Preservation stress resistance of melanin deficient conidia from Paecilomyces variotii and Penicillium roqueforti mutants generated via CRISPR/Cas9 genome editing

Fungal Biology and Biotechnology

Tom van den Brule¹  Jan Dijksterhuis¹  Jos Houbraken¹  Arthur F. J. Ram²  Pepijn P. P. Teunisse²  Sjoerd J. Seekles²  Han A. B. Wösten³  Maarten Punt³ 

[1] TIFN, Agro Business Park 82, 6708 PW, Wageningen, The Netherlands;Applied & Industrial Mycology, Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT, Utrecht, the Netherlands;TIFN, Agro Business Park 82, 6708 PW, Wageningen, The Netherlands;Department Molecular Microbiology and Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333BE, Leiden, The Netherlands;TIFN, Agro Business Park 82, 6708 PW, Wageningen, The Netherlands;Microbiology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands;

关键词: CRISPR/Cas9;    Cell factory;    Melanin;    Food spoilage;    Food spoiling fungi;    Polyketide synthase;    Conidia;    Aspergillus niger;    Penicillium roqueforti;    Paecilomyces variotii;   

DOI  :  10.1186/s40694-021-00111-w

来源: Springer

【 摘 要 】

BackgroundThe filamentous fungi Paecilomyces variotii and Penicillium roqueforti are prevalent food spoilers and are of interest as potential future cell factories. A functional CRISPR/Cas9 genome editing system would be beneficial for biotechnological advances as well as future (genetic) research in P. variotii and P. roqueforti.ResultsHere we describe the successful implementation of an efficient AMA1-based CRISPR/Cas9 genome editing system developed for Aspergillus niger in P. variotii and P. roqueforti in order to create melanin deficient strains. Additionally, kusA− mutant strains with a disrupted non-homologous end-joining repair mechanism were created to further optimize and facilitate efficient genome editing in these species. The effect of melanin on the resistance of conidia against the food preservation stressors heat and UV-C radiation was assessed by comparing wild-type and melanin deficient mutant conidia.ConclusionsOur findings show the successful use of CRISPR/Cas9 genome editing and its high efficiency in P. variotii and P. roqueforti in both wild-type strains as well as kusA− mutant background strains. Additionally, we observed that melanin deficient conidia of three food spoiling fungi were not altered in their heat resistance. However, melanin deficient conidia had increased sensitivity towards UV-C radiation.

【 授权许可】

CC BY