The development of an improved PCR-based marker system for Sw-5, an important TSWV resistance gene of tomato

Sw-5 is an important disease resistance gene of tomato, providing broad resistance to Tomato spotted wilt virus (TSWV). A cleaved amplified polymorphic sequence (CAPS) marker, closely linked to the gene, has been reported. Although the Sw-5 locus has been characterised, a gene-specific marker has not been developed. This paper presents a PCR-based marker-system that consists of the co-amplification of a dominant marker representing the Sw-5 gene sequence, and the modified CAPS marker as a positive control and indicator of genotype.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Tospoviruses - An Australian perspective

The detection, distribution, molecular and biological properties, vector relations and control of tospoviruses present in Australia, including Tomato spotted wilt virus (TSWV), Capsicum chlorosis virus (CaCV) and Iris yellow spot virus (IYSV), are reviewed. TSWV occurs throughout Australia where it has caused serious sporadic epidemics since it was first described in the 1920s. The frequency and distribution of outbreaks has increased in the 1990s, with the arrival and dispersal of the western flower thrips (Frankliniella occidentalis) being one factor favouring this situation. The crops most frequently and severely affected are capsicum, lettuce, tomato, potato and several species of ornamentals. Minimal differences were found between the nucleocapsid (N) gene amino acid sequences of Australian isolates and these were most closely related to a clade of northern European isolates. CaCV was first detected in Australia in 1999 and is most closely related to Watermelon silver mottle virus, a serogroup IV tospovirus. The natural hosts include capsicum, tomato, peanut and Hoya spp. The virus also occurs in Thailand and Taiwan. IYSV was first found in Australia in 2003, infecting onion and leek, with the distribution in three States suggesting that the virus has been present for some time.

Field isolates of Tomato spotted wilt virus overcoming resistance in capsicum in Australia

In 2002 at Virginia, South Australia, capsicum cultivars having the Tsw resistance gene against Tomato spotted wilt virus (TSWV) developed symptoms typical of TSWV infection and several glasshouse-grown crops were almost 100% infected. Samples reacted with TSWV antibodies in ELISA. Virus isolates from infected plants induced severe systemic symptoms, rather than a hypersensitive reaction, when inoculated onto capsicum cultivars and Capsicum chinense genotypes ( PI 152225 and PI 159236) that carry the Tsw resistance gene. Isolates virulent towards the Tsw gene had molecular and biological properties very similar to standard TSWV isolates, including a hypersensitive reaction in Sw-5 (TSWV-resistant) tomato genotypes. Tsw-virulent isolates were found during surveys at Virginia in 2002 and 2004 in both TSWV-resistant and susceptible cultivars of capsicum and tomato.

Management of virus diseases in vegetables

• To undertake an audit of management systems used for tomato spotted wilt virus (TSWV) in greenhouse and field production with the aim of improving disease management determining knowledge gaps in virus-vector relationships. • To investigate the basis for the development of resistance breaking strains of TSWV in capsicums and apply this to virus management in capsicums. • To further develop effective virus management systems in vegetable cucurbit crops. Aspects to be investigated include value of barrier crops, non-insecticide products and cultivar tolerance to virus. • To further develop and assess the adoption and impact of integrated viral disease management systems in field grown and protected cropping systems as part of the vegetable industry development plan.

Application methods and dosages of thiamethoxam in thrips control on tomato plants

O tripes Frankliniella schultzei Trybom (Thysanoptera: Thripidae) é um dos mais importantes vetores de tomato spotted wilt tospovirus (TSWV) na cultura do tomate no Brasil. Dois métodos de aplicação do inseticida thiamethoxam foram comparados no controle de tripes em tomate. O experimento foi conduzido em delineamento de blocos ao acaso com sete tratamentos e quatro repetições. Utilizou-se um pulverizador costal manual com lança simples provida de ponta de pulverização com jato cônico vazio JD 14-2 ou modificada, com aplicador tipo esguicho, em substituição à ponta de pulverização. Dosagens de 150 e 200 g i.a. ha-1 foram usadas em única aplicação com esguicho e 50 g i.a. ha-1 em pulverizações semanais, iniciadas aos 48 dias após a semeadura. A eficiência do inseticida em teste foi comparada com diafenthiuron (400 g i.a. ha-1), profenofos + cypermethrin (320+32 g i.a. ha-1, respectivamente) e methamidophos (60 g i.a. 100 L-1 de água), aplicados em pulverização. Não houve diferença estatística entre os métodos de aplicação e doses com thiamethoxam no controle de F. schultzei aos 24 dias após a aplicação, sendo vantajosa a realização de uma única aplicação das maiores dosagens de thiamethoxam com esguicho em comparação às aplicações semanais da menor dosagem em pulverização. A eficiência de controle do tripes com o inseticida thiamethoxam variou de 93 a 95%, nos diferentes métodos e dosagens em teste. Diafenthiuron e profenofos + cypermethrin apresentaram eficiência menor (78 e 88%, respectivamente), porém foram superiores às obtidas com methamidophos (71%). Os produtos e doses utilizados não causaram fitotoxicidade às plantas de tomate.

Incidência e caracterização dos vírus infectando orquídeas no Estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modelización espacial de huevos de Thrips (Thysanoptera: Frankliniella occidentalis) en tomate de cáscara por medio de técnicas geoestadísticas

El tomate de cáscara (Physalis ixocarpa Brot.) es un cultivo alimenticio de gran importancia económica en México. Sin embargo, es afectado por diversas plagas y enfermedades tales como los Thrips (Thysanoptera: Frankliniella occidentalis) y el virus de la marchitez manchada del tomate (TSWV) que llegan a causar hasta un 80% de pérdidas. El objetivo del presente trabajo fue modelizar la distribución espacial de huevos de Thrips mediante técnicas geoestadísticas y obtener, en consecuencia, mapas de incidencia por medio del Kriging. Se georreferenciaron 121 puntos de muestreo en cada una de las parcelas comerciales de los municipios de Luvianos, Jocotitlán e Ixtlahuaca, a través del método de transectos en tres etapas fenológicas del cultivo. Se contabilizó el número de huevos de Thrips en cada punto de muestreo. Los resultados mostraron que las poblaciones de huevos deThrips presentan una distribución agregada, identificándose varios centros de conglomeración a través de los mapas obtenidos. Los semivariogramas obtenidos de la distribución espacial se ajustaron principalmente a los modelos gaussianos y esféricos. La distribución de huevos de Thrips se presentó en centros de agregación dentro de las parcelas estudiadas, lo cual permitirá establecer estrategias y medidas de control o mitigación en términos de sitios específicos de infestación de huevos de Thrips.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy (R) or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to I infected in 800 samples with pepper but never detecting more than I infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Predictive Models for Tomato Spotted Wilt Virus Spread Dynamics, Considering Frankliniella occidentalis Specific Life Processes as Influenced by the Virus

Several models have been studied on predictive epidemics of arthropod vectored plant viruses in an attempt to bring understanding to the complex but specific relationship between the three cornered pathosystem (virus, vector and host plant), as well as their interactions with the environment. A large body of studies mainly focuses on weather based models as management tool for monitoring pests and diseases, with very few incorporating the contribution of vector's life processes in the disease dynamics, which is an essential aspect when mitigating virus incidences in a crop stand. In this study, we hypothesized that the multiplication and spread of tomato spotted wilt virus (TSWV) in a crop stand is strongly related to its influences on Frankliniella occidentalis preferential behavior and life expectancy. Model dynamics of important aspects in disease development within TSWV-F. occidentalis-host plant interactions were developed, focusing on F. occidentalis' life processes as influenced by TSWV. The results show that the influence of TSWV on F. occidentalis preferential behaviour leads to an estimated increase in relative acquisition rate of the virus, and up to 33% increase in transmission rate to healthy plants. Also, increased life expectancy; which relates to improved fitness, is dependent on the virus induced preferential behaviour, consequently promoting multiplication and spread of the virus in a crop stand. The development of vector-based models could further help in elucidating the role of tri-trophic interactions in agricultural disease systems. Use of the model to examine the components of the disease process could also boost our understanding on how specific epidemiological characteristics interact to cause diseases in crops. With this level of understanding we can efficiently develop more precise control strategies for the virus and the vector.