Development and use of a switchgrass (Panicum virgatum L.) transformation pipeline by the BioEnergy Science Center to evaluate plants for reduced cell wall recalcitrance Mike Himmel

Richard S. Nelson, C. Neal Stewart, Jiqing Gou, Susan Holladay, Lina Gallego-Giraldo, Amy Flanagan, David G.J. Mann, Hiroshi Hisano, Wegi A. Wuddineh, Charleson R. Poovaiah, Avinash Srivastava, Ajaya K. Biswal, Hui Shen, Luis L. Escamilla-Treviño, Jiading Yang, C. Frank Hardin, Rangaraj Nandakumar, Chunxiang Fu, Jiyi Zhang, Xirong XiaoRyan Percifield, Fang Chen, Jeffrey L. Bennetzen, Michael Udvardi, Mitra Mazarei, Richard A. Dixon, Zeng Yu Wang, Yuhong Tang, Debra Mohnen, Brian H. Davison

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: The mission of the BioEnergy Science Center (BESC) was to enable efficient lignocellulosic-based biofuel production. One BESC goal was to decrease poplar and switchgrass biomass recalcitrance to biofuel conversion while not affecting plant growth. A transformation pipeline (TP), to express transgenes or transgene fragments (constructs) in these feedstocks with the goal of understanding and decreasing recalcitrance, was considered essential for this goal. Centralized data storage for access by BESC members and later the public also was essential. Results: A BESC committee was established to codify procedures to evaluate and accept genes into the TP. A laboratory information management system (LIMS) was organized to catalog constructs, plant lines and results from their analyses. One hundred twenty-eight constructs were accepted into the TP for expression in switchgrass in the first 5 years of BESC. Here we provide information on 53 of these constructs and the BESC TP process. Eleven of the constructs could not be cloned into an expression vector for transformation. Of the remaining constructs, 22 modified expression of the gene target. Transgenic lines representing some constructs displayed decreased recalcitrance in the field and publications describing these results are tabulated here. Transcript levels of target genes and detailed wall analyses from transgenic lines expressing six additional tabulated constructs aimed toward modifying expression of genes associated with wall structure (xyloglucan and lignin components) are provided. Altered expression of xyloglucan endotransglucosylase/hydrolases did not modify lignin content in transgenic plants. Simultaneous silencing of two hydroxycinnamoyl CoA:shikimate hydroxycinnamoyl transferases was necessary to decrease G and S lignin monomer and total lignin contents, but this reduced plant growth. Conclusions: A TP to produce plants with decreased recalcitrance and a LIMS for data compilation from these plants were created. While many genes accepted into the TP resulted in transgenic switchgrass without modified lignin or biomass content, a group of genes with potential to improve lignocellulosic biofuel yields was identified. Results from transgenic lines targeting xyloglucan and lignin structure provide examples of the types of information available on switchgrass lines produced within BESC. This report supplies useful information when developing coordinated, large-scale, multi-institutional reverse genetic pipelines to improve crop traits.

Original languageEnglish
Article number309
JournalBiotechnology for Biofuels
Volume10
Issue number1
DOIs
Publication statusPublished - Dec 22 2017

Keywords

  • Bioenergy
  • Cell wall
  • Ethanol
  • HCT
  • Lignin
  • Lignocellulosic feedstocks
  • Recalcitrance
  • Reverse genetics
  • Transformation pipeline
  • XTH

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • Energy(all)
  • Management, Monitoring, Policy and Law

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    Nelson, R. S., Stewart, C. N., Gou, J., Holladay, S., Gallego-Giraldo, L., Flanagan, A., Mann, D. G. J., Hisano, H., Wuddineh, W. A., Poovaiah, C. R., Srivastava, A., Biswal, A. K., Shen, H., Escamilla-Treviño, L. L., Yang, J., Hardin, C. F., Nandakumar, R., Fu, C., Zhang, J., ... Davison, B. H. (2017). Development and use of a switchgrass (Panicum virgatum L.) transformation pipeline by the BioEnergy Science Center to evaluate plants for reduced cell wall recalcitrance Mike Himmel. Biotechnology for Biofuels, 10(1), [309]. https://doi.org/10.1186/s13068-017-0991-x