Rapid Assessment of Lignin Content and Structure in Switchgrass (Panicum virgatum L.) Grown Under Different Environmental Conditions

Switchgrass (Panicum virgatum L.) is a candidate feedstock in bioenergy, and plant breeding and molecular genetic strategies are being used to improve germplasm. In order to assess these subsequent modifications, baseline biomass compositional data are needed in a relevant variety of environments. In this study, switchgrass cv. Alamo was grown in the field, greenhouse, and growth chamber and harvested into individual leaf and stem tissue components. These components were analyzed with pyrolysis vapor analysis using molecular beam mass spectrometry, Fourier transform infrared, and standard wet chemistry methods to characterize and compare the composition among the different growth environments. The details of lignin content, S/G ratios, and degree of cross-linked lignin are discussed. Multivariate approaches such as projection to latent structures regression found a very strong correlation between the lignin content obtained by standard wet chemistry methods and the two high throughput techniques employed to rapidly assess lignin in potential switchgrass candidates. The models were tested on unknown samples and verified by wet chemistry. The similar lignin content found by the two methods shows that both approaches are capable of determining lignin content in biomass in a matter of minutes.

[1]  R. Dixon,et al.  The biosynthesis of monolignols: a "metabolic grid", or independent pathways to guaiacyl and syringyl units? , 2001, Phytochemistry.

[2]  Thomas A. Milne,et al.  Direct mass-spectrometric studies of the pyrolysis of carbonaceous fuels: III. Primary pyrolysis of lignin , 1986 .

[3]  Mark F. Davis,et al.  Assessment of Populus wood chemistry following the introduction of a Bt toxin gene. , 2006, Tree physiology.

[4]  F. Hileman,et al.  Pyrolysis mass spectrometry of recent and fossil biomaterials: Compendium and atlas , 1982 .

[5]  J. Frisvad,et al.  Using direct electrospray mass spectrometry in taxonomy and secondary metabolite profiling of crude fungal extracts , 1996 .

[6]  E. Shin,et al.  Characterizing biomatrix materials using pyrolysis molecular beam mass spectrometer and pattern recognition , 2003 .

[7]  R. Zhong,et al.  Essential role of caffeoyl coenzyme A O-methyltransferase in lignin biosynthesis in woody poplar plants. , 2000, Plant physiology.

[8]  Mark F. Davis,et al.  Two high-throughput techniques for determining wood properties as part of a molecular genetics analysis of hybrid poplar and loblolly pine , 1999 .

[9]  O. Schmidt,et al.  Monitoring of chemical changes in white-rot degraded beech wood by pyrolysis—gas chromatography and Fourier-transform infrared spectroscopy , 1991 .

[10]  Mariam B. Sticklen,et al.  Plant genetic engineering for biofuel production: towards affordable cellulosic ethanol , 2010, Nature Reviews Genetics.

[11]  Akwasi A. Boateng,et al.  Biomass Yield and Biofuel Quality of Switchgrass Harvested in Fall or Spring , 2006 .

[12]  T. Sonoda,et al.  Quantitative analysis of detailed lignin monomer composition by pyrolysis-gas chromatography combined with preliminary acetylation of the samples. , 2001, Analytical chemistry.

[13]  Robert A. Graybosch,et al.  Opportunities and roadblocks in utilizing forages and small grains for liquid fuels , 2008, Journal of Industrial Microbiology & Biotechnology.

[14]  Thomas A. Milne,et al.  Molecular characterization of the pyrolysis of biomass , 1987 .

[15]  Zengyu Wang,et al.  Lignin deposition and associated changes in anatomy, enzyme activity, gene expression, and ruminal degradability in stems of tall fescue at different developmental stages. , 2002, Journal of agricultural and food chemistry.

[16]  K. Law,et al.  Fibre morphology and soda-sulphite pulping of switchgrass. , 2001, Bioresource Technology.

[17]  D. Meier,et al.  Pyrolysis-Gas Chromatography-Mass Spectrometry , 1992 .

[18]  N. Carpita,et al.  A rapid method to screen for cell-wall mutants using discriminant analysis of Fourier transform infrared spectra. , 1998, The Plant journal : for cell and molecular biology.

[19]  C. Saiz-Jimenez,et al.  Pyrolysis-gas chromatography-mass spectrometry of isolated, synthetic and degraded lignins , 1984 .

[20]  Gautam Sarath,et al.  Internode structure and cell wall composition in maturing tillers of switchgrass (Panicum virgatum. L). , 2007, Bioresource technology.

[21]  D B Kell,et al.  Rapid identification of urinary tract infection bacteria using hyperspectral whole-organism fingerprinting and artificial neural networks. , 1998, Microbiology.

[22]  L. Lynd,et al.  How biotech can transform biofuels , 2008, Nature Biotechnology.

[23]  B. V. Conger,et al.  Agrobacterium‐Mediated Genetic Transformation of Switchgrass , 2002 .

[24]  Mark F. Davis,et al.  Within tree variability of lignin composition in Populus , 2008, Wood Science and Technology.

[25]  R. Perrin,et al.  Net energy of cellulosic ethanol from switchgrass , 2008, Proceedings of the National Academy of Sciences.

[26]  Robert B. Mitchell,et al.  Chemical composition and response to dilute-acid pretreatment and enzymatic saccharification of alfalfa, reed canarygrass, and switchgrass , 2006 .

[27]  H. Pereira,et al.  Determination of tree to tree variation in syringyl/guaiacyl ratio of Eucalyptus globulus wood lignin by analytical pyrolysis , 1999 .

[28]  B. V. Baar Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry. , 2000 .

[29]  C. Y. Ward,et al.  Biomass production from selected herbaceous species in the southeastern USA , 1989 .

[30]  T. Kondo,et al.  Quantitative analysis for the cellulose I alpha crystalline phase in developing wood cell walls. , 1999, International journal of biological macromolecules.

[31]  Yi Li,et al.  FT-IR imaging and pyrolysis-molecular beam mass spectrometry: new tools to investigate wood tissues , 2005, Wood Science and Technology.

[32]  H. Pereira,et al.  Influence of tree eccentric growth on syringyl/guaiacyl ratio in Eucalyptus globulus wood lignin assessed by analytical pyrolysis , 2001 .

[33]  Derek Stewart,et al.  Fourier-transform infrared and Raman spectroscopic evidence for the incorporation of cinnamaldehydes into the lignin of transgenic tobacco (Nicotiana tabacum L.) plants with reduced expression of cinnamyl alcohol dehydrogenase , 1997, Planta.

[34]  Joshua S Yuan,et al.  Plants to power: bioenergy to fuel the future. , 2008, Trends in plant science.

[35]  K. Vogel,et al.  Genotypic variability in mineral composition of switchgrass. , 2009, Bioresource technology.

[36]  D. Meier,et al.  Studies on isolated lignins and lignins in woody materials by pyrolysis-gas chromatography-mass spectrometry and off-line pyrolysis-gas chromatography with flame ionization detection , 1987 .

[37]  V. R. Tolbert,et al.  High-value renewable energy from prairie grasses. , 2002, Environmental science & technology.

[38]  Thomas A. Milne,et al.  Direct mass-spectrometric studies of the pyrolysis of carbonaceous fuels: I. A flame-pyrolysis molecular-beam sampling technique , 1983 .

[39]  B. Conger,et al.  Plant Regeneraton from Callus Cultures of Switchgrass , 1994 .

[40]  J. Kadla,et al.  Comparison of morphological and chemical properties between juvenile wood and compression wood of loblolly pine , 2005 .

[41]  G. A. Jung,et al.  Chemical Composition of Parenchyma and Sclerenchyma Cell Walls Isolated from Orchardgrass and Switchgrass , 1991 .

[42]  K. Alexandrova,et al.  Micropropagation of switchgrass by node culture. , 1996, Crop science.

[43]  K. Vogel,et al.  Lignification of switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii) plant parts during maturation and its effect on fibre degradability , 1992 .

[44]  H. Richards,et al.  Construction of a GFP-BAR plasmid and its use for switchgrass transformation , 2001, Plant Cell Reports.

[45]  Barry Goodell,et al.  Use of NIR and pyrolysis-MBMS coupled with multivariate analysis for detecting the chemical changes associated with brown-rot biodegradation of spruce wood. , 2002, FEMS microbiology letters.

[46]  T. Kondo,et al.  Quantitative analysis for the cellulose Iα crystalline phase in developing wood cell walls , 1999 .

[47]  Masato Yoshida,et al.  Relation between growth stress and lignin concentration in the cell wall: Ultraviolet microscopic spectral analysis , 1998, Journal of Wood Science.

[48]  D. I. Bransby,et al.  Biomass yield, composition and production costs for eight switchgrass varieties in Alabama , 1991 .

[49]  K. Alexandrova,et al.  In Vitro Development of Inflorescences from Switchgrass Nodal Segments , 1996 .

[50]  Thomas A. Milne,et al.  Molecular characterization of the pyrolysis of biomass. 2. Applications , 1987 .

[51]  Jeffrey F. Pedersen,et al.  Evaluation of a Filter Bag System for NDF, ADF, and IVDMD Forage Analysis , 1999 .

[52]  B. Conger,et al.  In vitro culture of switchgrass: Influence of 2,4-d and picloram in combination with benzyladenine on callus initiation and regeneration , 2004, Plant Cell, Tissue and Organ Culture.

[53]  A. Gutiérrez,et al.  Identification of residual lignin markers in eucalypt kraft pulps by Py–GC/MS , 2001 .

[54]  K. Vogel,et al.  Analysis of expressed sequence tags and the identification of associated short tandem repeats in switchgrass , 2005, Theoretical and Applied Genetics.

[55]  K. Moore,et al.  Describing and Quantifying Growth Stages of Perennial Forage Grasses , 1991 .

[56]  F. Agblevor,et al.  Molecular-beam mass-spectrometric analysis of lignocellulosic materials: I. Herbaceous biomass , 1994 .