PVX-potyvirus synergistic infections differentially alter microRNA accumulation in Nicotiana benthamiana.

In comparison to single infections, co-infection of Nicotiana benthamiana with Potato virus X (PVX) and Potato virus Y (PVY) or Plum pox virus (PPV), resulted in increased systemic symptoms (synergism in pathology). Previous studies have shown that virus infections affected the accumulation of various microRNAs (miRNAs) and miRNA target genes. Our studies revealed that double infection by PVX and PVY or PPV that produced the most severe symptoms in N. benthamiana altered accumulation of miR156, 171, 398, and 168, and/or their target transcripts to a greater extent or in a different direction than single infections that produced milder symptoms. These findings indicate a differential effect on miRNA metabolism of the combined infection by two unrelated plant viruses, which may account in part for the severe symptoms caused by PVX/potyvirus-associated synergisms.

Transcriptional changes and oxidative stress associated with the synergistic interaction between Potato virus X and Potato virus Y and their relationship with symptom expression.

Many virus diseases of economic importance to agriculture result from mixtures of different pathogens invading the host at a given time. This contrasts with the relatively scarce studies available on the molecular events associated with virus-host interactions in mixed infections. Compared with single infections, co-infection of Nicotiana benthamiana with Potato virus X (PVX) and Potato virus Y (PVY) resulted in increased systemic symptoms (synergism) that led to necrosis of the newly emerging leaves and death of the plant. A comparative transcriptional analysis was undertaken to identify quantitative and qualitative differences in gene expression during this synergistic infection and correlate these changes with the severe symptoms it caused. Global transcription profiles of doubly infected leaves were compared with those from singly infected leaves using gene ontology enrichment analysis and metabolic pathway annotator software. Functional gene categories altered by the double infection comprise suites of genes regulated coordinately, which are associated with chloroplast functions (downregulated), protein synthesis and degradation (upregulated), carbohydrate metabolism (upregulated), and response to biotic stimulus and stress (upregulated). The expressions of reactive oxygen species-generating enzymes as well as several mitogen-activated protein kinases were also significantly induced. Accordingly, synergistic infection induced a severe oxidative stress in N. benthamiana leaves, as judged by increases in lipid peroxidation and by the generation of superoxide radicals in chloroplasts, which correlated with the misregulation of antioxidative genes in microarray data. Interestingly, expression of genes encoding oxylipin biosynthesis was uniquely upregulated by the synergistic infection. Virus-induced gene silencing of alpha-dioxygenase1 delayed cell death during PVX-PVY infection.

Stereospecific synthesis of polyfunctionalized carbacephams induced by titanocene(III) chloride.

Enantiomerically pure N-substituted epoxyalkene-2-azetidinones reacted with titanocene monochloride to give stereospecifically polyfunctionalized bicyclic beta-lactams. Four isomeric epoxyaldehydes 2 reacted with TiCp2Cl to give exclusively the respective carbacephams 7 while under the same reaction conditions the epoxyesters 1, which are more hindered for an intramolecular addition, gave the cyclization products 6 (only two isomers) and/or the elimination products 5 (all isomers).