Methodologies for probing the metatranscriptome of grassland soil
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Since 1905, the Department of Agricultural Engineering, now the Department of Agricultural and Biosystems Engineering (ABE), has been a leader in providing engineering solutions to agricultural problems in the United States and the world. The department’s original mission was to mechanize agriculture. That mission has evolved to encompass a global view of the entire food production system–the wise management of natural resources in the production, processing, storage, handling, and use of food fiber and other biological products.
History
In 1905 Agricultural Engineering was recognized as a subdivision of the Department of Agronomy, and in 1907 it was recognized as a unique department. It was renamed the Department of Agricultural and Biosystems Engineering in 1990. The department merged with the Department of Industrial Education and Technology in 2004.
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1905–present
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- Department of Agricultural Engineering (1907–1990)
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- College of Agriculture and Life Sciences (parent college)
- College of Engineering (parent college)
- Department of Industrial Education and Technology, (merged, 2004)
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Abstract
Metatranscriptomics provides an opportunity to identify active microbes and expressed genes in complex soil communities in response to particular conditions. Currently, there are a limited number of soil metatranscriptome studies to provide guidance for using this approach in this challenging matrix. Hence, we evaluated the technical challenges of applying soil metatranscriptomics to a highly diverse, low activity natural system. We used a non-targeted rRNA removal approach, duplex nuclease specific (DSN) normalization, to generate a metatranscriptomic library from field collected soil supporting a perennial grass, Miscanthus x giganteus (a biofuel crop), and evaluated its ability to provide insight into its active community members and their expressed protein-coding genes. We also evaluated various bioinformatics approaches for analyzing our soil metatranscriptome, including annotation of unassembled transcripts, de novo assembly, and aligning reads to known genomes. Further, we evaluated various databases for their ability to provide annotations for our metatranscriptome. Overall, our results emphasize that low activity, highly genetically diverse and relatively stable microbiomes, like soil, requires very deep sequencing to sample the transcriptome beyond the common core functions. We identified several key areas that metatranscriptomic analyses will benefit from including increased rRNA removal, assembly of short read transcripts, and more relevant reference bases while providing a priority set of expressed genes for functional assessment.
Comments
This is the accepted manuscript of an article published in Journal of Microbiological Methods, 131 (December 2016): 122-129, http://dx.doi.org/10.1016/j.mimet.2016.10.018.