Location: Crops Pathology and Genetics ResearchTitle: A genetic screen for bioluminescence genes in the fungus Armillaria mellea, through the use of Agrobacterium tumefaciens-mediated random insertional mutagenesis
|POOLE, VIRGINIA - Middle Tennessee State University|
|ROBERTSON, BRIAN - Middle Tennessee State University|
|BERGEMANN, SARAH - Middle Tennessee State University|
Submitted to: Mycological Society of America
Publication Type: Abstract Only
Publication Acceptance Date: 6/30/2016
Publication Date: 8/1/2016
Citation: Poole, V., Robertson, B.J., Baumgartner, K., Bergemann, S. 2016. A genetic screen for bioluminescence genes in the fungus Armillaria mellea, through the use of Agrobacterium tumefaciens-mediated random insertional mutagenesis. Mycological Society of America Abstracts. #218. ipmb.berkeley.edu/sites/default/files/MSA%202016%20abstracts%20and%20index.pdf.
Technical Abstract: Bioluminescence is reported from 71 saprobic species of fungi from four, distant lineages in the order Agaricales. Analyses of the fungal luminescent chemistry shows that all four lineages share a functionally conserved substrate and luciferase, indicating that the bioluminescent pathway is likely conserved throughout Basidiomycota; however, the genes encoding for bioluminescence are unknown. The goal of the current study is to uncover the genes responsible for bioluminescence in Armillaria mellea. This species is an ideal candidate for bioluminescence research because it luminesces at a high, consistent magnitude, fruits in vitro, and mycelia and basidiospores can be easily transformed with foreign DNA. Transformations are achieved using Agrobacterium-mediated insertional mutagenesis to transfer an engineered Ti plasmid (T-DNA) with a hygromycin resistance selective marker. Isolates are cultured on 1% MEA+hygromycin and screened for luminescence to identify A. mellea transformants that lack the luminescent phenotype. To date, approximately one hundred fifty A. mellea transformants have been screened for luminescence and a single haploid isolate lacking the luminescent phenotype was identified. The goal of the study is to characterize many of the genes involved in bioluminescence, and to understand the origin and evolution of this physiological mechanism in the Agaricales.