PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 16499607

A highly efficient polyethylene glycol-mediated transformation method for mushrooms.

Gang Li·Ruixue Li·Qiuyun Liu·Qiang Wang·Min Chen·Baojian Li
View original source ↗Export metadata ↓

Abstract

A highly efficient transformation system mediated by polyethylene glycol was developed for the cultivated mushroom Pleurotus ostreatus. Eighty to 180 integrative and stable-resistant colonies appeared per mug of DNA per 10(7) viable protoplasts in a transformation experiment with the hygromycin B phosphotransferase gene (hph), which is about 40-1800 times higher than that previously reported in P. ostreatus. One hundred to 150 transformants emitting green fluorescence were observed per mug of DNA per 10(7) viable protoplasts in a transformation with the green fluorescent protein gene, but green fluorescence disappeared 30 h after transformation, suggesting that the green fluorescent protein gene was only transiently expressed in P. ostreatus. Plasmid pAN7-1 was also transferred into two important cultivated mushrooms, Ganoderma lucidum and Lentinus edodes, and 120-150 and 85-100 transformants per mug of DNA per 10(7) viable protoplasts were obtained, respectively, which is seven to 38 times and 24-28 times greater than previously reported. These data indicate that this new polyethylene glycol-mediated transformation procedure is highly efficient for mushrooms, and could be a useful tool in mushroom improvement by gene engineering.

Explore related subjects

Drug Resistance, FungalGenes, ReporterGreen Fluorescent ProteinsPhosphotransferases (Alcohol Group Acceptor)PlasmidsPleurotusPolyethylene GlycolsProtoplastsReishiShiitake MushroomsTransformation, GeneticJournal ArticleResearch Support, Non-U.S. Gov't

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Gang Li, Ruixue Li, Qiuyun Liu, Qiang Wang, Min Chen, Baojian Li. 2006. A highly efficient polyethylene glycol-mediated transformation method for mushrooms.. https://doi.org/10.1111/j.1574-6968.2006.00110.x

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2006

Identification of F-box proteins that are involved in resistance to methylmercury in Saccharomyces cerevisiae.

We searched for F-box proteins that might be related to the mechanism that protects Saccharomyces cerevisiae against the toxic effects of methylmercury. We found that overexpression of Hrt3 and of Ylr224w rendered yeast cells resistant to methylmercury. Yeast cells that overexpressed Hrt3 and Ylr224w were barely resistant to methylmercury in the presence of a proteasome inhibitor. Our results suggest the existence of some protein(s) that enhances the toxicity of methylmercury in yeast cells and, also, that overexpression of Hrt3 or Ylr224w can confer resistance to methylmercury by enhancing the polyubiquitination of this protein(s) and its degradation in proteasomes.

Drug Resistance, Fungal↗
PubMed2006

Evaluating the performance of chemical control in the presence of resistant pathogens.

Resistance to chemical control is a major impediment to combating many socially and economically important diseases. Theoretical and experimental studies have shown that reducing the intensity of treatment can slow, or even prevent, the invasion of resistance, yet reducing treatment levels also results in a net increase in disease severity. Clearly there is a need to identify control strategies that balance the conflicting aims of resistance management and disease suppression. Using a mathematical model for the dynamics of fungicide resistance in crop pathogens, we present a broadly applicable measure of the performance of chemical control in the presence of resistant pathogen strains. We illustrate how to optimise fungicide performance with respect to the intensity of treatment as a function of the duration of treatment and the fitness of the resistant strain. We find that in the short term, fungicide performance is optimised at high levels of treatment despite rapid selection for resistance, while the long-term optimum performance is achieved when treatment renders the fungicide-sensitive and fungicide-resistant pathogens equally fit. We further present evidence that under prescribed conditions, the ratio of dose size and frequency, and the fungicide mode of action, can have a significant effect on fungicide performance.

Drug Resistance, Fungal↗