Enrichment of perforate septal pore caps from the basidiomycetous fungus Rhizoctonia solani by combined use of French press, isopycnic centrifugation, and Triton X-100

Kenneth G.A. van Driel, Arend F. van Peer, Han A.B. Wösten, Arie J. Verkleij, Teun Boekhout, Wally H. Müller*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

6 Citations (Scopus)

Abstract

Septal pore caps occur in many filamentous basidiomycetes located at both sides of the dolipore septum and are at their base connected to the endoplasmic reticulum. The septal pore cap ultrastructure has been described extensively by the use of electron microscopy, but its composition and function are not yet known. To enable biochemical and functional analyses in the future, we here describe an enrichment method for perforate septal pore caps from Rhizoctonia solani. Our method is based on the combined use of French press and isopycnic centrifugation, using a discontinuous sucrose gradient followed by a treatment with Triton X-100. Enrichment was monitored by the use of scanning electron microscopy and transmission electron microscopy. Using the same isolation method, smaller septal pore caps were isolated from two other basidiomycetes as well. Furthermore, we showed pore-occluding material co-purified with the septal pore caps. This observation supports the hypothesis that septal pore caps play a key role in the plugging process of the septal pores in filamentous basidiomycetes.

Original languageEnglish
Pages (from-to)298-304
Number of pages7
JournalJournal of Microbiological Methods
Volume71
Issue number3
DOIs
Publication statusPublished - 1 Dec 2007
Externally publishedYes

Keywords

  • Basidiomycete
  • Electron microscopy
  • Fungi
  • Isolation
  • Isopycnic centrifugation
  • Organelle
  • Rhizoctonia solani
  • Septal pore cap
  • Triton X-100

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