Vectors Of Plant Viruses Research Articles

Species Diversity of Thrips on Cotton

Exploratory surveys were conducted to study the diversity of thrips species on cotton at Coimbatore,Tamil Nadu. Taxonomic studies revealed the presence of three species, viz. Scirtothrips dorsalis Hood,Thrips palmi Karny and Thrips tabaci Lindeman on leaves and four species, viz. Thrips florum Schmutz,Thrips hawaiiensis (Morgan), Frankliniella schultzei (Trybom) and Thrips parvispinus (Karny) in flowers.T. parvispinus is one of the notorious pest species of South East Asia and also a serious pest of quarantineimportance. F. schultzei, S. dorsalis, T. palmi and T. tabaci are notorious pests as well as vectors of plantviruses. Since these species may attain major pest status, the report demands regular monitoring andsurveys in cotton.

Silencing the Autophagy-Related Genes ATG3 and ATG9 Promotes SRBSDV Propagation and Transmission in Sogatella furcifera.

Simple SummaryPlant viruses are mostly transmitted by insects and cause severe damage to crops. The southern rice black-streaked dwarf virus is a rice virus exclusively transmitted by a planthopper, Sogatella furcifera. Autophagy is usually initiated as an adaptive response for cell survival in unfavorable conditions, such as pathogen invasion. The invasion of the virus in the vector may elicit autophagy and may subsequently function to suppress or promote virus propagation and transmission in the insect vector. Here, we annotated two key autophagy-related genes, SfATG3 and SfATG9, in the planthopper vector and found that the genes are conserved among various insects. Interestingly, exposure of the vector to the virus activated the expression of SfATG3 and SfATG9, and silencing SfATG3 or SfATG9 promoted propagation and transmission rates of the virus. These results indicate that the expression of autophagy-related genes is initiated in the vector following exposure to the virus and that autophagy-related genes functions to suppress SRBSDV propagation and transmission.Autophagy plays diverse roles in the interaction among pathogen, vector, and host. In the plant virus and insect vector system, autophagy can be an antiviral/pro-viral factor to suppress/promote virus propagation and transmission. Here, we report the antiviral role of autophagy-related genes ATG3 and ATG9 in the white-backed planthopper (Sogatella furcifera) during the process of transmitting the southern rice black-streaked dwarf virus (SRBSDV). In this study, we annotated two autophagy-related genes, SfATG3 and SfATG9, from the female S. furcifera transcriptome. The cDNA of SfATG3 and SfATG9 comprised an open reading frame (ORF) of 999 bp and 2295 bp that encodes a protein of 332 and 764 amino acid residues, respectively. SfATG3 has two conserved domains and SfATG9 has one conserved domain. In S. furcifera females exposed to SRBSDV, expression of autophagy-related genes was significantly activated and shared similar temporal patterns to those of SRBSDV S9-1 and S10, all peaking at 4 d post viral exposure. Silencing the expression of SfATG3 and SfATG9 promoted SRBSDV propagation and transmission. This study provides evidence for the first time that S. furcifera autophagy-related genes ATG3 and ATG9 play an antiviral role to suppress SRBSDV propagation and transmission.

Drought as a modulator of plant–virus–vector interactions: Effects on symptom expression, plant immunity and vector behaviour

AbstractPlant–virus interactions are affected by environmental conditions that determine plant vulnerability to pathogens and the population dynamics of insect vectors. We hypothesize that drought enhances horizontal transmission by dampening the basal immunity of plants, which triggers symptom expression and vector manipulation. The potato yellow vein virus (PYVV) causes potato yellow vein disease (PYVD), a re‐emerging epidemic of potato crops in South America, and is transmitted horizontally by the greenhouse whitefly (GWF), Trialeurodes vaporariorum (Hemiptera: Aleyrodidae), or vertically through infected seed tubers. We investigated the role of drought and temperature as modulators of PYVD symptom expression, plant immune response, and vector survival, development and host preference. We found that drought induced symptom expression, suppressed the salicylic acid pathway and increased PYVV replication. GWF survival was reduced on PYVV‐infected hosts and development was slowest when they fed on plants with PYVD symptoms, which also triggered adults’ attraction to PYVV‐infected plants. However, adults previously fed on infected plants showed the opposite effect, being more attracted to PYVV‐free plants. We propose a theoretical model that explains the role of drought in modulating potato–PYVV–GWF interactions and provides new insights into plant–virus–vector coevolution.

Induction of resistance to Myzus persicae-nicotianae in Cucumber mosaic virus infected tobacco plants using silencing of CMV-2b gene

Aphids such as tobacco aphid Myzus persicae-nicotianae, are among the most important plant viral vectors and plant viruses encode genes to interact with their vectors. Cucumber mosaic virus (CMV) encodes 2b protein as a suppressor of plant immune and it plays a vital role in CMV accumulation and susceptibility to aphid vectors. In this study, the resistance of tobacco plants (Nicotiana tabacum) to M. p. nicotianae was evaluated by silencing of 2b in CMV-infected plants. However, the pFGC-C.h silencing gene construct was transiently expressed using Agrobacterium tumefacience, LBA 4404 in tobacco leaves, and four days later, the plants were mechanically inoculated by CMV (Kurdistan isolate), and then, 15 days post-inoculation 1 nonviruliferous aphid was placed on each leaf for evaluation of resistance to M. p. nicotianae. To evaluate the tobacco plants resistance and susceptibility to M. p. nicotianae, the number of aphids existent per tobacco leaf, life table and, demographic parameters were recorded and used as a comparison indicator. The obtained results were analyzed using the age-stage, two-sex life table. The highest number of aphids was recorded on the control CMV-infected plants, while the lowest number on CMV infected leaves expressing CMV-2b silencing construct (pFGC-C.h). The obtained data revealed the lowest rate for all of intrinsic rate of natural increase (rm) (0.246/day), the rate of reproduction (r0) (17.04 females/generation), and finite rate of increase (λ) (1.279/day), on the pFGC-C.h treatment. The maximum generation time (T) (11.834 days) was observed on (V) treatment. However, the collected data revealed induction of resistance to tobacco aphids by silencing of CMV-2b in CMV infected plants.

Virus Infection and Host Plant Suitability Affect Feeding Behaviors of Cannabis Aphid (Hemiptera: Aphididae), a Newly Described Vector of Potato Virus Y.

Aphids are the most prolific vectors of plant viruses resulting in significant yield losses to crops worldwide. Potato virus Y (PVY) is transmitted in a non-persistent manner by 65 species of aphids. With the increasing acreage of hemp (Cannabis sativa L.) (Rosales: Cannabaceae) in the United States, we were interested to know if the cannabis aphid (Phorodon cannabis Passerini) (Hemiptera: Aphididae) is a potential vector of PVY. Here, we conduct transmission assays and utilize the electrical penetration graph (EPG) technique to determine whether cannabis aphids can transmit PVY to hemp (host) and potato (non-host) (Solanum tuberosum L.) (Solanales: Solanaceace). We show for the first time that the cannabis aphid is an efficient vector of PVY to both hemp (96% transmission rate) and potato (91%) using cohorts of aphids. In contrast, individual aphids transmitted the virus more efficiently to hemp (63%) compared to potato (19%). During the initial 15 min of EPG recordings, aphids demonstrated lower number and time spent performing intracellular punctures on potato compared to hemp, which may in part explain low virus transmission to potato using individual aphids. During the entire 8-hour recording, viruliferous aphids spent less time ingesting phloem compared to non-viruliferous aphids on hemp. This reduced host acceptance could potentially cause viruliferous aphids to disperse thereby increasing virus transmission. Overall, our study shows that cannabis aphid is an efficient vector of PVY, and that virus infection and host plant suitability affect feeding behaviors of the cannabis aphid in ways which may increase virus transmission.

Effector-mediated plant-virus-vector interactions.

Hemipterans (such as aphids, whiteflies, and leafhoppers) are some of the most devastating insect pests due to the numerous plant pathogens they transmit as vectors, which are primarily viral. Over the past decade, tremendous progress has been made in broadening our understanding of plant–virus–vector interactions, yet on the molecular level, viruses and vectors have typically been studied in isolation of each other until recently. From that work, it is clear that both hemipteran vectors and viruses use effectors to manipulate host physiology and successfully colonize a plant and that co-evolutionary dynamics have resulted in effective host immune responses, as well as diverse mechanisms of counterattack by both challengers. In this review, we focus on advances in effector-mediated plant–virus–vector interactions and the underlying mechanisms. We propose that molecular synergisms in vector–virus interactions occur in cases where both the virus and vector benefit from the interaction (mutualism). To support this view, we show that mutualisms are common in virus–vector interactions and that virus and vector effectors target conserved mechanisms of plant immunity, including plant transcription factors, and plant protein degradation pathways. Finally, we outline ways to identify true effector synergisms in the future and propose future research directions concerning the roles effectors play in plant–virus–vector interactions.

Plant viral vectors: an strategy for transient expression of recombinant proteins in plants

Plants have been studied as bioreactors for over 30 years. Their advantages over other expression methods arouse interest in this heterologous protein expression system. Over the years, several methods of plant transformation were developed, and among them, the transient expression system - that occurs temporarily and without integration into the host genome. The use of viral vectors for this type of expression stands out, and the viral genome can be “full-length” or “deconstructed”. Some genome modifications in the “deconstructed” form help searching for efficient protein expression, and several types of viruses are studied to develop these vectors. This review reports different types of viral vectors, especially those particularly suitable for the development of effective vectors of fast and high yield recombinant protein production, based on Tobamoviruses, Potexviruses, Geminiviruses and Comoviruses.

A reproducible and sensitive method for generating high-quality transcriptomes from single whitefly salivary glands and other low-input tissues.

Transcriptomic studies are an important tool for understanding the molecular pathways underlying host plant use by agricultural pests, including vectors of damaging plant pathogens. Thus far, bulk RNA-Seq has been the main approach for non-model insects. This method relies on pooling large numbers of whole organisms or hundreds of individually dissected organs. The latter approach is logistically challenging, may introduce artifacts of handling and storage, and is not compatible with biological replication. Here, we tested an approach to generate transcriptomes of individual salivary glands and other low-input body tissues from whiteflies (Bemisia tabaci MEAM1), which are major vectors of plant viruses. By comparing our outputs to published bulk RNA-Seq datasets for whole whitefly bodies and pools of salivary glands, we demonstrate that this approach recovers similar numbers of transcripts relative to bulk RNA-Seq in a tissue-specific manner, and for some metrics, exceeds performance of bulk tissue RNA-Seq. Libraries generated from individual salivary glands also yielded additional novel transcripts not identified in pooled salivary gland datasets, and had hundreds of enriched transcripts when compared with whole head tissues. Overall, our study demonstrates that it is feasible to produce high quality, replicated transcriptomes of whitefly salivary glands and other low-input tissues. We anticipate that our approach will expand hypothesis-driven research on salivary glands of whiteflies and other Hemiptera, thus enabling novel control strategies to disrupt feeding and virus transmission.

Discovery of a widespread presence bunyavirus that may have symbiont-like relationships with different species of aphids.

Aphids are important agricultural pests, vectors of many plant viruses and have sophisticated relationships with symbiotic microorganisms. Abundant asymptomatic RNA viruses have been reported in aphids due to the application of RNA-seq, but aphid-virus interactions remain unclear. Bunyavirales is the most abundant RNA virus order, which can infect mammals, arthropods, and plants. However, many bunyaviruses have specific hosts, such as insects. Here, we discovered 18 viruses from 10 aphid species by RNA-seq. Importantly, a widespread presence bunyavirus, Aphid bunyavirus 1 (ABV-1), was determined to have a wide host range, infecting and replicating in all 10 tested aphid species. ABV-1 may be transmitted horizontally during feeding on plant leaves and vertically through reproduction. In a comparison of the physiological parameters of ABV-1high and ABV-1low strains of pea aphid, higher ABV-1 titers reduced the total nymphal duration and induced the reproduction. Moreover, viral titer significantly affected the lipid and protein contents in pea aphids. In summary, we proposed that ABV-1 may have stable symbiont-like relationships with aphids, and these observations may provide a new direction for studying bunyaviruses in aphids and establishing a model for virus-aphid interactions.

Notes on the embryological development of the Brevipalpus yothersi (Acari: Tenuipalpidae)

Mites of the Brevipalpus genus are economically important due to their role as vectors of plant viruses. Among them, B. yothersi is well studied because it is the recognized main vector of the citrus leprosis virus C (CiLV-C), the causal agent of citrus leprosis, which is one of the most damaging viral diseases in the citrus crops. However, studies of several aspects of the biology of B. yothersi are still scarce, including its embryology. We performed a descriptive embryonic development study of B. yothersi, utilizing light and scanning electron microscope to obtain biological information of this particular phytovirus vector. Distinct events were recorded, such as centering of the nucleus, mitotic divisions for primitive body formation and appendages, as well as the presence of channels inside the egg and their possible association with the respiratory tract. Here we provide the first notes about embryonic development of this important mite vector in the Americas.

Modelling interference between vectors of non-persistently transmitted plant viruses to identify effective control strategies.

Aphids are the primary vector of plant viruses. Transient aphids, which probe several plants per day, are considered to be the principal vectors of non-persistently transmitted (NPT) viruses. However, resident aphids, which can complete their life cycle on a single host and are affected by agronomic practices, can transmit NPT viruses as well. Moreover, they can interfere both directly and indirectly with transient aphids, eventually shaping plant disease dynamics. By means of an epidemiological model, originally accounting for ecological principles and agronomic practices, we explore the consequences of fertilization and irrigation, pesticide deployment and roguing of infected plants on the spread of viral diseases in crops. Our results indicate that the spread of NPT viruses can be i) both reduced or increased by fertilization and irrigation, depending on whether the interference is direct or indirect; ii) counter-intuitively increased by pesticide application and iii) reduced by roguing infected plants. We show that a better understanding of vectors' interactions would enhance our understanding of disease transmission, supporting the development of disease management strategies.

NLR1 is a strong candidate for the Rm3 dominant green peach aphid (Myzus persicae) resistance trait in peach.

The green peach aphid (GPA), Myzus persicae, is a polyphagous, sap-sucking aphid and a vector of many plant viruses. In peach, Prunus persica, three individual dominant GPA resistance loci have been genetically defined (Rm1-3), but knowledge of the underlying genes is limited. In this study, we focused on the Rm3 locus. Bulk segregant analysis (BSA) mapping in segregating progeny populations delimited Rm3 to an interval spanning 160kb containing 21 genes on chromosome 1. RNA-seq data provided no evidence of candidate genes, but chromosomal structural variations were predicted around a nucleotide-binding site-leucine-rich repeat (NLR) gene (ppa000596m) within the Rm3 fine-mapping interval. Following bacterial artificial chromosome (BAC) library construction for a GPA-resistant peach cultivar and the sequencing of three target BAC clones, a chromosomal structural variation encompassing two novel TIR-NLR-class disease resistance (R) protein-coding genes was identified, and the expressed NLR gene (NLR1) was identified as a candidate for M. persicae resistance. Consistent with its proposed role in controlling GPA resistance, NLR1 was only expressed in the leaves of resistant peach phenotypes. A molecular marker that was designed based on the NLR1 sequence co-segregated with the GPA-resistant phenotype in four segregating populations, 162 peach cultivars, and 14 wild relatives, demonstrating the dominant inheritance of the Rm3 locus. Our findings can be exploited to facilitate future breeding for GPA-resistance in peach.

First Report of Sugarcane Mosaic Virus in Zea mays L. in Ukraine

Maize viral diseases especially maize dwarf mosaic disease (MDMD), which is caused by potyviruses, lead to significant crop losses worldwide. Aim. The aim of this work was to identify the causal agent of mosaic symptoms, observed on maize plants during 2018—2020 in Kyiv region. Methods. Enzyme-linked immunosorbent assay in the DAS-ELISA modification using commercial Loewe Biochemica test systems for Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus (SCMV), Wheat streak mosaic virus (WSMV) were applied to identify the causal agent of maize disease in collected samples. Transmission electron microscopy was used in order to direct viral particle visualisation. Aphids, which are natural vectors of plant viruses, were found on diseased plants. Results. Plants with typical mosaic symptoms were observed in corn crops of the Kyiv region in early June 2018. The pathogen was transmitted by mechanical inoculation to maize and sweet maize plants with the manifestation of mosaic symptoms. Electron microscopy of the sap from diseased plants revealed the presence of flexible filamentous virions 750 nm long and 13 nm in diameter, typical for the genus Potyvirus. In August, mosaic symptoms and aphids Rhopalosiphum padi were found on previously healthy plants in the same maize crop. In 2020, in the same sown area, maize plants were free of viral infection during inspection in June, but a re-inspection in September revealed mosaic symptoms on maize crop and the presence of aphids in the leaf axils. The presence of SCMV in maize samples collected in June and August/September 2018 and 2020, as well as in inoculated maize and sweet maize plants, was confirmed by ELISA using a commercial test system. The obtained data allow suggesting that Rhopalosiphum padi is a natural vector of SCMV in agrocenoses of Ukraine. It should be noted that co-infection with MDMV and WSMV in the affected plants was not detected. Conclusions. This study presents the first report of SCMV in maize in Ukraine.

Pathogenic viruses and viral vectors in plants: A comprehensive review

Viruses are extremely small obligate molecular parasites that can be seen only with an electron microscope. In plant pathology there are the organisms named plant viral vectors whose task is to transmit pathogens. Transmission of vector can be mechanical and insect that present two pathways which allow the spreading of plant virus particles. Plant virologists with other association in molecular biology and plant pathology nominated top 10 viruses based on scientific and economic importance. This paper reviews the current standing of most known antiviral defense mechanism called RNA silencing and most known molecular diagnostic method in plant pathology, polymerase chain reaction (PCR).

Tiny Flies: A Mighty Pest That Threatens Agricultural Productivity-A Case for Next-Generation Control Strategies of Whiteflies.

Simple SummaryDespite being a pest of global importance, effective management of whiteflies by the implication of environmentally friendly approaches is still a far-reaching task. In this review, we have tried to bring the readers’ attention to next-generation control strategies such as RNA interference and genetic modifications of plants for the expression of anti-whitefly proteins. These strategies offer huge promise to provide an effective and sustainable solution to the problem of whiteflies, either in isolation or in combination with other widely used practices under the regimes of integrated pest management. Focus has also been given to advanced technologies such as nanotechnology and genome editing, with promising prospects for field applications. The importance, applicability, and demand of these technologies for the control of whiteflies have been highlighted. We have also attempted to present the holistic picture of challenges in the path of commercial application of these promising technologies. To underline the pest status of whiteflies concisely, we have enlisted all economically important species of the pest along with their host plants/crops across the world. A comprehensive list of various insecticides of chemical, microbial, and botanical origin, applied in the field for the control of sweetpotato whitefly along with their resistance status, ecotoxicities, and effects on biological control agents, has been provided for readers.Whiteflies are a group of universally occurring insects that are considered to be a serious pest in their own way for causing both direct and indirect damages to crops. A few of them serve as vectors of plant viruses that are detrimental to the crop in question and cause an actual loss in productivity. A lot of attention is focused on pest control measures under the umbrella of IPM. In this review, we attempt to summarize the existing literature on how and why whiteflies are a serious concern for agriculture and society. We reviewed why there could be a need for fresh insight into the ways and means with which the pest can be combated. Here, we have emphasized next-generation strategies based on macromolecules, i.e., RNA interference and genetic engineering (for the expression of anti-whitefly proteins), as these strategies possess the greatest scope for research and improvement in the future. Recent scientific efforts based on nanotechnology and genome editing, which seem to offer great potential for whitefly/crop pest control, have been discussed. Comprehensive apprehensions related to obstacles in the path of taking lab-ready technologies into the farmers’ field have also been highlighted. Although the use of RNAi, GM crops, nanotechnologies, for the control of whiteflies needs to be evaluated in the field, there is an emerging range of possible applications with promising prospects for the control of these tiny flies that are mighty pests.

Distribution and Genetic Variability of Bemisia tabaci Cryptic Species (Hemiptera: Aleyrodidae) in Italy.

Simple SummaryBemisia tabaci is a key pest of horticultural, fibre and ornamental crops, primarily as a vector of plant viruses. This whitefly is considered as a complex of morphologically indistinguishable species that differ in several biological traits. In Italy, the Mediterranean (MED) and Middle East–Asia Minor 1 (MEAM1) species are known to be present in southern regions as well as in Sicily and Sardinia, where they have been responsible for economically important yield losses to horticultural crops since the 1990s. Recent reports of infestations in some areas of central Italy prompted a reassessment of the species distribution in the country, and several sites have been inspected at different central and southern regions and in the main islands. The survey confirmed that B. tabaci is nowadays established in central Italy even at more northern latitudes than those noticed before, with MED that is clearly prevailing on MEAM1 species throughout the country. The extensive presence of B. tabaci in Italy causes new concerns for the growing number of horticultural and ornamental productive sites that are potentially endangered by whitefly-transmitted viruses. The prevalence of the MED species is even more worrying for its ability to rapidly develop insecticide resistances.Bemisia tabaci is a key pest of horticultural, fibre and ornamental crops worldwide, primarily as a vector of plant viruses. In Italy, B. tabaci has established since the 1980s–1990s in southern regions as well as in Sicily and Sardinia. Recent reports of infestations in some areas of central Italy prompted a new survey to assess the whitefly distribution in the country as well as to update the species and haplotype composition of the populations present in southern Italy and in the main islands. The survey confirmed that B. tabaci is nowadays established in central Italy even at more northern latitudes than those noticed before. Most of the specimens collected throughout the country belonged to the Mediterranean (MED) species. The MEDQ1 and Q2 haplogroups were prevailing in open-field and greenhouse cultivations, respectively, except in Sardinia where only Q1 specimens were found on a wide range of crops and weeds. Population genetics analyses showed that several MEDQ1 haplotypes currently occur in Italy and their distribution is unrelated to evident temporal and geographic trends, except for a new genetic variant which seems to have originated in Sardinia. The MED species is known to better adapt to insecticide treatments and high temperatures, and its northward spread in Italy may have been favoured by the intensive agricultural practices and steady increase in both winter and summer temperatures occurring in the last few decades. The extensive presence of B. tabaci in Italy proves that a strict surveillance for possible new outbreaks of whitefly-transmitted viruses should be addressed to a range of sites that are expanding northwards.

Identification of candidate insect vectors of Grapevine red blotch virus by means of an artificial feeding diet

The ability of an insect to vector a plant virus has traditionally been determined through controlled greenhouse transmission experiments that are notoriously resource- and time-consuming and of unreliable outcome. Therefore, before committing to these protracted experiments, it is important that researchers identify biologically viable candidate vectors. This study presents an approach for identifying potential insect vectors of circulative plant viruses that significantly reduces the time-to-results while also reducing costs and required resources. This artificial transmission protocol, using a buffered sucrose solution as the virus recipient in place of a living plant, also eliminates plant-to-plant variation in susceptibility to infection, allowing assessment of virus–insect compatibility required for successful transmission of persistent viruses. To validate this approach, species of hemipteran insects, including leafhoppers, froghoppers, aphids, sharpshooters, and treehoppers, were subjected to an artificial feeding system to determine their ability to vector the emerging Grapevine red blotch virus (GRBV) (family: Geminiviridae). Test insects were allowed to feed on a potted grapevine infected with GRBV for 3 days and then transferred to tubes containing the sucrose solution partitioned by a thinly stretched Parafilm™ membrane. After 3 days of feeding through the membrane, viruliferous test insects were stored for species identification and the sucrose solutions were tested by conventional polymerase chain reaction for the presence of GRBV DNA. Out of all the insects tested (n = 395) only nine treehoppers from two different species successfully transmitted GRBV to the sucrose solutions, indicating a high likelihood of vector capability, to be validated by greenhouse or field experiments.

Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts

Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts

Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts.

Most plant viruses in nature are transmitted from one plant to another by hemipteran insects. A high population density of the vector insects that are highly efficient at virus transmission plays a key role in virus epidemics in fields. Studying virus-insect vector interactions can advance our understanding of virus transmission and epidemics with the aim of designing novel strategies to control plant viruses and their vector insects. Immunofluorescence labeling has been widely used to analyze interactions between pathogens and hosts and is used here in the white-backed planthopper (WBPH, Sogatella furcifera), which efficiently transmits the southern rice black streaked dwarf virus (SRBSDV, genus Fijivirus, family Reoviridae), to locate the virions and insect proteins in the midgut epithelial cells. Using laser scanning confocal microscopy, we studied the morphological characteristics of midgut epithelial cells, cellular localization of insect proteins, and the colocalization of virions and an insect protein. This protocol can be used to study virus activities in insects, functions of insect proteins, and interactions between virus and vector insect.

Diversity and infectivity of the RNA virome among different cryptic species of an agriculturally important insect vector: whitefly Bemisia tabaci

A large number of insect-specific viruses (ISVs) have recently been discovered, mostly from hematophagous insect vectors because of their medical importance, but little attention has been paid to important plant virus vectors such as the whitefly Bemisia tabaci, which exists as a complex of cryptic species. Public SRA datasets of B. tabaci and newly generated transcriptomes of three Chinese populations are here comprehensively investigated to characterize the whitefly viromes of different cryptic species. Twenty novel ISVs were confidently identified, mostly associated with a particular cryptic species while different cryptic species harbored one or more core ISVs. Microinjection experiments showed that some ISVs might cross-infect between the two invasive whitefly cryptic species, Middle East Asia Minor 1 (MEAM1) and Mediterranean (MED), but others appeared to have a more restricted host range, reflecting the possibility of distinct long-term coevolution of these ISVs and whitefly hosts. Moreover, analysis of the profiles of virus-derived small-interfering RNAs indicated that some of the ISVs can successfully replicate in whitefly and the antiviral RNAi pathway of B. tabaci is actively involved in response to ISV infections. Our study provides a comprehensive analysis of the RNA virome, the distinct relationships and cross-cryptic species infectivity of ISVs in an agriculturally important insect vector.