Advertisement

Planta

, Volume 249, Issue 5, pp 1465–1475 | Cite as

Production of deuterated biomass by cultivation of Lemna minor (duckweed) in D2O

  • Barbara R. EvansEmail author
  • Marcus Foston
  • Hugh M. O’Neill
  • David Reeves
  • Caroline Rempe
  • Kathi McGrath
  • Arthur J. Ragauskas
  • Brian H. Davison
Original Article
  • 281 Downloads

Abstract

Main conclusion

Common duckweed Lemna minor was cultivated in 50% D2O to produce biomass with 50–60% deuterium incorporation containing cellulose with degree of polymerization close (85%) to that of H2O-grown controls.

The small aquatic plant duckweed, particularly the genus Lemna, widely used for toxicity testing, has been proposed as a potential source of biomass for conversion into biofuels as well as a platform for production of pharmaceuticals and specialty chemicals. Ability to produce deuterium-substituted duckweed can potentially extend the range of useful products as well as assist process improvement. Cultivation of these plants under deuterating conditions was previously been reported to require addition of kinetin to induce growth and was hampered by anomalies in cellular morphology and protein metabolism. Here, we report the production of biomass with 50–60% deuterium incorporation by long-term photoheterotrophic growth of common duckweed Lemna minor in 50% D2O with 0.5% glucose. L. minor grown in 50% D2O without addition of kinetin exhibited a lag phase twice that of H2O-grown controls, before start of log phase growth at 40% of control rates. Compared to continuous white fluorescent light, growth rates increased fivefold for H2O and twofold for 50% D2O when plants were illuminated at higher intensity with a metal halide lamp and a diurnal cycle of 12-h light/12-h dark. Deuterium incorporation was determined by a combination of 1H and 2H nuclear magnetic resonance (NMR) to be 40–60%. The cellulose from the deuterated plants had an average-number degree of polymerization (DPn) and polydispersity index (PDI) close to that of H2O-grown controls, while Klason lignin content was reduced. The only major gross morphological change noted was root inhibition.

Keywords

Lemna minor Duckweed Biomass Deuteration Cellulose Nuclear magnetic resonance 

Notes

Acknowledgements

This research was supported by the U. S. Department of Energy, Office of Science, through the Genomic Science Program, Office of Biological and Environmental Research, under Contract FWP ERKP752. The research at Oak Ridge National Laboratory’s Center for Structural Molecular Biology (CSMB) was supported by the U. S. Department of Energy, Office of Science, through the Office of Biological and Environmental Research under Contract FWP ERKP291, using facilities supported by the Office of Basic Energy Sciences, U. S. Department of Energy. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U. S. Department of Energy under Contract DE-AC05-00OR22725. D. Reeves was supported by a U. S. Department of Energy Higher Education Research Experience internship managed by Oak Ridge Institute of Science and Education. C. Rempe was supported by a Department of Energy Science Undergraduate Laboratory Internship and Higher Education Research Experience internship managed by Oak Ridge Institute of Science and Education. K. McGrath was supported by the DOE Academies Creating Teacher Scientists (ACTS) summer 2010 program.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

References

  1. Bali G, Foston MB, O’Neill HM, Evans BR, He J, Ragauskas AJ (2013) The effect of deuterium incorporation on the structure of bacterial cellulose. Carbohydr Res 374:82–88Google Scholar
  2. Bergmann BA, Cheng J, Classen J, Stomp A-M (2000) In vitro selection of duckweed geographical isolates for potential use in swine lagoon effluent renovation. Bioresour Technol 73:13–20Google Scholar
  3. Blake MI, Crane FA, Uphaus RA, Katz JJ (1968) Effect of heavy water on the germination of a number of species of seeds. Planta 78:35–38Google Scholar
  4. Blazey EB, McClure JW (1968) The distribution and taxonomic significance of lignin in the Lemnaceae. Am J Bot 55:1240–1245Google Scholar
  5. Bornkamm R (1966) A seasonal rhythm of growth in Lemna minor L. Planta 69:178–186PubMedGoogle Scholar
  6. Borucki B, von Stetten D, Seibeck S, Lamparter T, Michael N, Mroginski MA, Otto H, Murgida DH, Heyn MP, Hildebrandt P (2005) Light-induced proton release of phytochrome is coupled to the transient deprotonation of the tetrapyrrole chromophore. J Biol Chem 280:34358–34364Google Scholar
  7. Brain RA, Solomon KR (2007) A protocol for conducting 7-day daily renewal tests with Lemna gibba. Nat Protoc 2:979–987PubMedGoogle Scholar
  8. Butt HI, Yang Z, Gong Q, Chen E, Wang X, Zhao G, Ge X, Zhang X, Li F (2017) GaMYB85, an R2R3 MYB gene, in transgenic Arabidopsis plays an important role in drought tolerance. BMC Plant Biol 17:142.  https://doi.org/10.1186/s12870-017-1078-3 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Chandra S, Bhaduri SK, Sardar D (1991) Chemical characterization of pressed fibrous residues of four aquatic weeds. Aquat Bot 42:81–85Google Scholar
  10. Cooke RJ, Davies DD (1980) General characteristics of normal and stress-enhanced protein degradation in Lemna minor (duckweed). Biochem J 192:499–506PubMedPubMedCentralGoogle Scholar
  11. Cooke RJ, Grego S, Oliver J, Davies DD (1979a) The effect of deuterium oxide on protein turnover in Lemna minor. Planta 146:229–236PubMedGoogle Scholar
  12. Cooke RJ, Oliver J, Davies DD (1979b) Stress and protein turnover in Lemna minor. Plant Physiol 64:1109–1113PubMedPubMedCentralGoogle Scholar
  13. Cooke RJ, Grego S, Roberts K, Davies DD (1980) The mechanism of deuterium oxide-induced protein degradation in Lemna minor. Planta 148:374–380PubMedGoogle Scholar
  14. Cope BT, Bose S, Crespi HL, Katz JJ (1965) Growth of Lemna in H2O–D2O mixtures: enhancement by kinetin. Bot Gaz 126:214–221Google Scholar
  15. Cox KM, Sterling JD, Regan JT, Gadaska JR, Frantz KK, Peele CG, Black A, Passmore D, Moldovan-Loomis C, Srinivasan M, Cuison S, Cardarelli PM, Dickey LF (2006) Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat Biotechnol 24:1591–1597PubMedGoogle Scholar
  16. De Coursey TE, Cherny VV (1997) Deuterium isotope effects on permeation and gating of proton channels in rat alveolar epithelium. J Gen Physiol 169:415–434Google Scholar
  17. DeWitt SH, Maryanoff BE (2018) Deuterated drug molecules: focus on FDA-approved deutetrabenazine. Biochemistry 57:472–473PubMedGoogle Scholar
  18. Evans BR, Shah R (2015) Development of approaches for deuterium labeling in plants. In: Kelman Z (ed) Methods in enzymology: volume 565 isotope labeling of biomolecules. Elsevier, Oxford, pp 213–243Google Scholar
  19. Evans B, Bali G, Reeves D, O’Neill H, Sun Q, Shah R, Ragauskas A (2014) Effect of D2O on growth properties and chemical structure of annual ryegrass (Lolium multiflorum). J Agric Food Chem 62:2592–2604Google Scholar
  20. Evans BR, Bali G, Foston M, Ragauskas AJ, O’Neill H, Shah R, McGaughey J, Reeves D, Rempe CS, Davison BH (2015) Production of deuterated switchgrass by hydroponic cultivation. Planta 242:215–222Google Scholar
  21. Evans BR, Bali G, Ragauskas A, Shah R, O’Neill H, Howard C, Lavenhouse F, Ramirez D, Weston K, Ramey K, Cangemi V, Kinney B, Partee C, Ware T, Davison B (2017) Alleopathic effects of exogenous phenylalanine: a comparison of four monocot species. Planta 246:673–685Google Scholar
  22. Firsov A, Tarasenko I, Mitiouchkina T, Shaloiko L, Kozlov O, Vinokurov L, Rasskazova E, Murashev A, Vainstein A, Dolgov S (2018) Expression and immunogenicity of M2e peptide of avian influenza virus H5N1 fused to ricin toxin B chain produced in duckweed plants. Front Chem 6:22.  https://doi.org/10.3389/fchem.2018.00022 CrossRefPubMedPubMedCentralGoogle Scholar
  23. Foston MB, McGaughey J, O’Neill H, Evans BR, Ragauskas AJ (2012) Deuterium incorporation in biomass cell wall components by NMR analysis. Analyst 137:1090–1093Google Scholar
  24. Foston M, Ragauskas AJ (2010) Changes in lignocellulosic supramolecular and ultrastructure during dilute acid pretreatment of populus and switchgrass. Biomass Bioenergy 34(12):1885–1895Google Scholar
  25. Gasdaska J, Spencer D, Dickey L (2003) Advantages of therapeutic protein production in the aquatic plant Lemna. Bioprocess J 3:50–56Google Scholar
  26. Halford B (2016) The deuterium switcheroo. Chem Eng News 94:32–36Google Scholar
  27. Joshi R, Anwar K, Das P, Singla-Pareek SL, Pareek A (2017) Overview of methods for assessing salinity and drought tolerance of transgenic wheat lines. In: Bhalla PL, Singh MB (eds) Wheat biotechnology. Springer, New York, pp 83–95Google Scholar
  28. Körner S, Vermaat JE, Veenstra S (2003) The capacity of duckweed to treat wastewater: ecological consideration for a sound design. J Environ Qual 32:1583–1590Google Scholar
  29. Langan P, Evans BR, Foston M, Heller WT, O’Neill HM, Petridis L, Pingali SV, Ragauskas AJ, Smith JC, Davison B (2012) Neutron technologies for bioenergy research. Ind Biotechnol 8:209–216Google Scholar
  30. Liu C, Dai Z, Sun H (2017) Potential of duckweed (Lemna minor) for removal of nitrogen and phosphorus from water under salt stress. J Environ Manag 187:497–503Google Scholar
  31. Löppert H, Kronberger W, Kandeler R (1978) Phytochrome-mediated changes in the membrane potential of subepidermal cells of Lemna paucicostata 6746. Planta 138:133–136PubMedGoogle Scholar
  32. McClure JW (1975) The applicability of polyphenolic data to systematic problems in the Lemnaceae. Aquat Bot 1:395–405Google Scholar
  33. Moody M, Miller J (2005) Lemna minor growth inhibition test. In: Blaise C, Férard J-F (eds) Small scale freshwater toxicity investigations. Springer, Amsterdam, pp 271–298Google Scholar
  34. Novacky A, Ullrich-Eberius CI, Lüttge U (1978) Membrane potential changes during transport of hexoses in Lemna gibba G1. Planta 138:263–270PubMedGoogle Scholar
  35. Porath D, Hepher B, Koton A (1979) Duckweed as an aquatic crop: evaluation of clones for aquaculture. Aquat Bot 7:273–278Google Scholar
  36. Porra R (2002) The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. Photosynth Res 73:149–156PubMedGoogle Scholar
  37. Reid GSG (1997) Carbohydrate metabolism: structural carbohydrates. In: Dey PM, Harborne JB (eds) Plant biochemistry. Academic Press, San Diego, CA, USA, London, UK, pp 205–235Google Scholar
  38. Sarkar HK, Song PS (1981) Phototransformation and dark reversion of phytochrome in deuterium oxide. Biochemistry 20:4315–4320PubMedGoogle Scholar
  39. Schmidt C (2017) First deuterated drug approved. Nat Biotechnol 35:493–494PubMedGoogle Scholar
  40. Siegel SM, Halpern LA, Giumaro C (1964) Germination and seedling growth of winter rye in deuterium oxide. Nature 201:1244–1245Google Scholar
  41. Singh A, Jha SK, Bagri J, Pandey GK (2015) ABA inducible rice protein phosphatase 2C confers ABA insensitivity and abiotic stress tolerance in arabidopsis. PLoS One 10(4):e0125168.  https://doi.org/10.1371/journal.pone.0125168 CrossRefPubMedPubMedCentralGoogle Scholar
  42. Stomp A-M (2005) The duckweeds. A valuable plant for biomanufacturing. Biotechnol Annu Rev 11:69–99Google Scholar
  43. Trewavas A (1970) The turnover of nucleic acids in Lemna minor. Plant Physiol 45:742–751PubMedPubMedCentralGoogle Scholar
  44. Waber J, Sakai WS (1974) Effect of growth in 99.8% deuterium oxide on ultrastructure of winter rye. Plant Physiol 53:128–130PubMedPubMedCentralGoogle Scholar
  45. Yakir D, De Niro MJ (1990) Oxygen and hydrogen isotope fractionation during cellulose metabolism in Lemna gibba L. Plant Physiol 23:325–332Google Scholar
  46. Zhao X, Elliston A, Collins SRA, Moates GK, Coleman MJ, Waldron KW (2012) Enzymatic saccharification of duckweed (Lemna minor) biomass without thermophysical pretreatment. Biomass Bioenerg 47:352–361Google Scholar
  47. Zhao X, Moates GK, Wellner N, Collins SRA, Coleman MJ, Waldron KW (2014) Chemical characterization and analysis of the cell wall polysaccharides of duckweed (Lemna minor). Carbohydr Polym 111:410–418Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Chemical Sciences DivisionOak Ridge National LaboratoryOak RidgeUSA
  2. 2.Institute of Paper Science and Technology, School of Chemistry and BiochemistryGeorgia Institute of TechnologyAtlantaUSA
  3. 3.Biology and Soft Matter DivisionOak Ridge National LaboratoryOak RidgeUSA
  4. 4.Biosciences DivisionOak Ridge National LaboratoryOak RidgeUSA
  5. 5.Bredesen Center for Interdisciplinary Research and Graduate EducationUniversity of TennesseeKnoxvilleUSA
  6. 6.Department of Energy, Environmental and Chemical EngineeringWashington UniversitySt. LouisUSA
  7. 7.School of Genome Science and Technology, F337 Walters Life ScienceUniversity of TennesseeKnoxvilleUSA
  8. 8.Sierra Vista High SchoolLas VegasUSA
  9. 9.Department of Chemical and Biomolecular EngineeringUniversity of TennesseeKnoxvilleUSA

Personalised recommendations