Dölfors, Fredrik
- Department of Plant Biology, Swedish University of Agricultural Sciences
Research article2022Peer reviewedOpen access
Dolfors, Fredrik; Holmquist, Louise; Tzelepis, Georgios; Dixelius, Christina
Rhizoctonia solani is a soil-borne fungus, which rarely produces any spores in culture. Hence, all inoculation procedures are based on mycelia, often as a coat on cereal kernels, placed in close vicinity to the plant to be infected. In this protocol, an inoculation method is described where the fungus is first allowed to infest a perlite-maize flour substrate for 10 days, followed by thorough soil mixing to generate uniform fungal distribution. Pre-grown seedlings are then replanted in the infested soil. Plant materials can be harvested, five (sugar beet) and ten days (Arabidopsis) post infection, followed by a rapid cleaning step ahead of any nucleic acid preparation. Commercial DNA or RNA extraction kits can be used or, if higher DNA yield is required, a CTAB extraction method. Our purpose was to develop a reliable and reproducible protocol to determine the infection levels in planta upon infection with R. solani. This protocol is less laborious compared to previous ones, improves the consistency of plant infection, reproducibility between experiments, and suits both a root crop and Arabidopsis.
Arabidopsis thaliana; Beta vulgaris; DNA; Rhizoctonia solani; RNA
Bio-protocol
2022, Volume: 12, number: 2, article number: e4300Publisher: BIO-PROTOCOL
Botany
DOI: https://doi.org/10.21769/BioProtoc.4300
https://res.slu.se/id/publ/123318