1. |
2. Detection of plant viruses using nanopore highthroughput sequencing : validation reportAnja Pecman, Veronika Bukvič, Ana Vučurović, Irena Bajde, Jakob Brodarič, Nataša Mehle, Denis Kutnjak, 2023, final research report Keywords: diagnostics, method validation, nanopores, high-throughput sequencing Published in DiRROS: 02.09.2024; Views: 86; Downloads: 1393
Full text (6,04 MB) This document has many files! More... |
3. |
4. TESTING OF CHRYSANTHEMUM STEM NECROSIS VIRUS AND OTHER AMERICAN CLADE 1 TOSPOVIRUSES BY RT-PCR : validation reportNataša Mehle, Irena Bajde, Jakob Brodarič, Ana Vučurović, 2024, final research report Abstract: Detection of Chrysanthemum stem necrosis virus (CSNV) and other tospoviruses of American
clade 1. Keywords: CSNV, orthotospovirses, diagnostics, method validation, EURL-Virology Published in DiRROS: 02.09.2024; Views: 98; Downloads: 33
Full text (808,74 KB) This document has many files! More... |
5. Development and validation of a one-step reverse transcription real-time PCR assay for simultaneous detection and identification of tomato mottle mosaic virus and tomato brown rugose fruit virusAntonio Tiberini, Ariana Manglli, Anna Taglienti, Ana Vučurović, Jakob Brodarič, Luca Ferretti, Marta Luigi, Andrea Gentili, Nataša Mehle, 2022, original scientific article Abstract: Tobamovirus species represent a threat to solanaceous crops worldwide, due to their extreme stability and because they are seed borne. In particular, recent outbreaks of tomato brown rugose fruit virus in tomato and pepper crops led to the establishment of prompt control measures, and the need for reliable diagnosis was urged. Another member of the genus, tomato mottle mosaic virus, has recently gained attention due to reports in different continents and its common features with tomato brown rugose fruit virus. In this study, a new real-time RT-PCR detection system was developed for tomato brown rugose fruit virus and tomato mottle mosaic virus on tomato leaves and seeds using TaqMan chemistry. This test was designed to detect tomato mottle mosaic virus by amplifying the movement protein gene in a duplex assay with the tomato brown rugose fruit virus target on the CP-3’NTR region, which was previously validated as a single assay. The performance of this test was evaluated, displaying analytical sensitivity 10−5–10−6-fold dilution for seeds and leaves, respectively, and good analytical specificity, repeatability, and reproducibility. Using the newly developed and validated test, tomato brown rugose fruit virus detection was 100% concordant with previously performed analyses on 106 official samples collected in 2021 from different continents. Keywords: real-time PCR, tomato mottle mosaic virus, tomato brown rugose fruit virus, leaves detection, seeds detections, performance criteria Published in DiRROS: 16.07.2024; Views: 196; Downloads: 181
Full text (1,99 MB) This document has many files! More... |
6. Tomato brown rugose fruit virus in aqueous environments : survival and significance of water-mediated transmissionNataša Mehle, Katarina Bačnik, Irena Bajde, Jakob Brodarič, Adrian Fox, Ion Gutiérrez-Aguirre, Miha Kitek, Denis Kutnjak, Yue Lin Loh, Olivera Maksimović, Maja Ravnikar, Elise Vogel, Christine Vos, Ana Vučurović, 2023, original scientific article Abstract: Tomato brown rugose fruit virus (ToBRFV) has recently emerged as a major disease of tomatoes and peppers. ToBRFV is a seed- and contact-transmitted virus. In Slovenia, ToBRFV RNA was detected in samples of wastewater, river, and water used to irrigate plants. Even though the source of detected RNA could not be clearly established, this raised the question of the significance of the detection of ToBRFV in water samples and experimental studies were performed to address this question. The data presented here confirm that the release of virus particles from the roots of infected plants is a source of infectious ToBRFV particles in water and that the virus can remain infective up to four weeks in water stored at room temperature, while its RNA can be detected for much longer. These data also indicate that irrigation with ToBRFV-contaminated water can lead to plant infection. In addition, it has been shown that ToBRFV circulated in drain water in commercial tomato greenhouses from other European countries and that an outbreak of ToBRFV can be detected by regular monitoring of drain water. A simple method for concentrating ToBRFV from water samples and a comparison of the sensitivity of different methods, including the determination of the highest ToBRFV dilution still capable of infecting test plants, were also investigated. The results of our studies fill the knowledge gaps in the epidemiology and diagnosis of ToBRFV, by studying the role of water-mediated transmission, and provide a reliable risk assessment to identify critical points for monitoring and control. Keywords: tomato brown rugose fruit virus, tomato, hydroponics, water-linked epidemiology, survival Published in DiRROS: 12.07.2024; Views: 193; Downloads: 196
Full text (3,00 MB) This document has many files! More... |
7. Epidemiology of Flavescence dorée and hazelnut decline in Slovenia : geographical distribution and genetic diversity of the associated 16SrV phytoplasmasZala Kogej Zwitter, Gabrijel Seljak, Tjaša Jakomin, Jakob Brodarič, Ana Vučurović, Sandra Pedemay, Pascal Salar, Sylvie Malembic-Maher, Xavier Foissac, Nataša Mehle, 2023, original scientific article Abstract: Flavescence dorée (FD) phytoplasma from 16SrV-C and -D subgroups cause severe damage to grapevines throughout Europe. This phytoplasma is transmitted from grapevine to grapevine by the sap-sucking leafhopper Scaphoideus titanus. European black alder and clematis serve as perennial plant reservoirs for 16SrV-C phytoplasma strains, and their host range has recently been extended to hazelnuts. In Slovenia, hazelnut orchards are declining due to 16SrV phytoplasma infections, where large populations of the non-autochthonous leafhopper Orientus ishidae have been observed. To better characterise the phytoplasma-induced decline of hazelnut and possible transmission fluxes between these orchards and grapevine, genetic diversity of 16SrV phytoplasmas in grapevine, hazelnut and leafhoppers was monitored from 2017 to 2022. The nucleotide sequence analysis was based on the map gene. The most prevalent map genotype in grapevine in all wine-growing regions of Slovenia was M54, which accounted for 84 % of the 176 grapevines tested. Besides M54, other epidemic genotypes with lower frequency were M38 (6 %), M51 (3 %), M50 (2 %) and M122 (1 %). M38, M50 and M122 were also detected in infected cultivated hazelnuts and in specimens of O. ishidae leafhopper caught in declining hazelnut orchards. It suggests that this polyphagous vector could be responsible for phytoplasma infection in hazelnut orchards and possibly for some phytoplasma exchanges between hazelnuts and grapevine. We hereby describe new genotypes: M158 in grapevine as well as four never reported genotypes M159 to M162 in hazelnut. Of these four genotypes in hazelnut, one (M160) was also detected in O. ishidae. Analysis of additional genes of the new genotypes allowed us to assign them to the VmpA-III cluster, which corresponds to the 16SrV-C strains previously shown to be compatible with S. titanus transmission. Keywords: phytoplasmas, haselnuts, grapevine, Flavescence dorée, epidemiology, plant disease Published in DiRROS: 26.07.2023; Views: 710; Downloads: 539
Full text (3,82 MB) This document has many files! More... |