Rhizoctonia solani Infection Assay of Young Sugar Beet and Arabidopsis plantlets

Fredrik Dölfors; Louise Holmquist; Georgios Tzelepis; Christina Dixelius

doi:10.21769/BioProtoc.4300

Rhizoctonia solani Infection Assay of Young Sugar Beet and Arabidopsis plantlets

Bio Protoc. 2022 Jan 20;12(2):e4300. doi: 10.21769/BioProtoc.4300.

Authors

Fredrik Dölfors¹, Louise Holmquist^{1

2}, Georgios Tzelepis¹, Christina Dixelius¹

Affiliations

¹ Swedish University of Agricultural Sciences, Department of Plant Biology, Uppsala BioCenter, Linnean Center for Plant Biology, P.O. Box 7080, S-75007 Uppsala, Sweden.
² DLF Beet Seed, Säbyholmsvägen 24, 26191 Landskrona, Sweden.

Abstract

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. Graphic abstract: Plant infectionInoculation of R. solaniPreparation of inoculumPCR analysis Overview of the R. solani infection procedure.

Keywords: Arabidopsis thaliana; Beta vulgaris; DNA; RNA; Rhizoctonia solani.