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Detekce a identifikace virů pomocí sekvenování nové generace (NGS)
PODRÁBSKÁ, Kateřina
Next generation sequencing is a modern method applied in plant virology for sensitive detection of previously characterized and novel pathogens without any preceding knowledge of them. In this study three novel and two already described viruses were detected by de novo assembly of Illumina single-end reads ( Hi-Seq 2500 system) from total poly(A) enriched RNA of diseased red clover (Trifolium pratense) and indicator plant (Nicotiana occidentalis 37B). The complete genomic sequence of novel Red clover carlavirus A (RCCA) was determined from Illumina reads, 5´, 3´ RACE, cloning, RT-PCR and Sanger sequencing. The presence of RCCV was also confirmed in mechanically inoculated tobacco plant.
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Optimized biological method for the detection of sensitivity of red clover varieties to white clover mosaic virus (WClMV) and red clover mottle virus (RCMV) infection in the climatic conditions of the Czech Republic
Jakešová, H. ; Fránová, Jana
Red clover is a very important leguminous plant used in crop rotations on arable land, in temporary as well as permanent clover and grass stands. The persistence of clovers is negatively influenced by pests and diseases that attack the root and shoot parts of the plant. In the course of studies into the presence of viral diseases (2007–2011), the most frequently detected viruses were White clover mosaic virus, (WClMV, genus Potexvirus, family Alphaflexiviridae) and Red clover mottle virus (RCMV, genus Comovirus, family Secoviridae). The two viruses were observed in populations of clover throughout the Czech Republic. RCMV significantly reduced the yield and stand persistence of red clover. Given that no direct protection has yet been developed against the viruses, plant breeding directed to enhancing resistance to both WClMV and RCMV offers good prospects. Both viruses are mechanically transmissible.
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Identification of filamentous virus which infected red clover (\kur{Trifolium pratense})
BEČKOVÁ, Martina
Samples of red clover plants characteristic with dwarf growth were investigated with transmission electron microscopy. Most frequent virus particles observed there were filamentous ones of 300 to 800 nm long. Nucleic acid was isolated, transcribed and amplified using PCR with potexvirus- and potyvirus-specific primers 353G1, 353G2 (Petrzik, K., unpublished data), P9502, P0502 (Revers et al., 1999) and Poty2/P4 (Gibbs and Mackenzie, 1997). Successful amplification with potexvirus-specific primers, sequencing and comparison with GenBank sequences revealed White clover mosaic virus (WClMV) there. Coat protein gene of this virus was obtained with amplification and compared with complete sequence of WClMV from GenBank. O-strain of WClMV was identified.
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