Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs

Konstantina Katsarou; Nikoleta Kryovrysanaki; Kriton Kalantidis

doi:10.1007/978-1-0716-1464-8_24

Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs

Methods Mol Biol. 2022:2316:287-312. doi: 10.1007/978-1-0716-1464-8_24.

Authors

Konstantina Katsarou^{1

2}, Nikoleta Kryovrysanaki^{1

2}, Kriton Kalantidis^{3

4}

Affiliations

¹ Biology Department, University of Crete, Heraklion, Greece.
² IMBB/FoRTH, Heraklion, Greece.
³ Biology Department, University of Crete, Heraklion, Greece. kriton@imbb.forth.gr.
⁴ IMBB/FoRTH, Heraklion, Greece. kriton@imbb.forth.gr.

PMID: 34845703
DOI: 10.1007/978-1-0716-1464-8_24

Abstract

Viroids are considered the most minimalistic group of pathogens. Despite their presumed inability to encode for proteins, viroids induce several diseases in plants of primary economic importance. The production of viroid derived siRNAs (vd-siRNAs) of 21-24 nt, accompanies viroid infections in plants and results from the activation of the RNA silencing mechanism and specifically the function of Dicer endonucleases. A comprehensive set of experiments for the study and thorough analysis of viroid-infected plants has been developed. Here we present a detailed experimental plan including optimized protocols for plant infection by agroinfiltration, RNA extraction, and northern blot hybridization for the detection of both viroid genomic RNA and vd-siRNAs.

Keywords: Agroinfiltration; Northern blot; RNA extraction; Viroid; vd-siRNAs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Northern
Plant Diseases / genetics
Plants
RNA Interference
RNA, Double-Stranded
RNA, Small Interfering / genetics
RNA, Viral / genetics
Viroids* / genetics

Substances

RNA, Double-Stranded
RNA, Small Interfering
RNA, Viral