Isolation of RNA from plants rich in secondary metabolites using commercial kits often results in contaminated preparations which are not suitable for downstream applications. Although many specific protocols appropriate for plants with a high content of phenolics, anthocyanins and polysaccharides have been developed, these are often expensive, time consuming and not applicable to different types of tissues. This study presents a simple and efficient modification of RNA extraction from different types of tissues using two commercial reagent kits. By simple improvement, we routinely obtained high-quality RNA of the following plants: the blackcurrant bush, black chokeberry bush, pear tree, apricot tree, apple tree, hardy kiwi, tangerine tree, highbush blueberry and cranberry plant.
Prune dwarf virus (PDV) isolates have been investigated for genetic diversity. Full-length nucleotide and amino acid sequences of viral coat protein from 23 isolates collected from different stone fruit trees (sour and sweet cherry trees, wild cherry tree, plum tree, almond tree, peach tree) and different countries (Poland, Italy, Germany, USA, Israel) were analysed and compared to 57 others available in GenBank. Comparison of all sequenced virus isolates revealed diversity of 86-100 % at nucleotide level and 79-100 % at amino acid level. The ratio of non-synonymous to synonymous polymorphic sites indicated that purifying selection dominated in case of PDV. However, six codons showed to be under strong positive selection, including the codon located inside the structure involved in RNAbinding activity.
Prune dwarf virus (PDV), a worldwide pathogen of stone fruit trees, has many isolates with different biological, serological and molecular properties. Monoclonal antibodies (MAbs) to the Prunus mahaleb isolate of PDV were used to investigate the serological variability of virus isolates, by TAS-ELISA (triple antibody sandwich enzyme-linked immunosorbent assay). The twenty-two PDV isolates from Germany (1), Italy (7), Poland (13) and the USA (1) were characterised against eight single MAbs. The virus showed high serological variability. Analysis of the MAbs reaction allowed for the identification of 13 serogroups.
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