ABSTRACT
Double-stranded RNA and total RNA purified from sour cherry leaves (Prunus cerasus, cv. Amarelka Chvalkovicka) was analyzed by high-throughput sequencing. BLAST annotation identified contigs with homology to several already known cherry-infecting viruses (prune dwarf virus, prunus necrotic ringspot virus, prunus virus F, little cherry virus 1) as well as contigs with sequences more distantly related to those of members of the family Betaflexiviridae and in particular to prunus virus T of the genus Tepovirus. The full genome sequence of a putative virus (6,847 nucleotides [nt]; GenBank no. MT090966) was assembled and completed at the genome ends. The genome has a typical tepovirus organization, containing three overlapping open reading frames (ORFs), encoding a replication-associated protein, a movement protein and a capsid protein, respectively. Both its genome organization and its phylogenetic relationships show that the virus belongs to the genus Tepovirus, but considering the species demarcation criteria for the family Betaflexiviridae, it appears to represent a novel virus species, and we propose the name "cherry virus T" (ChVT) for this virus.
Subject(s)
Flexiviridae/genetics , Flexiviridae/isolation & purification , Genome, Viral , Plant Diseases/virology , Prunus avium/virology , Base Sequence , Flexiviridae/classification , High-Throughput Nucleotide Sequencing , Open Reading Frames , Phylogeny , Whole Genome SequencingABSTRACT
An investigation was conducted to improve the biological detection of pear blister canker viroid (PBCVd), which over a period of 2 to 3 years induces pear blister canker disease on the perry pear (Pyrus communis) cv. A20. PBCVd was not detected by dot blot hybridization or bioassay in any of the tested species of Amelanchier, Aronia, Cotoneaster, Crataegus, and Pyracantha. However, some species of Chaenomeles, Cydonia, and Sorbus, five out of 13 species of Malus, 15 Pyrus species, and 16 commercial pear cultivars were susceptible to PBCVd, although none developed symptoms. Only in perry pear seedlings did approximately 5% of the population react to infection with pure PBCVd strains by developing petiole, leaf, or bark necrosis 2 to 3 years (cv. A20) or 3 to 5 months (cv. Fieudière) after inoculation. The selected Fieud 37 and Fieud 110 clones are proposed as PBCVd indicators to replace A20. Results from bioassays on the indicators and from detection by a PBCVd-cRNA probe were essentially in agreement except for some Malus species.
ABSTRACT
Studies conducted over the last 10 years have revealed that the disease caused by the apple scar skin viroid (ASSVd) is extremely rare in Europe. ASSVd was detected by molecular hybridization and indexing in field plots on the apple indicators Starkrimson and Indo, which showed symptoms of dapple apple disease within 2 years, and rough scarred skin within 3 years, respectively. Results from both approaches were in agreement. In an attempt to improve the biological detection of ASSVd, the Japanese PK13 isolate was inoculated to 4 Prunus, 13 Malus, 17 Pyrus, and 17 other pomaceous species. All the species tested of the Malus, Pyrus, Sorbus, Chaenomeles, Cydonia, and Pyronia genera were susceptible to ASSVd based upon back indexing and hybridization, but none developed leaf or bark symptoms during a 2-year period. The viroid was not detected in the tested members of genera Amelanchier, Aronia, Cotoneaster, Crataegus, Prunus, and Pyracantha. Symptoms on fruit of 42 commercial apple cultivars experimentally inoculated with ASSVd fell into five groups ranging from inconspicuous spots to severely scarred skin and cracking. ASSVd was eliminated from most of the infected apple plants when they were subjected to a dormant stage followed by thermotherapy and shoot tip grafting. Analysis of more than 400 apple seedlings, originated from Starkrimson and Indo fruits with typical ASSVd symptoms, showed that there is little or no seed transmission of this viroid. However, ASSVd was transmitted at a low rate under field conditions to adjacent trees.