干旱区生态环境知识资源中心(Arid): An advanced genetic toolkit for exploring the biology of the rock-inhabiting black fungus Knufia petricola

Arid

DOI	10.1038/s41598-020-79120-5
	An advanced genetic toolkit for exploring the biology of the rock-inhabiting black fungus Knufia petricola
	Voigt, Oliver; Knabe, Nicole; Nitsche, Sarah; Erdmann, Eileen A.; Schumacher, Julia; Gorbushina, Anna A.
通讯作者	Schumacher, J (corresponding author), Bundesanstalt Mat Forsch & Prufung BAM, Dept Mat & Environm 4, D-12205 Berlin, Germany.
来源期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
出版年	2020
卷号	10 期号:1
英文摘要	Microcolonial black fungi are a group of ascomycetes that exhibit high stress tolerance, yeast-like growth and constitutive melanin formation. They dominate a range of hostile natural and man-made environments, from desert rocks and salterns to dishwashers, roofs and solar panels. Due to their slow growth and a lack of genetic tools, the underlying mechanisms of black fungi's phenotypic traits have remained largely unexplored. We chose to address this gap by genetically engineering the rock-inhabiting fungus Knufia petricola (Eurotiomycetes, Chaetothyriales), a species that exhibits all characteristics of black fungi. A cell biological approach was taken by generating K. petricola strains expressing green or red fluorescent protein variants. By applying: (1) traditional gene replacement; (2) gene editing and replacement via plasmid-based or ribonucleoprotein (RNP)-based CRISPR/Cas9, and (3) silencing by RNA interference (RNAi), we constructed mutants in the pathways leading to melanin, carotenoids, uracil and adenine. Stable single and double mutants were generated with homologous recombination (HR) rates up to 100%. Efficient, partially cloning-free strategies to mutate multiple genes with or without resistance cassettes were developed. This state-of-the-art genetic toolkit, together with the annotated genome sequence of strain A95, firmly established K. petricola as a model for exploring microcolonial black fungi.
类型	Article
语种	英语
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000603656600010
WOS关键词	MODEL MICROCOLONIAL FUNGUS ; FILAMENTOUS FUNGI ; CRISPR-CAS9 ; CRISPR/CAS9 ; EXTRACTION ; PCR ; TRANSFORMATION ; SARCINOMYCES ; EVOLUTION ; YEASTS
WOS类目	Multidisciplinary Sciences
WOS研究方向	Science & Technology - Other Topics
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/349185
作者单位	[Voigt, Oliver; Knabe, Nicole; Nitsche, Sarah; Erdmann, Eileen A.; Schumacher, Julia; Gorbushina, Anna A.] Bundesanstalt Mat Forsch & Prufung BAM, Dept Mat & Environm 4, D-12205 Berlin, Germany; [Gorbushina, Anna A.] Free Univ Berlin, Dept Earth Sci, D-12249 Berlin, Germany; [Erdmann, Eileen A.; Gorbushina, Anna A.] Free Univ Berlin, Dept Biol, Chem, Pharm, D-14195 Berlin, Germany
推荐引用方式 GB/T 7714	Voigt, Oliver,Knabe, Nicole,Nitsche, Sarah,et al. An advanced genetic toolkit for exploring the biology of the rock-inhabiting black fungus Knufia petricola[J],2020,10(1).
APA	Voigt, Oliver,Knabe, Nicole,Nitsche, Sarah,Erdmann, Eileen A.,Schumacher, Julia,&Gorbushina, Anna A..(2020).An advanced genetic toolkit for exploring the biology of the rock-inhabiting black fungus Knufia petricola.SCIENTIFIC REPORTS,10(1).
MLA	Voigt, Oliver,et al."An advanced genetic toolkit for exploring the biology of the rock-inhabiting black fungus Knufia petricola".SCIENTIFIC REPORTS 10.1(2020).