High-throughput sequencing (HTS), more specifically RNA sequencing of plant tissues, has become an indispensable tool for plant virologists to detect and identify plant viruses. During the data analysis step, plant virologists typically compare the obtained sequences to reference virus databases. In this way, they are neglecting sequences without homologies to viruses, which usually represent the majority of sequencing reads.
View Article and Find Full Text PDFHigh-throughput sequencing (HTS) technologies have become indispensable tools assisting plant virus diagnostics and research thanks to their ability to detect any plant virus in a sample without prior knowledge. As HTS technologies are heavily relying on bioinformatics analysis of the huge amount of generated sequences, it is of utmost importance that researchers can rely on efficient and reliable bioinformatic tools and can understand the principles, advantages, and disadvantages of the tools used. Here, we present a critical overview of the steps involved in HTS as employed for plant virus detection and virome characterization.
View Article and Find Full Text PDFHigh throughput sequencing of total RNA isolated from symptomatic leaves of a sweet cherry tree ( cv. 0900 Ziraat) from Turkey identified a new member of the genus designated cherry virus Turkey (CVTR). The presence of the virus was confirmed by electron microscopy and overlapping RT-PCR for sequencing its whole-genome.
View Article and Find Full Text PDFFive isolates of a new member of the family , tentatively named blackcurrant leafroll-associated virus 1 (BcLRaV-1), were identified in the currant. The 17-kb-long genome codes for 10 putative proteins. The replication-associated polyprotein has several functional domains, including papain-like proteases, methyltransferase, Zemlya, helicase, and RNA-dependent RNA polymerase.
View Article and Find Full Text PDFThe increased incidence of the crown gall disease caused by Agrobacterium tumefaciens has long been associated with activities of root-knot nematodes (Meloidogyne spp.). Pot experiments on tomato were designed to assess plant vitality, nematode reproduction, and crown gall incidence in combined infection with Agrobacterium and Meloidogyne spp.
View Article and Find Full Text PDFGenomic DNA of blueberry red ringspot virus (genus Soymovirus, family Caulimoviridae) from highbush blueberry plants growing for years in the Czech Republic and Slovenia was sequenced. The circular dsDNA genomes consist of 8,303 and 8,299 nt, respectively, and contain eight open reading frames (ORFs) longer than 100 amino acids. The European isolates are 90% to 99% identical in aa sequences of distinct proteins.
View Article and Find Full Text PDFAssays based on real-time PCR (TaqMan) that can detect a number of viroids in the genus Pospiviroid have been developed and evaluated. The assays are designed for detecting viroids from tomato leaf material but detection from other solanaceous hosts of these viroids has been confirmed. These methods have been validated by nine laboratories and comprise a reliable set of assays for the detection of CEVd, TASVd, CLVd and a generic assay which will detect the six viroids of concern to European tomato growers: PSTVd, TCDVd, CEVd, CLVd, TASVd and CSVd.
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