Improved vectors for Agrobacterium mediated genetic manipulation of Hypholoma spp. and other homobasidiomycetes

Suhad A.A. Al-Salihi, Thomas A. Scott, Andy M. Bailey, Gary D. Foster

Research output: Contribution to journalArticle (Academic Journal)peer-review

4 Citations (Scopus)
422 Downloads (Pure)

Abstract

Abstract The basidiomycete fungi Hypholoma fasciculare and H. sublateritium are both prolific producers of sesquiterpenes and triterpenes, some of which have relevant pharmaceutical properties. Although H. sublateritium has been transformed in the past, the low reported efficiencies highlighted the need for establishing an effective simple transformation system for these valuable species. We have optimized Agrobacterium tumefaciens-mediated transformation through testing various parameters in these two Hypholoma species, showing that a mixture of homogenized mycelia and Agrobacterium (strain LBA4404) co-cultivated for 84 h at 25 °C is optimal for efficient transformation in these basidiomycetes. This study also reveals the requirements for transgene expression, with the first report of GFP expression in these Hypholoma, the need for an intron for such transgene expression, and further demonstrates the functionality of the expression vector by its use in Clitopilus passeckerianus. This development of transformation system and expression constructs, can facilitate further genetic investigation such as gene functionality in these fungi.
Original languageEnglish
Pages (from-to)4-9
Number of pages6
JournalJournal of Microbiological Methods
Volume142
Early online date24 Aug 2017
DOIs
Publication statusPublished - 1 Nov 2017

Keywords

  • Agaricales
  • Agrobacterium
  • Hypholoma promoter
  • GFP

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