Volume 14 Part 1 Article 8
Title: Ballistics for the delivery of transforming DNA to mushrooms
Authors: A.J. Moore, M.R. Challen, T.J. Elliott and R.J. Warner
Ballistic (biolistic®, particle bombardment) delivery of DNA has enabled the transformation of several intractable plant species and the application of this technology to microbial systems has been steadily increasing. The ballistic transformation of yeasts and some lower fungi indicate that the technology has potential for use with mushrooms. As a variety of tissue types can be bombarded the technology should prove useful for mushrooms that are difficult to transform by protoplast mediated methods. A novel ballistics device (gene gun) has been developed that is cheap to make and easy to use. This gun, which is powered by compressed air, has been evaluated using a yeast and two ink-cap mushrooms Coprinus bilanatus and Coprinus cinereus. Auxotrophic host cells were bombarded with tungsten microcarriers coated with plasmid DNAs containing the appropriate wild type gene. Transformants restored to prototrophy were recovered from experiments with all three species. Changing several gun related parameters including discharge pressure, target distance, and size of tungsten microcarrier did not enhance the numbers of transformants recovered. The gene gun was used to bombard lamellae and mycelial tissue from the mushroom Agaricus bisporus with a variety of plasmid vectors. Putative phleomycin resistant transformants were recovered.Please login to download the PDF for this proceeding.