Background: Botrytis cinerea is a haploid necrotrophic ascomycete which is responsible for 'grey mold' disease in more than 200 plant species. Broad molecular research has been conducted on this pathogen in recent years, resulting in the sequencing of two strains, which has generated a wealth of information toward developing additional tools for molecular transcriptome, proteome and secretome investigations. Nonetheless, transformation protocols have remained a significant bottleneck for this pathogen, hindering functional analysis research in many labs. Results: In this study, we tested three different transformation methods for B. cinerea: electroporation, air-pressure-mediated and sclerotium-mediated transformation. We demonstrate successful transformation with three different DNA constructs using both air-pressure- and sclerotium-mediated transformation. Conclusions: These transformation methods, which are fast, simple and reproducible, can expedite functional gene analysis of B. cinerea.
Bibliographical noteFunding Information:
This paper is contribution no. 616-11 from the Agricultural Research Organization, the Volcani Center, Bet Dagan, Israel. The study was funded by research grant no. VWZN2556 from the Niedersachsen-Israel Fund to M. L. and by research grant nos. IS-3947-06 to A.L. and IS-4210-09 to M. L. from BARD, the United States-Israel Binational Agricultural Research and Development Fund. We thank Prof. Oded Yarden for hosting part of the work in his laboratory which was funded by United States-Israel Binational Agricultural Research and Development Fund No. US-4414-11C. We want to thank Dr. Herve Huet and Dr. Aviv Dombrovsky for their approval and help in operating the Bim-Lab instrument.
- Botrytis cinerea
- Sclerotinia sclerotiorum