Host plant resistance against tomato spotted wilt virus in peanut (Arachis hypogaea) and its impact on susceptibility to the virus, virus population genetics, and vector feeding behavior and survival.

Tomato spotted wilt virus (TSWV) severely affects peanut production in the southeastern United States. Breeding efforts over the last three decades resulted in the release of numerous peanut genotypes with field resistance to TSWV. The degree of field resistance in these genotypes has steadily increased over time, with recently released genotypes exhibiting a higher degree of field resistance than older genotypes. However, most new genotypes have never been evaluated in the greenhouse or laboratory against TSWV or thrips, and the mechanism of resistance is unknown. In this study, TSWV-resistant and -susceptible genotypes were subjected to TSWV mechanical inoculation. The incidence of TSWV infection was 71.7 to 87.2%. Estimation of TSWV nucleocapsid (N) gene copies did not reveal significant differences between resistant and susceptible genotypes. Parsimony and principal component analyses of N gene nucleotide sequences revealed inconsistent differences between virus isolates collected from resistant and susceptible genotypes and between old (collected in 1998) and new (2010) isolates. Amino acid sequence analyses indicated consistent differences between old and new isolates. In addition, we found evidence for overabundance of nonsynonymous substitutions. However, there was no evidence for positive selection. Purifying selection, population expansion, and differentiation seem to have influenced the TSWV populations temporally rather than positive selection induced by host resistance. Choice and no-choice tests indicated that resistant and susceptible genotypes differentially affected thrips feeding and survival. Thrips feeding and survival were suppressed on some resistant genotypes compared with susceptible genotypes. These findings reveal how TSWV resistance in peanut could influence evolution, epidemiology, and management of TSWV.

Second generation peanut genotypes resistant to thrips-transmitted tomato spotted wilt virus exhibit tolerance rather than true resistance and differentially affect thrips fitness.

Spotted wilt disease caused by Tomato spotted wilt virus (TSWV) (family Bunyaviridae; genus Tospovirus) is a major constraint to peanut (Arachis hypogaea L.) production in the southeastern United States. Reducing yield losses to TSWV has heavily relied on planting genotypes that reduce the incidence of spotted wilt disease. However, mechanisms conferring resistance to TSWV have not been identified in these genotypes. Furthermore, no information is available on how these genotypes influence thrips fitness. In this study, we investigated the effects of newly released peanut genotypes (Georganic, GA-06G, Tifguard, and NC94022) with field resistance to TSWV and a susceptible genotype (Georgia Green) on tobacco thrips, Frankliniella fusca (Hinds), fitness, and TSWV incidence. Thrips-mediated transmission resulted in TSWV infection in both TSWV-resistant and susceptible genotypes and they exhibited typical TSWV symptoms. However, some resistant genotypes had reduced viral loads (fewer TSWV N-gene copies) than the susceptible genotype. F. fusca larvae acquired TSWV from resistant and susceptible genotypes indicating that resistant genotypes also can serve as inoculum sources. Unlike resistant genotypes in other crops that produce local lesions (hypersensitive reaction) upon TSWV infection, widespread symptom development was noticed in peanut genotypes. Results indicated that the observed field resistance in peanut genotypes could be because of tolerance. Further, fitness studies revealed some, but not substantial, differences in thrips adult emergence rates and developmental time between resistant and susceptible genotypes. Thrips head capsule length and width were not different when reared on different genotypes.

Transmission of Iris yellow spot virus by Frankliniella fusca and Thrips tabaci (Thysanoptera: Thripidae).

Thrips-transmitted Iris yellow spot virus (IYSV) (Family Bunyaviridae, Genus Tospovirus) affects onion production in the United States and worldwide. The presence of IYSV in Georgia was confirmed in 2003. Two important thrips species that transmit tospoviruses, the onion thrips (Thrips tabaci (Lindeman)) and the tobacco thrips (Frankliniella fusca (Hinds)) are known to infest onion in Georgia. However, T. tabaci is the only confirmed vector of IYSV. Experiments were conducted to test the vector status of F. fusca in comparison with T. tabaci. F. fusca and T. tabaci larvae and adults reared on IYSV-infected hosts were tested with antiserum specific to the nonstructural protein of IYSV through an antigen coated plate ELISA. The detection rates for F. fusca larvae and adults were 4.5 and 5.1%, respectively, and for T. tabaci larvae and adults they were 20.0 and 24.0%, respectively, indicating that both F. fusca and T. tabaci can transmit IYSV. Further, transmission efficiencies of F. fusca and T. tabaci were evaluated by using an indicator host, lisianthus (Eustoma russellianum (Salisbury)). Both F. fusca and T. tabaci transmitted IYSV at 18.3 and 76.6%, respectively. Results confirmed that F. fusca also can transmit IYSV but at a lower efficiency than T. tabaci. To attest if low vector competency of our laboratory-reared F. fusca population affected its IYSV transmission capability, a Tomato spotted wilt virus (Family Bunyaviridae, Genus Tospovirus) transmission experiment was conducted. F. fusca transmitted Tomato spotted wilt virus at a competent rate (90%) suggesting that the transmission efficiency of a competent thrips vector can widely vary between two closely related viruses.

Evaluation of Lisianthus as an Indicator Host for Iris yellow spot virus.

Iris yellow spot virus (IYSV) can severely affect onion production. IYSV is transmitted by the onion thrips, Thrips tabaci. However, information on IYSV-thrips-onion interactions is limited due to the difficulty associated with infecting onion plants experimentally. Lisianthus (Eustoma russellianum) was used as an indicator host to study mechanical transmission of IYSV, IYSV transmission by T. tabaci, IYSV distribution in the host plant, and the effect of temperature on IYSV symptom expression. Mechanical inoculation tests from IYSV-infected onion plants to noninfected lisianthus plants resulted in a mean transmission rate of 82.5 ± 6.9% (mean ± standard error), and from IYSV-infected lisianthus plants to noninfected lisianthus plants resulted in a mean transmission rate of 89.2 ± 7.1%. T. tabaci adults transmitted IYSV at a rate of 80.0 ± 8.3% from infected onion plants to noninfected lisianthus plants. To assess IYSV distribution in infected lisianthus plants, leaf sections, stems, and roots were tested by enzyme-linked immunosorbent assay (ELISA). All the plant parts tested positive for IYSV, but not on every plant assayed. Alternating night and day temperatures of 18 and 23°C, 25 and 30°C, and 30 and 37°C were evaluated for the effects on IYSV symptom expression. More severe symptoms developed on inoculated plants incubated at the 18 and 23°C or 25 and 30°C temperature regimes than at the 30 and 37°C regime, and symptoms were observed earliest on plants incubated at the 25 and 30°C temperature regime compared to the other temperature regimes.

Specific and Spillover Effects on Vectors Following Infection of Two RNA Viruses in Pepper Plants.

Mixed infection of plant viruses is ubiquitous in nature and can affect virus-plant-vector interactions differently than single virus infection. While several studies have examined virus-virus interactions involving mixed virus infection, relatively few have examined effects of mixed virus infection on vector preference and fitness, especially when multiple vectors are involved. This study explored how single and mixed viral infection of a non-persistently transmitted cucumber mosaic virus (CMV) and propagative and persistently-transmitted tomato spotted wilt orthotospovirus (TSWV) in pepper, Capsicum annum L., influenced the preference and fitness of their vectors, the green peach aphid, Myzus persicae (Sulzer), and the tobacco thrips, Frankliniella fusca (Hinds), respectively. In general, mixed infected plants exhibited severe symptoms compared with individually infected plants. An antagonistic interaction between the two viruses was observed when CMV titer was reduced following mixed infection with TSWV in comparison with the single infection. TSWV titer did not differ between single and mixed infection. Myzus persicae settling preference and median developmental were not significantly different between CMV and/or TSWV-infected and non-infected plants. Moreover, M. persicae fecundity did not differ between CMV-infected and non-infected pepper plants. However, M. persicae fecundity was substantially greater on TSWV-infected plants than non-infected plants. Myzus persicae fecundity on mixed-infected plants was significantly lower than on singly-infected and non-infected plants. Frankliniella fusca fecundity was higher on CMV and/or TSWV-infected pepper plants than non-infected pepper plants. Furthermore, F. fusca-induced feeding damage was higher on TSWV-infected than on CMV-infected, mixed-infected, or non-infected pepper plants. Overall, our results indicate that the effects of mixed virus infection on vectors were not different from those observed following single virus infection. Virus-induced host phenotype-modulated effects were realized on both specific and non-specific vectors, suggesting crosstalk involving all vectors and viruses in this pathosystem. The driving forces of these interactions need to be further examined. The effects of interactions between two viruses and two vectors towards epidemics of one or both viruses also need to be examined.

Effects of Thrips Density, Mode of Inoculation, and Plant Age on Tomato Spotted Wilt Virus Transmission in Peanut Plants.

Spotted wilt caused by tomato spotted wilt virus (TSWV; family Bunyaviridae; genus Tospovirus) is a serious disease of peanut (Arachis hypogaea L.) in the southeastern United States. Peanut genotypes with field resistance to TSWV are effective in suppressing spotted wilt. All commercially available genotypes with field resistance to TSWV were developed through conventional breeding. As a part of the breeding process, peanut genotypes are regularly screened under field situations. Despite numerous advantages associated with field screening, it is often limited by inconsistent vector (thrips) and TSWV pressure. A greenhouse transmission protocol would aid in thorough screening of selected genotypes and conserve time. In this study, various parameters associated with TSWV transmission, including tobacco thrips, Frankliniella fusca (Hinds) density, mode of inoculation, and plant age, were evaluated. Greater incidences of TSWV infection were obtained with thrips-mediated inoculation when compared with mechanical inoculation. TSWV inoculation with three, five, and 10 thrips resulted in greater incidences of TSWV infection in plants than inoculation with one thrips. However, incidences of TSWV infection did not vary between plants inoculated with three, five, and 10 viruliferous thrips. With both thrips-mediated and mechanical inoculation methods, incidences of TSWV infection in 1-wk-old plants were greater than in 4-wk-old plants. TSWV copy numbers, as determined by qPCR, also decreased with plant age. Results suggest that using at least three thrips per plant and 1- to 2-wk-old plants would maximize TSWV infection in inoculated plants.