Cassava common mosaic disease (CCMD) has been reported in all regions where cassava is grown in the Americas and the causal agent, Cassava common mosaic virus (CsCMV), has been identified as a mechanically transmitted potexvirus (Alphaflexiviridae). In Argentina, cassava is grown mainly in the northeast (NEA) region that shares borders with Brazil and Paraguay. Increasing incidences of CCMD were observed during the years 2014 to 2016 associated with severe leaf mosaic symptoms and yield reductions where the occurrence of CsCMV was confirmed by RT‐PCR and sequencing. In this work, the virus has been successfully purified and a double‐antibody sandwich (DAS‐) ELISA test has been developed from an Argentinean isolate of CsCMV to extend the diagnostics of the disease. A collection of 726 samples was screened and CsCMV was detected with 100% prevalence in the NEA region. Additional co‐infecting viruses were detected in some plants (64.4%); in these, CCMD symptoms correlated with CsCMV only, although more severe symptoms could be observed in mixed infected plants. Sequence analysis of the conserved RdRp domain showed a wider diversity of CsCMV isolates. Interestingly, a separate phylogenetic cluster was formed by isolates from the NEA region that only shared 77.1% to 80.3% nucleotide identity with the other clusters. These results indicate the presence of mixed strains occurring in the NEA region and suggest the presence of geographically distinct strains of CsCMV in South America.
The effectiveness of Pichia ohmeri and Saccharomyces cerevisiae in the biodegradation of patulin was evaluated in vitro. Patulin is a toxin produced by Penicillium expansum, the predominant fungal contaminant in post-harvest apple. The biodegradation experiment was carried out in culture medium (Yeast Medium broth, YM) and commercial apple juice. These substrates were artificially contaminated with patulin previously produced by P. expansum strain 2 in malt extract broth and purified over a silica gel column. The YM broth was inoculated with P. ohmeri 158 with proved anti-P. expansum activity, whereas the apple juice was inoculated with dried Saccharomyces cerevisiae cells. The residual patulin in contaminated substrates was determined by reversed-phase HPLC. P. ohmeri 158 in YM broth degraded over 83% of the initial 223 µg (8.92 µg/ml) of patulin after incubation at 25 °C for two days under static conditions; after five days of incubation, this percentage was greater than 99%, and patulin levels fell below the limit of detection after 15 days. In the apple juices inoculated with 0.25 g/l of commercial dried S. cerevisiae cells (corresponding 1.8 x 107 cells/ml), 96% of patulin was degraded (initial contamination of 4.5 µg/ml of patulin) after 143 hours of incubation at 25 °C under static conditions. However, 90% degradation occurred when the juice was contaminated with 7.0 µg/ml under the same conditions, indicating that the biodegradation rate is concentrationdependent. The effective biodegradation of patulin using P. ohmeri 158 and S. cerevisiae demonstrates a promising application for innocuous yeast isolated from natural microbiota in the biological control, which can prevent both fruit spoilage and P. expansum mycotoxin contamination.
Complete nucleotide (nt) and deduced amino acid sequences of two onion yellow dwarf virus (OYDV) isolates showing mild and severe symptoms in onion but being unable to infect garlic were determined. The genomes consisted of 10,459 and 10,461 nt (without the 3' poly(A) tail) and were 92.2 % identical. Comparison of their whole genomes, polyproteins and P1, HC-Pro, P3, CI, VPg and NIa-Pro regions with those of garlic isolates previously identified as OYDV gave percentage values below that proposed as the molecular threshold for potyvirus species demarcation. This and the striking differences in host range between onion and garlic isolates suggest that they represent different virus species.
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