Abstract

RNA silencing (RS) is a conserved eukaryotic mechanism acting as anti-viral immune systemin plants. Small interfering RNAs (siRNAs) are RS effectors accumulating in virus-infected tissuesas immune system components, providing target specificity for post-transcriptional degradation ofinvading RNAs. Virus-derived siRNAs (vsiRNAs, 21-24 nt) are abundant in infected plants, withcertain regions ("hot spots") more represented than others.Two isolates of Potato virus Y (PVY), PVYC-to and PVY-SON41, yield very differentdisease phenotypes on tomato (Solanum lycopersicum), the former inducing leaf distortions, thelatter mild symptoms. We applied in silico and molecular approaches to identify PVY vsiRNAssuppressing host mRNAs by sequence complementarity and RS-based suppression, inducingdysfunctional processes upon infection. Aim was to explore differential expression of host-targetingvsiRNA, as responsible of isolates differences.A computational pipeline retrieved 21nt vsiRNAs from PVY isolates, complementary topredicted mRNAs (Solgenomics, ITAG2.3). Two PVYC-to genome regions showed specificsecondary structures absent in PVY-SON41, accounting for vsiRNAs accumulation. Two tomatotranscript lists were obtained, perfectly or imperfectly (one or two mismatches) complementary tovsiRNA computed for either PVY isolates, putative targets of vsiRNA-driven RS suppression.Targets common to both isolates were discarded, leaving a gene list representing only PVYC-totargets. Quantitative RT-PCR of transcription factors (e.g. NAC, MYB, TCP, HD-ZIP, MADS-boxfamilies) active in leaf development and morphogenesis showed differential expression uponPVYC-to and PVY-SON41 infections. Lower isolate-specific mRNA accumulation suggested RSdrivenpost-transcriptional regulation, confirming a link between symptoms and vsiRNAs-mediatedRS. Results from vsiRNAs deep sequencing from infected leaves are also discussed.

Autore Pugliese

• Finetti Sialer Mariella Matilde , Catalano Domenico , Cillo Fabrizio

Tutti gli autori

• Finetti-Sialer M.M.; Catalano D.; Cillo F.

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Anno di pubblicazione

2013

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Settori ERC

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