Viruliferous Whiteflies Research Articles

Resistance to Three Distinct Begomovirus Species in the Agronomical Superior Tropical Pumpkin Line AVPU1426 Developed at the World Vegetable Center

The Squash Leaf Curl China Virus (SLCCNV) and Tomato Leaf Curl New Delhi Virus (ToLCNDV) are species of Begomovirus (whitefly-vectored Geminiviridae) and cause serious damage to the cucurbit crops of the genus Cucurbita in the areas of South and Southeast Asia, across Asia, the Middle East and the Mediterranean, respectively. Cucurbita moschata’s inbred line AVPU1426, developed at the World Vegetable Center (WorldVeg) from a Bangladeshi landrace through pedigree selection, was observed to be resistant to both begomoviruses in field tests conducted at the WorldVeg Research and Training Station, Kasetsart University, Kamphaeng Saen, Thailand, which is a hotspot for these viruses. When AVPU1426 was tested for reaction to inoculation by viruliferous whiteflies with Squash Leaf Curl Philippines Virus (Taiwan strain) (SLCPV-TW) in the screen net-house at WorldVeg headquarters in Taiwan, it showed good resistance, though SLCPV-TW DNA-A could be detected in all inoculated plants, indicating that it did not show immunity to this virus. The objective of this study was to validate the resistance to SLCCNV and ToLCNDV in AVPU1426 by using whitefly-mediated inoculations to determine the mode of inheritance of the resistance. The results showed that AVPU1426 was resistant to SLCCNV and ToLCNDV. Upon crossing AVPU1426 with a susceptible check variety, Waltham Butternut, the resistance to both begomoviruses was observed to be conferred by a single recessive gene. This open-pollinated pumpkin line AVPU1426 bears flat round, mottled green immature fruits with yellow flesh in mature fruit. The fruit yield of AVPU1426 (20.74 t/ha−1) was comparable to ‘Rajah’ (18.61 t/ha−1), a recently released commercial F1 hybrid of East-West Seed (EWS). The fruit of AVPU1426 were estimated to have a good average β-carotene content (1.57 mg/100 g fresh weight). This line is a good source to breed pumpkins resistant to the three begomoviruses.

Competitive Interactions Between Whitefly- and Aphid-Transmitted Poleroviruses Within the Plant Host and the Insect Vectors.

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.

Virus Vector Relationship of Yellow Mosaic Virus and Whitefly, Bemisia tabaci (Gennadius) in Soybean

Background: Whitefly is one of the most destructive sucking pest in the tropical and subtropical regions of the world and causing significant crop losses directly by sucking sap from the plants and indirectly through the transmission of viral diseases specifically caused by the genus Begomovirus. The Begomovirus species viz., Mungbean yellow mosaic India virus (MYMIV) and Mungbean yellow mosaic virus (MYMV) are causing yellow mosaic virus disease in soybean, which is transmitted by whiteflies. The disease accounts to 30-70 per cent yield loss and increases up to 80 - 100 per cent during severe incidence. Hence, there is a need for development of integrated pest management strategies against disease and whiteflies, for this the knowledge on virus-vector relationship is required. But, the studies on biological relationship of yellow mosaic virus disease and whitefly in soybean are scarce. At this juncture, considering the importance of disease in soybean, the present investigation was carried out to know the virus -vector relationship of the YMV and whitefly in soybean.Methods: The experiment on virus-vector relationship of yellow mosaic virus and whitefly in soybean was conducted at Department of Entomology, College of Agriculture, G. B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand during 2016-17. The data on number of whiteflies per plant, acquisition and inoculation access feeding period and pre and post starvation period required for effective transmission of virus was recorded.Result: A single viruliferous whitefly was able to transmit virus and ten viruliferous whiteflies per plant were required for cent per cent transmission of virus. The minimum acquisition access and inoculation access feeding periods required for virus transmission was 0.25h (15 min) each; while the 100 per cent virus transmission was recorded with acquisition and inoculation period of 12h, each. The per cent transmission was increased with the increase of acquisition and inoculation periods. The rate of transmission was positively correlated with pre-acquisition starvation period and negatively correlated with post- acquisition starvation period.

Population structure of whitefly (Bemisia tabaci) and the link between vector dynamics and seasonal incidence of yellow mosaic disease in blackgram (Vigna mungo)

AbstractThe hemipteran pest silverleaf whitefly, Bemisia tabaci (Gennadius) (Aleyrodidae), is of high economic importance, as it infests a wide range of agricultural and horticultural crops. The pest is regarded as a cryptic species complex. In the present study, the cryptic species composition and viruliferous status of B. tabaci in association with yellow mosaic disease (YMD), caused by Mungbean yellow mosaic virus (MYMV), in blackgram [Vigna mungo (L.) Hepper (Fabaceae)], were assessed over three cropping seasons, viz., Rabi, summer, and Kharif, at three locations in Tamil Nadu, India. This study identified three cryptic B. tabaci species: (1) Asia I, (2) Asia II‐5, and (3) Asia II‐8. Asia II‐8 was the dominant species in all three seasons across locations. Despite the absence of YMD symptoms in many plants, up to 73% of whiteflies sampled from YMD‐resistant and susceptible blackgram cultivars were MYMV‐viruliferous. This suggests that the YMD incidence of blackgram does not directly associate with viruliferous whitefly populations in the field.

The roles of entomopathogenic fungal infection of viruliferous whiteflies in controlling tomato yellow leaf curl virus

Efforts to control plant viral diseases have mainly focused on chemical control of their vectors and the development of plant resistance. However, the development of pesticide resistance in many vectors has meant that other vector control options such as the utilization of entomopathogens are receiving increasing attention. Entomopathogens may have the added benefit of suppressing viruses in vectors as well as killing the vectors, although this effect needs further testing under field conditions. Here, we pursue this idea by focusing on the entomopathogenic fungusIsaria javanica, isolated from Q type Bemisia tabaci (whitefly), which is considered the main vector of Tomato yellow leaf curl virus (TYLCV). We also provide insights into potential mechanisms involved in fungus-virus interactions.When nymphs and adults of viruliferous Q whiteflies were infected by I. javanica, there was a significant decrease in the amount of virus they carried. Isaria javanica fermentation (at 2 × 106 spores/mL) caused degradation of the CP capsid protein of TYLCV, with the supernatant from the fermentation showing higher degradation activity than the precipitate. The fungal infection was associated with an increase in plant height and a decrease in the disease index of tomato plants attacked by viruliferous Q type whiteflies. These findings indicate that entomopathogenic I. javanica fungi isolated from Q type whiteflies not only play an insecticidal role but also act as a plant virus suppressor that decreases the amount of TYLCV in the insect vector.

Foliar application of systemic insecticides disrupts feeding behavior of the whitefly Bemisia tabaci MEAM1 and the transmission of tomato chlorosis virus in potato plants

The whiteflies of the Bemisia tabaci (Genn.) complex (Hemiptera: Aleyrodidae) are serious agricultural pests that cause severe losses to vegetable, ornamental and fiber crops, including potato plants, mainly as a vector of economically important viruses. Among the most important viruses affecting potato is tomato chlorosis virus (ToCV) (Closteroviridae: Crinivirus), which is semi-persistently transmitted by whiteflies of the genera Bemisia and Trialeurodes. Chemical control is the main method used to manage B. tabaci and ToCV; however, this whitefly is resistant to most commercially available insecticides, and some products may not effectively prevent the vector stylet activities associated with virus transmission. The effective management of B. tabaci is crucial to reduce the spread of vector-borne diseases, and to minimize economic losses. We evaluated the effects of the foliar spraying with the systemic insecticides acetamiprid, flupyradifurone and cyantraniliprole on the probing behavior of non-viruliferous and ToCV-viruliferous B. tabaci MEAM1 and ToCV transmission in potato plants. To evaluate ToCV transmission in greenhouse conditions, viruliferous whiteflies were released on potato plants at different time points (3, 24 and 72 h and 7 days) after insecticide spraying. The EPG assay showed that at 3 h after insecticide application, the probing behavior differed, depending mainly on the state of the insects (viruliferous or not), whereas 72 h after application, the probing behavior differed only on plants treated with acetamiprid and flupyradifurone, for both viruliferous and non-viruliferous whiteflies. ToCV transmission was reduced mainly in plants treated with flupyradifurone and acetamiprid, likely as a result of phloem activity disruption.

Field Screen and Genotyping of Phaseolus vulgaris against Two Begomoviruses in Georgia, USA.

Simple SummarySnap bean (Phaseolus vulgaris) production and quality have been negatively impacted by two whitefly-transmitted begomoviruses: cucurbit leaf crumple virus (CuLCrV) and sida golden mosaic Florida virus (SiGMFV), which often appear as a mixed infection in Georgia. However, there is no information available in terms of resistance to these two viruses in commercial cultivars/genotypes. Hence, commercially available snap bean varieties/genotypes (n = 84 in 2018; n = 80 in 2019; most of the genotypes were common in both years (with a few exceptions) were screened in two field seasons of 2018 and 2019. We also included two commonly grown Lima bean (Phaseolus lunatus) varieties in our field screening. As a result of this screening, we identified twenty Phaseolus genotypes with high-to-moderate levels of resistance and twenty-one genotypes with high levels of susceptibility. While there were differences among the Phaseolus spp. in severity of viral symptoms, suggesting differential susceptibility to viruses (CuLCrV and SiGMFV) and potential field resistance, the resistance mechanism is yet to be characterized. However, based on the greenhouse evaluation with two genotypes-each (susceptible vs. resistant) exposed to viruliferous whiteflies infected with CuLCrV and SiGMFV, we observed that the susceptible genotypes accumulated higher copy numbers of both viruses and displayed severe crumple severity compared to the resistant genotypes, indicating that resistant might potentially be against the virus complex than against the whiteflies. Adult whitefly counts differed among the Phaseolus spp. in both the years, indicating variability in host preference. We further sequenced 82 genotypes (80 snap bean and two Lima bean) to unravel the variations within the genomes. Genome sequencing followed by bioinformatic analyses revealed a considerable number of sequence variants, single nucleotide polymorphisms (SNPs), and insertions and deletions (InDels) in the genomes. Considering the variations in disease response and the underlying variations in the sequenced genomes, it can be speculated that some of the phenotypic variations (against CuLCrV and SiGMFV) could be due to a high level of genomic variation in the host. Future genome-wide association studies with the identified genomic variants may shed some light on this.The production and quality of Phaseolus vulgaris (snap bean) have been negatively impacted by leaf crumple disease caused by two whitefly-transmitted begomoviruses: cucurbit leaf crumple virus (CuLCrV) and sida golden mosaic Florida virus (SiGMFV), which often appear as a mixed infection in Georgia. Host resistance is the most economical management strategy against whitefly-transmitted viruses. Currently, information is not available with respect to resistance to these two viruses in commercial cultivars. In two field seasons (2018 and 2019), we screened Phaseolus spp. genotypes (n = 84 in 2018; n = 80 in 2019; most of the genotypes were common in both years with a few exceptions) for resistance against CuLCrV and/or SiGMFV. We also included two commonly grown Lima bean (Phaseolus lunatus) varieties in our field screening. Twenty Phaseolus spp. genotypes with high to moderate-levels of resistance (disease severity ranging from 5%–50%) to CuLCrV and/or SiGMFV were identified. Twenty-one Phaseolus spp. genotypes were found to be highly susceptible with a disease severity of ≥66%. Furthermore, based on the greenhouse evaluation with two genotypes-each (two susceptible and two resistant; identified in field screen) exposed to viruliferous whiteflies infected with CuLCrV and SiGMFV, we observed that the susceptible genotypes accumulated higher copy numbers of both viruses and displayed severe crumple severity compared to the resistant genotypes, indicating that resistance might potentially be against the virus complex rather than against the whiteflies. Adult whitefly counts differed significantly among Phaseolus genotypes in both years. The whole genome of these Phaseolus spp. [snap bean (n = 82); Lima bean (n = 2)] genotypes was sequenced and genetic variability among them was identified. Over 900 giga-base (Gb) of filtered data were generated and >88% of the resulting data were mapped to the reference genome, and SNP and Indel variants in Phaseolus spp. genotypes were obtained. A total of 645,729 SNPs and 68,713 Indels, including 30,169 insertions and 38,543 deletions, were identified, which were distributed in 11 chromosomes with chromosome 02 harboring the maximum number of variants. This phenotypic and genotypic information will be helpful in genome-wide association studies that will aid in identifying the genetic basis of resistance to these begomoviruses in Phaseolus spp.

Interplay of stress responses to carbamazepine treatment, whitefly infestation and virus infection in tomato plants

Reclaimed wastewater is increasingly used to irrigate agriculture crops. We have previously shown that carbamazepine (CBZ), an anticonvulsant human medication, not entirely discarded during wastewater purification, induces a stress response in tomatoes grown with roots bathing in CBZ-containing water. Induction of stress-related osmoprotectants (sugars, amino acids, proteins) was conspicuous in CBZ-treated seedlings. Here, tomato seedlings were grown in pots watered with increasing concentrations of CBZ. Soluble sugars effectively reacted to CBZ in both leaves and roots. However, the induction of stress-related amino acids and proteins was relevant in roots, but insignificant in leaves. Therefore, roots may be the site where CBZ stress is exerted. Moreover, roots may protect the whole plant from the pharmaceutical. Tomato crops endure biotic stresses, caused by whitefly (Bemisia tabaci) infestation and by the tomato yellow leaf curl begomovirus (TYLCV) they vector. The interplay between CBZ, TYLCV infection and B. tabaci was studied in tomato. Whiteflies preferred CBZ-treated plantlets than control plants, which may be due to increased amount of sugars in leaves. The increased amount of viruliferous whiteflies on CBZ-treated plants is expected to be accompanied by a rise in virus amounts. In fact, CBZ caused a reduction of TYLCV amounts. CBZ-dependent activation of autophagy degradation may explain this decrease in virus amounts. TYLCV infection mitigates the activation of stress markers associated with CBZ treatment. Altogether, CBZ in the water used to irrigate tomatoes grown in pots causes a relatively weak plant stress response, but is definitively sensed by insect and by virus.

Interplay of Cucurbit Yellow Stunting Disorder Virus With Cucurbit Chlorotic Yellows Virus and Transmission Dynamics by Bemisia tabaci MED.

Cucurbit chlorotic yellows virus (CCYV) and cucurbit yellow stunting disorder virus (CYSDV) are two closely related criniviruses that often coinfect cucurbits and are associated with cucurbit yellows disease. Both viruses are distributed worldwide and are transmitted in a semipersistent manner by the whitefly vectors Bemisia tabaci MED or MEAM1. The major goal of this study was to provide insight into the interaction of CCYV and CYSDV in cucumber and to study the effect on transmission by B. tabaci MED. The titers of both viruses were estimated in single- and dually infected cucumber plants via reverse transcription PCR assays. In mixed infections, the accumulation of both viruses was significantly decreased. When B. tabaci MED adults were placed on cucumber infected with both viruses, their simultaneous transmission efficiency was significantly higher, whereas transmission efficiency of each individual virus was low. Moreover, nonviruliferous whiteflies preferentially settled on crinivirus-infected cucumber plants, whereas viruliferous whiteflies were attracted by healthy cucumber plants. Finally, the titer of both viruses was calculated in five commercial cucumber hybrids, followed by subsequent transmission experiments. Our results show that although the titers of CYSDV and CCYV were significantly lower in mixed infections in cucumbers, their simultaneous transmission increased.

Solanum pseudocapsicum: Wild source of resistance to Chilli leaf curl disease

The present investigation was undertaken to screen Solanum pseudocapsicum, against chilli leaf curl disease (ChiLCD) which is the most destructive virus disease in terms of incidence and yield loss in chilli peppers. The screening was done both under natural epiphytotic condition and through challenge inoculation using viruliferous whiteflies. The genus, which is perennial in nature and has a life span of about 10 years, was found free of infection under field screening even after four years of continuous exposure to high disease pressure. Under challenge inoculation, S. pseudocapsicum was found asymptomatic upon infection with ChiLCV and ToLCNDV, the two pre-dominant begomovirus prevalent in the study area. S. pseudocapsicum plants exposed to leaf curl disease under field condition was analysed for presence of ChiLCV through PCR and was found to show presence of virus indicating it to be a symptomless carrier of the disease. S. pseudocapsicum has proven to be a source of resistance against ChiLCD.

Mesta yellow vein mosaic virus: application of loop-mediated isothermal amplification method to study efficiency of acquisition, retention and transmission by Bemisia tabaci (Hemiptera: Aleyrodidae) in Kenaf

Whitefly is an important pest and vector of many plant begomoviruses. Investigations were carried out to understand the acquisition, retention, transmission, and gender basis transmission efficiency of Mesta yellow vein mosaic virus (MeYVMV) by whitefly. The minimum acquisition access feeding period (AAFP) was 0.10 h which resulted in 11.1% infected plants. Based on LAMP assay and yellowing symptoms, 100% virus acquisition was achieved by whitefly with an exposure of 18 to 48 h. Virus was transmitted in tested plants up to 88.8% after 1 to 10 days retention time of viruliferous whiteflies, whereas no symptoms were observed after 35 days. Fifteen minutes of exposure to viruliferous whiteflies on healthy plants resulted in 11.1% transmission. Cent percent transmission of virus was achieved in plants with 12 h exposure period. Highest virus transmission efficiency of 55.60% was observed in female whitefly compared to 33.3% in single male whitefly. Present study shall help to understand the interaction of insect-plant–virus relationship, epidemiological knowledge, and formulation of management strategies against virus and its vector.

Characterization of Local and Systemic Impact of Whitefly (Bemisia tabaci) Feeding and Whitefly-Transmitted Tomato Mottle Virus Infection on Tomato Leaves by Comprehensive Proteomics

Tomato mottle virus (ToMoV) is a single-stranded DNA (ssDNA) begomovirus transmitted to solanaceous crops by the whitefly species complex (Bemisia tabaci), causing stunted growth, leaf mottling, and reduced yield. Using a genetic repertoire of seven genes, ToMoV pathogenesis includes the manipulation of multiple plant biological processes to circumvent antiviral defenses. To further understand the effects of whitefly feeding and whitefly-transmitted ToMoV infection on tomato plants (Solanum lycopersicum ‘Florida Lanai’), we generated comprehensive protein profiles of leaves subjected to feeding by either viruliferous whiteflies harboring ToMoV, or non-viruliferous whiteflies, or a no-feeding control. The effects of whitefly feeding and ToMoV infection were measured both locally and systemically by sampling either a mature leaf directly from the site of clip-cage confined whitefly feeding, or from a newly formed leaf 10 days post feeding (dpf). At 3 dpf, tomato’s response to ToMoV included proteins associated with translation initiation and elongation as well as plasmodesmata dynamics. In contrast, systemic impacts of ToMoV on younger leaves 10 dpf were more pronounced and included a virus-specific change in plant proteins associated with mRNA maturation and export, RNA-dependent DNA methylation, and other antiviral plant processes. Our analysis supports previous findings and provides novel insight into tomato’s local and systemic response to whitefly feeding and ToMoV infection.

Differential expression profiling of defense related genes for Leaf Curl Virus (ChiLCV) in resistant and susceptible genotypes of Chiili

The present investigation was undertaken to study the expression of eight defense related genes in leaf curl resistant line DLS-Sel-10 and susceptible line Phule Mukta after different days post inoculation (dpi) with viruliferous white flies to understand their role in resistance to chilli leaf curl virus (ChiLCV). The expression level of Ca PPO, Ca AsPer, Ca ATP/ADP and CaTopoII was observed to be higher in resistant genotype DLS-Sel-10 than the susceptible Phule Mukta at all the time points studied. Expression of CaNBS-LRR increased up to 12 dpi while that of Ca Thionin, and Ca SKP1 increased up to 24 dpi in the resistant line, thereafter it started declining. The CaSPI expression did not show any specific pattern in both the test plants. The heat map clustered all the genes under study into two major clusters based on their expression profiles, one comprising CaAsPer, Ca-Thionin, CaATP/ADP transporter, CaPPO and CaTopoII while other group comprised CaSKPI, Ca NBS and CaSPI. The challenge inoculation of the test genotypes also revealed that viral titre increased at a much slower rate in DLS-Sel-10 than Phule Mukta, suggesting thereby that DLS-Sel-10 is resisting the accumulation of ChiLCV and has a more active defense machinery than Phule Mukta.

Transmission of the Bean-Associated Cytorhabdovirus by the Whitefly Bemisia tabaci MEAM1.

The knowledge of genomic data of new plant viruses is increasing exponentially; however, some aspects of their biology, such as vectors and host range, remain mostly unknown. This information is crucial for the understanding of virus–plant interactions, control strategies, and mechanisms to prevent outbreaks. Typically, rhabdoviruses infect monocot and dicot plants and are vectored in nature by hemipteran sap-sucking insects, including aphids, leafhoppers, and planthoppers. However, several strains of a potentially whitefly-transmitted virus, papaya cytorhabdovirus, were recently described: (i) bean-associated cytorhabdovirus (BaCV) in Brazil, (ii) papaya virus E (PpVE) in Ecuador, and (iii) citrus-associated rhabdovirus (CiaRV) in China. Here, we examine the potential of the Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) to transmit BaCV, its morphological and cytopathological characteristics, and assess the incidence of BaCV across bean producing areas in Brazil. Our results show that BaCV is efficiently transmitted, in experimental conditions, by B. tabaci MEAM1 to bean cultivars, and with lower efficiency to cowpea and soybean. Moreover, we detected BaCV RNA in viruliferous whiteflies but we were unable to visualize viral particles or viroplasm in the whitefly tissues. BaCV could not be singly isolated for pathogenicity tests, identification of the induced symptoms, and the transmission assay. BaCV was detected in five out of the seven states in Brazil included in our study, suggesting that it is widely distributed throughout bean producing areas in the country. This is the first report of a whitefly-transmitted rhabdovirus.

Foliar Spraying of Tomato Plants with Systemic Insecticides: Effects on Feeding Behavior, Mortality and Oviposition of Bemisia tabaci (Hemiptera: Aleyrodidae) and Inoculation Efficiency of Tomato Chlorosis Virus.

Simple SummaryThe whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) causes serious losses to vegetable, ornamental and fiber crops, including tomato plants, mainly as a vector of economically important viruses. Among the most important viruses affecting tomato is the tomato chlorosis virus (ToCV) (Closteroviridae: Crinivirus), which is semi-persistently transmitted by whiteflies. Effective management of this pest is crucial to reduce the spread of vector-borne diseases and to reduce crop damage and losses. In this study we evaluated the effect of systemic insecticides (cyantraniliprole, acetamiprid and flupyradifurone) on the feeding behavior, mortality and oviposition of B. tabaci MEAM1 and their ability to interfere with the inoculation of ToCV in tomato plants. Our findings indicate that systemic insecticides cause high mortality when compared to untreated plants. Also, we found that flupyradifurone affects stylet activities of B. tabaci and significantly reduce phloem ingestion, a behavior that is closely linked to the transmission of ToCV. Overall, our findings indicate that flupyradifurone may contribute to management of this pest and ToCV in tomato crops.Tomato chlorosis virus (ToCV) is a phloem-limited crinivirus transmitted by whiteflies and seriously affects tomato crops worldwide. As with most vector-borne viral diseases, no cure is available, and the virus is managed primarily by the control of the vector. This study determined the effects of the foliar spraying with the insecticides, acetamiprid, flupyradifurone and cyantraniliprole, on the feeding behavior, mortality, oviposition and transmission efficiency of ToCV by B. tabaci MEAM1 in tomato plants. To evaluate mortality, oviposition and ToCV transmission in greenhouse conditions, viruliferous whiteflies were released on insecticide-treated plants at different time points (3, 24 and 72 h; 7 and 14 days) after spraying. Insect mortality was higher on plants treated with insecticides; however, only cyantraniliprole and flupyradifurone differed from them in all time points. The electrical penetration graph (DC-EPG) technique was used to monitor stylet activities of viruliferous B. tabaci in tomato plants 72 h after insecticide application. Only flupyradifurone affected the stylet activities of B. tabaci, reducing the number and duration of intracellular punctures (pd) and ingestion of phloem sap (E2), a behavior that possibly resulted in the lower percentage of ToCV transmission in this treatment (0–60%) in relation to the control treatment (60–90%) over the periods evaluated. Our results indicate that flupyradifurone may contribute to management of this pest and ToCV in tomato crops.

Apoptotic neurodegeneration in whitefly promotes the spread of TYLCV.

The mechanism by which plant viruses manipulate the behavior of insect vectors has largely been described as indirect manipulation through modifications of the host plant. However, little is known about the direct interaction of the plant virus on the nervous system of its insect vector, and the substantial behavioral effect on virus transmission. Using a system consisting of a Tomato yellow leaf curl virus (TYLCV) and its insect vector whitefly, we found that TYLCV caused caspase-dependent apoptotic neurodegeneration with severe vacuolar neuropathological lesions in the brain of viruliferous whitefly by inducing a putative inflammatory signaling cascade of innate immunity. The sensory defects caused by neurodegeneration removed the steady preference of whitefly for virus-infected plants, thereby enhancing the probability of the virus to enter uninfected hosts, and eventually benefit TYLCV spread among the plant community. These findings provide a neuromechanism for virus transmission to modify its associated insect vector behavior.

Evaluation of disease severity caused by Bean golden mosaic virus in different bean cultivars

Common bean (Phaseolus vulgaris L.) can be affected by several viral diseases, including golden mosaic caused by the Bean golden mosaic virus (BGMV). When infected by BGMV, bean plants undergo major physiological changes, leading to a reduction or complete loss of productivity. This study aimed to evaluate and compare seven bean cultivars and lineages, as well as the legume calopo (Calopogonium mucunoides Desv.), for the severity of golden mosaic symptoms and BGMV titre following inoculation. The study was conducted in the greenhouse under controlled environmental conditions. Plants were inoculated with viruliferous whiteflies [Bemisia tabaci (Gennadius)], and evaluated for severity of disease symptoms at 7, 10, 15, 20, and 25 days after inoculation (DAI). The virus titre was measured by quantitative PCR (qPCR) analysis with primers specific for the virus coat protein gene. All bean genotypes were symptomatic for BGMV infection, but calopo was not. There were differences among the tested cultivars and lineages for the expression of symptoms. Differences in BGMV titre also were observed among the cultivars at 10 and 25 DAI. The bean cultivar ‘Tangará’ had a greater viral titre but exhibited lower symptom severity than the other cultivars. Calapo was tolerant to BGMV.

Virus-virus interactions in a plant host and in a hemipteran vector: Implications for vector fitness and virus epidemics

Mixed virus infection in host plants can differentially alter the plant phenotype, influence vector fitness, and affect virus acquisition and inoculation by vectors than single-virus infection. Vector acquisition of multiple viruses from multiple host plants could also differentially affect vector fitness and virus inoculation than acquisition of one virus. Whitefly-virus pathosystems in the southern United States include both the above-stated facets. For the first facet, this study examined the effects of single and mixed infection of cucurbit leaf crumple virus (CuLCrV, a begomovirus) and cucurbit yellow stunting disorder virus (CYSDV, a crinivirus) infecting squash on whitefly (Bemisia tabaci Gennadius MEAM1) host preference and fitness. Mixed infection of CuLCrV and CYSDV in squash plants severely altered their phenotype than single infection. The CYSDV load was reduced in mixed-infected squash plants than in singly-infected plants. Consequently, whiteflies acquired reduced amounts of CYSDV from mixed-infected plants than singly-infected plants. No differences in CuLCrV load were found between singly- and mixed-infected squash plants, and acquisition of CuLCrV by whiteflies did not vary between singly- and mixed-infected squash plants. Both singly- and mixed-infected plants similarly affected whitefly preference, wherein non-viruliferous and viruliferous (CuLCrV and/or CYSDV) whiteflies preferred non-infected plants over infected plants. The fitness study involving viruliferous and non-viruliferous whiteflies revealed no differences in developmental time and fecundity. For the second facet, this study evaluated the effects of individual or combined acquisition of tomato-infecting tomato yellow leaf curl virus (TYLCV, a begomovirus) and squash-infecting CuLCrV on whitefly host preference and fitness. Whiteflies that acquired both CuLCrV and TYLCV had significantly lower CuLCrV load than whiteflies that acquired CuLCrV alone, whereas TYLCV load remained unaltered when acquired individually or in conjunction with CuLCrV. Whitefly preference was not affected following individual or combined virus acquisition. Viruliferous (CuLCrV and/or TYLCV) whiteflies preferred to settle on non-infected tomato and squash plants. The mere presence of CuLCrV and/or TYLCV in whiteflies did not affect their fitness. Taken together, these results indicate that mixed infection of viruses in host plants and acquisition of multiple viruses by the vector could have implications for virus accumulation, virus acquisition, vector preference, and epidemics that sometimes are different from single-virus infection or acquisition.

Transmission and host range of Horsegram yellow mosaic virus (HgYMV) causing common bean (Phaseolus vulgaris L.) yellowing disease in Sri Lanka

The common bean (Phaseolus vulgaris L.) yellowing disease (BYD) is caused by the Horsegram yellow mosaic virus (HgYMV), which is a begomovirus transmitted by whiteflies (Bemisia tabaci Genn.). It is a serious threat to the common bean cultivation in Sri Lanka. Despite the fact that the disease has been affecting bean production for a long time, its etiology and virus vector relationships are not understood well. Therefore, the present work was carried out to identify major aspects of the etiology of the virus. Initially a survey was conducted to determine the prevalence of the virus in representative bean growing locations in Sri Lanka, followed by a host range study using 54 plant species. Characteristics of virus-vector relationships were determined by conducting a series of virus transmission tests under glass-house conditions. The survey results revealed the incidence of BYD at some time or the other in most of the sampled localities, which are scattered around all four major districts where the common bean is grown. Through the host range study, it was demonstrated that Ageratum sp., Glycine max Merrill., Macrotyloma uniflorum (Lam.) Verdc., and Phaseolus lunatus L. are susceptible to HgYMV, indicating a narrow host range. In addition to the symptomatology, the successful entry of the virus following inoculation was confirmed through PCR using degenerate primers. Relative transmission efficiency was found to be 96.7 %, which corresponds to 0.24 of the estimated probability of transmission through a single whitefly (p*). The virus could be transmitted through probing by a single viruliferous whitefly with an incidence of 36.7 % (p* = 0.37). The minimum acquisition access period and inoculation access period were 20 min in each case and the respective disease incidence values were 30.0 % (p* = 0.04) and 36.7 % (p* = 0.05). The whiteflies could retain the virus in an active state for up to 9 days.

Amplifier Hosts May Play an Essential Role in Tomato Begomovirus Epidemics in Brazil.

Current control of tomato golden mosaic disease, caused in Brazil predominantly by tomato severe rugose virus (ToSRV), is dependent on both, planting resistant/tolerant hybrids and intensive insecticide sprays (two to three per week) for controlling Bemisia tabaci, the vector of ToSRV. Resistant hybrids only confer moderate resistance to infection by ToSRV and some tolerance to the disease. Insecticide sprays, although widely used, have failed in most tomato production areas in Brazil, as they are unable to reduce primary spread, i.e., infection caused by the influx of viruliferous whiteflies coming from external sources of inoculum. Severe epidemics are recurrently observed in some tomato fields in several Brazilian regions, which prompted us to postulate the existence in the agroecosystem, in some places and time, of amplifier hosts that provide the necessary force of infection for epidemics to occur, even in the absence of secondary spread in the target crop. Amplifier hosts are ideally asymptomatic, occur in high density near the target crop, and support growth of both virus and vector. Soybean and common bean are potential amplifier hosts for begomovirus in tomato crops. Our results support the hypothesis that soybean plants may play an important role as an amplifier host of ToSRV for tomato crops in the field, although this does not seem to be a frequent phenomenon. Successful amplification will depend on several factors, including the soybean cultivar, the soybean stage of development at the moment of infection, the ToSRV isolate, and the perfect synchrony between the beginning of a soybean field and the end of a ToSRV-infected crop, and, later, between the senescence of the ToSRV-infected soybean plants and the new tomato crop. The concept of amplifier hosts has been widely used in ecology of zoonoses but, to our knowledge, has never been used in botanical epidemiology.