Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Development and field assessment of transgenic hybrid switchgrass for improved biofuel traits

  • 58 Accesses


Development of commercially relevant bioenergy switchgrass cultivars requires reducing recalcitrance for bioprocessing without compromising biomass yield. Low-lignin transgenic switchgrass has been produced via down-regulation of caffeic acid O-methyltransferase (COMT), a lignin biosynthetic enzyme, or by over-expression of the MYB4 transcription factor, a repressor of the lignin biosynthetic pathway. The aim of this study was to evaluate parents and selected hybrids obtained from COMT and MYB4 hybrid families under field conditions for agronomic performance and biomass quality. Plant height, width, number of tillers, dry weight, cell wall composition including lignin content, and sugar release were measured after the establishment year (2014) and the second growing season (2015). For COMT hybrids, biomass yield of the transgenic hybrids was similar to or greater than the wild-type parents selected for high biomass. Lignin content of COMT transgenic hybrids was reduced by 10%, S/G ratio decreased by 27%, and sugar release increased between 20% and 44% compared to their wild-type parents. These results indicate that hybridization of COMT with a high-yielding locally selected genotype resulted in both improved agronomic performance and enhanced biomass quality in the offspring. On the other hand, the MYB transgenic hybrid showed a 10% reduction in biomass yield compared with its wild-type parent in year 1, but not in year 2. The lignin S/G ratio was not reduced in MYB transgenic hybrids, nor was sugar release increased. These data indicate that the MYB transgene may not be suitable for an agronomic setting. More testing is needed of transgenic and wild-type, high-biomass selections for use as breeding parents. These results show that combining low-lignin transgenic switchgrass with a breeding and selection program for biomass yield will allow for the deployment of effective transgenes in high-yielding genetic backgrounds.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. Alexander LW, Haynes ER, Burris J, Jackson S, Stewart CN Jr (2014) Cultural treatments for accelerated growth and flowering of Panicum virgatum. Biofuels 5:771–780

  2. Balan VS, Kumar S, Bals B, Chundawat S, Jin M, Dale B (2012) Biochemical and thermochemical conversion of switchgrass to biofuels. In: Monti A (ed) Switchgrass: a valuable biomass crop for energy. Springer, London, pp 153–185

  3. Baxter HL, Mazarei M, Labbe N, Kline LM, Cheng Q, Windham MT, Mann DGJ, Fu C, Ziebell A, Sykes RW, Rodriguez M, Davis MF, Mielenz JR, Dixon RA, Wang Z-Y, Stewart CN Jr (2014) Two-year field analysis of reduced recalcitrance transgenic switchgrass. Plant Biotechnol J 12:914–924

  4. Baxter HL, Poovaiah CR, Yee KL, Mazarei M, Rodriguez M, Thompson OA, Shen H, Turner GB, Decker SR, Sykes RW, Chen F, Davis MF, Mielenz JR, Davison BH, Dixon RA, Stewart CN Jr (2015) Field evaluation of transgenic switchgrass plants overexpressing PvMYB4 for reduced biomass recalcitrance. BioEnergy Res 8:910–921

  5. Baxter HL, Alexander LW, Mazarei M, Haynes E, Turner GB, Sykes RW, Decker SR, Davis MF, Dixon RA, Wang Z-Y, Stewart CN Jr (2016) Hybridization of downregulated-COMT transgenic switchgrass lines with field-selected switchgrass for improved biomass traits. Euphytica 209:341–355

  6. Bhandari HS, Walker DW, Bouton JH (2014) Effects of ecotypes and morphotypes in feedstock composition of switchgrass (Panicum virgatum L.). GCB Bioenergy 6:26–34

  7. Boateng AA, Hicks KB, Vogel KP (2006) Pyrolysis of switchgrass (Panicum virgatum) harvested at several stages of maturity. J Analytical Appl Pyrolysis 75:55–64

  8. Casler MD, Vogel KP (2014) Selection of biomass yield in upland, lowland, and hybrid switchgrass. Crop Sci 54:626–636

  9. Casler MD, Buxton DR, Vogel KP (2002) Genetic modification of lignin concentration affects fitness of perennial herbaceous plants. Theory Appl Genet 104:127–131

  10. Cassida KA, Muir JP, Hussey MA, Read JC, Venuto BC, Ocumpaugh WR (2005) Biofuel component concentrations and yields of switchgrass in south central US environments. Crop Sci 45:682–692

  11. Chang VS, Holtzapple MT (2000) Fundamental factors affecting biomass enzymatic reactivity. Appl Biochem Biotechnol 84:5–37

  12. Chen F, Dixon R (2007) Lignin modification improves fermentable sugar yields in biofuel production. Nat Biotechnol 25:759–761

  13. Costich DE, Friebe B, Sheehan MJ, Casler MD, Buckler ES (2010) Genome-size variation in switchgrass (Panicum virgatum): flow cytometry and cytology reveal rampant aneuploidy. Plant Gen 3:130–141

  14. Energy independence and security act (2007) Public Law 110–140, http://www.gpo.gov/fdsys/pkg/BILLS-110hr6enr/pdf/BILLS-110hr6enr.pdf

  15. Fu C, Mielenz JR, Xiao X, Ge Y, Hamilton CY, Rodriguez M, Chen F, Foston M, Ragauskas A, Bouton J, Dixon RA, Wang Z-Y (2011) Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass. Proc Natl Acad Sci USA 108:3803–3808

  16. Goldenberg J, Guardabassi P (2010) The potential for first-generation ethanol production from sugarcane. Biofuels, Bioprod Biorefin 4:17–41

  17. Hardin CF, Fu C, Hisano H, Xiao X, Shen H, Stewart CN, Wang ZY (2013) Standardization of switchgrass sample collection for cell wall and biomass trait analysis. Bioenergy Res 6:755–762

  18. Himmel ME, Ding S-Y, Johnson DK, Adney WS, Nimlos MR, Brady JW, Foust TD (2007) Biomass recalcitrance: engineering plants and enzymes for biofuels production. Science 315:804–807

  19. Hopkins AA, Vogel KP, Moore KJ, Johnson KD, Carlson IT (1995) Genotype effects and genotype by environment interactions for traits of elite switchgrass populations. Crop Sci 35:125–132

  20. Hu Z, Sykes R, Davis MF, Brummer EC, Ragauskas AJ (2010) Chemical profiles of switchgrass. Bioresource Technol 101:3253–3257

  21. King ZR, Bray AL, LaFayette PR, Parrott WA (2014) Biolistic transformation of elite genotypes of switchgrass (Panicum virgatum L.). Plant Cell Rep 33:313–322

  22. Kwit C, Moon HS, Warwick S, Stewart CN Jr (2011) Transgene introgression in crop relatives: molecular evidence and mitigation strategies. Trends Biotechnol 29:284–291

  23. McLaughlin SB, Walsh ME (1998) Evaluating environmental consequences of producing herbaceous crops for bioenergy. Biomass Bioenergy 14:317–324

  24. Moon HS, Abercrombie LL, Eda S, Blanvillain R, Thomson JG, Ow DW, Stewart CN Jr (2011) Transgene excision in pollen using codon optimized serine resolvase CinH-RS2 site-specific recombination system. Plant Mol Biol 75:621–631

  25. Moore KJ, Moser LW, Vogel KP, Waller SS, Johnson BE, Pedersen JF (1991) Describing and quantifying growth stages of perennial forage grasses. Agron J 83:1073

  26. Parrish DJ, Fike JH (2005) The biology and agronomy of switchgrass for biofuels. Crit Rev Plant Sci 24:423–459

  27. Pedersen JF, Vogel KP, Funnell DL (2005) Impact of reduced lignin on plant fitness. Crop Sci 45:812–819

  28. Poovaiah CR, Mazarei M, Decker SR, Turner GB, Sykes RW, Davis MF, Stewart CN (2015) Transgenic switchgrass (Panicum virgatum L.) biomass is increased by overexpression of switchgrass sucrose synthase (PvSUS1). Biotechnol J 10:552–563

  29. Price DL, Casler MD (2013) Predictive relationships between plant morphological traits and biomass yield in switchgrass. Crop Sci 54:637–645

  30. Sanderson MA, Reed RL, McLaughlin SB, Wullschleger SD, Conger BV, Parrish DJ, Wolf DD, Taliaferro C, Hopkins AA, Ocumpaugh WR, Hussey MA, Read JC, Tischler CR (1996) Switchgrass as a sustainable bioenergy crop. Bioresour Technol 56:83–93

  31. Sanderson MA, Alder PR, Boateng AA, Casler MD, Sarath G (2006) Switchgrass as a biofuels feedstock in the USA. Can J Plant Sci 86:1315–1325

  32. Serba DD, Sykes RW, Gjersing EL, Decker SR, Daverdin G, Devos KM, Brummer EC, Saha MC (2016) Cell wall composition and underlying QTL in an F1 pseudo-testcross population of switchgrass. Bioenerg Res 9:836–850

  33. Shen H, Fu CX, Xiao XR, Ray T, Tang YH, Wang ZY, Chen F (2009) Developmental control of lignification in stems of lowland switchgrass variety Alamo and the effects on saccharification efficiency. Bioenerg Res 2:233–245

  34. Shen H, He X, Poovaiah CR, Wuddineh WA, Ma J, Mann DG, Wang H, Jackson L, Tang Y, Stewart CN Jr, Chen F, Dixon RA (2012) Functional characterization of the switchgrass (Panicum virgatum) R2R3-MYB transcription factor PvMYB4 for improvement of lignocellulosic feedstocks. New Phytol 193:121–136

  35. Shen H, Poovaiah CR, Ziebell A, Tschaplinski TJ, Pattathil S, Gjersing E, Engle EL, Katahira R, Pu Y, Sykes R, Chen F, Ragauskas A, Mielenz JR, Hahn MG, Davis M, Stewart CN Jr, Dixon RA (2013) Enhanced characteristics of genetically modified switchgrass (Panicum virgatum L.) for high biofuel production. Biotechnol Biofuels 6:71

  36. Thammasouk K, Tandjo D, Penner MH (1997) Influence of extractives on the analysis of herbaceous biomass. J Agri Food Chem 45:437–443

  37. Wright L, Turhollow A (2010) Switchgrass selection as a “model” bioenergy crop: a history of the process. Biomass Bioenerg 34:851–868

  38. Wudinneh WA, Mazarei M, Zhang J-Y, Poovaiah CR, Mann DGJ, Ziebell A, Sykes RW, Davis MF, Udvardi MK, Stewart CN Jr (2015) Identification and overexpression of gibberellin 2-oxidase (GA2ox) in switchgrass (Panicum virgatum L.) for improved plant architecture and reduced biomass recalcitrance. Plant Biotechnol J 13:636–647

  39. Wudinneh WA, Mazarei M, Zhang J-Y, Turner GB, Sykes RW, Decker SR, Davis MF, Udvardi MK, Stewart CN Jr (2016) Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement. Fron Plant Sci 7:520

  40. Xu B, Escamilla-Trevino LL, Sathitsuksanoh N, Shen Z, Shen H, Zhang YH, Dixon RA, Zhao B (2011) Silencing of 4-coumarate:coenzyme a ligase in switchgrass leads to reduced lignin content and improved fermentable sugar yields for biofuel production. New Phytol 192:611–625

Download references


We thank Fred Allen, Hem Bhandari, and Ken Goddard for providing the Tennessee field accessions of switchgrass field selections. Richard Dixon provided MYB4 parent plants. We also thank Hayley Rideout, Ben Wolfe, Marcus Laxton, and the UT field staff for assistance with data collection, preparing samples for cell wall characterization and general field maintenance, and Reggie Millwood for assistance with the USDA APHIS BRS permit regulations. This work was supported by the Agriculture and Food Research Initiative (United States Department of Agriculture) and Southeastern Partnership for Integrated Biomass and Supply Systems (The IBSS Partnership), and enabled by the Bioenergy Science Center. The Bioenergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

Author information

Correspondence to Lisa Alexander.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 30 kb)

Supplementary material 2 (DOCX 35 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Alexander, L., Hatcher, C., Mazarei, M. et al. Development and field assessment of transgenic hybrid switchgrass for improved biofuel traits. Euphytica 216, 25 (2020). https://doi.org/10.1007/s10681-020-2558-3

Download citation


  • Panicum virgatum
  • Biomass
  • Lignin
  • Recalcitrance
  • Renewable energy