Abstract
Armillaria mellea is a significant pathogen that causes
Armillaria root disease on numerous hosts in forests, gardens and
agricultural environments worldwide. Using a yeast-adapted pCAMBIA0380 Agrobacterium vector, we have constructed a series of vectors for transformation of A. mellea,
assembled using yeast-based recombination methods. These have been
designed to allow easy exchange of promoters and inclusion of introns.
The vectors were first tested by transformation into basidiomycete Clitopilus passeckerianus to ascertain vector functionality then used to transform A. mellea. We show that heterologous promoters from the basidiomycetes Agaricus bisporus and Phanerochaete chrysosporium
that were used successfully to control the hygromycin resistance
cassette were not able to support expression of mRFP or GFP in A. mellea. The endogenous A. mellea gpd
promoter delivered efficient expression, and we show that inclusion of
an intron was also required for transgene expression. GFP and mRFP
expression was stable in mycelia and fluorescence was visible in
transgenic fruiting bodies and GFP was detectable in planta. Use of these vectors has been successful in giving expression of the fluorescent proteins GFP and mRFP in A. mellea, providing an additional molecular tool for this pathogen.
Original language | English |
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Article number | 29226 |
Number of pages | 24 |
Journal | Scientific Reports |
Volume | 6 |
DOIs | |
Publication status | Published - 7 Jul 2016 |
Keywords
- Honey fungus
- Agrobacterium
- Basidiomycete
- Fluorescent protein
- Transformation
- Clitopilus passeckerianus