title: Transcriptomic analysis of lignocellulosic biomass degradation by the anaerobic fungal isolate Orpinomyces sp. strain C1A.
abstract: BACKGROUND
  Anaerobic fungi reside in the rumen and alimentary tract of herbivores where they play an important role in the digestion of ingested plant biomass. The anaerobic fungal isolate Orpinomyces sp. strain C1A is an efficient biomass degrader, capable of simultaneous saccharification and fermentation of the cellulosic and hemicellulosic fractions in multiple types of lignocellulosic biomass. To understand the mechanistic and regulatory basis of biomass deconstruction in anaerobic fungi, we analyzed the transcriptomic profiles of C1A when grown on four different types of lignocellulosic biomass (alfalfa, energy cane, corn stover, and sorghum) versus a soluble sugar monomer (glucose).
  
  
  RESULTS
  A total of 468.2 million reads (70.2 Gb) were generated and assembled into 27,506 distinct transcripts. CAZyme transcripts identified included 385, 246, and 44 transcripts belonging to 44, 13, and 8 different glycoside hydrolases (GH), carbohydrate esterases, and polysaccharide lyases families, respectively. Examination of CAZyme transcriptional patterns indicates that strain C1A constitutively transcribes a high baseline level of CAZyme transcripts on glucose. Although growth on lignocellulosic biomass substrates was associated with a significant increase in transcriptional levels in few GH families, including the highly transcribed GH1 β-glucosidase, GH6 cellobiohydrolase, and GH9 endoglucanase, the transcriptional levels of the majority of CAZyme families and transcripts were not significantly altered in glucose-grown versus lignocellulosic biomass-grown cultures. Further, strain C1A co-transcribes multiple functionally redundant enzymes for cellulose and hemicellulose saccharification that are mechanistically and structurally distinct. Analysis of fungal dockerin domain-containing transcripts strongly suggests that anaerobic fungal cellulosomes represent distinct catalytic units capable of independently attacking and converting intact plant fibers to sugar monomers.
  
  
  CONCLUSIONS
  Collectively, these results demonstrate that strain C1A achieves fast, effective biomass degradation by the simultaneous employment of a wide array of constitutively-transcribed cellulosome-bound and free enzymes with considerable functional overlap. We argue that the utilization of this indiscriminate strategy could be justified by the evolutionary history of anaerobic fungi, as well as their functional role within their natural habitat in the herbivorous gut.
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK USA.
authors.forename: M B
authors.initials: MB
authors.lastname: Couger
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK USA.
authors.forename: Noha H
authors.initials: NH
authors.lastname: Youssef
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK USA ; Department of Biology and Health Sciences, McNeese State University, Lake Charles, LA USA.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Struchtemeyer
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK USA.
authors.forename: Audra S
authors.initials: AS
authors.lastname: Liggenstoffer
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK USA.
authors.forename: Mostafa S
authors.identifier: 0000-0002-1067-1647
authors.initials: MS
authors.lastname: Elshahed
keywords: Anaerobic gut fungi; Cellulosome; Lignocellulosic biomass; Transcriptomics