-limited bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) has resulted in tremendous losses and the death of millions of trees worldwide. CLas is transmitted by the Asian citrus psyllid Diaphorina citri. The closely-related bacteria ‘Candidatus Liberibacter solanacearum’ (CLso), associated with vegetative disorders in carrots, is transmitted by the carrot psyllid Bactericera trigonica. A promising approach to prevent the transmission of these pathogens is to interfere with the vector-pathogen interactions, but our understanding of these processes is limited. It was recently reported that CLas induced changes in the nuclear architecture, and activated programmed cell death, in D. citri midgut cells. Here, we used electron and fluorescent microscopy and show that CLas induces the formation of endoplasmic reticulum (ER)-associated bodies. The bacterium recruits those ER structures into Liberibacter containing vacuoles (LCVs), in which bacterial cells seem to propagate. ER- associated LCV formation was unique to CLas, as we could not detect these bodies in B. trigonica infected with CLso. ER recruitment is hypothesized to generate a safe replicative body to escape cellular immune responses in the insect gut. Understanding the molecular interactions that undelay these responses will open new opportunities for controlling CLas.

Whitefly (Bemisia tabaci) is an important insect pest, causing severe damage to agricultural crops. The pest was recorded in a commercial rosemary (Rosmarinus officinalis, Lamiaceae) field, colonizing rosemary variety (var.) '2', but not '11'. A series of field and controlled laboratory choice bioassays confirmed the observed phenomenon. Mature potted plants of the two varieties were randomly organized in a lemon verbena (Lippia citrodora) and lemon grass (Cymbopogon spp.) fields. Seven days later var. '2' was significantly more colonized by whiteflies than var. '11'. Under lab conditions, whiteflies were significantly more attracted to var. '2' plantlets than to var. '11' following choice bioassays. Furthermore, cotton plants dipped in an essential oil emulsion of var. '2' had significantly greater colonization than cotton plants dipped in the essential oil emulsion of var. '11'. Similar results were obtained in 'plant-plant', 'plant-no plant' as well as, 'essential oil-essential oil' choice bioassay designs. Analyses of the essential oils of the two varieties identified a set of common and unique volatiles in each variety. Among these volatiles were β-caryophyllene and limonene, two compounds known to be associated with plant-insect interactions. The attraction of B. tabaci to pure (>95%) β-caryophyllene and limonene using a range of concentrations was examined in vitro by choice bioassays. The compounds were attractive to the insect at moderate concentration, but not at the lowest or highest concentrations used, where the insect was not attracted or repelled, respectively. Limonene attracted the insects at rates that were 10-fold lower than β-caryophyllene. The results emphasized the role of host plant volatiles in shaping the structure of B. tabaci populations in nature and in agricultural systems, and provided insights into the factors that contribute to the development of insect populations with unique characteristics. The results could also serve for future development of bio-pesticides and in breeding programs.

The sweetpotato whitefly Bemisia tabaci is a highly destructive agricultural and ornamental crop pest. It damages host plants through both phloem feeding and vectoring plant pathogens. Introductions of B. tabaci are difficult to quarantine and eradicate because of its high reproductive rates, broad host plant range, and insecticide resistance. A total of 791 Gb of raw DNA sequence from whole genome shotgun sequencing, and 13 BAC pooling libraries were generated by Illumina sequencing using different combinations of mate-pair and pair-end libraries. Assembly gave a final genome with a scaffold N50 of 437 kb, and a total length of 658 Mb. Annotation of repetitive elements and coding regions resulted in 265.0 Mb TEs (40.3%) and 20 786 protein-coding genes with putative gene family expansions, respectively. Phylogenetic analysis based on orthologs across 14 arthropod taxa suggested that MED/Q is clustered into a hemipteran clade containing A. pisum and is a sister lineage to a clade containing both R. prolixus and N. lugens. Genome completeness, as estimated using the CEGMA and Benchmarking Universal Single-Copy Orthologs pipelines, reached 96% and 79%. These MED/Q genomic resources lay a foundation for future ‘pan-genomic’ comparisons of invasive vs. noninvasive, invasive vs. invasive, and native vs. exotic Bemisia, which, in return, will open up new avenues of investigation into whitefly biology, evolution, and management.

BACKGROUND The whitefly Bemisia tabaci is a major cosmopolitan pest and comprises a complex of more than 36 cryptic species that cause serious damage to agricultural crops worldwide. In this study, the Mediterranean species of B. tabaci, formerly known as Q biotype, was identified for the first time in Brazil. RESULTS Adult B. tabaci were collected from different localities and hosts from Rio Grande do Sul, the southernmost state of the country that borders Uruguay and Argentina. Partial sequencing of the mitochondrial cytochrome oxidase I (mtCOI) gene indicated that B. tabaci MED species appears to be restricted to the province of Barra do Quarai, infesting Capsicum annuum cultivated in greenhouses and Ipomoea batatas in open fields. The partial mtCOI sequences obtained shared 100% nucleotide identity with reference sequences for the Q biotype reported from Uruguay. The secondary endosymbionts Hamiltonella and Cardinium were detected by PCR in the new identified MED species from Brazil, similarly to the Q biotype from Uruguay. CONCLUSION Our results indicate the presence of the MED species in Brazil. The close monitoring of this new identified species in the southern region of Brazil is essential to avoid its geographical expansion to more important agricultural areas in the country. © 2014 Society of Chemical Industry

Mealybugs have a haplodiploid reproduction system, with paternal genome elimination (PGE); the males are diploid soon after fertilization, but during embryogenesis, the male paternal set of chromosomes becomes heterochromatic (HC) and therefore inactive. Previous studies have suggested that paternal genes can be passed on from mealybug males to their sons, but not necessarily by any son, to the next generation. We employed crosses between two mealybug species--Planococcus ficus (Signoret) and Planococcus citri (Risso)--and between two populations of P. ficus, which differ in their mode of pheromone attraction, in order to demonstrate paternal inheritance from males to F2 through F1 male hybrids. Two traits were monitored through three generations: mode of male pheromone attraction (pherotype) and sequences of the internal transcribed spacer 2 (ITS2) gene segment (genotype). Our results demonstrate that paternal inheritance in mealybugs can occur from males to their F2 offspring, through F1 males (paternal line). F2 backcrossed hybrid males expressed paternal pherotypes and ITS2 genotypes although their mother originated through a maternal population. Further results revealed other, hitherto unknown, aspects of inheritance in mealybugs, such as that hybridization between the two species caused absence of paternal traits in F2 hybrid females produced by F1 hybrid females. Furthermore, hybridization between the two species raised the question of whether unattracted males have any role in the interactions between P. ficus and P. citri.

Lineages of the generalist hemipteran herbivore Myzus persicae (green peach aphid) that have expanded their host range to include tobacco often have elevated nicotine tolerance. The tobacco-adapted M. persicae lineage used in this study was able to reproduce on nicotine-containing artificial diets at concentrations that were 15-fold higher than those that were lethal to a non-adapted M. persicae lineage. Fecundity of the nicotine-tolerant M. persicae lineage was increased by 100 μM nicotine in artificial diet, suggesting that this otherwise toxic alkaloid can serve as a feeding stimulant at low concentrations. This lineage also was pre-adapted to growth on tobacco, exhibiting no drop in fecundity when it was moved onto tobacco from a different host plant. Although growth of the non-tobacco-adapted M. persicae lineage improved after three generations on tobacco, this higher reproductive rate was not associated with increased nicotine tolerance. M. persicae gene expression microarrays were used to identify transcripts that are up-regulated in response to nicotine in the tobacco-adapted lineage. Induced expression was found for CYP6CY3, which detoxifies nicotine in M. persicae, other genes encoding known classes of detoxifying enzymes, as well as genes encoding secreted M. persicae salivary proteins.

Begomoviruses are a group of icosahedral single stranded DNA viruses exclusively transmitted by the sweet potato whitefly Bemisia tabaci in a persistent, circulative manner. In this mode of transmission, begomoviruses are acquired by their insect vector as intact virions from the plant phloem, move along the food canal, foregut and esophagus and reach the midgut where they are absorbed into the hemolymph via the filter chamber. The filter chamber is the site where most of the ingested food is filtered, and the first site where the majority of begomoviruses appear to be translocated into the hemolymph via unknown proteins or receptors. Transport from the filter chamber to the hemolymph is aided by a Heat Shock Protein 70. Virus particles not translocated across the filter chamber circulate in the midgut loop but it is not known whether absorption into the hemolymph occurs along this loop. Localization studies have confirmed that begomoviruses are not associated with the hindgut and absorption of virions in this organ is unlikely. In the hemolymph, virions have been shown to interact with a GroEL chaperone produced by the whitefly's endosymbiontic bacteria for ensuring their safe journey to the salivary glands. Virions penetrate the primary salivary glands via unknown proteins or receptors and are transported and secreted outside the whitefly to the plant with salivary secretions. Several recent studies have demonstrated the implications of insect and endosymbiont proteins such as the heat shock protein 70 and the bacterial GroEL protein, in the transmission of begomoviruses by B. tabaci. Additional studies attempting to identify other proteins that aid or interact with begomoviruses along their circulation pathway in the whitefly are reviewed in this paper.

Numerous animal and plant viruses are transmitted by arthropod vectors in a persistent, circulative manner. Tomato yellow leaf curl virus (TYLCV) is transmitted by the sweet potato whitefly Bemisia tabaci . We report here that infection with Rickettsia spp., a facultative endosymbiont of whiteflies, altered TYLCV- B. tabaci interactions. A B. tabaci strain infected with Rickettsia acquired more TYLCV from infected plants, retained the virus longer, and exhibited nearly double the transmission efficiency compared to an uninfected B. tabaci strain with the same genetic background. Temporal and spatial antagonistic relationships were discovered between Rickettsia and TYLCV within the whitefly. In different time course experiments, the levels of virus and Rickettsia within the insect were inversely correlated. Fluorescence in situ hybridization analysis of Rickettsia -infected midguts provided evidence for niche exclusion between Rickettsia and TYLCV. In particular, high levels of the bacterium in the midgut resulted in higher virus concentrations in the filter chamber, a favored site for virus translocation along the transmission pathway, whereas low levels of Rickettsia in the midgut resulted in an even distribution of the virus. Taken together, these results indicate that Rickettsia , by infecting the midgut, increases TYLCV transmission efficacy, adding further insights into the complex association between persistent plant viruses, their insect vectors, and microorganism tenants that reside within these insects. IMPORTANCE Interest in bacterial endosymbionts in arthropods and many aspects of their host biology in agricultural and human health systems has been increasing. A recent and relevant studied example is the influence of Wolbachia on dengue virus transmission by mosquitoes. In parallel with our recently studied whitefly- Rickettsia -TYLCV system, other studies have shown that dengue virus levels in the mosquito vector are inversely correlated with bacterial load. Our work here presents evidence of unifying principles between vectors of plant and animal viruses in a role for endosymbionts in manipulating vector biology and pathogen transmission. Our results demonstrate the influence of an interesting and prominent bacterial endosymbiont in Bemisia tabaci in TYLCV transmission, a worldwide disease infecting tomatoes. Besides its agricultural importance, this system provides interesting insights into Bemisia interaction with these newly discovered endosymbionts.

BACKGROUND: Plant defensive metabolites such as nicotine can provide barriers to host-range expansion by generalist herbivores. Nicotine is one of the most abundant and toxic plant secondary metabolites in nature and is defined by high toxicity to plant-feeding insects. There is significant variation in nicotine tolerance among Bemisia tabaci (tobacco whitefly) isolates. Some nicotine-tolerant B. tabaci strains can consume 40-fold higher nicotine levels than susceptible strains, and also show cross-resistance to neonicotinoid insecticides. In this study, biological and molecular assays were used to investigate the responses of B. tabaci strains that differ in their ability to tolerate dietary nicotine. RESULTS: Egg laying and honeydew secretion bioassays as well as gene expression microarrays were used to measure B. tabaci biological parameters and gene transcripts misregulated in response to nicotine in resistant and susceptible strains. The resistant B. tabaci strain laid significantly fewer eggs and excreted more honeydew on a tobacco strain with high levels of nicotine, suggesting a fitness cost effect. The molecular response was drastic in the susceptible strain, while the resistant strain exhibited moderate response. Higher expression of the previously identified CYP6CM1 P450 monooxygenase gene related to the resistance to neonicotinoids, as well as other P450s and metabolic genes, was identified in the resistant and susceptible strains after exposure to nicotine. CONCLUSIONS: Nicotine is a very toxic plant natural compound, and its mode of action resembles that of synthetic neonicotinoids. The biological and molecular responses observed in this study suggest that nicotine may play an important role in providing barriers for host-plant expansion by generalists, and may act as a natural factor that contributes to the development of insect populations resistant to synthetic pesticides. © 2014 Society of Chemical Industry Supporting information may be found in the online version of this article.