Spotted Wilt Virus Infection Research Articles

Dehydration and tomato spotted wilt virus infection combine to alter feeding and survival parameters for the western flower thrips, Frankliniella occidentalis

Dehydration and tomato spotted wilt virus (TSWV) infection substantially impact the feeding of western flower thrips, Frankliniella occidentalis. Until now, the dynamics between these biotic and abiotic stresses have not been examined for thrips. Here, we report water balance characteristics and changes in other biological parameters during infection with TSWV for the western flower thrips. There were no apparent differences in water balance parameters during TSWV infection of male or female thrips. Our results show that, although water balance characteristics of western flower thrips are minimally impacted by TSWV infection, the increase in feeding and activity when dehydration and TSWV are combined suggests that virus transmission could be increased under periods of drought. Importantly, survival and progeny generation were impaired during TSWV infection and dehydration bouts. The negative impact on survival and reproduction suggests that the interactions between TSWV infection and dehydration will likely reduce thrips populations. The opposite effects of dehydration on feeding/activity and survival/reproduction for virus infected thrips suggest the impact of vectorial capacity will likely be minor for TSWV transmission. As water stress significantly impacts insect-plant-virus dynamics, these studies highlight that all interactions and effects need to be measured to understand thrips-TSWV interactions in their role as viral vector to plants.

Genome-wide characterization of the tomato UDP-glycosyltransferase gene family and functional identification of SlUDPGT52 in drought tolerance

Abiotic stresses are major factors constraining the growth, development and productivity of tomato (Solanum lycopersicum), the most cultivated vegetable crop worldwide. Uridine diphosphate glycosyltransferases (UDPGTs or UGTs) are essential enzymes that utilize 5-uridine diphosphate as a glycosyl donor molecule to facilitate the catalysis of glycosylation reactions across diverse substrates, thereby playing a pivotal role in conferring abiotic stress tolerance. Currently, there is a limited understanding of the structure and functions of the UDPGT gene family in tomato. In this work, 106 members of the SlUDPGT gene family were identified through in silico analysis, besides, their protein sequence properties, phylogenetic relationships, gene structure, chromosomal distribution, cis-acting elements, tissue expression and hormone- and stress-induced expression were comprehensively investigated. The expression of representative SlUDPGTs under abiotic stress and exogenous hormone treatments, including salt, polyethylene glycol, methyl viologen, gibberellic acid, jasmonic acid, abscisic acid and brassinolide, was investigated through qRT‒PCR analysis. Numerous cis-acting elements linked to stress and hormone signaling were present in the promoter regions of SlUDPGTs. According to microarray data, most SlUDPGT genes were responsive to hormones and abiotic stresses, while certain SlUDPGTs were specifically differentially expressed under Botrytis cinerea and tomato spotted wilt virus infection. Additionally, diverse expression profiles of SlUDPGTs were observed in various tissues and developmental stages. Furthermore, CRISPR/Cas9-mediated knockout of SlUDPGT52 led to enhanced drought tolerance due to enhanced reactive oxygen species (ROS) scavenging. These findings lay the foundations for the future functional characterization of specific UDPGT gene family members, assisting the biotechnology-mediated improvement of tomato and other horticultural crops.

HMG-like DSP1 is a damage signal to mediate the western flower thrips, Frankliniella occidentalis, immune responses to tomato spotted wilt virus infection

Tomato spotted wilt virus (TSWV) causes a serious plant disease and is transmitted by specific thrips including the western flower thrips, Frankliniella occidentalis. The persistent and circulative virus transmission suggests an induction of immune defenses in the thrips. We investigated the immune responses of F. occidentalis to TSWV infection. Immunofluorescence assay demonstrated viral infection in the larval midguts at early stage and subsequent propagation to the salivary gland in adults. In the larval midgut, TSWV infection led to the release of DSP1, a damage-associated molecular pattern, from the gut epithelium into the hemolymph. DSP1 up-regulated PLA2 activity, which would lead to biosynthesis of eicosanoids that activate cellular and humoral immune responses. Phenoloxidase (PO) activity was enhanced following induction of PO and its activating protease gene expressions. Antimicrobial peptide genes and dual oxidase, which produces reactive oxygen species, were induced by the viral infection. Expression of four caspase genes increased and TUNEL assay confirmed apoptosis in the larval midgut after the virus infection. These immune responses to viral infection were significantly suppressed by the inhibition of DSP1 release. We infer that TSWV infection induces F. occidentalis immune responses, which are activated by the release of DSP1 from the infection foci within midguts.

Three-dimensional analysis of membrane structures associated with tomato spotted wilt virus infection.

To study viral infection, the direct structural visualization of the viral life cycle consisting of virus attachment, entry, replication, assemblyand transport is essential. Although conventional electron microscopy (EM) has been extremely helpful in the investigation of virus-host cell interactions, three-dimensional (3D) EM not only provides important information at the nanometer resolution, but can also create 3D maps of large volumes, even entire virus-infected cells. Here, we determined the ultrastructural details of tomato spotted wilt virus (TSWV)-infected plant cells using focused ion beam scanning EM (FIB-SEM). The viral morphogenesis and dynamic transformation of paired parallel membranes (PPMs) were analyzed. The endoplasmic reticulum (ER) membrane network consisting of tubules and sheets was related to viral intracellular trafficking and virion storage. Abundant lipid-like bodies, clustering mitochondria, cell membrane tubules, and myelin-like bodies were likely associated with viral infection. Additionally, connecting structures between neighboring cells were found only in infected plant tissues and showed the characteristics of tubular structure. These novel connections that formed continuously in the cell wall or were wrapped by the cell membranes of neighboring cells appeared frequently in the large-scale 3D model, suggesting additional strategies for viral trafficking that were difficult to distinguish using conventional EM.

Sex-biased proteomic response to tomato spotted wilt virus infection of the salivary glands of Frankliniella occidentalis, the western flower thrips

Successful transmission of tomato spotted wilt virus (TSWV) by Frankliniella occidentalis requires robust infection of the salivary glands (SGs) and virus delivery to plants during salivation. Feeding behavior and transmission efficiency are sexually-dimorphic traits of this thrips vector species. Proteins secreted from male and female SG tissues, and the effect of TSWV infection on the thrips SG proteome are unknown. To begin to discern thrips factors that facilitate virus infection of SGs and transmission by F. occidentalis, we used gel- and label-free quantitative and qualitative proteomics to address two hypotheses: (i) TSWV infection modifies the composition and/or abundance of SG-expressed proteins in adults; and (ii) TSWV has a differential effect on the male and female SG proteome and secreted saliva. Our study revealed a sex-biased SG proteome for F. occidentalis, and TSWV infection modulated the SG proteome in a sex-dependent manner as evident by the number, differential abundance, identities and generalized roles of the proteins. Male SGs exhibited a larger proteomic response to the virus than female SGs. Intracellular processes modulated by TSWV in males indicated perturbation of SG cytoskeletal networks and cell-cell interactions, i.e., basement membrane (BM) and extracellular matrix (ECM) proteins, and subcellular processes consistent with a metabolic slow-down under infection. Several differentially-abundant proteins in infected male SGs play critical roles in viral life cycles of other host-virus pathosystems. In females, TSWV modulated processes consistent with tissue integrity and active translational and transcriptional regulation. A core set of proteins known for their roles in plant cell-wall degradation and protein metabolism were identified in saliva of both sexes, regardless of virus infection status. Saliva proteins secreted by TSWV-infected adults indicated energy generation, consumption and protein turnover, with an enrichment of cytoskeletal/BM/ECM proteins and tricarboxylic acid cycle proteins in male and female saliva, respectively. The nonstructural TSWV protein NSs - a multifunctional viral effector protein reported to target plant defenses against TSWV and thrips - was identified in female saliva. This study represents the first description of the SG proteome and secretome of a thysanopteran and provides many candidate proteins to further unravel the complex interplay between the virus, insect vector, and plant host.

Integration of transcriptomics and network analysis reveals co-expressed genes in Frankliniella occidentalis larval guts that respond to tomato spotted wilt virus infection

BackgroundThe gut is the first barrier to infection by viruses that are internally borne and transmitted persistently by arthropod vectors to plant and animal hosts. Tomato spotted wilt virus (TSWV), a plant-pathogenic virus, is transmitted exclusively by thrips vectors in a circulative-propagative manner. Frankliniella occidentalis (western flower thrips), the principal thrips vector of TSWV, is transmission-competent only if the virus is acquired by young larvae. To begin to understand the larval gut response to TSWV infection and accumulation, a genome-assisted, transcriptomic analysis of F. occidentalis gut tissues of first (early L1) and second (early L2 and late L2) instar larvae was conducted using RNA-Seq to identify differentially-expressed transcripts (DETs) in response to TSWV compared to non-exposed cohorts.ResultsThe larval gut responded in a developmental stage-dependent manner, with the majority of DETs (71%) associated with the early L1 stage at a time when virus infection is limited to the midgut epithelium. Provisional annotations of these DETs inferred roles in digestion and absorption, insect innate immunity, and detoxification. Weighted gene co-expression network analysis using all assembled transcripts of the gut transcriptome revealed eight gene modules that distinguish larval development. Intra-module interaction network analysis of the three most DET-enriched modules revealed ten central hub genes. Droplet digital PCR-expression analyses of select network hub and connecting genes revealed temporal changes in gut expression during and post exposure to TSWV.ConclusionsThese findings expand our understanding of the developmentally-mediated interaction between thrips vectors and orthotospoviruses, and provide opportunities for probing pathways for biomarkers of thrips vector competence.

NbTMP14 Is Involved in Tomato Spotted Wilt Virus Infection and Symptom Development by Interaction with the Viral NSm Protein.

Tomato spotted wilt virus (TSWV) is one of the most destructive plant viruses, causing severe losses in many important crops worldwide. The non-structural protein NSm of TSWV is a viral movement protein that induces viral symptoms. However, the molecular mechanisms by which NSm contributes to symptom development are unclear. Here, we present evidence that NSm directly interacts with Nicotiana benthamiana chloroplast thylakoid membrane protein TMP14 (NbTMP14) by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. The interaction between NSm and NbTMP14 led to the translocation of the NbTMP14 protein from the chloroplast to the cytoplasm in TSWV-infected plants, and overexpressing NSm decreased NbTMP14 mRNA accumulation. In addition, abnormal chloroplasts and starch accumulation were observed in TSWV-infected plants. Silencing of NbTMP14 by TRV VIGS also showed similar results to those of TSWV-infected plants. Overexpressing NbTMP14 in transgenic N. benthamiana plants impeded TSWV infection, and silencing NbTMP14 in N. benthamiana plants increased disease symptom severity and virus accumulation. To our knowledge, this is the first report showing that the plant chloroplast TMP14 protein is involved in viral infection. Knowledge of the interaction between NSm and NbTMP14 advances our understanding of the molecular mechanisms underlying TSWV symptom development and infection.

Sixty-One Years Following Registration, Phorate Applied In-Furrow at Planting Suppresses Development of Late Leaf Spot on Peanut.

Late and early leaf spot are caused by Nothopassalora personata and Passalora arachidicola, respectively, and are damaging diseases of peanut (Arachis hypogaea L.) capable of defoliation and yield loss. Management of these diseases is most effective through the integration of tactics that reduce starting inoculum and prevent infection. The insecticide phorate was first registered in 1959 and has been used in peanut production for decades in-furrow at planting to suppress thrips. Phorate further provides significant suppression of Tomato spotted wilt virus infection beyond suppression of its thrips vector alone by activating defense-related responses in the peanut plant. From six experiments conducted from 2017 to 2019 in Blackville, SC, Reddick, FL, and Quincy, FL, significantly less leaf spot defoliation was exhibited on peanuts treated with phorate in-furrow at planting (26%) compared with nontreated checks (48%). In-season fungicides were excluded from five of the experiments, whereas the 2018 Quincy, FL, experiment included eight applications on a 15-day interval. Across individual experiments, significant suppression of defoliation caused by late leaf spot was observed from 64 to 147 days after planting. Although more variable within location-years, pod yield following phorate treatment was overall significantly greater than for nontreated peanut (2,330 compared with 2,030 kg/ha; P = 0.0794). The consistent defoliation suppression potential was estimated to confer an average potential net economic yield savings of $90 to $120 per hectare under analogous leaf spot defoliation. To our knowledge, these are the first data in the 61 years since its registration demonstrating significant suppression of leaf spot on peanut following application of phorate in-furrow at planting. Results support phorate use in peanut as an effective and economical tactic to incorporate to manage late and early leaf spot infections and development of fungicide resistance.

Developmentally regulated Arabidopsis thaliana susceptibility totomato spotted wilt virus infection.

Tomato spotted wilt virus (TSWV) is one of the most devastating plant viruses and often causes severe crop losses worldwide. Generally, mature plants become more resistant to pathogens, known as adult plant resistance. In this study, we demonstrated a new phenomenon involving developmentally regulated susceptibility of Arabidopsis thaliana to TSWV. We found that Arabidopsis plants become more susceptible to TSWV as plants mature. Most young 3‐week‐old Arabidopsis were not infected by TSWV. Infection of TSWV in 4‐, 5‐, and 6‐week‐old Arabidopsis increased from 9%, 21%, and 25%, respectively, to 100% in 7‐ to 8‐week‐old Arabidopsis plants. Different isolates of TSWV and different tospoviruses show a low rate of infection in young Arabidopsis but a high rate in mature plants. When Arabidopsis dcl2/3/4 or rdr1/2/6 mutant plants were inoculated with TSWV, similar results as observed for the wild‐type Arabidopsis plants were obtained. A cell‐to‐cell movement assay showed that the intercellular movement efficiency of TSWV NSm:GFP fusion was significantly higher in 8‐week‐old Arabidopsis leaves compared with 4‐week‐old Arabidopsis leaves. Moreover, the expression levels of pectin methylesterase and β‐1,3‐glucanase, which play critical roles in macromolecule cell‐to‐cell trafficking, were significantly up‐regulated in 8‐week‐old Arabidopsis leaves compared with 4‐week‐old Arabidopsis leaves during TSWV infection. To date, this mature plant susceptibility to pathogen infections has rarely been investigated. Thus, the findings presented here should advance our knowledge on the developmentally regulated mature host susceptibility to plant virus infection.

Early detection of tomato spotted wilt virus infection in tobacco using the hyperspectral imaging technique and machine learning algorithms

The hyperspectral imaging technique was used for the non-destructive detection of tomato spotted wilt virus (TSWV) infection in tobacco at an early stage. Spectra ranging from 400 to 1000 nm with 128 bands from inoculated and healthy tobacco plants were analyzed by using three wavelength selection methods (successive projections algorithm (SPA), boosted regression tree (BRT), and genetic algorithm (GA)), and four machine learning (ML) techniques (boosted regression tree (BRT), support vector machine (SVM), random forest (RF), and classification and regression tress (CART)). The results indicated that the models built by the BRT algorithm using the wavelengths selected by SPA as the input variables obtained the best outcome for the 10-fold cross-validation with the mean overall accuracy of 85.2% and area under receiver operating curve (AUC) of 0.932. The band selection results and variable contribution analysis in BRT modeling jointly showed that the near-infrared (NIR) spectral region is informative and important for the differentiation of infected and healthy tobacco leaves. Different stages of post-inoculation were split according to the molecular identification and visual observation. The classification results at different stages indicated that the hyperspectral imaging data combined with ML methods and wavelength selection algorithms can be used for the early detection of TSWV in tobacco, both at the presymptomatic stage and during the period before the systematic infection can be detected by the molecular identification approach.

Transcriptome changes associated with Tomato spotted wilt virus infection in various life stages of its thrips vector, Frankliniella fusca (Hinds).

Persistent propagative viruses maintain intricate interactions with their arthropod vectors. In this study, we investigated the transcriptome-level responses associated with a persistent propagative phytovirus infection in various life stages of its vector using an Illumina HiSeq sequencing platform. The pathosystem components included a Tospovirus, Tomato spotted wilt virus (TSWV), its insect vector, Frankliniella fusca (Hinds), and a plant host, Arachis hypogaea (L.). We assembled (de novo) reads from three developmental stage groups of virus-exposed and non-virus-exposed F. fusca into one transcriptome consisting of 72 366 contigs and identified 1161 differentially expressed (DE) contigs. The number of DE contigs was greatest in adults (female) (562) when compared with larvae (first and second instars) (395) and pupae (pre- and pupae) (204). Upregulated contigs in virus-exposed thrips had blastx annotations associated with intracellular transport and virus replication. Upregulated contigs were also assigned blastx annotations associated with immune responses, including apoptosis and phagocytosis. In virus-exposed larvae, Blast2GO analysis identified functional groups, such as multicellular development with downregulated contigs, while reproduction, embryo development and growth were identified with upregulated contigs in virus-exposed adults. This study provides insights into differences in transcriptome-level responses modulated by TSWV in various life stages of an important vector, F. fusca.

From immunity to susceptibility: virus resistance induced in tomato by a silenced transgene is lost as TGS overcomes PTGS

Tomato line 30.4 was obtained engineering the nucleocapsid (N) gene of tomato spotted wilt virus into plant genome, and immunity to tomato spotted wilt virus infection of its self-pollinated homozygous progeny was observed. Despite the presence of a high amount of transgenic transcripts, transgenic proteins have not been detected, suggesting a mechanism of resistance mediated by RNA. In the present study, we identify post-transcriptional gene silencing as the main mechanism of resistance, which is able to spread systemically through grafting, and show that the line 30.4 resistant plants produce both 24 and 21-22nt N-gene specific siRNA classes. The transgenic locus in chromosome 4 shows complex multiple insertions of four T-DNA copies in various orientations, all with 3' end deletions in the terminator and part of the N gene. However, for three of them, polyadenylated transcripts are produced, due to flanking tomato genome sequences acting as alternative terminators. Interestingly, starting at the fifth generation after the transformation event, some individual plants show a tomato spotted wilt virus-susceptible phenotype. The change is associated with the disappearance of transgene-specific transcripts and siRNAs, and with hyper-methylation of the transgene, which proceeds gradually through the generations. Once it reaches a critical threshold, the shift from post-transcriptional gene silencing to transcriptional silencing of the transgene eliminates the previously well established virus resistance.

Tomato Spotted Wilt Virus Infection Reduces the Fitness of a Nonvector Herbivore on Pepper

Plant pathogens and insect herbivores often share hosts under natural conditions. Hence, pathogen-induced changes in a host plant can affect the herbivore and vice versa. Even though plant viruses are ubiquitous in the field, little is known about plant-mediated interactions between viruses and nonvector herbivores. Here we tested whether the performance of the sweet potato whitefly, Bemisia tabaci (Gennadius) biotype Q, was altered when raised on pepper infected with Tomato spotted wilt virus (TSWV). TSWV infection reduced B. tabaci fecundity and longevity and increased B. tabaci developmental time but did not affect the insect's survival or female body lengths. Our results demonstrate that TSWV infection can decrease the fitness of B. tabaci biotype Q on pepper plants.

Response of antioxidative enzymes to long-term Tomato spotted wilt virus infection and virus elimination by meristem-tip culture in two Impatiens species

Ornamental plants Impatiens hawkerii and Impatiens walleriana are susceptible to Tomato spotted wilt virus infection that can be eradicated by meristem-tip propagation. Comparison of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) profiles of healthy, infected and virus-eliminated plants showed that long-term infection did not cause major changes in I. hawkerii, but caused induction of several APX and GPX isoforms, increase in Mn-SOD and APX and a decrease in CAT activity in I. walleriana. This is the first report on the effects of virus elimination by meristem-tip propagation on the antioxidative metabolism in plants.

Utilizing immunoassays to determine systemic tomato spotted wilt virus infection for elucidating field resistance in peanut

Tomato spotted wilt disease was compared in three peanut cultivars, SunOleic 95R, Southern Runner (S. Runner) and DP-1, at two planting dates (April and May) and row spacings (7.6 and 15.2 cm) in a 2-year study in Florida. Tomato spotted wilt virus (TSWV) was detected directly using an enzyme-linked immuno-sorbent assay (ELISA) in both leaf and root crown tissues throughout the growing season to determine the timing and percentage of infected plants. Under low disease pressure, in harvested samples, more April-planted peanuts were infected with TSWV than May-planted. TSWV infection was not affected by plant spacings regardless of disease pressure. Under low disease pressure, from 60 days after planting (DAP), the most susceptible cultivar, SunOleic 95R, had a significantly higher percentage of plants with TSWV compared to the more resistant cultivars. Under high disease pressure, TSWV infection was detected earlier (30 DAP), and there was a clear separation of cultivars, with SunOleic 95R showing the highest infection (75%) followed by S. Runner (55%) and then DP-1 (20%). A higher incidence of TSWV in root crowns compared to leaves was observed for all cultivars. A delayed accumulation of TSWV in a cultivar was a reliable indicator of resistance. The field resistance manifested by peanut may be due to factors that decrease TSWV systemic spread resulting in slower TSWV accumulation in root crowns.

First Report of Tomato spotted wilt virus Infection of Tomatillo in Georgia.

During the 2000 spring season, tomatillo (Physalis ixocarpa) plants showing chlorotic streaks on leaves were observed in an experimental plot of the University of Georgia's Coastal Plain Experiment Station in Tift County, GA. Leaf samples from 192 plants were collected. These included plants that had chlorotic streaks and those without obvious symptoms. Samples were tested by ELISA using a commercially available Tomato spotted wilt virus (TSWV) detection kit (Agdia Inc., Elkhart, IN). TSWV was found in 10 samples that had chlorotic streaks on leaves, and the remaining plants with no obvious symptoms were negative for TSWV. Infected plants were found in both cultivars, Verde Puebla and Toma Verde. The presence of the virus had no apparent effect on plant size or fruit appearance. TSWV infection of the ELISA-positive samples was further verified by immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) (1). The primer pair (5'-ATGTCTAAGGTTAAGCTC-3' and 5' TTAAGCAAGTTCTGTGAG-3') represented the first and last 18 bases of the coding region of the nucleocapsid gene of TSWV, respectively, and produced approximately 800-bp PCR product (1). IC-RT-PCR gave a single DNA band of expected size and no amplification was found in the uninfected control. This is the first report of TSWV on tomatillo in Georgia. Reference: (1) R. K. Jain et al. Plant Dis. 82:900, 1998.

Effect of plant age at time of infection by tomato spotted wilt tospovirus on the yield of field-grown tomato

Naturally infected tomato plants that expressed tomato spotted wilt virus symptoms at 24, 38, 45, 60, 67, and 74 days after transplanting were monitored for production in an experimental crop grown in the open from May to September in northeastern Spain. Plants were tagged, tested for tomato spotted wilt virus infection by enzyme-linked immunosorbent assay, and data on symptom expression and yield were individually recorded. Plants that developed symptoms at 24, 38, or 45 days after transplanting yielded significantly less and produced fewer and smaller tomatoes than those that developed symptoms at 60, 67, and 74 days after transplanting. These later infected plants showed similar patterns of production with maximum yields between 27 July and 17 August, when most fruit was harvested. Production components such as fruit number per plant, yield of mature fruit per plant, or fruit weight increased the older the plants were when first symptoms were exhibited. However, marketable fruit production was drastically decreased by tomato spotted wilt virus infection, due to abnormal ripening of mature fruit in infected plants. Little and no significantly different amounts of marketable fruit were produced, irrespectively of plant age at time of symptom expression. Implications for spotted wilt management in tomato are discussed.

A protoplast system for studying tomato spotted wilt virus infection.

A plant protoplast system for studying tomato spotted wilt tospovirus (TSWV) infection was established and tested. Using polyethylene glycol-mediated inoculation with highly infectious TSWV particles, generally 50% or more of Nicotiana rustica protoplasts were infected. In these cells viral RNA and viral protein synthesis became detectable at 16 h post-inoculation (p.i.) and continued at least until 90 h p.i. Both the structural viral proteins [nucleoprotein (N) and the envelope glycoproteins G1 and G2] and the nonstructural viral proteins NSs and NSm accumulated to amounts sufficient for detection and immunocytological analysis. Local lesion tests on petunia leaves and electron microscopical analysis confirmed the production of mature, infectious virus particles, underlining the conclusion that a full infection cycle was completed in this system. Upon inoculation of Vigna unguiculata (cowpea) protoplasts with TSWV particles, comparable proportions of infected cells and amounts of NSs, NSm and N protein were obtained, but much lower amounts of viral glycoproteins were detected than in N. rustica protoplasts, and progeny virus particles were less abundant. With the N. rustica-based protoplast system, a powerful synchronized single-cell infection system has now become available for more precise in vivo studies of the processes occurring during tospovirus infection.

Engineered resistance to tomato spotted wilt virus in transgenic peanut expressing the viral nucleocapsid gene

The nucleocapsid gene of tomato spotted wilt virus Hawaiian L isolate in a sense orientation, and the GUS and NPTII marker genes, were introduced into peanut (Arachis hypogaea cv. New Mexico Valencia A) using Agrobacterium-mediated transformation. Modifications to a previously defined transformation protocol reduced the time required for production of transformed peanut plants. Transgenes were stably integrated into the peanut genome and transmitted to progeny. RNA expression and production of nucleocapsid protein in transgenic peanut were observed. Progeny of transgenic peanut plants expressing the nucleocapsid gene showed a 10- to 15-day delay in symptom development after mechanical inoculations with the donor isolate of tomato spotted wilt virus. All transgenic plants were protected from systemic tomato spotted wilt virus infection. Inoculated non-transformed control plants and plants transformed with a gene cassette not containing the nucleocapsid gene became systemically infected and displayed typical tomato spotted wilt virus symptoms. These results demonstrate that protection against tomato spotted wilt virus can be achieved in transgenic peanut plants by expression of the sense RNA of the tomato spotted wilt virus nucleocapsid gene

First Report of Tomato Spotted Wilt Virus Infection ofFicusSpecies in Spain

First Report of Tomato Spotted Wilt Virus Infection of<i>Ficus</i>Species in Spain