Advertisement

BioEnergy Research

, Volume 8, Issue 3, pp 922–933 | Cite as

A Novel Delivery System for the Root Symbiotic Fungus, Sebacina vermifera, and Consequent Biomass Enhancement of Low Lignin COMT Switchgrass Lines

  • Prasun Ray
  • Takako Ishiga
  • Stephen R. Decker
  • Geoffrey B. Turner
  • Kelly D. Craven
Article

Abstract

Sebacina vermifera (MAFF-305830) is a mycorrhizal fungus originally isolated from the roots of orchids that we have previously shown to be tremendously beneficial in enhancing biomass yield and drought tolerance in switchgrass, an important bioenergy crop for cellulosic ethanol production in the United States. Towards this end, we have developed a bentonite clay particle-based delivery system for mass production and dissemination of S. vermifera for large-scale field trials. A greenhouse-based experiment was conducted to evaluate this novel delivery method for biomass enhancement of wild type and transgenic, low lignin (COMT down-regulated) switchgrass lines compared to an efficient in vitro colonization method. S. vermifera colonization enhanced plant biomass regardless of delivery method, although the percentage of fungal biomass in planta increased with the clay-based delivery system. Further, we found that release of some clay minerals in solution was enhanced in the presence of S. vermifera, while others were seemingly reduced. Intriguingly, the presence of S. vermifera has little or no impact on cell wall composition, including lignification. This research is the first report documenting the development of a bentonite clay particle-based delivery system for mass production of any symbiotic microbe and suggests that S. vermifera can be packaged with a mineral composite and effectively delivered to a target host plant.

Keywords

Switchgrass Mycorrhizae Sebacina COMT 

Notes

Acknowledgments

S. vermifera (MAFF-305830) used in this study was obtained from the National Institute of Agro-biological Sciences, Tsukuba, Ibaraki, Japan. The COMT lines used in this study were provided by Chunxiang Fu and Zeng-Yu Wang, Forage Improvement Division, The Samuel Roberts Noble Foundation. We thank Crissa Doeppke, Melissa Glenn, Kimberly Mazza, Logan Schuster, and Kevin Cowley in NREL for their efforts in preparing samples for the HTP recalcitrance pipeline; Erica Gjersing Robert Sykes and Mark Davis in NREL for cell wall composition analysis; David Huhman for ion chromatography; Stacy Allen for qRT-PCR; Jin Nakashima for assistance with SEM and confocal microscopy; Stephen L. Webb for assistance with statistical analysis; and Myoung-Hwan Chi, Blue Stewart, Colleen Elles, and Amanda Hammon for greenhouse assistance. This work was supported by the Bioenergy Science Center, a US Department of Energy Bioenergy Research Center, through the Office of Biological and Environmental Research in the DOE Office of Science.

Conflict of Interest

All the authors in this manuscript declare no conflict of interests inherent to this submission.

Supplementary material

12155_2015_9636_MOESM1_ESM.doc (1.2 mb)
ESM 1 (DOC 1190 kb)

References

  1. 1.
    Oberwinkler F, Riess K, Bauer R, Selosse MA, Weiss M, Garnica S, Zuccaro A (2013) Enigmatic Sebacinales. Mycol Prog 12(1):1–27. doi: 10.1007/s11557-012-0880-4 CrossRefGoogle Scholar
  2. 2.
    DeMars B, Boerner R (1996) Vesicular arbuscular mycorrhizal development in the Brassicaceae in relation to plant life span. Flora 191Google Scholar
  3. 3.
    Selosse MA, Bauer R, Moyersoen B (2002) Basal hymenomycetes belonging to the Sebacinaceae are ectomycorrhizal on temperate deciduous trees. New Phytol 155(1):183–195. doi: 10.1046/j.1469-8137.2002.00442.x CrossRefGoogle Scholar
  4. 4.
    Weiss M, Sykorova Z, Garnica S, Riess K, Martos F, Krause C, Oberwinkler F, Bauer R, Redecker D (2011) Sebacinales everywhere: previously overlooked ubiquitous fungal endophytes. PLoS ONE 6(2):e16793. doi: 10.1371/journal.pone.0016793 PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Warcup J (1988) Mycorrhizal associations of isolates of Sebacina vermifera. New Phytol 110(2):227–231CrossRefGoogle Scholar
  6. 6.
    Ghimire SR, Charlton ND, Craven KD (2009) The mycorrhizal fungus, Sebacina vermifera, enhances seed germination and biomass production in switchgrass (Panicum virgatum L). Bioenergy Res 2(1–2):51–58CrossRefGoogle Scholar
  7. 7.
    Ghimire SR, Craven KD (2011) Enhancement of switchgrass (Panicum virgatum L.) biomass production under drought conditions by the ectomycorrhizal fungus Sebacina vermifera. Appl Environ Microbiol 77(19):7063–7067. doi: 10.1128/AEM.05225-11 PubMedCentralCrossRefPubMedGoogle Scholar
  8. 8.
    McLaughlin SB, Adams Kszos L (2005) Development of switchgrass (Panicum virgatum) as a bioenergy feedstock in the United States. Biomass Bioenergy 28(6):515–535CrossRefGoogle Scholar
  9. 9.
    Wang MQ (2001) Development and use of GREET 1.6 fuel-cycle model for transportation fuels and vehicle technologies. Argonne National Lab., IL (United States). Funding organisation: US Department of Energy (United States)Google Scholar
  10. 10.
    Farrell AE, Plevin RJ, Turner BT, Jones AD, O’hare M, Kammen DM (2006) Ethanol can contribute to energy and environmental goals. Science 311(5760):506–508CrossRefPubMedGoogle Scholar
  11. 11.
    Schmer MR, Vogel KP, Mitchell RB, Perrin RK (2008) Net energy of cellulosic ethanol from switchgrass. Proc Natl Acad Sci 105(2):464–469PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Aden A, Foust T (2009) Technoeconomic analysis of the dilute sulfuric acid and enzymatic hydrolysis process for the conversion of corn stover to ethanol. Cellulose 16(4):535–545CrossRefGoogle Scholar
  13. 13.
    Abramson M, Shoseyov O, Shani Z (2010) Plant cell wall reconstruction toward improved lignocellulosic production and processability. Plant Sci 178(2):61–72CrossRefGoogle Scholar
  14. 14.
    Fu C, Mielenz JR, Xiao X, Ge Y, Hamilton CY, Rodriguez M Jr, Chen F, Foston M, Ragauskas A, Bouton J, Dixon RA, Wang ZY (2011) Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass. Proc Natl Acad Sci U S A 108(9):3803–3808. doi: 10.1073/pnas.1100310108 PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Malusá E, Sas-Paszt L, Ciesielska J (2012) Technologies for beneficial microorganisms inocula used as biofertilizers. Sci World J 2012Google Scholar
  16. 16.
    Heijnen C, Hok-A-Hin C, Van Veen J (1992) Improvements to the use of bentonite clay as a protective agent, increasing survival levels of bacteria introduced into soil. Soil Biol Biochem 24(6):533–538CrossRefGoogle Scholar
  17. 17.
    Marx DH (1969) The influence of ectotrophic mycorrhizal fungi on the resistance of pine roots to pathogenic infections. I. Antagonism of mycorrhizal fungi to root pathogenic fungi and soil bacteria. Phytopathology 59:153–163Google Scholar
  18. 18.
    Charlton ND, Shoji J-Y, Ghimire SR, Nakashima J, Craven KD (2012) Deletion of the fungal gene soft disrupts mutualistic symbiosis between the grass endophyte Epichloë festucae and the host plant. Eukaryot Cell 11(12):1463–1471PubMedCentralCrossRefPubMedGoogle Scholar
  19. 19.
    Young C, Bryant M, Christensen M, Tapper B, Bryan G, Scott B (2005) Molecular cloning and genetic analysis of a symbiosis-expressed gene cluster for lolitrem biosynthesis from a mutualistic endophyte of perennial ryegrass. Mol Gen Genomics 274(1):13–29CrossRefGoogle Scholar
  20. 20.
    Sykes R, Kodrzycki B, Tuskan G, Foutz K, Davis M (2008) Within tree variability of lignin composition in Populus. Wood Sci Technol 42(8):649–661CrossRefGoogle Scholar
  21. 21.
    Selig MJ, Tucker MP, Sykes RW, Reichel KL, Brunecky R, Himmel ME, Davis MF, Decker SR (2010) Original research: Lignocellulose recalcitrance screening by integrated high-throughput hydrothermal pretreatment and enzymatic saccharification. Ind Biotechnol 6(2):104–111CrossRefGoogle Scholar
  22. 22.
    Decker SR, Brunecky R, Tucker MP, Himmel ME, Selig MJ (2009) High-throughput screening techniques for biomass conversion. Bioenergy Res 2(4):179–192CrossRefGoogle Scholar
  23. 23.
    Decker SR, Carlile M, Selig MJ, Doeppke C, Davis M, Sykes R, Turner G, Ziebell A (2012) Reducing the effect of variable starch levels in biomass recalcitrance screening. In: Biomass conversion. Springer, pp 181–195Google Scholar
  24. 24.
    Davison BH, Drescher SR, Tuskan GA, Davis MF, Nghiem NP (2006) Variation of S/G ratio and lignin content in a Populus family influences the release of xylose by dilute acid hydrolysis. Appl Biochem Biotechnol 130(1–3):427–435. doi: 10.1385/Abab:130:1:427 CrossRefGoogle Scholar
  25. 25.
    Rossi MJ, Furigo A, Oliveira VL (2007) Inoculant production of ectomycorrhizal fungi by solid and submerged fermentations. Food Technol Biotechnol 45(3):277Google Scholar
  26. 26.
    Schwartz MW, Hoeksema JD, Gehring CA, Johnson NC, Klironomos JN, Abbott LK, Pringle A (2006) The promise and the potential consequences of the global transport of mycorrhizal fungal inoculum. Ecol Lett 9(5):501–515CrossRefPubMedGoogle Scholar
  27. 27.
    Vogel KP, Jung H-JG (2001) Genetic modification of herbaceous plants for feed and fuel. Crit Rev Plant Sci 20(1):15–49CrossRefGoogle Scholar
  28. 28.
    Huang Y, Rickerl D, Kephart K (1996) Recovery of deep-point injected soil nitrogen-15 by switchgrass, alfalfa, ineffective alfalfa, and corn. J Environ Qual 25(6):1394–1400CrossRefGoogle Scholar
  29. 29.
    Schultz R, Collettil J, Isenhart T, Simpkins W, Mize C, Thompson M (1995) Design and placement of a multi-species riparian buffer strip system. Agrofor Syst 29(3):201–226CrossRefGoogle Scholar
  30. 30.
    Ma Z, Wood C, Bransby D Management and soil influence on switchgrass carbon sequestration and biomass accumulation. In: Agronomy abstracts, ASA, Madison, WI, 1996. p 32Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Prasun Ray
    • 1
  • Takako Ishiga
    • 1
  • Stephen R. Decker
    • 2
  • Geoffrey B. Turner
    • 2
  • Kelly D. Craven
    • 1
  1. 1.Plant Biology DivisionThe Samuel Roberts Noble FoundationArdmoreUSA
  2. 2.Biosciences CenterNational Renewable Energy LaboratoryGoldenUSA

Personalised recommendations