Location: Bioenergy ResearchTitle: Transformation and electrophoretic karyotyping of Coniochaeta ligniaria NRRL30616 Author
|Skory, Christopher - Chris|
Submitted to: Current Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/24/2010
Publication Date: 1/11/2011
Publication URL: http://hdl.handle.net/10113/49654
Citation: Nichols, N.N., Szynkarek, M., Skory, C.D., Gorsich, S.W., Lopez, M.J., Guisado, G.M., Nichols, W.A. 2011. Transformation and electrophoretic karyotyping of Coniochaeta ligniaria NRRL30616. Current Genetics. 57(3):169-175. Interpretive Summary: These experiments laid the groundwork for making a useful microbe even more useful. This fungus, called Coniochaeta ligniaria, has the native ability to withstand the harsh milieu that results when fibrous biomass is prepared for use in a process called pretreatment. The strain has already proven useful as a way to clean up pretreated biomass by metabolizing toxic compounds, resulting in a cleaner sugar stream that can be converted to fuel ethanol in a second step. Availability of genetic tools for this fungus could allow its use to make other chemicals from biomass. In this study, appropriate selectable genetic markers were identified, and basic mutagenesis and transformation methods were developed. Tools for genetic engineering may allow use of this microbe as a platform for conversion of biomass sugars to valuable products, which would be of interest to producers of fuels and chemicals.
Technical Abstract: This study undertook initial characterization of the genetic system of Coniochaeta ligniaria NRRL30616, an Actinomycete with utility for conversion of biomass sugars to fuels and chemicals. Transformation of C. ligniaria using hygromycin as a dominant selectable marker was achieved using protoplasts generated by including cells in 1% (v/v) beta-mercaptoethanol, followed by cell-wall-digesting enzymes. Twelve chromosomes with an estimated total size of 28.2 Mb were detected in C. ligniaria. The gas chromatography content of chromosomal DNA and of coding regions from cDNA sequences was 49.2% and 51.9%, respectively.