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Olivier T.,Walloon Agricultural Research Center | Sveikauskas V.,Phytosanitary Research Laboratory | Demonty E.,Walloon Agricultural Research Center | De Jonghe K.,Belgium Institute for Agricultural and Fisheries Research | And 11 more authors.
European Journal of Plant Pathology | Year: 2016

Four pospiviroid detection methods consisting of a pair of RT-PCR (ANSES 1-2), a pair of real time RT-PCR (Botermans 1-2) and two single RT-PCR methods (Luigi and Olivier) were evaluated for their relative accuracy, diagnostic sensitivity, diagnostic specificity, analytical sensitivity and reproducibility through a comparison in eight laboratories. All detection methods were tested on 13 tomato leaf and nine tomato seed samples as well as on 100-fold dilutions of the corresponding RNA extracts. Two different RNA extraction kits and three combinations of RNA extraction and RT-PCR kits were assessed. According to the statistical analysis of the results, ANSES 1-2 method provided the best performance criteria whatever the matrix consisted of, while the relative accuracies of Botermans 1-2 and Olivier methods were not significantly different from ANSES 1-2 for seed and leaf samples respectively. The results also showed that the relative accuracy of the Luigi and Olivier methods significantly increased when primer concentrations were raised to 1 μM, the latter method giving the best relative accuracy of the test in these conditions. No statistical differences were observed between accuracies obtained for the two extraction kits or the three combinations of extraction and RT-PCR kits used. ANSES 1-2 method is proposed as international standard for the generic detection of pospiviroids on tomato leaves and seeds. © 2015, Koninklijke Nederlandse Planteziektenkundige Vereniging.

Faggioli F.,Italian Agricultural Research Council | Luigi M.,Italian Agricultural Research Council | Sveikauskas V.,Phytosanitary Research Laboratory | Olivier T.,Walloon Agricultural Research Center | And 5 more authors.
European Journal of Plant Pathology | Year: 2015

Greenhouse experiments were conducted to investigate the importance of tomato seeds as a route of pospiviroid dissemination. More than 7000 seeds were collected from tomato plants mechanically inoculated with Tomato apical stunt viroid, Citrus exocortis viroid, Columnea latent viroid and Potato spindle tuber viroid. Fruits and part of the seeds were found to be 100 % infected by the inoculated pospiviroid by means of RT-PCR testing. Much of the rest of the seeds were sown and produced about 4700 seedlings. None of the seedlings were found positive for the respective pospiviroid. This shows that pospiviroid seed transmission in tomato is very rare. © 2015, Koninklijke Nederlandse Planteziektenkundige Vereniging.

Olivier T.,Walloon Agricultural Research Center | Sveikauskas V.,Phytosanitary Research Laboratory | Grausgruber-Groger S.,Austrian Agency for Health and Food Safety AGES | Virscek Marn M.,Agricultural Institute of Slovenia | And 4 more authors.
Crop Protection | Year: 2015

Viroids are RNA molecules which are harmful to numerous cultivated plant species and considered, for some of them, as quarantine organisms. Because these pathogens are readily transmitted by contact, effective disinfection measures to clean the cultivating tools, machineries or facilities have to be found and approved. To this end, five disinfectants: Virkon®, Hyprelva™ SL, Jet 5 ®, MENNO® clean and Virocid™, were evaluated for their effectiveness against Potato spindle tuber viroid (PSTVd). The virucids were assessed on dried droplets and in aqueous solutions of PSTVd infected tomato sap. For both experimental setups, the infectiousness of the sap and disinfectant mixtures was checked by inoculation on tomato seedlings followed by RT-PCRs. The experiments involving aqueous solutions, which allowed a discrimination of tested virucids, were carried out in 5 laboratories. In these experiments, MENNO® clean, the only approved product for PSTVd in several European countries, was not found to be effective at the minimum manufacturer's recommended contact time or at an intermediate contact time for the minimum manufacturer's recommended concentration. Conversely, the four other commercial disinfectants: Virocid™ Hyprelva™ SL; Virkon® and Jet 5®, were not found to be significantly different from the bleach control. However, in the experiments performed on dried sap droplets, the effectiveness of the five commercial disinfectants decreased substantially. © 2014 Elsevier Ltd.

Ostrauskas H.,Phytosanitary Research Laboratory | Ostrauskas H.,Nature Research Center | Ivinskis P.,Nature Research Center | Buda V.,Nature Research Center | Buda V.,Vilnius University
Acta Zoologica Lituanica | Year: 2010

A survey for the North American spruce budworm (Choristoneura fumiferana (Clemens, 1865)) was carried out using sticky traps baited with a synthetic sex pheromone blend from 2002 to 2004. The traps were deployed at 90 localities throughout Lithuania: in parks, botanical gardens, forest trees nurseries and forest lots, where non-native coniferous trees were planted. No C. fumiferana were found, but a total of 88 other moth species were trapped and identified. Acleris ferrugana (Denis & Schiffermüller, 1775) was the most frequently found species. Other species of trapped moths revealed new information on the Lithuanian fauna. Merrifieldia leucodactyla (Denis & Schiffermüller, 1775) moths, a new species for the Lithuanian fauna, were caught in six localities. Seven new localities were recorded for the occurrence of the following very rare species: Diplodoma laichartingella Goeze, 1783, Depressaria (Depressaria) sordidatella Tengström, 1848, Elachista (Elachista) consortella Stainton, 1851, Mompha (Mompha) sturnipennella (Treitschke, 1833), Bryotropha senectella (Zeller, 1839), Dichelia histrionana (Frölich, 1828). 63 new localities were recorded for the occurrence of 16 moth species which are rare in Lithuania.

Ostrauskas H.,Phytosanitary Research Laboratory | Ivinskis P.,Nature Research Center
Acta Zoologica Lituanica | Year: 2010

Two samples of tomato with damaged fruit imported from Spain were collected from Vilnius vegetable warehouses on 20 May 2009. Only one live Tuta absoluta larva was detected, and it was reared to pupa. Later (during July-September 2009) sticky traps with synthetic sex pheromone blend were used to search for these moths in greenhouses where tomatoes were grown, and in the vicinity of warehouses in which tomato fruit were temporary stored. The tomato pinworm T. absoluta was detected at three sites. The species status in Lithuania: distributed in greenhouses, first records. Five other random moth species were trapped.

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