Spectroscopy and atomic force microscopy of biomass.

PubMed

Tetard, L; Passian, A; Farahi, R H; Kalluri, U C; Davison, B H; Thundat, T

2010-05-01

Scanning probe microscopy has emerged as a powerful approach to a broader understanding of the molecular architecture of cell walls, which may shed light on the challenge of efficient cellulosic ethanol production. We have obtained preliminary images of both Populus and switchgrass samples using atomic force microscopy (AFM). The results show distinctive features that are shared by switchgrass and Populus. These features may be attributable to the lignocellulosic cell wall composition, as the collected images exhibit the characteristic macromolecular globule structures attributable to the lignocellulosic systems. Using both AFM and a single case of mode synthesizing atomic force microscopy (MSAFM) to characterize Populus, we obtained images that clearly show the cell wall structure. The results are of importance in providing a better understanding of the characteristic features of both mature cells as well as developing plant cells. In addition, we present spectroscopic investigation of the same samples.

Time course field analysis of COMT-downregulated switchgrass: Lignification, recalcitrance, and rust susceptibility

DOE PAGES

Baxter, Holly L.; Mazarei, Mitra; Fu, Chunxiang; ...

2016-05-18

Modifying plant cell walls by manipulating lignin biosynthesis can improve biofuel yields from lignocellulosic crops. For example, transgenic switchgrass lines with downregulated expression of caffeic acid O-methyltransferase, a lignin biosynthetic enzyme, produce up to 38% more ethanol than controls. The aim of the present study was to understand cell wall lignification over the second and third growing seasons of COMT-downregulated field-grown switchgrass. COMT gene expression, lignification, and cell wall recalcitrance were assayed for two independent transgenic lines at monthly intervals. Switchgrass rust (Puccinia emaculata) incidence was also tracked across the seasons. Trends in lignification over time differed between the 2more » years. In 2012, sampling was initiated in mid-growing season on reproductive-stage plants and there was little variation in the lignin content of all lines (COMT-downregulated and control) over time. COMT-downregulated lines maintained 11-16% less lignin, 33-40% lower S/G (syringyl-to-guaiacyl) ratios, and 15-42% higher sugar release relative to controls for all time points. In 2013, sampling was initiated earlier in the season on elongation-stage plants and the lignin content of all lines steadily increased over time, while sugar release expectedly decreased. S/G ratios increased in non-transgenic control plants as biomass accumulated over the season, while remaining relatively stable across the season in the COMT-downregulated lines. Differences in cell wall chemistry between transgenic and non-transgenic lines were not apparent until plants transitioned to reproductive growth in mid-season, after which the cell walls of COMT-downregulated plants exhibited phenotypes consistent with what was observed in 2012. There were no differences in rust damage between transgenics and controls at any time point. Finally, these results provide relevant fundamental insights into the process of lignification in a maturing field-grown biofuel feedstock with downregulated lignin biosynthesis.« less

Time course field analysis of COMT-downregulated switchgrass: Lignification, recalcitrance, and rust susceptibility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baxter, Holly L.; Mazarei, Mitra; Fu, Chunxiang

Modifying plant cell walls by manipulating lignin biosynthesis can improve biofuel yields from lignocellulosic crops. For example, transgenic switchgrass lines with downregulated expression of caffeic acid O-methyltransferase, a lignin biosynthetic enzyme, produce up to 38% more ethanol than controls. The aim of the present study was to understand cell wall lignification over the second and third growing seasons of COMT-downregulated field-grown switchgrass. COMT gene expression, lignification, and cell wall recalcitrance were assayed for two independent transgenic lines at monthly intervals. Switchgrass rust (Puccinia emaculata) incidence was also tracked across the seasons. Trends in lignification over time differed between the 2more » years. In 2012, sampling was initiated in mid-growing season on reproductive-stage plants and there was little variation in the lignin content of all lines (COMT-downregulated and control) over time. COMT-downregulated lines maintained 11-16% less lignin, 33-40% lower S/G (syringyl-to-guaiacyl) ratios, and 15-42% higher sugar release relative to controls for all time points. In 2013, sampling was initiated earlier in the season on elongation-stage plants and the lignin content of all lines steadily increased over time, while sugar release expectedly decreased. S/G ratios increased in non-transgenic control plants as biomass accumulated over the season, while remaining relatively stable across the season in the COMT-downregulated lines. Differences in cell wall chemistry between transgenic and non-transgenic lines were not apparent until plants transitioned to reproductive growth in mid-season, after which the cell walls of COMT-downregulated plants exhibited phenotypes consistent with what was observed in 2012. There were no differences in rust damage between transgenics and controls at any time point. Finally, these results provide relevant fundamental insights into the process of lignification in a maturing field-grown biofuel feedstock with downregulated lignin biosynthesis.« less

Selection signatures in four lignin genes from switchgrass populations divergently selected for in vitro dry matter digestibility

USDA-ARS?s Scientific Manuscript database

Switchgrass is undergoing development as a dedicated cellulosic bioenergy crop. Fermentation of lignocellulosic biomass to ethanol in a bioenergy system, or to volatile fatty acids in a livestock production system, is strongly and negatively influenced by lignification of cell walls. This study dete...

Sugar Release and Growth of Biofuel Crops are Improved by Downregulation of Pectin Biosynthesis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Donohoe, Bryon S; Sykes, Robert W; Gjersing, Erica L

Cell walls in crops and trees have been engineered for production of biofuels and commodity chemicals, but engineered varieties often fail multi-year field trials and are not commercialized. We engineered reduced expression of a pectin biosynthesis gene (Galacturonosyltransferase 4, GAUT4) in switchgrass and poplar, and find that this improves biomass yields and sugar release from biomass processing. Both traits were maintained in a 3-year field trial of GAUT4-knockdown switchgrass, with up to sevenfold increased saccharification and ethanol production and sixfold increased biomass yield compared with control plants. We show that GAUT4 is an a-1,4-galacturonosyltransferase that synthesizes homogalacturonan (HG). Downregulation ofmore » GAUT4 reduces HG and rhamnogalacturonan II (RGII), reduces wall calcium and boron, and increases extractability of cell wall sugars. Decreased recalcitrance in biomass processing and increased growth are likely due to reduced HG and RGII cross-linking in the cell wall.« less

Agave proves to be a low recalcitrant lignocellulosic feedstock for biofuels production on semi-arid lands.

PubMed

Li, Hongjia; Pattathil, Sivakumar; Foston, Marcus B; Ding, Shi-You; Kumar, Rajeev; Gao, Xiadi; Mittal, Ashutosh; Yarbrough, John M; Himmel, Michael E; Ragauskas, Arthur J; Hahn, Michael G; Wyman, Charles E

2014-01-01

Agave, which is well known for tequila and other liquor production in Mexico, has recently gained attention because of its attractive potential to launch sustainable bioenergy feedstock solutions for semi-arid and arid lands. It was previously found that agave cell walls contain low lignin and relatively diverse non-cellulosic polysaccharides, suggesting unique recalcitrant features when compared to conventional C4 and C3 plants. Here, we report sugar release data from fungal enzymatic hydrolysis of non-pretreated and hydrothermally pretreated biomass that shows agave to be much less recalcitrant to deconstruction than poplar or switchgrass. In fact, non-pretreated agave has a sugar release five to eight times greater than that of poplar wood and switchgrass . Meanwhile, state of the art techniques including glycome profiling, nuclear magnetic resonance (NMR), Simon's Stain, confocal laser scanning microscopy and so forth, were applied to measure interactions of non-cellulosic wall components, cell wall hydrophilicity, and enzyme accessibility to identify key structural features that make agave cell walls less resistant to biological deconstruction when compared to poplar and switchgrass. This study systematically evaluated the recalcitrant features of agave plants towards biofuels applications. The results show that not only does agave present great promise for feeding biorefineries on semi-arid and arid lands, but also show the value of studying agave's low recalcitrance for developments in improving cellulosic energy crops.

Comparison of four glycosyl residue composition methods for effectiveness in detecting sugars from cell walls of dicot and grass tissues.

PubMed

Biswal, Ajaya K; Tan, Li; Atmodjo, Melani A; DeMartini, Jaclyn; Gelineo-Albersheim, Ivana; Hunt, Kimberly; Black, Ian M; Mohanty, Sushree S; Ryno, David; Wyman, Charles E; Mohnen, Debra

2017-01-01

The effective use of plant biomass for biofuel and bioproduct production requires a comprehensive glycosyl residue composition analysis to understand the different cell wall polysaccharides present in the different biomass sources. Here we compared four methods side-by-side for their ability to measure the neutral and acidic sugar composition of cell walls from herbaceous, grass, and woody model plants and bioenergy feedstocks. Arabidopsis, Populus , rice, and switchgrass leaf cell walls, as well as cell walls from Populus wood, rice stems, and switchgrass tillers, were analyzed by (1) gas chromatography-mass spectrometry (GC-MS) of alditol acetates combined with a total uronic acid assay; (2) carbodiimide reduction of uronic acids followed by GC-MS of alditol acetates; (3) GC-MS of trimethylsilyl (TMS) derivatives; and (4) high-pressure, anion-exchange chromatography (HPAEC). All four methods gave comparable abundance ranking of the seven neutral sugars, and three of the methods were able to quantify unique acidic sugars. The TMS, HPAEC, and carbodiimide methods provided comparable quantitative results for the specific neutral and acidic sugar content of the biomass, with the TMS method providing slightly greater yield of specific acidic sugars and high total sugar yields. The alditol acetate method, while providing comparable information on the major neutral sugars, did not provide the requisite quantitative information on the specific acidic sugars in plant biomass. Thus, the alditol acetate method is the least informative of the four methods. This work provides a side-by-side comparison of the efficacy of four different established glycosyl residue composition analysis methods in the analysis of the glycosyl residue composition of cell walls from both dicot (Arabidopsis and Populus ) and grass (rice and switchgrass) species. Both primary wall-enriched leaf tissues and secondary wall-enriched wood/stem tissues were analyzed for mol% and mass yield of the non-cellulosic sugars. The TMS, HPAEC, and carbodiimide methods were shown to provide comparable quantitative data on the nine neutral and acidic sugars present in all plant cell walls.

Agave proves to be a low recalcitrant lignocellulosic feedstock for biofuels production on semi-arid lands

PubMed Central

2014-01-01

Background Agave, which is well known for tequila and other liquor production in Mexico, has recently gained attention because of its attractive potential to launch sustainable bioenergy feedstock solutions for semi-arid and arid lands. It was previously found that agave cell walls contain low lignin and relatively diverse non-cellulosic polysaccharides, suggesting unique recalcitrant features when compared to conventional C4 and C3 plants. Results Here, we report sugar release data from fungal enzymatic hydrolysis of non-pretreated and hydrothermally pretreated biomass that shows agave to be much less recalcitrant to deconstruction than poplar or switchgrass. In fact, non-pretreated agave has a sugar release five to eight times greater than that of poplar wood and switchgrass . Meanwhile, state of the art techniques including glycome profiling, nuclear magnetic resonance (NMR), Simon’s Stain, confocal laser scanning microscopy and so forth, were applied to measure interactions of non-cellulosic wall components, cell wall hydrophilicity, and enzyme accessibility to identify key structural features that make agave cell walls less resistant to biological deconstruction when compared to poplar and switchgrass. Conclusions This study systematically evaluated the recalcitrant features of agave plants towards biofuels applications. The results show that not only does agave present great promise for feeding biorefineries on semi-arid and arid lands, but also show the value of studying agave’s low recalcitrance for developments in improving cellulosic energy crops. PMID:24708685

Revealing the transcriptomic complexity of switchgrass by PacBio long-read sequencing.

PubMed

Zuo, Chunman; Blow, Matthew; Sreedasyam, Avinash; Kuo, Rita C; Ramamoorthy, Govindarajan Kunde; Torres-Jerez, Ivone; Li, Guifen; Wang, Mei; Dilworth, David; Barry, Kerrie; Udvardi, Michael; Schmutz, Jeremy; Tang, Yuhong; Xu, Ying

2018-01-01

Switchgrass ( Panicum virgatum L.) is an important bioenergy crop widely used for lignocellulosic research. While extensive transcriptomic analyses have been conducted on this species using short read-based sequencing techniques, very little has been reliably derived regarding alternatively spliced (AS) transcripts. We present an analysis of transcriptomes of six switchgrass tissue types pooled together, sequenced using Pacific Biosciences (PacBio) single-molecular long-read technology. Our analysis identified 105,419 unique transcripts covering 43,570 known genes and 8795 previously unknown genes. 45,168 are novel transcripts of known genes. A total of 60,096 AS transcripts are identified, 45,628 being novel. We have also predicted 1549 transcripts of genes involved in cell wall construction and remodeling, 639 being novel transcripts of known cell wall genes. Most of the predicted transcripts are validated against Illumina-based short reads. Specifically, 96% of the splice junction sites in all the unique transcripts are validated by at least five Illumina reads. Comparisons between genes derived from our identified transcripts and the current genome annotation revealed that among the gene set predicted by both analyses, 16,640 have different exon-intron structures. Overall, substantial amount of new information is derived from the PacBio RNA data regarding both the transcriptome and the genome of switchgrass.

Surface and ultrastructural characterization of raw and pretreated switchgrass.

PubMed

Donohoe, Bryon S; Vinzant, Todd B; Elander, Richard T; Pallapolu, Venkata Ramesh; Lee, Y Y; Garlock, Rebecca J; Balan, Venkatesh; Dale, Bruce E; Kim, Youngmi; Mosier, Nathan S; Ladisch, Michael R; Falls, Matthew; Holtzapple, Mark T; Sierra-Ramirez, Rocio; Shi, Jian; Ebrik, Mirvat A; Redmond, Tim; Yang, Bin; Wyman, Charles E; Hames, Bonnie; Thomas, Steve; Warner, Ryan E

2011-12-01

The US Department of Energy-funded Biomass Refining CAFI (Consortium for Applied Fundamentals and Innovation) project has developed leading pretreatment technologies for application to switchgrass and has evaluated their effectiveness in recovering sugars from the coupled operations of pretreatment and enzymatic hydrolysis. Key chemical and physical characteristics have been determined for pretreated switchgrass samples. Several analytical microscopy approaches utilizing instruments in the Biomass Surface Characterization Laboratory (BSCL) at the National Renewable Energy Laboratory (NREL) have been applied to untreated and CAFI-pretreated switchgrass samples. The results of this work have shown that each of the CAFI pretreatment approaches on switchgrass result in different structural impacts at the plant tissue, cellular, and cell wall levels. Some of these structural changes can be related to changes in chemical composition upon pretreatment. There are also apparently different structural mechanisms that are responsible for achieving the highest enzymatic hydrolysis sugar yields. Copyright © 2011. Published by Elsevier Ltd.

Field-grown miR156 transgenic switchgrass reproduction, yield, global gene expression analysis, and bioconfinement

DOE Office of Scientific and Technical Information (OSTI.GOV)

Johnson, Chelsea R.; Millwood, Reginald J.; Tang, Yuhong

Genetic engineering has been effective in altering cell walls for biofuel production in the bioenergy crop, switchgrass (Panicum virgatum). However, regulatory issues arising from gene flow may prevent commercialization of engineered switchgrass in the eastern United States where the species is native. And, depending on its expression level, microRNA156 (miR156) can reduce, delay, or eliminate flowering, which may serve to decrease transgene flow. Here, in this unique field study of transgenic switchgrass that was permitted to flower, two low (T14 and T35) and two medium (T27 and T37) miR156-overexpressing 'Alamo' lines with the transgene under the control of the constitutivemore » maize (Zea mays) ubiquitin 1 promoter, along with nontransgenic control plants, were grown in eastern Tennessee over two seasons.« less

Field-grown miR156 transgenic switchgrass reproduction, yield, global gene expression analysis, and bioconfinement

DOE PAGES

Johnson, Chelsea R.; Millwood, Reginald J.; Tang, Yuhong; ...

2017-11-30

Genetic engineering has been effective in altering cell walls for biofuel production in the bioenergy crop, switchgrass (Panicum virgatum). However, regulatory issues arising from gene flow may prevent commercialization of engineered switchgrass in the eastern United States where the species is native. And, depending on its expression level, microRNA156 (miR156) can reduce, delay, or eliminate flowering, which may serve to decrease transgene flow. Here, in this unique field study of transgenic switchgrass that was permitted to flower, two low (T14 and T35) and two medium (T27 and T37) miR156-overexpressing 'Alamo' lines with the transgene under the control of the constitutivemore » maize (Zea mays) ubiquitin 1 promoter, along with nontransgenic control plants, were grown in eastern Tennessee over two seasons.« less

Production of deuterated switchgrass by hydroponic cultivation

DOE PAGES

Evans, Barbara R.; Bali, Garima; Foston, Marcus B.; ...

2015-04-21

Deuterium enrichment of biological materials can potential enable expanded experimental use of small angle neutron scattering (SANS) to investigate molecular structural transitions of complex systems such as plant cell walls. Two key advances have been made that facilitate cultivation of switchgrass, an important forage and biofuel crop, for controlled isotopic enrichment: (1) perfusion system with individual chambers and (2) hydroponic growth from tiller cuttings. Plants were grown and maintained for several months with periodic harvest. Photosynthetic activity was monitored by measurement of CO 2 in outflow from the growth chambers. Plant morphology and composition appeared normal compared to matched controlsmore » grown with H 2O. Using this improved method, gram quantities of switchgrass leaves and stems were produced by continuous hydroponic cultivation using growth medium consisting of basal mineral salts in 50% D 2O. Deuterium incorporation was confirmed by detection of the O-D and C-D stretching peaks with FTIR and quantified by 1H- and 2H-NMR. Lastly, this capability to produce deuterated lignocellulosic biomass under controlled conditions will enhance investigation of cell wall structure and its deconstruction by neutron scattering and NMR techniques.« less

Cell Wall Composition and Underlying QTL in an F1 Pseudo-Testcross Population of Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Serba, Desalegn D.; Sykes, Robert W.; Gjersing, Erica L.

Natural genetic variation for reduced recalcitrance can be used to improve switchgrass for biofuel production. A full-sib switchgrass mapping population developed by crossing a lowland genotype, AP13, and upland genotype, VS16, was evaluated at three locations (Ardmore and Burneyville, OK and Watkinsville, GA). Biomass harvested after senescence in 2009 and 2010 was evaluated at the National Renewable Energy Laboratory (NREL) for sugar release using enzymatic hydrolysis and for lignin content and syringyl/guaiacyl lignin monomer (S/G) ratio using pyrolysis molecular beam mass spectrometry (py-MBMS). Glucose and xylose release ranged from 120 to 313 and 123 to 263 mg g -1, respectively,more » while lignin content ranged from 19 to 27% of the dry biomass. Statistically significant differences were observed among the genotypes and the environments for the cell wall composition traits. Regression analysis showed that a unit increase in lignin content reduced total sugar release by an average of 10 mg g -1. Quantitative trait loci (QTL) analysis detected 9 genomic regions underlying sugar release and 14 for lignin content. The phenotypic variation explained by the individual QTL identified for sugar release ranged from 4.5 to 9.4 and for lignin content from 3.8 to 11.1%. Mapping of the QTL regions to the switchgrass genome sequence (v1.1) found that some of the QTL colocalized with genes involved in carbohydrate processing and metabolism, plant development, defense systems, and transcription factors. Finally, the markers associated with QTL can be implemented in breeding programs to efficiently develop improved switchgrass cultivars for biofuel production.« less

Cell Wall Composition and Underlying QTL in an F1 Pseudo-Testcross Population of Switchgrass

DOE PAGES

Serba, Desalegn D.; Sykes, Robert W.; Gjersing, Erica L.; ...

2016-04-23

Natural genetic variation for reduced recalcitrance can be used to improve switchgrass for biofuel production. A full-sib switchgrass mapping population developed by crossing a lowland genotype, AP13, and upland genotype, VS16, was evaluated at three locations (Ardmore and Burneyville, OK and Watkinsville, GA). Biomass harvested after senescence in 2009 and 2010 was evaluated at the National Renewable Energy Laboratory (NREL) for sugar release using enzymatic hydrolysis and for lignin content and syringyl/guaiacyl lignin monomer (S/G) ratio using pyrolysis molecular beam mass spectrometry (py-MBMS). Glucose and xylose release ranged from 120 to 313 and 123 to 263 mg g -1, respectively,more » while lignin content ranged from 19 to 27% of the dry biomass. Statistically significant differences were observed among the genotypes and the environments for the cell wall composition traits. Regression analysis showed that a unit increase in lignin content reduced total sugar release by an average of 10 mg g -1. Quantitative trait loci (QTL) analysis detected 9 genomic regions underlying sugar release and 14 for lignin content. The phenotypic variation explained by the individual QTL identified for sugar release ranged from 4.5 to 9.4 and for lignin content from 3.8 to 11.1%. Mapping of the QTL regions to the switchgrass genome sequence (v1.1) found that some of the QTL colocalized with genes involved in carbohydrate processing and metabolism, plant development, defense systems, and transcription factors. Finally, the markers associated with QTL can be implemented in breeding programs to efficiently develop improved switchgrass cultivars for biofuel production.« less

Identification and overexpression of a knotted1-like transcription factor in switchgrass ( Panicum virgatum L.) for lignocellulosic feedstock improvement

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wuddineh, Wegi A.; Mazarei, Mitra; Zhang, Ji -Yi

High biomass production and wide adaptation has made switchgrass ( Panicum virgatum L.) an important candidate lignocellulosic bioenergy crop. One major limitation of this and other lignocellulosic feedstocks is the recalcitrance of complex carbohydrates to hydrolysis for conversion to biofuels. Lignin is the major contributor to recalcitrance as it limits the accessibility of cell wall carbohydrates to enzymatic breakdown into fermentable sugars. Therefore, genetic manipulation of the lignin biosynthesis pathway is one strategy to reduce recalcitrance. Here, we identified a switchgrass Knotted1 transcription factor, PvKN1, with the aim of genetically engineering switchgrass for reduced biomass recalcitrance for biofuel production. Genemore » expression of the endogenous PvKN1 gene was observed to be highest in young inflorescences and stems. Ectopic overexpression of PvKN1 in switchgrass altered growth, especially in early developmental stages. Transgenic lines had reduced expression of most lignin biosynthetic genes accompanied by a reduction in lignin content suggesting the involvement of PvKN1 in the broad regulation of the lignin biosynthesis pathway. Moreover, the reduced expression of the Gibberellin 20-oxidase (GA20ox) gene in tandem with the increased expression of Gibberellin 2-oxidase (GA2ox) genes in transgenic PvKN1 lines suggest that PvKN1 may exert regulatory effects via modulation of GA signaling. Furthermore, overexpression of PvKN1 altered the expression of cellulose and hemicellulose biosynthetic genes and increased sugar release efficiency in transgenic lines. Our findings demonstrated that switchgrass PvKN1 is a putative ortholog of maize KN1 that is linked to plant lignification and cell wall and development traits as a major regulatory gene. Therefore, targeted overexpression of PvKN1 in bioenergy feedstocks may provide one feasible strategy for reducing biomass recalcitrance and simultaneously improving plant growth characteristics.« less

Identification and overexpression of a knotted1-like transcription factor in switchgrass ( Panicum virgatum L.) for lignocellulosic feedstock improvement

DOE PAGES

Wuddineh, Wegi A.; Mazarei, Mitra; Zhang, Ji -Yi; ...

2016-04-28

High biomass production and wide adaptation has made switchgrass ( Panicum virgatum L.) an important candidate lignocellulosic bioenergy crop. One major limitation of this and other lignocellulosic feedstocks is the recalcitrance of complex carbohydrates to hydrolysis for conversion to biofuels. Lignin is the major contributor to recalcitrance as it limits the accessibility of cell wall carbohydrates to enzymatic breakdown into fermentable sugars. Therefore, genetic manipulation of the lignin biosynthesis pathway is one strategy to reduce recalcitrance. Here, we identified a switchgrass Knotted1 transcription factor, PvKN1, with the aim of genetically engineering switchgrass for reduced biomass recalcitrance for biofuel production. Genemore » expression of the endogenous PvKN1 gene was observed to be highest in young inflorescences and stems. Ectopic overexpression of PvKN1 in switchgrass altered growth, especially in early developmental stages. Transgenic lines had reduced expression of most lignin biosynthetic genes accompanied by a reduction in lignin content suggesting the involvement of PvKN1 in the broad regulation of the lignin biosynthesis pathway. Moreover, the reduced expression of the Gibberellin 20-oxidase (GA20ox) gene in tandem with the increased expression of Gibberellin 2-oxidase (GA2ox) genes in transgenic PvKN1 lines suggest that PvKN1 may exert regulatory effects via modulation of GA signaling. Furthermore, overexpression of PvKN1 altered the expression of cellulose and hemicellulose biosynthetic genes and increased sugar release efficiency in transgenic lines. Our findings demonstrated that switchgrass PvKN1 is a putative ortholog of maize KN1 that is linked to plant lignification and cell wall and development traits as a major regulatory gene. Therefore, targeted overexpression of PvKN1 in bioenergy feedstocks may provide one feasible strategy for reducing biomass recalcitrance and simultaneously improving plant growth characteristics.« less

Switchgrass and Giant Miscanthus Biomass and Theoretical Ethanol Production from Reclaimed Mine Lands

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wolfrum, Edward J; Scagline-Mellor, Steffany; Griggs, Thomas

Switchgrass (Panicum virgatum L.) and giant miscanthus (Miscanthus x giganteus Greef & Deuter ex Hodkinson & Renvoize) are productive on marginal lands in the eastern USA, but their productivity and composition have not been compared on mine lands. Our objectives were to compare biomass production, composition, and theoretical ethanol yield (TEY) and production (TEP) of these grasses on a reclaimed mined site. Following 25 years of herbaceous cover, vegetation was killed and plots of switchgrass cultivars Kanlow and BoMaster and miscanthus lines Illinois and MBX-002 were planted in five replications. Annual switchgrass and miscanthus yields averaged 5.8 and 8.9 Mgmore » dry matter ha-1, respectively, during 2011 to 2015. Cell wall carbohydrate composition was analyzed via near-infrared reflectance spectroscopy with models based on switchgrass or mixed herbaceous samples including switchgrass and miscanthus. Concentrations were higher for glucan and lower for xylan in miscanthus than in switchgrass but TEY did not differ (453 and 450 L Mg-1, respectively). In response to biomass production, total ethanol production was greater for miscanthus than for switchgrass (5594 vs 3699 L ha-1), did not differ between Kanlow and BoMaster switchgrass (3880 and 3517 L ha-1, respectively), and was higher for MBX-002 than for Illinois miscanthus (6496 vs 4692 L ha-1). Relative to the mixed feedstocks model, the switchgrass model slightly underpredicted glucan and slightly overpredicted xylan concentrations. Estimated TEY was slightly lower from the switchgrass model but both models distinguished genotype, year, and interaction effects similarly. Biomass productivity and TEP were similar to those from agricultural sites with marginal soils.« less

Monitoring Process Streams Towards Understanding Ionic Liquid Pretreatment of Switchgrass and Corn Stover

USDA-ARS?s Scientific Manuscript database

Pretreatment of Biomass is essential for breaking apart highly ordered and crystalline plant cell walls and loosening the lignin and hemicellulose conjugation to cellulose microfibrils, thereby facilitating enzyme accessibility and adsorption and reducing cotsts of downstream saccharification proces...

Evaluation of parameters affecting switchgrass tissue culture: toward a consolidated procedure for Agrobacterium-mediated transformation of switchgrass (Panicum virgatum)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Chien-Yuan; Donohoe, Bryon S.; Ahuja, Neha

Switchgrass (Panicum virgatum), a robust perennial C4-type grass, has been evaluated and designated as a model bioenergy crop by the U.S. DOE and USDA. Conventional breeding of switchgrass biomass is difficult because it displays self-incompatible hindrance. Therefore, direct genetic modifications of switchgrass have been considered the more effective approach to tailor switchgrass with traits of interest. Successful transformations have demonstrated increased biomass yields, reduction in the recalcitrance of cell walls and enhanced saccharification efficiency. Several tissue culture protocols have been previously described to produce transgenic switchgrass lines using different nutrient-based media, co-cultivation approaches, and antibiotic strengths for selection. After evaluatingmore » the published protocols, we consolidated these approaches and optimized the process to develop a more efficient protocol for producing transgenic switchgrass. First, seed sterilization was optimized, which led to a 20% increase in yield of induced calluses. Second, we have selected a N 6 macronutrient/B 5 micronutrient (NB)-based medium for callus induction from mature seeds of the Alamo cultivar, and chose a Murashige and Skoog-based medium to regenerate both Type I and Type II calluses. Third, Agrobacterium-mediated transformation was adopted that resulted in 50-100% positive regenerated transformants after three rounds (2 weeks/round) of selection with antibiotic. Genomic DNA PCR, RT-PCR, Southern blot, visualization of the red fluorescent protein and histochemical β-glucuronidase (GUS) staining were conducted to confirm the positive switchgrass transformants. The optimized methods developed here provide an improved strategy to promote the production and selection of callus and generation of transgenic switchgrass lines. The process for switchgrass transformation has been evaluated and consolidated to devise an improved approach for transgenic switchgrass production. With the optimization of seed sterilization, callus induction, and regeneration steps, a reliable and effective protocol is established to facilitate switchgrass engineering.« less

Evaluation of parameters affecting switchgrass tissue culture: toward a consolidated procedure for Agrobacterium-mediated transformation of switchgrass (Panicum virgatum)

DOE PAGES

Lin, Chien-Yuan; Donohoe, Bryon S.; Ahuja, Neha; ...

2017-12-19

Switchgrass (Panicum virgatum), a robust perennial C4-type grass, has been evaluated and designated as a model bioenergy crop by the U.S. DOE and USDA. Conventional breeding of switchgrass biomass is difficult because it displays self-incompatible hindrance. Therefore, direct genetic modifications of switchgrass have been considered the more effective approach to tailor switchgrass with traits of interest. Successful transformations have demonstrated increased biomass yields, reduction in the recalcitrance of cell walls and enhanced saccharification efficiency. Several tissue culture protocols have been previously described to produce transgenic switchgrass lines using different nutrient-based media, co-cultivation approaches, and antibiotic strengths for selection. After evaluatingmore » the published protocols, we consolidated these approaches and optimized the process to develop a more efficient protocol for producing transgenic switchgrass. First, seed sterilization was optimized, which led to a 20% increase in yield of induced calluses. Second, we have selected a N 6 macronutrient/B 5 micronutrient (NB)-based medium for callus induction from mature seeds of the Alamo cultivar, and chose a Murashige and Skoog-based medium to regenerate both Type I and Type II calluses. Third, Agrobacterium-mediated transformation was adopted that resulted in 50-100% positive regenerated transformants after three rounds (2 weeks/round) of selection with antibiotic. Genomic DNA PCR, RT-PCR, Southern blot, visualization of the red fluorescent protein and histochemical β-glucuronidase (GUS) staining were conducted to confirm the positive switchgrass transformants. The optimized methods developed here provide an improved strategy to promote the production and selection of callus and generation of transgenic switchgrass lines. The process for switchgrass transformation has been evaluated and consolidated to devise an improved approach for transgenic switchgrass production. With the optimization of seed sterilization, callus induction, and regeneration steps, a reliable and effective protocol is established to facilitate switchgrass engineering.« less

Breakdown of cell wall nanostructure in dilute acid pretreated biomass.

PubMed

Pingali, Sai Venkatesh; Urban, Volker S; Heller, William T; McGaughey, Joseph; O'Neill, Hugh; Foston, Marcus; Myles, Dean A; Ragauskas, Arthur; Evans, Barbara R

2010-09-13

The generation of bioethanol from lignocellulosic biomass holds great promise for renewable and clean energy production. A better understanding of the complex mechanisms of lignocellulose breakdown during various pretreatment methods is needed to realize this potential in a cost and energy efficient way. Here we use small-angle neutron scattering (SANS) to characterize morphological changes in switchgrass lignocellulose across molecular to submicrometer length scales resulting from the industrially relevant dilute acid pretreatment method. Our results demonstrate that dilute acid pretreatment increases the cross-sectional radius of the crystalline cellulose fibril. This change is accompanied by removal of hemicellulose and the formation of R(g) ∼ 135 A lignin aggregates. The structural signature of smooth cell wall surfaces is observed at length scales larger than 1000 A, and it remains remarkably invariable during pretreatment. This study elucidates the interplay of the different biomolecular components in the breakdown process of switchgrass by dilute acid pretreatment. The results are important for the development of efficient strategies of biomass to biofuel conversion.

Quantitative trait loci for cell wall composition traits measured using near-infrared spectroscopy in the model C4 perennial grass Panicum hallii

DOE PAGES

Milano, Elizabeth R.; Payne, Courtney E.; Wolfrum, Edward J.; ...

2018-02-03

Biofuels derived from lignocellulosic plant material are an important component of current renewable energy strategies. Improvement efforts in biofuel feedstock crops have been primarily focused on increasing biomass yield with less consideration for tissue quality or composition. Four primary components found in the plant cell wall contribute to the overall quality of plant tissue and conversion characteristics, cellulose and hemicellulose polysaccharides are the primary targets for fuel conversion, while lignin and ash provide structure and defense. We explore the genetic architecture of tissue characteristics using a quantitative trait loci (QTL) mapping approach in Panicum hallii, a model lignocellulosic grass system.more » Diversity in the mapping population was generated by crossing xeric and mesic varietals, comparative to northern upland and southern lowland ecotypes in switchgrass. We use near-infrared spectroscopy with a primary analytical method to create a P. hallii specific calibration model to quickly quantify cell wall components. Ash, lignin, glucan, and xylan comprise 68% of total dry biomass in P. hallii: comparable to other feedstocks. We identified 14 QTL and one epistatic interaction across these four cell wall traits and found almost half of the QTL to localize to a single linkage group. Panicum hallii serves as the genomic model for its close relative and emerging biofuel crop, switchgrass (P. virgatum). We used high throughput phenotyping to map genomic regions that impact natural variation in leaf tissue composition. Understanding the genetic architecture of tissue traits in a tractable model grass system will lead to a better understanding of cell wall structure as well as provide genomic resources for bioenergy crop breeding programs.« less

Quantitative trait loci for cell wall composition traits measured using near-infrared spectroscopy in the model C4 perennial grass Panicum hallii

DOE Office of Scientific and Technical Information (OSTI.GOV)

Milano, Elizabeth R.; Payne, Courtney E.; Wolfrum, Edward J.

Biofuels derived from lignocellulosic plant material are an important component of current renewable energy strategies. Improvement efforts in biofuel feedstock crops have been primarily focused on increasing biomass yield with less consideration for tissue quality or composition. Four primary components found in the plant cell wall contribute to the overall quality of plant tissue and conversion characteristics, cellulose and hemicellulose polysaccharides are the primary targets for fuel conversion, while lignin and ash provide structure and defense. We explore the genetic architecture of tissue characteristics using a quantitative trait loci (QTL) mapping approach in Panicum hallii, a model lignocellulosic grass system.more » Diversity in the mapping population was generated by crossing xeric and mesic varietals, comparative to northern upland and southern lowland ecotypes in switchgrass. We use near-infrared spectroscopy with a primary analytical method to create a P. hallii specific calibration model to quickly quantify cell wall components. Ash, lignin, glucan, and xylan comprise 68% of total dry biomass in P. hallii: comparable to other feedstocks. We identified 14 QTL and one epistatic interaction across these four cell wall traits and found almost half of the QTL to localize to a single linkage group. Panicum hallii serves as the genomic model for its close relative and emerging biofuel crop, switchgrass (P. virgatum). We used high throughput phenotyping to map genomic regions that impact natural variation in leaf tissue composition. Understanding the genetic architecture of tissue traits in a tractable model grass system will lead to a better understanding of cell wall structure as well as provide genomic resources for bioenergy crop breeding programs.« less

Chemical depolymerization of switchgrass xylan with oligosaccharide product analysis by HPAEC-PAD and mass spectrometry

USDA-ARS?s Scientific Manuscript database

Xylan is a barrier to enzymatic hydrolysis of plant cell walls. It is well accepted that the xylan layer needs to be removed to efficiently hydrolyze cellulose and consequently pretreatment conditions are in part optimized for maximal xylan depolymerization or displacement. Xylan consists of a lon...

Transgenic switchgrass ( Panicum virgatum L.) targeted for reduced recalcitrance to bioconversion: A two-year comparative analysis of field-grown lines modified for target gene or genetic element expression

DOE PAGES

Dumitrache, Alexandru; Natzke, Jace; Rodriguez, Jr., Miguel; ...

2016-11-18

Five different types of transgenic ( GAUT4, miRNA, MYB4, COMT and FPGS) Panicum virgatum L. (switchgrass) were grown in a field in Knoxville, Tenn., USA over two consecutive years between 2011 and 2015 in separate experiments. Clonal replicates were established (year-one) and produced much greater biomass during the second year. After each growing season the above ground biomass was analyzed for cell wall sugars and for recalcitrance to enzymatic digestibility, and biofuel using a separate hydrolysis and fermentation (SHF) screen. Here, each transgenic event and control had more glucan, xylan and less ethanol (g/g basis) from the second year ofmore » growth relative to the first year plants. There was no correlation between plant carbohydrate content and biofuel production. In each of cell wall-targeted transgenics, GAUT4, MYB4, COMT and FPGS, the second year of growth resulted in increased carbohydrate abundance (up to 12%) and reduced recalcitrance through higher ethanol yields (up to 21%) over the non-transgenic control plants.« less

Transgenic switchgrass ( Panicum virgatum L.) targeted for reduced recalcitrance to bioconversion: A two-year comparative analysis of field-grown lines modified for target gene or genetic element expression

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dumitrache, Alexandru; Natzke, Jace; Rodriguez, Jr., Miguel

Five different types of transgenic ( GAUT4, miRNA, MYB4, COMT and FPGS) Panicum virgatum L. (switchgrass) were grown in a field in Knoxville, Tenn., USA over two consecutive years between 2011 and 2015 in separate experiments. Clonal replicates were established (year-one) and produced much greater biomass during the second year. After each growing season the above ground biomass was analyzed for cell wall sugars and for recalcitrance to enzymatic digestibility, and biofuel using a separate hydrolysis and fermentation (SHF) screen. Here, each transgenic event and control had more glucan, xylan and less ethanol (g/g basis) from the second year ofmore » growth relative to the first year plants. There was no correlation between plant carbohydrate content and biofuel production. In each of cell wall-targeted transgenics, GAUT4, MYB4, COMT and FPGS, the second year of growth resulted in increased carbohydrate abundance (up to 12%) and reduced recalcitrance through higher ethanol yields (up to 21%) over the non-transgenic control plants.« less

Selective chemical oxidation and depolymerization of switchgrass (Panicum virgatum L.) xylan with oligosaccharide product analysis by mass spectrometry

USDA-ARS?s Scientific Manuscript database

Xylan is a barrier to enzymatic hydrolysis of plant cell walls. It is well accepted that the xylan layer needs to be removed to efficiently hydrolyze cellulose and consequently pretreatment conditions are in part optimized for maximal xylan depolymerization or displacement. Xylan consists of a long ...

Targeted discovery of glycoside hydrolases from a switchgrass-adapted compost community

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allgaier, M.; Reddy, A.; Park, J. I.

2009-11-15

Development of cellulosic biofuels from non-food crops is currently an area of intense research interest. Tailoring depolymerizing enzymes to particular feedstocks and pretreatment conditions is one promising avenue of research in this area. Here we added a green-waste compost inoculum to switchgrass (Panicum virgatum) and simulated thermophilic composting in a bioreactor to select for a switchgrass-adapted community and to facilitate targeted discovery of glycoside hydrolases. Small-subunit (SSU) rRNA-based community profiles revealed that the microbial community changed dramatically between the initial and switchgrass-adapted compost (SAC) with some bacterial populations being enriched over 20-fold. We obtained 225 Mbp of 454-titanium pyrosequence datamore » from the SAC community and conservatively identified 800 genes encoding glycoside hydrolase domains that were biased toward depolymerizing grass cell wall components. Of these, {approx}10% were putative cellulases mostly belonging to families GH5 and GH9. We synthesized two SAC GH9 genes with codon optimization for heterologous expression in Escherichia coli and observed activity for one on carboxymethyl cellulose. The active GH9 enzyme has a temperature optimum of 50 C and pH range of 5.5 to 8 consistent with the composting conditions applied. We demonstrate that microbial communities adapt to switchgrass decomposition using simulated composting condition and that full-length genes can be identified from complex metagenomic sequence data, synthesized and expressed resulting in active enzyme.« less

Targeted Discovery of Glycoside Hydrolases from a Switchgrass-Adapted Compost Community

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reddy, Amitha; Allgaier, Martin; Park, Joshua I.

2011-05-11

Development of cellulosic biofuels from non-food crops is currently an area of intense research interest. Tailoring depolymerizing enzymes to particular feedstocks and pretreatment conditions is one promising avenue of research in this area. Here we added a green-waste compost inoculum to switchgrass (Panicum virgatum) and simulated thermophilic composting in a bioreactor to select for a switchgrass-adapted community and to facilitate targeted discovery of glycoside hydrolases. Smallsubunit (SSU) rRNA-based community profiles revealed that the microbial community changed dramatically between the initial and switchgrass-adapted compost (SAC) with some bacterial populations being enriched over 20-fold. We obtained 225 Mbp of 454-titanium pyrosequence datamore » from the SAC community and conservatively identified 800 genes encoding glycoside hydrolase domains that were biased toward depolymerizing grass cell wall components. Of these, ,10percent were putative cellulasesmostly belonging to families GH5 and GH9. We synthesized two SAC GH9 genes with codon optimization for heterologous expression in Escherichia coli and observed activity for one on carboxymethyl cellulose. The active GH9 enzyme has a temperature optimum of 50uC and pH range of 5.5 to 8 consistent with the composting conditions applied. We demonstrate that microbial communities adapt to switchgrass decomposition using simulated composting condition and that full-length genes can be identified from complex metagenomic sequence data, synthesized and expressed resulting in active enzyme.« less

Selection signatures in four lignin genes from switchgrass populations divergently selected for in vitro dry matter digestibility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Shiyu; Kaeppler, Shawn M.; Vogel, Kenneth P.

Switchgrass is undergoing development as a dedicated cellulosic bioenergy crop. Fermentation of lignocellulosic biomass to ethanol in a bioenergy system or to volatile fatty acids in a livestock production system is strongly and negatively influenced by lignification of cell walls. This study detects specific loci that exhibit selection signatures across switchgrass breeding populations that differ in in vitro dry matter digestibility (IVDMD), ethanol yield, and lignin concentration. Allele frequency changes in candidate genes were used to detect loci under selection. Out of the 183 polymorphisms identified in the four candidate genes, twenty-five loci in the intron regions and four locimore » in coding regions were found to display a selection signature. All loci in the coding regions are synonymous substitutions. Selection in both directions were observed on polymorphisms that appeared to be under selection. Genetic diversity and linkage disequilibrium within the candidate genes were low. The recurrent divergent selection caused excessive moderate allele frequencies in the cycle 3 reduced lignin population as compared to the base population. As a result, this study provides valuable insight on genetic changes occurring in short-term selection in the polyploid populations, and discovered potential markers for breeding switchgrass with improved biomass quality.« less

Selection signatures in four lignin genes from switchgrass populations divergently selected for in vitro dry matter digestibility

DOE PAGES

Chen, Shiyu; Kaeppler, Shawn M.; Vogel, Kenneth P.; ...

2016-11-28

Switchgrass is undergoing development as a dedicated cellulosic bioenergy crop. Fermentation of lignocellulosic biomass to ethanol in a bioenergy system or to volatile fatty acids in a livestock production system is strongly and negatively influenced by lignification of cell walls. This study detects specific loci that exhibit selection signatures across switchgrass breeding populations that differ in in vitro dry matter digestibility (IVDMD), ethanol yield, and lignin concentration. Allele frequency changes in candidate genes were used to detect loci under selection. Out of the 183 polymorphisms identified in the four candidate genes, twenty-five loci in the intron regions and four locimore » in coding regions were found to display a selection signature. All loci in the coding regions are synonymous substitutions. Selection in both directions were observed on polymorphisms that appeared to be under selection. Genetic diversity and linkage disequilibrium within the candidate genes were low. The recurrent divergent selection caused excessive moderate allele frequencies in the cycle 3 reduced lignin population as compared to the base population. As a result, this study provides valuable insight on genetic changes occurring in short-term selection in the polyploid populations, and discovered potential markers for breeding switchgrass with improved biomass quality.« less

Co-firing switchgrass in a 50 MW pulverized coal boiler

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ragland, K.W.; Aerts, D.J.; Weiss, C.

1996-12-31

Switchgrass is being co-fired with pulverized coal in a 50 MW wall-fired, radiant boiler at MG&E`s Blount Street generating station. Shredded switchgrass is fed to a hammermill onto a live bottom storage bunker. Twin screw augers move the switchgrass onto a belt conveyor which leads to a rotary air lock valve and through a pressurized pipe to the boiler where it is injected into the furnace at two points between first and second level coal burners. The main objective of the project is to evaluate the boiler performance, slagging behavior, and emissions of the co-fired switchgrass at replacement of ratesmore » of up to 20% by mass (13% by heat input). Initial co-firing tests to examine fuel handling and feeding, combustion behavior, boiler response and emissions are favorable. In November a 100 hour co-fire test is planned.« less

Field Evaluation of Transgenic Switchgrass Plants Overexpressing PvMYB4 for Reduced Biomass Recalcitrance

DOE PAGES

Baxter, Holly L.; Poovaiah, Charleson R.; Yee, Kelsey L.; ...

2015-01-07

High biomass yields and minimal agronomic input requirements have made switchgrass, Panicum virgatum L., a leading candidate lignocellulosic bioenergy crop. Large-scale lignocellulosic biofuel production from such crops is limited by the difficulty to deconstruct cell walls into fermentable sugars: the recalcitrance problem. In this study, we assessed the field performance of switchgrass plants overexpressing the switchgrass MYB 4 ( PvMYB4) transcription factor gene. PvMYB 4 transgenic switchgrass can have great lignin reduction, which commensurately increases sugar release and biofuel production. Our results over two growing seasons showed that one transgenic event (out of eight) had important gains in both biofuelmore » (32% more) and biomass (63% more) at the end of the second growing season relative to non-transgenic controls. These gains represent a doubling of biofuel production per hectare, which is the highest gain reported from any field-grown modified feedstock. In contrast to this transgenic event, which had relatively low ectopic overexpression of the transgene, five of the eight transgenic events planted did not survive the first field winter. The dead plants were all high-overexpressing events that performed well in the earlier greenhouse studies. Disease susceptibility was not compromised in any transgenic events over the field experiments. These results demonstrate the power of modifying the expression of an endogenous transcription factor to improve biofuel and biomass simultaneously, and also highlight the importance of field studies for "sorting" transgenic events. In conclusion, further research is needed to develop strategies for fine-tuning temporal-spatial transgene expression in feedstocks to optimize desired phenotypes.« less

Field Evaluation of Transgenic Switchgrass Plants Overexpressing PvMYB4 for Reduced Biomass Recalcitrance

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baxter, Holly L.; Poovaiah, Charleson R.; Yee, Kelsey L.

High biomass yields and minimal agronomic input requirements have made switchgrass, Panicum virgatum L., a leading candidate lignocellulosic bioenergy crop. Large-scale lignocellulosic biofuel production from such crops is limited by the difficulty to deconstruct cell walls into fermentable sugars: the recalcitrance problem. In this study, we assessed the field performance of switchgrass plants overexpressing the switchgrass MYB 4 ( PvMYB4) transcription factor gene. PvMYB 4 transgenic switchgrass can have great lignin reduction, which commensurately increases sugar release and biofuel production. Our results over two growing seasons showed that one transgenic event (out of eight) had important gains in both biofuelmore » (32% more) and biomass (63% more) at the end of the second growing season relative to non-transgenic controls. These gains represent a doubling of biofuel production per hectare, which is the highest gain reported from any field-grown modified feedstock. In contrast to this transgenic event, which had relatively low ectopic overexpression of the transgene, five of the eight transgenic events planted did not survive the first field winter. The dead plants were all high-overexpressing events that performed well in the earlier greenhouse studies. Disease susceptibility was not compromised in any transgenic events over the field experiments. These results demonstrate the power of modifying the expression of an endogenous transcription factor to improve biofuel and biomass simultaneously, and also highlight the importance of field studies for "sorting" transgenic events. In conclusion, further research is needed to develop strategies for fine-tuning temporal-spatial transgene expression in feedstocks to optimize desired phenotypes.« less

Using The Corngrass1 Gene To Enhance The Biofuel Properties Of Crop Plants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hake, Sarah; Chuck, George

2015-10-29

The development of novel plant germplasm is vital to addressing our increasing bioenergy demands. The major hurdle to digesting plant biomass is the complex structure of the cell walls, the substrate of fermentation. Plant cell walls are inaccessible matrices of macromolecules that are polymerized with lignin, making fermentation difficult. Overcoming this hurdle is a major goal toward developing usable bioenergy crop plants. Our project seeks to enhance the biofuel properties of perennial grass species using the Corngrass1 (Cg1) gene and its targets. Dominant maize Cg1 mutants produce increased biomass by continuously initiating extra axillary meristems and leaves. We cloned Cg1more » and showed that its phenotype is caused by over expression of a unique miR156 microRNA gene that negatively regulates SPL transcription factors. We transferred the Cg1 phenotype to other plants by expressing the gene behind constitutive promoters in four different species, including the monocots, Brachypodium and switchgrass, and dicots, Arabidopsis and poplar. All transformants displayed a similar range of phenotypes, including increased biomass from extended leaf production, and increased vegetative branching. Field grown switchgrass transformants showed that overall lignin content was reduced, the ratio of glucans to xylans was increased, and surprisingly, that starch levels were greatly increased. The goals of this project are to control the tissue and temporal expression of Cg1 by using different promoters to drive its expression, elucidate the function of the SPL targets of Cg1 by generating gain and loss of function alleles, and isolate downstream targets of select SPL genes using deep sequencing and chromatin immunoprecipitation. We believe it is possible to control biomass accumulation, cell wall properties, and sugar levels through manipulation of either the Cg1 gene and/or its SPL targets.« less

Efficient biomass pretreatment using ionic liquids derived from lignin and hemicellulose

PubMed Central

Socha, Aaron M.; Parthasarathi, Ramakrishnan; Shi, Jian; Pattathil, Sivakumar; Whyte, Dorian; Bergeron, Maxime; George, Anthe; Tran, Kim; Stavila, Vitalie; Venkatachalam, Sivasankari; Hahn, Michael G.; Simmons, Blake A.; Singh, Seema

2014-01-01

Ionic liquids (ILs), solvents composed entirely of paired ions, have been used in a variety of process chemistry and renewable energy applications. Imidazolium-based ILs effectively dissolve biomass and represent a remarkable platform for biomass pretreatment. Although efficient, imidazolium cations are expensive and thus limited in their large-scale industrial deployment. To replace imidazolium-based ILs with those derived from renewable sources, we synthesized a series of tertiary amine-based ILs from aromatic aldehydes derived from lignin and hemicellulose, the major by-products of lignocellulosic biofuel production. Compositional analysis of switchgrass pretreated with ILs derived from vanillin, p-anisaldehyde, and furfural confirmed their efficacy. Enzymatic hydrolysis of pretreated switchgrass allowed for direct comparison of sugar yields and lignin removal between biomass-derived ILs and 1-ethyl-3-methylimidazolium acetate. Although the rate of cellulose hydrolysis for switchgrass pretreated with biomass-derived ILs was slightly slower than that of 1-ethyl-3-methylimidazolium acetate, 90–95% glucose and 70–75% xylose yields were obtained for these samples after 72-h incubation. Molecular modeling was used to compare IL solvent parameters with experimentally obtained compositional analysis data. Effective pretreatment of lignocellulose was further investigated by powder X-ray diffraction and glycome profiling of switchgrass cell walls. These studies showed different cellulose structural changes and differences in hemicellulose epitopes between switchgrass pretreatments with the aforementioned ILs. Our concept of deriving ILs from lignocellulosic biomass shows significant potential for the realization of a “closed-loop” process for future lignocellulosic biorefineries and has far-reaching economic impacts for other IL-based process technology currently using ILs synthesized from petroleum sources. PMID:25136131

Study of traits and recalcitrance reduction of field-grown COMT down-regulated switchgrass

DOE PAGES

Li, Mi; Pu, Yunqiao; Yoo, Chang Geun; ...

2017-01-03

The native recalcitrance of plants hinders the biomass conversion process using current biorefinery techniques. Down-regulation of the caffeic acid O-methyltransferase (COMT) gene in the lignin biosynthesis pathway of switchgrass reduced the thermochemical and biochemical conversion recalcitrance of biomass. Due to potential environmental influences on lignin biosynthesis and deposition, studying the consequences of physicochemical changes in field-grown plants without pretreatment is essential to evaluate the performance of lignin-altered plants. In this study, we determined the chemical composition, cellulose crystallinity and the degree of its polymerization, molecular weight of hemicellulose, and cellulose accessibility of cell walls in order to better understand themore » fundamental features of why biomass is recalcitrant to conversion without pretreatment. The most important is to investigate whether traits and features are stable in the dynamics of field environmental effects over multiple years.« less

Computational inference of the structure and regulation of the lignin pathway in Panicum virgatum

DOE PAGES

Faraji, Mojdeh; Fonseca, Luis L.; Escamilla-Treviño, Luis; ...

2015-09-17

Switchgrass is a prime target for biofuel production from inedible plant parts and has been the subject of numerous investigations in recent years. Yet, one of the main obstacles to effective biofuel production remains to be the major problem of recalcitrance. Recalcitrance emerges in part from the 3-D structure of lignin as a polymer in the secondary cell wall. Lignin limits accessibility of the sugars in the cellulose and hemicellulose polymers to enzymes and ultimately decreases ethanol yield. Monolignols, the building blocks of lignin polymers, are synthesized in the cytosol and translocated to the plant cell wall, where they undergomore » polymerization. The biosynthetic pathway leading to monolignols in switchgrass is not completely known, and difficulties associated with in vivo measurements of these intermediates pose a challenge for a true understanding of the functioning of the pathway. In this study, a systems biological modeling approach is used to address this challenge and to elucidate the structure and regulation of the lignin pathway through a computational characterization of alternate candidate topologies. The analysis is based on experimental data characterizing stem and tiller tissue of four transgenic lines (knock-downs of genes coding for key enzymes in the pathway) as well as wild-type switchgrass plants. These data consist of the observed content and composition of monolignols. The possibility of a G-lignin specific metabolic channel associated with the production and degradation of coniferaldehyde is examined, and the results support previous findings from another plant species. The computational analysis suggests regulatory mechanisms of product inhibition and enzyme competition, which are well known in biochemistry, but so far had not been reported in switchgrass. By including these mechanisms, the pathway model is able to represent all observations. In conclusion, the results show that the presence of the coniferaldehyde channel is necessary and that product inhibition and competition over cinnamoyl-CoA-reductase (CCR1) are essential for matching the model to observed increases in H-lignin levels in 4-coumarate:CoA-ligase (4CL) knockdowns. Moreover, competition for 4-coumarate:CoA-ligase (4CL) is essential for matching the model to observed increases in the pathway metabolites in caffeic acid O-methyltransferase (COMT) knockdowns. As far as possible, the model was validated with independent data.« less

Identification and Characterization of Switchgrass Histone H3 and CENH3 Genes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miao, Jiamin; Frazier, Taylor; Huang, Linkai

Switchgrass is one of the most promising energy crops and only recently has been employed for biofuel production. The draft genome of switchgrass was recently released; however, relatively few switchgrass genes have been functionally characterized. CENH3, the major histone protein found in centromeres, along with canonical H3 and other histones, plays an important role in maintaining genome stability and integrity. Despite their importance, the histone H3 genes of switchgrass have remained largely uninvestigated. In this study, we identified 17 putative switchgrass histone H3 genes in silico. Of these genes, 15 showed strong homology to histone H3 genes including six H3.1more » genes, three H3.3 genes, four H3.3-like genes and two H3.1-like genes. The remaining two genes were found to be homologous to CENH3. RNA-seq data derived from lowland cultivar Alamo and upland cultivar Dacotah allowed us to identify SNPs in the histone H3 genes and compare their differential gene expression. Interestingly, we also found that overexpression of switchgrass histone H3 and CENH3 genes in N. benthamiana could trigger cell death of the transformed plant cells. Localization and deletion analyses of the histone H3 and CENH3 genes revealed that nuclear localization of the N-terminal tail is essential and sufficient for triggering the cell death phenotype. Lastly, our results deliver insight into the mechanisms underlying the histone-triggered cell death phenotype and provide a foundation for further studying the variations of the histone H3 and CENH3 genes in switchgrass.« less

Identification and Characterization of Switchgrass Histone H3 and CENH3 Genes

DOE PAGES

Miao, Jiamin; Frazier, Taylor; Huang, Linkai; ...

2016-07-12

Switchgrass is one of the most promising energy crops and only recently has been employed for biofuel production. The draft genome of switchgrass was recently released; however, relatively few switchgrass genes have been functionally characterized. CENH3, the major histone protein found in centromeres, along with canonical H3 and other histones, plays an important role in maintaining genome stability and integrity. Despite their importance, the histone H3 genes of switchgrass have remained largely uninvestigated. In this study, we identified 17 putative switchgrass histone H3 genes in silico. Of these genes, 15 showed strong homology to histone H3 genes including six H3.1more » genes, three H3.3 genes, four H3.3-like genes and two H3.1-like genes. The remaining two genes were found to be homologous to CENH3. RNA-seq data derived from lowland cultivar Alamo and upland cultivar Dacotah allowed us to identify SNPs in the histone H3 genes and compare their differential gene expression. Interestingly, we also found that overexpression of switchgrass histone H3 and CENH3 genes in N. benthamiana could trigger cell death of the transformed plant cells. Localization and deletion analyses of the histone H3 and CENH3 genes revealed that nuclear localization of the N-terminal tail is essential and sufficient for triggering the cell death phenotype. Lastly, our results deliver insight into the mechanisms underlying the histone-triggered cell death phenotype and provide a foundation for further studying the variations of the histone H3 and CENH3 genes in switchgrass.« less

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

PubMed Central

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.; Straub, Christopher T.; Compton, Scott H.; Conway, Jonathan M.; Lee, Laura L.; Ragauskas, Arthur J.; Davison, Brian H.; Adams, Michael W. W.

2017-01-01

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates to C. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(−) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15% (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance. IMPORTANCE Key to a microbial process for solubilization of plant biomass is the organism's access to the carbohydrate content of lignocellulose. Economically viable routes will characteristically minimize physical, chemical, and biological pretreatment such that microbial steps contribute to the greatest extent possible. Recently, transgenic versions of plants and trees have been developed with the intention of lowering the barrier to lignocellulose conversion, with particular focus on lignin content and composition. Here, the extremely thermophilic bacterium Caldicellulosiruptor bescii was used to solubilize natural and genetically modified switchgrass lines, with and without the aid of hydrothermal treatment. For lignocellulose conversion, it is clear that the microorganism, plant biomass substrate, and processing steps must all be considered simultaneously to achieve optimal results. Whether switchgrass lines engineered for low lignin or natural variants with desirable properties are used, conversion will depend on microbial access to crystalline cellulose in the plant cell wall. PMID:28625990

Pollen-mediated gene flow from transgenic to non-transgenic switchgrass (Panicum virgatum L.) in the field.

PubMed

Millwood, Reginald; Nageswara-Rao, Madhugiri; Ye, Rongjian; Terry-Emert, Ellie; Johnson, Chelsea R; Hanson, Micaha; Burris, Jason N; Kwit, Charles; Stewart, C Neal

2017-05-02

Switchgrass is C 4 perennial grass species that is being developed as a cellulosic bioenergy feedstock. It is wind-pollinated and considered to be an obligate outcrosser. Genetic engineering has been used to alter cell walls for more facile bioprocessing and biofuel yield. Gene flow from transgenic cultivars would likely be of regulatory concern. In this study we investigated pollen-mediated gene flow from transgenic to nontransgenic switchgrass in a 3-year field experiment performed in Oliver Springs, Tennessee, U.S.A. using a modified Nelder wheel design. The planted area (0.6 ha) contained sexually compatible pollen source and pollen receptor switchgrass plants. One hundred clonal switchgrass 'Alamo' plants transgenic for an orange-fluorescent protein (OFP) and hygromycin resistance were used as the pollen source; whole plants, including pollen, were orange-fluorescent. To assess pollen movement, pollen traps were placed at 10 m intervals from the pollen-source plot in the four cardinal directions extending to 20 m, 30 m, 30 m, and 100 m to the north, south, west, and east, respectively. To assess pollination rates, nontransgenic 'Alamo 2' switchgrass clones were planted in pairs adjacent to pollen traps. In the eastward direction there was a 98% decrease in OFP pollen grains from 10 to 100 m from the pollen-source plot (Poisson regression, F1,8 = 288.38, P < 0.0001). At the end of the second and third year, 1,820 F 1 seeds were collected from pollen recipient-plots of which 962 (52.9%) germinated and analyzed for their transgenic status. Transgenic progeny production detected in each pollen-recipient plot decreased with increased distance from the edge of the transgenic plot (Poisson regression, F1,15 = 12.98, P < 0.003). The frequency of transgenic progeny detected in the eastward plots (the direction of the prevailing wind) ranged from 79.2% at 10 m to 9.3% at 100 m. In these experiments we found transgenic pollen movement and hybridization rates to be inversely associated with distance. However, these data suggest pollen-mediated gene flow is likely to occur up to, at least, 100 m. This study gives baseline data useful to determine isolation distances and other management practices should transgenic switchgrass be grown commercially in relevant environments.

Extended light exposure increases stem digestibility and biomass production of switchgrass

PubMed Central

Zhao, Chunqiao; Hou, Xincun; Zhu, Yi; Yue, Yuesen; Wu, Juying

2017-01-01

Switchgrass is a photoperiod-sensitive energy grass suitable for growing in the marginal lands of China. We explored the effects of extended photoperiods of low-irradiance light (7 μmol·m-2·s-1, no effective photosynthesis) on the growth, the biomass dry weight, the biomass allocation, and, especially, the stem digestibility and cell wall characteristics of switchgrass. Two extended photoperiods (i.e., 18 and 24 h) were applied over Alamo. Extended light exposure (18 and 24 h) resulted in delayed heading and higher dry weights of vegetative organs (by 32.87 and 35.94%, respectively) at the expense of reducing the amount of sexual organs (by 40.05 and 50.87%, respectively). Compared to the control group (i.e., natural photoperiod), the yield of hexoses (% dry matter) in the stems after a direct enzymatic hydrolysis (DEH) treatment significantly increased (by 44.02 and 46.10%) for those groups irradiated during 18 and 24 h, respectively. Moreover, the yield of hexoses obtained via enzymatic hydrolysis increased after both basic (1% NaOH) and acid (1% H2SO4) pretreatments for the groups irradiated during 18 and 24 h. Additionally, low-irradiance light extension (LILE) significantly increased the content of non-structural carbohydrates (NSCs) while notably reducing the lignin content and the syringyl to guaiacyl (S/G) ratio. These structural changes were in part responsible for the observed improved stem digestibility. Remarkably, LILE significantly decreased the cellulose crystallinity index (CrI) of switchgrass by significantly increasing both the arabinose substitution degree in xylan and the content of ammonium oxalate-extractable uronic acids, both favoring cellulose digestibility. Despite this LILE technology is not applied to the cultivation of switchgrass on a large scale yet, we believe that the present work is important in that it reveals important relationships between extended day length irradiations and biomass production and quality. Additionally, this study paves the way for improving biomass production and digestibility via genetic modification of day length sensitive transcription factors or key structural genes in switchgrass leaves. PMID:29166649

Extended light exposure increases stem digestibility and biomass production of switchgrass.

PubMed

Zhao, Chunqiao; Fan, Xifeng; Hou, Xincun; Zhu, Yi; Yue, Yuesen; Wu, Juying

2017-01-01

Switchgrass is a photoperiod-sensitive energy grass suitable for growing in the marginal lands of China. We explored the effects of extended photoperiods of low-irradiance light (7 μmol·m-2·s-1, no effective photosynthesis) on the growth, the biomass dry weight, the biomass allocation, and, especially, the stem digestibility and cell wall characteristics of switchgrass. Two extended photoperiods (i.e., 18 and 24 h) were applied over Alamo. Extended light exposure (18 and 24 h) resulted in delayed heading and higher dry weights of vegetative organs (by 32.87 and 35.94%, respectively) at the expense of reducing the amount of sexual organs (by 40.05 and 50.87%, respectively). Compared to the control group (i.e., natural photoperiod), the yield of hexoses (% dry matter) in the stems after a direct enzymatic hydrolysis (DEH) treatment significantly increased (by 44.02 and 46.10%) for those groups irradiated during 18 and 24 h, respectively. Moreover, the yield of hexoses obtained via enzymatic hydrolysis increased after both basic (1% NaOH) and acid (1% H2SO4) pretreatments for the groups irradiated during 18 and 24 h. Additionally, low-irradiance light extension (LILE) significantly increased the content of non-structural carbohydrates (NSCs) while notably reducing the lignin content and the syringyl to guaiacyl (S/G) ratio. These structural changes were in part responsible for the observed improved stem digestibility. Remarkably, LILE significantly decreased the cellulose crystallinity index (CrI) of switchgrass by significantly increasing both the arabinose substitution degree in xylan and the content of ammonium oxalate-extractable uronic acids, both favoring cellulose digestibility. Despite this LILE technology is not applied to the cultivation of switchgrass on a large scale yet, we believe that the present work is important in that it reveals important relationships between extended day length irradiations and biomass production and quality. Additionally, this study paves the way for improving biomass production and digestibility via genetic modification of day length sensitive transcription factors or key structural genes in switchgrass leaves.

A Genomics Approach to Deciphering Lignin Biosynthesis in Switchgrass[W

PubMed Central

Shen, Hui; Mazarei, Mitra; Hisano, Hiroshi; Escamilla-Trevino, Luis; Fu, Chunxiang; Pu, Yunqiao; Rudis, Mary R.; Tang, Yuhong; Xiao, Xirong; Jackson, Lisa; Li, Guifen; Hernandez, Tim; Chen, Fang; Ragauskas, Arthur J.; Stewart, C. Neal; Wang, Zeng-Yu; Dixon, Richard A.

2013-01-01

It is necessary to overcome recalcitrance of the biomass to saccharification (sugar release) to make switchgrass (Panicum virgatum) economically viable as a feedstock for liquid biofuels. Lignin content correlates negatively with sugar release efficiency in switchgrass, but selecting the right gene candidates for engineering lignin biosynthesis in this tetraploid outcrossing species is not straightforward. To assist this endeavor, we have used an inducible switchgrass cell suspension system for studying lignin biosynthesis in response to exogenous brassinolide. By applying a combination of protein sequence phylogeny with whole-genome microarray analyses of induced cell cultures and developing stem internode sections, we have generated a list of candidate monolignol biosynthetic genes for switchgrass. Several genes that were strongly supported through our bioinformatics analysis as involved in lignin biosynthesis were confirmed by gene silencing studies, in which lignin levels were reduced as a result of targeting a single gene. However, candidate genes encoding enzymes involved in the early steps of the currently accepted monolignol biosynthesis pathway in dicots may have functionally redundant paralogues in switchgrass and therefore require further evaluation. This work provides a blueprint and resources for the systematic genome-wide study of the monolignol pathway in switchgrass, as well as other C4 monocot species. PMID:24285795

Opportunities and roadblocks in utilizing forages and small grains for liquid fuels.

PubMed

Sarath, Gautam; Mitchell, Robert B; Sattler, Scott E; Funnell, Deanna; Pedersen, Jeffery F; Graybosch, Robert A; Vogel, Kenneth P

2008-05-01

This review focuses on the potential advantages and disadvantages of forages such as switchgrass (Panicum virgatum), and two small grains: sorghum (Sorghum bicolor), and wheat (Triticum aesitvum), as feedstocks for biofuels. It highlights the synergy provided by applying what is known from forage digestibility and wheat and sorghum starch properties studies to the biofuels sector. Opportunities therefore, exist to improve biofuel qualities in these crops via genetics and agronomics. In contrast to cereal crops, switchgrass still retains tremendous exploitable genetic diversity, and can be specifically improved to fit a particular agronomic, management, and conversion platform. Combined with emerging studies on switchgrass genomics, conversion properties and management, the future for genetic modification of this species through conventional and molecular breeding strategies appear to be bright. The presence of brown-midrib mutations in sorghum that alter cell wall composition by reducing lignin and other attributes indicate that sorghum could serve as an important model species for C(4)-grasses. Utilization of the brown-midrib traits could lead to the development of forage and sweet sorghums as novel biomass crops. Additionally, wheat crop residue, and wheat and sorghum with improved starch content and composition represent alternate biofuel sources. However, the use of wheat starch as a biofuel is unlikely but its value as a model to study starch properties on biofuel yields holds significant promise.

Transgenic miR156 switchgrass in the field: growth, recalcitrance and rust susceptibility

DOE PAGES

Baxter, Holly L.; Mazarei, Mitra; Dumitrache, Alexandru; ...

2017-04-24

Sustainable utilization of lignocellulosic perennial grass feedstocks will be enabled by high biomass production and optimized cell wall chemistry for efficient conversion into biofuels. MicroRNAs are regulatory elements that modulate the expression of genes involved in various biological functions in plants, including growth and development. In greenhouse studies, overexpressing a microRNA (miR156) gene in switchgrass had dramatic effects on plant architecture and flowering, which appeared to be driven by transgene expression levels. High expressing lines were extremely dwarfed, whereas low and moderate-expressing lines had higher biomass yields, improved sugar release and delayed flowering. Four lines with moderate or low miR156more » overexpression from the prior greenhouse study were selected for a field experiment to assess the relationship between miR156 expression and biomass production over three years. We also analysed important bioenergy feedstock traits such as flowering, disease resistance, cell wall chemistry and biofuel production. Phenotypes of the transgenic lines were inconsistent between the greenhouse and the field as well as among different field growing seasons. One low expressing transgenic line consistently produced more biomass (25%–56%) than the control across all three seasons, which translated to the production of 30% more biofuel per plant during the final season. The other three transgenic lines produced less biomass than the control by the final season, and the two lines with moderate expression levels also exhibited altered disease susceptibilities. Results of this study emphasize the importance of performing multiyear field studies for plants with altered regulatory transgenes that target plant growth and development.« less

Transgenic miR156 switchgrass in the field: growth, recalcitrance and rust susceptibility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baxter, Holly L.; Mazarei, Mitra; Dumitrache, Alexandru

Sustainable utilization of lignocellulosic perennial grass feedstocks will be enabled by high biomass production and optimized cell wall chemistry for efficient conversion into biofuels. MicroRNAs are regulatory elements that modulate the expression of genes involved in various biological functions in plants, including growth and development. In greenhouse studies, overexpressing a microRNA (miR156) gene in switchgrass had dramatic effects on plant architecture and flowering, which appeared to be driven by transgene expression levels. High expressing lines were extremely dwarfed, whereas low and moderate-expressing lines had higher biomass yields, improved sugar release and delayed flowering. Four lines with moderate or low miR156more » overexpression from the prior greenhouse study were selected for a field experiment to assess the relationship between miR156 expression and biomass production over three years. We also analysed important bioenergy feedstock traits such as flowering, disease resistance, cell wall chemistry and biofuel production. Phenotypes of the transgenic lines were inconsistent between the greenhouse and the field as well as among different field growing seasons. One low expressing transgenic line consistently produced more biomass (25%–56%) than the control across all three seasons, which translated to the production of 30% more biofuel per plant during the final season. The other three transgenic lines produced less biomass than the control by the final season, and the two lines with moderate expression levels also exhibited altered disease susceptibilities. Results of this study emphasize the importance of performing multiyear field studies for plants with altered regulatory transgenes that target plant growth and development.« less

Identification and thermochemical analysis of high-lignin feedstocks for biofuel and biochemical production

PubMed Central

2011-01-01

Background Lignin is a highly abundant biopolymer synthesized by plants as a complex component of plant secondary cell walls. Efforts to utilize lignin-based bioproducts are needed. Results Herein we identify and characterize the composition and pyrolytic deconstruction characteristics of high-lignin feedstocks. Feedstocks displaying the highest levels of lignin were identified as drupe endocarp biomass arising as agricultural waste from horticultural crops. By performing pyrolysis coupled to gas chromatography-mass spectrometry, we characterized lignin-derived deconstruction products from endocarp biomass and compared these with switchgrass. By comparing individual pyrolytic products, we document higher amounts of acetic acid, 1-hydroxy-2-propanone, acetone and furfural in switchgrass compared to endocarp tissue, which is consistent with high holocellulose relative to lignin. By contrast, greater yields of lignin-based pyrolytic products such as phenol, 2-methoxyphenol, 2-methylphenol, 2-methoxy-4-methylphenol and 4-ethyl-2-methoxyphenol arising from drupe endocarp tissue are documented. Conclusions Differences in product yield, thermal decomposition rates and molecular species distribution among the feedstocks illustrate the potential of high-lignin endocarp feedstocks to generate valuable chemicals by thermochemical deconstruction. PMID:22018114

Translational Genomics for the Improvement of Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Carpita, Nicholas; McCann, Maureen

2014-05-07

Our objectives were to apply bioinformatics and high throughput sequencing technologies to identify and classify the genes involved in cell wall formation in maize and switchgrass. Targets for genetic modification were to be identified and cell wall materials isolated and assayed for enhanced performance in bioprocessing. We annotated and assembled over 750 maize genes into gene families predicted to function in cell wall biogenesis. Comparative genomics of maize, rice, and Arabidopsis sequences revealed differences in gene family structure. In addition, differences in expression between gene family members of Arabidopsis, maize and rice underscored the need for a grass-specific genetic modelmore » for functional analyses. A forward screen of mature leaves of field-grown maize lines by near-infrared spectroscopy yielded several dozen lines with heritable spectroscopic phenotypes, several of which near-infrared (nir) mutants had altered carbohydrate-lignin compositions. Our contributions to the maize genome sequencing effort built on knowledge of copy number variation showing that uneven gene losses between duplicated regions were involved in returning an ancient allotetraploid to a genetically diploid state. For example, although about 25% of all duplicated genes remain genome-wide, all of the cellulose synthase (CesA) homologs were retained. We showed that guaiacyl and syringyl lignin in lignocellulosic cell-wall materials from stems demonstrate a two-fold natural variation in content across a population of maize Intermated B73 x Mo7 (IBM) recombinant inbred lines, a maize Association Panel of 282 inbreds and landraces, and three populations of the maize Nested Association Mapping (NAM) recombinant inbred lines grown in three years. We then defined quantitative trait loci (QTL) for stem lignin content measured using pyrolysis molecular-beam mass spectrometry, and glucose and xylose yield measured using an enzymatic hydrolysis assay. Among five multi-year QTL for lignin abundance, two for 4-vinylphenol abundance, and four for glucose and/or xylose yield, not a single QTL for aromatic abundance and sugar yield was shared. A genome-wide association study (GWAS) for lignin abundance and sugar yield of the 282-member maize Association Panel provided candidate genes in the eleven QTL and showed that many other alleles impacting these traits exist in the broader pool of maize genetic diversity. The maize B73 and Mo17 genotypes exhibited surprisingly large differences in gene expression in developing stem tissues, suggesting certain regulatory elements can significantly enhance activity of biomass synthesis pathways. Candidate genes, identified by GWAS or by differential expression, include genes of cell-wall metabolism, transcription factors associated with vascularization and fiber formation, and components of cellular signaling pathways. Our work provides new insights and strategies beyond modification of lignin to enhance yields of biofuels from genetically tailored biomass.« less

Identification and molecular characterization of the switchgrass AP2/ERF transcription factor superfamily, and overexpression of PvERF001 for improvement of biomass characteristics for biofuel

DOE PAGES

Wuddineh, Wegi A.; Mazarei, Mitra; Turner, Geoffry B.; ...

2015-07-20

The APETALA2/ethylene response factor (AP2/ERF) superfamily of transcription factors (TFs) plays essential roles in the regulation of various growth and developmental programs including stress responses. Members of these TFs in other plant species have been implicated to play a role in the regulation of cell wall biosynthesis. Here, we identified a total of 207 AP2/ERF TF genes in the switchgrass genome and grouped into four gene families comprised of 25 AP2-, 121 ERF-, 55 DREB (dehydration responsive element binding)-, and 5 RAV (related to API3/VP) genes, as well as a singleton gene not fitting any of the above families. Themore » ERF and DREB subfamilies comprised seven and four distinct groups, respectively. Analysis of exon/intron structures of switchgrass AP2/ERF genes showed high diversity in the distribution of introns in AP2 genes versus a single or no intron in most genes in the ERF and RAV families. The majority of the subfamilies or groups within it were characterized by the presence of one or more specific conserved protein motifs. In silico functional analysis revealed that many genes in these families might be associated with the regulation of responses to environmental stimuli via transcriptional regulation of the response genes. Moreover, these genes had diverse endogenous expression patterns in switchgrass during seed germination, vegetative growth, flower development, and seed formation. Interestingly, several members of the ERF and DREB families were found to be highly expressed in plant tissues where active lignification occurs. These results provide vital resources to select candidate genes to potentially impart tolerance to environmental stress as well as reduced recalcitrance. Furthermore, overexpression of one of the ERF genes ( PvERF001) in switchgrass was associated with increased biomass yield and sugar release efficiency in transgenic lines, exemplifying the potential of these TFs in the development of lignocellulosic feedstocks with improved biomass characteristics for biofuels.« less

Optimizing on-farm pretreatment of perennial grasses for fuel ethanol production.

PubMed

Digman, Matthew F; Shinners, Kevin J; Casler, Michael D; Dien, Bruce S; Hatfield, Ronald D; Jung, Hans-Joachim G; Muck, Richard E; Weimer, Paul J

2010-07-01

Switchgrass (Panicum virgatum L.) and reed canarygrass (Phalaris arundinacea L.) were pretreated under ambient temperature and pressure with sulfuric acid and calcium hydroxide in separate experiments. Chemical loadings from 0 to 100g (kg DM)(-1) and durations of anaerobic storage from 0 to 180days were investigated by way of a central composite design at two moisture contents (40% or 60% w.b.). Pretreated and untreated samples were fermented to ethanol by Saccharomyces cerevisiae D5A in the presence of a commercially available cellulase (Celluclast 1.5L) and beta-glucosidase (Novozyme 188). Xylose levels were also measured following fermentation because xylose is not metabolized by S. cerevisiae. After sulfuric acid pretreatment and anaerobic storage, conversion of cell wall glucose to ethanol for reed canarygrass ranged from 22% to 83% whereas switchgrass conversions ranged from 16% to 46%. Pretreatment duration had a positive effect on conversion but was mitigated with increased chemical loadings. Conversions after calcium hydroxide pretreatment and anaerobic storage ranged from 21% to 55% and 18% to 54% for reed canarygrass and switchgrass, respectively. The efficacy of lime pretreatment was found to be highly dependent on moisture content. Moreover, pretreatment duration was only found to be significant for reed canarygrass. Although significant levels of acetate and lactate were observed in the biomass after storage, S. cerevisiae was not found to be inhibited at a 10% solids loading. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Overexpression of gene encoding the key enzyme involved in proline-biosynthesis (PuP5CS) to improve salt tolerance in switchgrass (Panicum virgatum L.).

PubMed

Guan, Cong; Huang, Yan-Hua; Cui, Xin; Liu, Si-Jia; Zhou, Yun-Zhuan; Zhang, Yun-Wei

2018-05-25

Genetic improvement through overexpressing PuP5CS in switchgrass is feasible for enhancing plant salt stress tolerance. Switchgrass (Panicum virgatum L.) has developed into a dedicated bioenergy crop. To improve the biomass production of switchgrass grown on different types of soil, abiotic stress tolerance traits are considered for its genetic improvement. Proline accumulation is a widespread response when plants are subjected to abiotic stresses such as drought, cold and salinity. In plants, P5CS gene encodes the key regulatory enzyme that plays a crucial role in proline biosynthesis. Here, we introduced the PuP5CS gene (from Puccinellia chinampoensis) into switchgrass by Agrobacterium-mediated transformation. Transgenic lines overexpressing the PuP5CS gene showed phenotypic advantages, in leaf width, internode diameter, internode length, tiller numbers and precocious flowering under normal conditions, and the transgenic lines displayed better regenerative capacity in forming more tillers after harvest. Moreover, the PuP5CS gene enhanced the salt tolerance of transgenic switchgrass by altering a wide range of physiological responses. In accordance with the physiological results, histological analysis of cross sections through the leaf blade showed that the areas of bulliform cells and bundle sheath cells were significantly increased in PuP5CS-overexpressing leaves. The expression levels of ROS scavenging-associated genes in transgenic plants were higher than in control plants under salt stress. The results show that genetic improvement through overexpressing PuP5CS in switchgrass is feasible for enhancing plant stress tolerance.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wuddineh, Wegi A.; Mazarei, Mitra; Turner, Geoffry B.

The APETALA2/ethylene response factor (AP2/ERF) superfamily of transcription factors (TFs) plays essential roles in the regulation of various growth and developmental programs including stress responses. Members of these TFs in other plant species have been implicated to play a role in the regulation of cell wall biosynthesis. Here, we identified a total of 207 AP2/ERF TF genes in the switchgrass genome and grouped into four gene families comprised of 25 AP2-, 121 ERF-, 55 DREB (dehydration responsive element binding)-, and 5 RAV (related to API3/VP) genes, as well as a singleton gene not fitting any of the above families. Themore » ERF and DREB subfamilies comprised seven and four distinct groups, respectively. Analysis of exon/intron structures of switchgrass AP2/ERF genes showed high diversity in the distribution of introns in AP2 genes versus a single or no intron in most genes in the ERF and RAV families. The majority of the subfamilies or groups within it were characterized by the presence of one or more specific conserved protein motifs. In silico functional analysis revealed that many genes in these families might be associated with the regulation of responses to environmental stimuli via transcriptional regulation of the response genes. Moreover, these genes had diverse endogenous expression patterns in switchgrass during seed germination, vegetative growth, flower development, and seed formation. Interestingly, several members of the ERF and DREB families were found to be highly expressed in plant tissues where active lignification occurs. These results provide vital resources to select candidate genes to potentially impart tolerance to environmental stress as well as reduced recalcitrance. Furthermore, overexpression of one of the ERF genes ( PvERF001) in switchgrass was associated with increased biomass yield and sugar release efficiency in transgenic lines, exemplifying the potential of these TFs in the development of lignocellulosic feedstocks with improved biomass characteristics for biofuels.« less

Investigation of Plant Cell Wall Properties: A Study of Contributions from the Nanoscale to the Macroscale Impacting Cell Wall Recalcitrance

NASA Astrophysics Data System (ADS)

Crowe, Jacob Dillon

Biochemical conversion of lignocellulosic biomass to fuel ethanol is one of a few challenging, yet opportune technologies that can reduce the consumption of petroleum-derived transportation fuels, while providing parallel reductions in greenhouse gas emissions. Biomass recalcitrance, or resistance to deconstruction, is a major technical challenge that limits effective conversion of biomass to fermentable sugars, often requiring a costly thermochemical pretreatment step to improve biomass deconstruction. Biomass recalcitrance is imparted largely by the secondary cell wall, a complex polymeric matrix of cell wall polysaccharides and aromatic heteropolymers, that provides structural stability to cells and enables plant upright growth. Polymers within the cell wall can vary both compositionally and structurally depending upon plant species and anatomical fraction, and have varied responses to thermochemical pretreatments. Cell wall properties impacting recalcitrance are still not well understood, and as a result, the goal of this dissertation is to investigate structural features of the cell wall contributing to recalcitrance (1) in diverse anatomical fractions of a single species, (2) in response to diverse pretreatments, and (3) resulting from genetic modification. In the first study, feedstock cell wall heterogeneity was investigated in anatomical (stem, leaf sheaths, and leaf blades) and internode fractions of switchgrass at varying tissue maturities. Lignin content was observed as the key contributor to recalcitrance in maturing stem tissues only, with non-cellulosic substituted glucuronoarabinoxylans and pectic polysaccharides contributing to cell wall recalcitrance in leaf sheath and leaf blades. Hydroxycinnamate (i.e., saponifiable p-coumarate and ferulate) content along with xylan and pectin extractability decreased with tissue maturity, suggesting lignification is only one component imparting maturity specific cell wall recalcitrance. In the second study, alkaline hydrogen peroxide and liquid hot water pretreatments were shown to alter structural properties impacting nanoscale porosity in corn stover. Delignification by alkaline hydrogen peroxide pretreatment decreased cell wall rigidity, with subsequent cell wall swelling resulting in increased nanoscale porosity and improved enzymatic hydrolysis compared to limited swelling and increased accessible surface areas observed in liquid hot water pretreated biomass. The volume accessible to a 90 A dextran probe within the cell wall was found to be positively correlated to both enzyme binding and glucose hydrolysis yields, indicating cell wall porosity is a key contributor to effective hydrolysis yields. In the third study, the effect of altered xylan content and structure was investigated in irregular xylem (irx) Arabidopsis thaliana mutants to understand the role xylan plays in secondary cell wall development and organization. Higher xylan extractability and lower cellulose crystallinity observed in irx9 and irx15 irx15-L mutants compared to wild type indicated altered xylan integration into the secondary cell wall. Nanoscale cell wall organization observed using multiple microscopy techniques was impacted to some extent in all irx mutants, with disorganized cellulose microfibril layers in sclerenchyma secondary cell walls likely resulting from irregular xylan structure and content. Irregular secondary cell wall microfibril layers showed heterogeneous nanomechanical properties compared to wild type, which translated to mechanical deficiencies observed in stem tensile tests. These results suggest nanoscale defects in cell wall strength can correspond to macroscale phenotypes.

Production of polyhydroxybutyrate in switchgrass

DOEpatents

Somleva, Mariya N.; Snell, Kristi D.; Beaulieu, Julie; Peoples, Oliver P.; Garrison, Bradley; Patterson, Nii

2013-07-16

Transgenic plants, plant material, and plant cells for synthesis of polyhydroxyalkanoates, preferably poly(3-hydroxybutyrate) (also referred to a as PHB) are provided. Preferred plants that can be genetically engineered to produce PHB include plants that do not normally produce storage products such as oils and carbohydrates, and plants that have a C.sub.4 NAD-malic enzyme photosynthetic pathway. Such plants also advantageously produce lignocellulosic biomass that can be converted into biofuels. An exemplary plant that can be genetically engineered to produce PHB and produce lignocellulosic biomass is switchgrass, Panicum virgatum L. A preferred cultivar of switchgrass is Alamo. Other suitable cultivars of switchgrass include but are not limited to Blackwell, Kanlow, Nebraska 28, Pathfinder, Cave-in-Rock, Shelter and Trailblazer.

Structural Transformation of Isolated Poplar and Switchgrass Lignins from Dilute Acid Pretreatment

DOE PAGES

Sun, Qining; Pu, Yunqiao; Meng, Xianzhi; ...

2015-08-27

A key step in conversion of cellulosic biomass into sustainable fuels and chemicals is thermochemical pretreatment to reduce plant cell wall recalcitrance. Obtaining an improved understanding of the fundamental chemistry of lignin, the most recalcitrant component of biomass, during pretreatment is critical to the continued development of renewable biofuel production. To examine the intrinsic chemistry of lignin during dilute acid pretreatment (DAP), lignin was isolated from poplar and switchgrass using a cellulolytic enzyme system and then treated under DAP conditions. These results highlight that lignin is subjected to depolymerization reactions within the first 2 min of dilute acid pretreatment andmore » these changes are accompanied by increased generation of aliphatic and phenolic hydroxyl groups of lignin. This is followed by a competing set of depolymerization and repolymerization reactions that lead to a decrease in the content of guaiacyl lignin units and an increase in condensed lignin units as the reaction residence time is extended beyond 5 min. Finally, we showed that a detailed comparison of changes in functional groups and molecular weights of cellulolytic enzyme lignins with different structural parameters, related to the recalcitrant properties of lignin, could be successfully altered during DAP conditions.« less

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates toC. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(–) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15%more » (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance.« less

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

DOE PAGES

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.; ...

2017-06-16

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates toC. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(–) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15%more » (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance.« less

Lignocellulose fermentation and residual solids characterization for senescent switchgrass fermentation by Clostridium thermocellum in the presence and absence of continuous in situ ball-milling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Balch, Michael L.; Holwerda, Evert K.; Davis, Mark F.

Milling during lignocellulosic fermentation, henceforth referred to as cotreatment, is investigated as an alternative to thermochemical pretreatment as a means of enhancing biological solubilization of lignocellulose. We investigate the impact of milling on soluble substrate fermentation by Clostridium thermocellum with comparison to yeast, document solubilization for fermentation of senescent switchgrass with and without ball milling, and characterize residual solids. Soluble substrate fermentation by C. thermocellum proceeded readily in the presence of continuous ball milling but was completely arrested for yeast. Total fractional carbohydrate solubilization achieved after fermentation of senescent switchgrass by C. thermocellum for 5 days was 0.45 without cotreatmentmore » or pretreatment, 0.81 with hydrothermal pretreatment (200 degrees C, 15 minutes, severity 4.2), and 0.88 with cotreatment. Acetate and ethanol were the main fermentation products, and were produced at similar ratios with and without cotreatment. Analysis of solid residues was undertaken using molecular beam mass spectrometry (PyMBMS) and solid-state nuclear magnetic resonance spectroscopy (NMR) in order to provide insight into changes in plant cell walls during processing via various modes. The structure of lignin present in residual solids remaining after fermentation with cotreatment appeared to change little, with substantially greater changes observed for hydrothermal pretreatment - particularly with respect to formation of C-C bonds. The observation of high solubilization with little apparent modification of the residue is consistent with cotreatment enhancing solubilization primarily by increasing the access of saccharolytic enzymes to the feedstock, and C. thermocellum being able to attack all the major linkages in cellulosic biomass provided that these linkages are accessible.« less

Deletion of a gene cluster encoding pectin degrading enzymes in Caldicellulosiruptor bescii reveals an important role for pectin in plant biomass recalcitrance

DOE PAGES

Chung, Daehwan; Pattathil, Sivakumar; Biswal, Ajaya K.; ...

2014-10-10

A major obstacle, and perhaps the most important economic barrier to the effective use of plant biomass for the production of fuels, chemicals, and bioproducts, is our current lack of knowledge of how to efficiently and effectively deconstruct wall polymers for their subsequent use as feedstocks. Plants represent the most desired source of renewable energy and hydrocarbons because they fix CO 2, making their use carbon neutral. Their biomass structure, however, is a barrier to deconstruction, and this is often referred to as recalcitrance. Members of the bacterial genus Caldicellulosiruptor have the ability to grow on unpretreated plant biomass andmore » thus provide an assay for plant deconstruction and biomass recalcitrance. Using recently developed genetic tools for manipulation of these bacteria, a deletion of a gene cluster encoding enzymes for pectin degradation was constructed, and the resulting mutant was reduced in its ability to grow on both dicot and grass biomass, but not on soluble sugars. The plant biomass from three phylogenetically diverse plants, Arabidopsis (a herbaceous dicot), switchgrass (a monocot grass), and poplar (a woody dicot), was used in these analyses. These biomass types have cell walls that are significantly different from each other in both structure and composition. While pectin is a relatively minor component of the grass and woody dicot substrates, the reduced growth of the mutant on all three biomass types provides direct evidence that pectin plays an important role in biomass recalcitrance. Glycome profiling of the plant material remaining after growth of the mutant on Arabidopsis biomass compared to the wild-type revealed differences in the rhamnogalacturonan I, homogalacturonan, arabinogalactan, and xylan profiles. In contrast, only minor differences were observed in the glycome profiles of the switchgrass and poplar biomass. In conclusion, the combination of microbial digestion and plant biomass analysis provides a new and important platform to identify plant wall structures whose presence reduces the ability of microbes to deconstruct plant walls and to identify enzymes that specifically deconstruct those structures.« less

Physicochemical structural changes of poplar and switchgrass during biomass pretreatment and enzymatic hydrolysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meng, Xianzhi; Sun, Qining; Kosa, Matyas

Converting lignocellulosics to simple sugars for second generation bioethanol is complicated due to biomass recalcitrance, and it requires a pretreatment stage prior to enzymatic hydrolysis. In this study, native, pretreated (acid and alkaline) and partially hydrolyzed poplar and switchgrass were characterized by using Simons’ staining for cellulose accessibility, GPC for degree of polymerization (DP), and FTIR for chemical structure of plant cell wall. The susceptibility of the pretreated biomass to enzymatic hydrolysis could not be easily predicted from differences in cellulose DP and accessibility. During hydrolysis, the most significant DP reduction occurred at the very beginning of hydrolysis, and themore » DP began to decrease at a significantly slower rate after this initial period, suggesting an existence of a synergistic action of endo- and exoglucanases that contribute to the occurrence of a “peeling off” mechanism. Cellulose accessibility was found to be increased at the beginning of hydrolysis, after reaching a maximum value then started to decrease. In conclusion, the fresh enzyme restart hydrolysis experiment along with the accessibility data indicated that the factors associated with the nature of enzyme such as irreversible nonspecific binding of cellulases by lignin and steric hindrance of enzymes should be responsible for the gradual slowing down of the reaction rate.« less

Physicochemical structural changes of poplar and switchgrass during biomass pretreatment and enzymatic hydrolysis

DOE PAGES

Meng, Xianzhi; Sun, Qining; Kosa, Matyas; ...

2016-07-27

Converting lignocellulosics to simple sugars for second generation bioethanol is complicated due to biomass recalcitrance, and it requires a pretreatment stage prior to enzymatic hydrolysis. In this study, native, pretreated (acid and alkaline) and partially hydrolyzed poplar and switchgrass were characterized by using Simons’ staining for cellulose accessibility, GPC for degree of polymerization (DP), and FTIR for chemical structure of plant cell wall. The susceptibility of the pretreated biomass to enzymatic hydrolysis could not be easily predicted from differences in cellulose DP and accessibility. During hydrolysis, the most significant DP reduction occurred at the very beginning of hydrolysis, and themore » DP began to decrease at a significantly slower rate after this initial period, suggesting an existence of a synergistic action of endo- and exoglucanases that contribute to the occurrence of a “peeling off” mechanism. Cellulose accessibility was found to be increased at the beginning of hydrolysis, after reaching a maximum value then started to decrease. In conclusion, the fresh enzyme restart hydrolysis experiment along with the accessibility data indicated that the factors associated with the nature of enzyme such as irreversible nonspecific binding of cellulases by lignin and steric hindrance of enzymes should be responsible for the gradual slowing down of the reaction rate.« less

Switchgrass cultivar EG1101

DOEpatents

Bouton, Joseph H; Wood, Donald T

2012-11-27

A switchgrass cultivar designated EG1101 is disclosed. Also disclosed are seeds of switchgrass cultivar EG1101, plants of switchgrass EG1101, plant parts of switchgrass cultivar EG1101 and methods for producing a switchgrass plant produced by crossing switchgrass cultivar EG1101 with itself or with another switchgrass variety. Methods are also described for producing a switchgrass plant containing in its genetic material one or more transgenes and to the transgenic switchgrass plants and plant parts produced by those methods. Switchgrass cultivars or breeding cultivars and plant parts derived from switchgrass variety EG1101, methods for producing other switchgrass cultivars, lines or plant parts derived from switchgrass cultivar EG1101 and the switchgrass plants, varieties, and their parts derived from use of those methods are described herein. Hybrid switchgrass seeds, plants and plant parts produced by crossing the cultivar EG1101 with another switchgrass cultivar are also described.

Switchgrass cultivar EG1102

DOEpatents

Bouton, Joseph H; Wood, Donald T

2012-11-20

A switchgrass cultivar designated EG1102 is disclosed. The invention relates to the seeds of switchgrass cultivar EG1102, to the plants of switchgrass EG1102, to plant parts of switchgrass cultivar EG1102 and to methods for producing a switchgrass plant produced by crossing switchgrass cultivar EG1102 with itself or with another switchgrass variety. The invention also relates to methods for producing a switchgrass plant containing in its genetic material one or more transgenes and to the transgenic switchgrass plants and plant parts produced by those methods. This invention also relates to switchgrass cultivars or breeding cultivars and plant parts derived from switchgrass variety EG1102, to methods for producing other switchgrass cultivars, lines or plant parts derived from switchgrass cultivar EG1102 and to the switchgrass plants, varieties, and their parts derived from use of those methods. The invention further relates to hybrid switchgrass seeds, plants and plant parts produced by crossing the cultivar EG1102 with another switchgrass cultivar.

Interaction of carbon nanohorns with plants: Uptake and biological effects

DOE PAGES

Lahiani, Mohamed H.; Chen, Jihua; Irin, Fahmida; ...

2014-10-07

Single-Walled Carbon Nanohorns (SWCNHs) are a unique carbon-based nanomaterial with promising application in different fields including, medicine, genetic engineering and horticulture. Here, we investigated the biological response of six crop species (barley, corn, rice, soybean, switchgrass, tomato) and tobacco cell culture to the exposure of SWCNHs. We found that SWCNHs can activate seed germination of selected crops and enhance growth of different organs of corn, tomato, rice and soybean. At cellular level, growth of tobacco cells was increased in response to exposure of SWCNHs (78% increase compared to control). Uptake of SWCNHs by exposed crops and tobacco cells was confirmedmore » by transmission electron microscopy (TEM) and quantified by microwave induced heating (MIH) technique. At genetic level, SWCNHs were able to affect expression of a number of tomato genes that are involved in stress responses, cellular responses and metabolic processes. Our conclusion is that SWCNHs can be used as plant growth regulators and have the potential for plant-related applications.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Mi; Pu, Yunqiao; Yoo, Chang Geun

The native recalcitrance of plants hinders the biomass conversion process using current biorefinery techniques. Down-regulation of the caffeic acid O-methyltransferase (COMT) gene in the lignin biosynthesis pathway of switchgrass reduced the thermochemical and biochemical conversion recalcitrance of biomass. Due to potential environmental influences on lignin biosynthesis and deposition, studying the consequences of physicochemical changes in field-grown plants without pretreatment is essential to evaluate the performance of lignin-altered plants. In this study, we determined the chemical composition, cellulose crystallinity and the degree of its polymerization, molecular weight of hemicellulose, and cellulose accessibility of cell walls in order to better understand themore » fundamental features of why biomass is recalcitrant to conversion without pretreatment. The most important is to investigate whether traits and features are stable in the dynamics of field environmental effects over multiple years.« less

Identifying Differences in Abiotic Stress Gene Networks between Lowland and Upland Ecotypes of Switchgrass (DE-SC0008338)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Childs, Kevin; Buell, Robin; Zhao, Bingyu

Switchgrass (Panicum virgatum) is a warm-season C4 grass that is a target lignocellulosic biofuel species for use in the United States due to its local adaption capabilities and high biomass accumulation. Two ecotypes of switchgrass have been described. Members of the lowland ecotype are taller, have narrower leaf blades and generate more biomass compared to individuals from the upland ecotype. Additionally, lowland plants are generally found in the southern United States while upland switchgrass is more typically present in the northern United States. These differences are important as it is envisioned that switchgrass for biofuel production will typically be grownmore » on marginal lands in the northern United States to supplement and diversify farmers' traditional crop incomes. While lowland switchgrass is more productive, it has poor winter survivability in northern latitudes where upland switchgrass is expected to be grown for biofuel use. Abiotic stresses likely to be encountered by switchgrass include drought and salinity. Despite initially being described as preferring wetter environments, members of the lowland ecotype have been characterized as being more drought tolerant than plants of the upland ecotype. Nonetheless, direct trials have indicated that variation for drought tolerance exists in both ecotypes, but prior to this project, only a relatively small number of switchgrass lines had been tested for drought responses. Similarly, switchgrass cultivars have not been widely tested for salt tolerance, but a few studies have shown that even mild salt stress can inhibit growth. The effects of drought and salt stress on plant growth are complex. Both drought and salinity affect the osmotic potential of plant cells and negatively affect plant growth due to reduced water potential and reduced photosynthesis that results from lower stomatal conductance of CO 2. Plants respond to drought and salt stress by activating genes that directly attempt to reduce the stress (e.g., transmembrane pumps that partition Na +) and mitigate the effects of the stress (e.g., synthesis of osmoprotectant metabolites and stress-related signaling compounds). Prior to the start of this project, no gene expression analysis had been performed on switchgrass under conditions of drought or salt stress, and therefore, relevant gene networks responding to drought and salt stress were unknown in switchgrass. In this project, we performed drought, salt and alkali-salt screens on 49 switchgrass cultivars (Liu et al 2014; Liu et al 2015; Hu et al 2015; Kim et al 2016). These experiments demonstrated that a wide range of variation exists within switchgrass for drought, salt and alkali-salt tolerance and that, while the lowland ecotype of switchgrass is often considered more tolerant of abiotic stresses, there are some upland switchgrass lines that are also very tolerant of drought, salt and alkali-salt stress. We also conducted drought and salt time course experiments with Alamo and Dacotah. We have identified modules of coexpressed genes that differentiate Alamo and Dacotah drought responses. We are continuing to analyze these results and plan to submit manuscripts describing this work in early 2017. In an effort to show how drought- and salt-related gene modules could be dissected, we generated transgenic switchgrass overexpressing either PvGTγ-1 or ZmDREB2. Increased expression of PvGTγ-1 does confer increased salt tolerance, and we were able to identify genes that are induced and suppressed by PvGTγ-1. Overexpression of ZmDREB2 increases drought tolerance in switchgrass. Analysis of the PvGTγ-1 and ZmDREB2 overexpression work is ongoing, and we plan to prepare manuscripts about these experiments for submission in early 2017.« less

Outer membrane vesicles from Fibrobacter succinogenes S85 contain an array of carbohydrate-active enzymes with versatile polysaccharide-degrading capacity.

PubMed

Arntzen, Magnus Ø; Várnai, Anikó; Mackie, Roderick I; Eijsink, Vincent G H; Pope, Phillip B

2017-07-01

Fibrobacter succinogenes is an anaerobic bacterium naturally colonising the rumen and cecum of herbivores where it utilizes an enigmatic mechanism to deconstruct cellulose into cellobiose and glucose, which serve as carbon sources for growth. Here, we illustrate that outer membrane vesicles (OMVs) released by F. succinogenes are enriched with carbohydrate-active enzymes and that intact OMVs were able to depolymerize a broad range of linear and branched hemicelluloses and pectin, despite the inability of F. succinogenes to utilize non-cellulosic (pentose) sugars for growth. We hypothesize that the degradative versatility of F. succinogenes OMVs is used to prime hydrolysis by destabilising the tight networks of polysaccharides intertwining cellulose in the plant cell wall, thus increasing accessibility of the target substrate for the host cell. This is supported by observations that OMV-pretreatment of the natural complex substrate switchgrass increased the catalytic efficiency of a commercial cellulose-degrading enzyme cocktail by 2.4-fold. We also show that the OMVs contain a putative multiprotein complex, including the fibro-slime protein previously found to be important in binding to crystalline cellulose. We hypothesize that this complex has a function in plant cell wall degradation, either by catalysing polysaccharide degradation itself, or by targeting the vesicles to plant biomass. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Antimicrobial and anti-inflammatory activity of switchgrass-derived extractives

DOE Office of Scientific and Technical Information (OSTI.GOV)

Labbe, Nicole; Ownley, Bonnie H.; Gwinn, Kimberly D.

Switchgrass is an increasingly important biofuel crop, but knowledge of switchgrass fungal pathogens is not extensive. The purpose of this research was to identify the fungal pathogens that decrease crop yield of switchgrass grown in Tennessee and to investigate a potential sustainable disease management strategy from a value-added by-product of the switchgrass biofuel conversion process. The specific objectives were 1) to identify and characterize prevalent fungal pathogens of switchgrass in Tennessee, 2) assess switchgrass seed produced in the United States for seedborne fungal pathogens, and 3) evaluate switchgrass extractives for antimicrobial activity against plant pathogens.

Estimating switchgrass productivity in the Great Plains using satellite vegetation index and site environmental variables

USGS Publications Warehouse

Gu, Yingxin; Wylie, Bruce K.; Howard, Daniel M.

2015-01-01

Switchgrass is being evaluated as a potential feedstock source for cellulosic biofuels and is being cultivated in several regions of the United States. The recent availability of switchgrass land cover maps derived from the National Agricultural Statistics Service cropland data layer for the conterminous United States provides an opportunity to assess the environmental conditions of switchgrass over large areas and across different geographic locations. The main goal of this study is to develop a data-driven multiple regression switchgrass productivity model and identify the optimal climate and environment conditions for the highly productive switchgrass in the Great Plains (GP). Environmental and climate variables used in the study include elevation, soil organic carbon, available water capacity, climate, and seasonal weather. Satellite-derived growing season averaged Normalized Difference Vegetation Index (GSN) was used as a proxy for switchgrass productivity. Multiple regression analyses indicate that there are strong correlations between site environmental variables and switchgrass productivity (r = 0.95). Sufficient precipitation and suitable temperature during the growing season (i.e., not too hot or too cold) are favorable for switchgrass growth. Elevation and soil characteristics (e.g., soil available water capacity) are also an important factor impacting switchgrass productivity. An anticipated switchgrass biomass productivity map for the entire GP based on site environmental and climate conditions and switchgrass productivity model was generated. Highly productive switchgrass areas are mainly located in the eastern part of the GP. Results from this study can help land managers and biofuel plant investors better understand the general environmental and climate conditions influencing switchgrass growth and make optimal land use decisions regarding switchgrass development in the GP.

Comparative cell-specific transcriptomics reveals differentiation of C4 photosynthesis pathways in switchgrass and other C4 lineages

PubMed Central

Rao, Xiaolan; Lu, Nan; Li, Guifen; Nakashima, Jin; Tang, Yuhong; Dixon, Richard A.

2016-01-01

Almost all C4 plants require the co-ordination of the adjacent and fully differentiated cell types, mesophyll (M) and bundle sheath (BS). The C4 photosynthetic pathway operates through two distinct subtypes based on how malate is decarboxylated in BS cells; through NAD-malic enzyme (NAD-ME) or NADP-malic enzyme (NADP-ME). The diverse or unique cell-specific molecular features of M and BS cells from separate C4 subtypes of independent lineages remain to be determined. We here provide an M/BS cell type-specific transcriptome data set from the monocot NAD-ME subtype switchgrass (Panicum virgatum). A comparative transcriptomics approach was then applied to compare the M/BS mRNA profiles of switchgrass, monocot NADP-ME subtype C4 plants maize and Setaria viridis, and dicot NAD-ME subtype Cleome gynandra. We evaluated the convergence in the transcript abundance of core components in C4 photosynthesis and transcription factors to establish Kranz anatomy, as well as gene distribution of biological functions, in these four independent C4 lineages. We also estimated the divergence between NAD-ME and NADP-ME subtypes of C4 photosynthesis in the two cell types within C4 species, including differences in genes encoding decarboxylating enzymes, aminotransferases, and metabolite transporters, and differences in the cell-specific functional enrichment of RNA regulation and protein biogenesis/homeostasis. We suggest that C4 plants of independent lineages in both monocots and dicots underwent convergent evolution to establish C4 photosynthesis, while distinct C4 subtypes also underwent divergent processes for the optimization of M and BS cell co-ordination. The comprehensive data sets in our study provide a basis for further research on evolution of C4 species. PMID:26896851

Switching to switchgrass: Pathways and consequences of bioenergy switchgrass entering the Midwestern landscape

NASA Astrophysics Data System (ADS)

Krohn, Brian

The US has the ambitious goal of producing 60 billion liters of cellulosic biofuel by 2022. Researchers and US Federal Agencies have identified switchgrass (Panicum virgatum L.) as a potential feedstock for next generation biofuels to help meet this goal because of its excellent agronomic and environmental characteristics. With national policy supporting the development of a switchgrass to bioenergy industry two key questions arise: 1) Under what economic and political conditions will switchgrass enter the landscape? 2) Where on the landscape will switchgrass be cultivated given varying economic and political conditions? The goal of this dissertation is to answer these questions by analyzing the adoption of switchgrass across the upper Midwestern US at a high spatial resolution (30m) under varying economic conditions. In the first chapter, I model switchgrass yields at a high resolution and find considerable variability in switchgrass yields across space, scale, time, and nitrogen management. Then in the second chapter, I use the spatial results from chapter one to challenge the assumption that low-input (unmanaged) switchgrass systems cannot compete economically with high-input (managed) switchgrass systems. Finally, in the third chapter, I evaluate the economic and land quality conditions required for switchgrass to be competitive with a corn/soy rotation. I find that switchgrass can displace low-yielding corn/soy on environmentally sensitive land but, to be competitive, it requires economic support through payments for ecosystem services equal to 360 ha-1. With a total expenditure of 4.3 billion annually for ecosystem services, switchgrass could displace corn/soy on 12.2 million hectares of environmentally sensitive land and increase ethanol production above that from the existing corn by 20 billion liters. Thus, ecosystem services can be an effective means of meeting both bioenergy and environmental goals. Taking the three chapters in aggregate it is apparent that switchgrass faces many challenges before it will be adopted on the landscape and it is unlikely it will be adopted under traditional market pricing. However, switchgrass does have considerable potential to help meet the US's bioenergy and environmental goals through new mechanisms, such as payments for ecosystem services potentially coupled with low-input management systems.

Switchgrass for ethanol and lipid production

USDA-ARS?s Scientific Manuscript database

Switchgrass is being developed as a dedicated cellulosic biomass crop by the Agricultural Research Service (ARS). Switchgrass is a native prairie grass with high productivity and favorable agronomic traits. ARS researchers have recently released Liberty switchgrass, which has a 25-78% increase in bi...

ESTABLISHMENT AND EVALUATION OF SWITCHGRASS ON RECLAIMED MINE SOIL [English

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lang, David; Shankle, Brandon; Oswalt, Ernest

Switchgrass (Panicum virgatum L.) is a native warm season perennial grass that has productive potential of up to 20 Mg ha-1 of biomass and it persists for decades when harvested once per year. Switchgrass provides excellent ground cover and soil stabilization once established and contributes to soil sequestration of new carbon. Slow establishment on newly reclaimed soil, however, provides for significant erosive opportunities thereby requiring initial soil stabilization with a cover crop. Several planting options were evaluated on two topsoil substitute soils. The planting options included: 1) an existing stand of bermudagrass (Cynodon dactylon L.) that was killed with glyphosatemore » followed by disking in red oxidized topsoil substitute and prime farmland topsoil respread in 2007, 2) red oxidized topsoil substitute was seeded directly with switchgrass, 3) browntop millet (Panicum ramosum) was established with switchgrass, 4) or switchgrass was established in senescing browntop millet or wheat without tillage. Switchgrass was successfully established into a bermudagrass sod that had been killed with herbicides and disked as well as into a senescing stand of browntop millet or wheat. Significant soil erosion occurred on the disked area in 2008 leading to considerable repair work followed by planting wheat. Disked areas that did not erode had an excellent stand of switchgrass with 23.3 plants m-2 in November, 2008. Eroded areas replanted in April, 2009 into senescing wheat had 46 plants m-2 by July, 2009. The area planted directly into newly respread soil in May, 2009 was eroded severely by a 75 mm thunderstorm and was repaired, disked and replanted to switchgrass and browntop millet. Switchgrass seeded with browntop millet had a sparse switchgrass stand and was replanted to switchgrass in August, 2009. Rainfall volumes from August, 2009 to October, 2009 totaled 750 mm, but new erosion damage in areas successfully planted to switchgrass has been minimal.« less

Soil and variety effects on the energy and carbon balances of switchgrass-derived ethanol

USDA-ARS?s Scientific Manuscript database

This study examined the effects of soil and switchgrass variety on sustainability and eco-friendliness of switchgrass-based ethanol production. Using the Agricultural Land Management Alternatives with Numerical Assessment Criteria (ALMANAC) model, switchgrass biomass yields were simulated for severa...

Associations with flowering time, latitude, and climate in switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass is a North American perennial grass and emerging bioenergy feedstock, and increasing biomass yields will improve the economic viability of switchgrass as a bioenergy crop. Flowering time is an important determinant of biomass yields in switchgrass because the majority of biomass accumula...

Switchgrass for forage and bioenergy

USDA-ARS?s Scientific Manuscript database

Switchgrass is a native warm-season grass that has been used for hay, forage, and conservation purposes for decades and switchgrass research in Nebraska has been ongoing since 1936. Recently, switchgrass has been identified as a model perennial grass for bioenergy in the Great Plains and Midwest. Si...

Compatibility of switchgrass as an energy crop in farming systems of the southeastern USA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bransby, D.I.; Rodriguez-Kabana, R.; Sladden, S.E.

1993-12-31

The objective of this paper is to examine the compatibility of switchgrass as an energy crop in farming systems in the southeastern USA, relative to other regions. In particular, the issues addressed are (1) competition between switchgrass as an energy crop and existing farm enterprises, based primarily on economic returns, (2) complementarity between switchgrass and existing farm enterprises, and (3) environmental benefits. Because projected economic returns for switchgrass as an energy crop are highest in the Southeast, and returns from forestry and beef pastures (the major existing enterprises) are low, there is a very strong economic incentive in this region.more » In contrast, based on current information, economic viability of switchgrass as an energy crop in other regions appears doubtful. In addition, switchgrass in the southeastern USA would complement forage-livestock production, row crop production and wildlife and would provide several additional environmental benefits. It is concluded that the southeastern USA offers the greatest opportunity for developing switchgrass as an economically viable energy crop.« less

Natural cellulose fibers from switchgrass with tensile properties similar to cotton and linen.

PubMed

Reddy, Narendra; Yang, Yiqi

2007-08-01

We report the production and characteristics of natural cellulose fibers obtained from the leaves and stems of switchgrass. In this paper, the composition, structure and properties of fibers obtained from the leaves and stem of switchgrass have been studied in comparison to the common natural cellulose fibers, such as cotton, linen and kenaf. The leaves and stems of switchgrass have tensile properties intriguingly similar to that of linen and cotton, respectively. Fibers were obtained from the leaves and stems of switchgrass using a simple alkaline extraction and the structure and properties of the fibers were studied. Fibers obtained from switchgrass leaves have crystallinity of 51%, breaking tenacity of 5.5 g per denier (715 MPa) and breaking elongation of 2.2% whereas the corresponding values for fibers obtained from switchgrass stems are 46%, 2.7 g per denier and 6.8%, respectively. Switchgrass is a relatively easy to grow and high yield biomass crop that can be source to partially substitute the natural and synthetic fibers currently in use. We hope that this research will stimulate interests in using switchgrass as a novel fiber crop in addition to being promoted as a potential source for biofuels. (c) 2007 Wiley Periodicals, Inc.

Production and supply logistics of switchgrass as an energy feedstock

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a warm-season (C4), perennial grass that is native to the tallgrass ecoregion of North America (Figure 1). Historically, switchgrass has been used for summer forage, hay, ensiling, or in conservation plantings. At the end of the 20th century, switchgrass was de...

Identification, characterization, and gene expression analysis of nucleotide binding site (NB)-type resistance gene homologues in switchgrass

USDA-ARS?s Scientific Manuscript database

Background: Switchgrass (Panicum virgatum L.) is a warm-season perennial grass that can be used as a second generation bioenergy crop. However, foliar fungal pathogens, like switchgrass rust, have the potential to significantly reduce switchgrass biomass yield. Despite its importance as a prominent ...

Consolidated bioprocessing of transgenic switchgrass by an engineered and evolved Clostridium thermocellum strain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yee, Kelsey L; Rodriguez Jr, Miguel; Thompson, Olivia A

Background: Switchgrass is an abundant and dedicated bioenergy feedstock however its inherent recalcitrance is one of the economic hurdles for producing biofuels. The down-regulation of the caffeic acid O-methyl transferase (COMT) gene in the lignin pathway of switchgrass reduced lignin content and S/G ratio, and the transgenic lines showed improved fermentation yield with S. cerevisiae and C. thermocellum (ATCC 27405) in comparison to the wild-type switchgrass. Results: Here we examine the fermentation potential of the COMT transgenic switchgrass and its wild-type line, with an engineered and evolved Clostridium thermocellum (M1570) strain. The fermentation of the transgenic switchgrass had superior conversionmore » relative to the control line with an increase of 20% and ethanol was the primary metabolite accounting for 90% of the total metabolites measured by HPLC. Conclusions: The down-regulation of the COMT gene in switchgrass reduced recalcitrance and improved microbial bioconversion yield. Moreover, these results showed ethanol as the main fermentation metabolite produced by an engineered and evolved C. thermocellum strain grown on a transgenic switchgrass.« less

Characterization of Brachypodium distachyon as a nonhost model against switchgrass rust pathogen Puccinia emaculata.

PubMed

Gill, Upinder S; Uppalapati, Srinivasa R; Nakashima, Jin; Mysore, Kirankumar S

2015-05-08

Switchgrass rust, caused by Puccinia emaculata, is an important disease of switchgrass, a potential biofuel crop in the United States. In severe cases, switchgrass rust has the potential to significantly affect biomass yield. In an effort to identify novel sources of resistance against switchgrass rust, we explored nonhost resistance against P. emaculata by characterizing its interactions with six monocot nonhost plant species. We also studied the genetic variations for resistance among Brachypodium inbred accessions and the involvement of various defense pathways in nonhost resistance of Brachypodium. We characterized P. emaculata interactions with six monocot nonhost species and identified Brachypodium distachyon (Bd21) as a suitable nonhost model to study switchgrass rust. Interestingly, screening of Brachypodium accessions identified natural variations in resistance to switchgrass rust. Brachypodium inbred accessions Bd3-1 and Bd30-1 were identified as most and least resistant to switchgrass rust, respectively, when compared to tested accessions. Transcript profiling of defense-related genes indicated that the genes which were induced in Bd21after P. emaculata inoculation also had higher basal transcript abundance in Bd3-1 when compared to Bd30-1 and Bd21 indicating their potential involvement in nonhost resistance against switchgrass rust. In the present study, we identified Brachypodium as a suitable nonhost model to study switchgrass rust which exhibit type I nonhost resistance. Variations in resistance response were also observed among tested Brachypodium accessions. Brachypodium nonhost resistance against P. emaculata may involve various defense pathways as indicated by transcript profiling of defense related genes. Overall, this study provides a new avenue to utilize novel sources of nonhost resistance in Brachypodium against switchgrass rust.

Simultaneous Downregulation of MTHFR and COMT in Switchgrass Affects Plant Performance and Induces Lesion-Mimic Cell Death.

PubMed

Liu, Sijia; Fu, Chunxiang; Gou, Jiqing; Sun, Liang; Huhman, David; Zhang, Yunwei; Wang, Zeng-Yu

2017-01-01

Switchgrass ( Panicum virgatum ) has been developed into a model lignocellulosic bioenergy crop. Downregulation of caffeic acid O -methyltransferase (COMT), a key enzyme in lignin biosynthesis, has been shown to alter lignification and increase biofuel yield in switchgrass. Methylenetetrahydrofolate reductase (MTHFR) mediates C1 metabolism and provides methyl units consumed by COMT. It was predicted that co-silencing of MTHFR and COMT would impact lignification even more than either of the single genes. However, our results showed that strong downregulation of MTHFR in a COMT -deficient background led to altered plant growth and development, but no significant change in lignin content or composition was found when compared with COMT plants. Another unexpected finding was that the double MTHFR/COMT downregulated plants showed a novel lesion-mimic leaf phenotype. Molecular analyses revealed that the lesion-mimic phenotype was caused by the synergistic effect of MTHFR and COMT genes, with MTHFR playing a predominant role. Microarray analysis showed significant induction of genes related to oxidative and defense responses. The results demonstrated the lack of additive effects of MTHFR and COMT on lignification. Furthermore, this research revealed an unexpected role of the two genes in the modulation of lesion-mimic cell death as well as their synergistic effects on agronomic performance.

Simultaneous Downregulation of MTHFR and COMT in Switchgrass Affects Plant Performance and Induces Lesion-Mimic Cell Death

PubMed Central

Liu, Sijia; Fu, Chunxiang; Gou, Jiqing; Sun, Liang; Huhman, David; Zhang, Yunwei; Wang, Zeng-Yu

2017-01-01

Switchgrass (Panicum virgatum) has been developed into a model lignocellulosic bioenergy crop. Downregulation of caffeic acid O-methyltransferase (COMT), a key enzyme in lignin biosynthesis, has been shown to alter lignification and increase biofuel yield in switchgrass. Methylenetetrahydrofolate reductase (MTHFR) mediates C1 metabolism and provides methyl units consumed by COMT. It was predicted that co-silencing of MTHFR and COMT would impact lignification even more than either of the single genes. However, our results showed that strong downregulation of MTHFR in a COMT-deficient background led to altered plant growth and development, but no significant change in lignin content or composition was found when compared with COMT plants. Another unexpected finding was that the double MTHFR/COMT downregulated plants showed a novel lesion-mimic leaf phenotype. Molecular analyses revealed that the lesion-mimic phenotype was caused by the synergistic effect of MTHFR and COMT genes, with MTHFR playing a predominant role. Microarray analysis showed significant induction of genes related to oxidative and defense responses. The results demonstrated the lack of additive effects of MTHFR and COMT on lignification. Furthermore, this research revealed an unexpected role of the two genes in the modulation of lesion-mimic cell death as well as their synergistic effects on agronomic performance. PMID:28676804

Mild pyrolysis of P3HB/Switchgrass blends for the production of bio-oil enriched with crotonic acid

USDA-ARS?s Scientific Manuscript database

The mild pyrolysis of switchgrass/poly-3-hydroxybutyrate (P3HB) blends that mimic P3HB-producing switchgrass lines was studied in a pilot scale fluidized bed reactor with the goal of simultaneously producing crotonic acid and switchgrass-based bio-oil. Factors such as pyrolysis temperature, residenc...

Effects of Quality Composts and Other Organic Amendments and Their Humic and Fulvic Acid fractions on the Germination of Slickspot Peppergrass (Lepidium Papilliferum) and Switchgrass in Various Experimental Conditions

DTIC Science & Technology

2012-01-01

Figure 2. Effect of compost HAs at different concentrations on the number of germinated seeds of Shawnee switchgrass expressed as percentages of...and C/P20%) on the early growth of these switchgrass species. 2. Experimental 2.1. Germination N. 60 seeds of each switchgrass species were...the germination process. In this experiment, n. 60 seeds of each switchgrass species were previously soaked in distilled water for 12 h, successively

Identification, characterization, and gene expression analysis of nucleotide binding site (NB)-type resistance gene homologues in switchgrass

DOE PAGES

Frazier, Taylor P.; Palmer, Nathan A.; Xie, Fuliang; ...

2016-11-08

Switchgrass ( Panicum virgatum L.) is a warm-season perennial grass that can be used as a second generation bioenergy crop. However, foliar fungal pathogens, like switchgrass rust, have the potential to significantly reduce switchgrass biomass yield. Despite its importance as a prominent bioenergy crop, a genome-wide comprehensive analysis of NB-LRR disease resistance genes has yet to be performed in switchgrass. In this study, we used a homology-based computational approach to identify 1011 potential NB-LRR resistance gene homologs (RGHs) in the switchgrass genome (v 1.1). In addition, we identified 40 RGHs that potentially contain unique domains including major sperm protein domain,more » jacalin-like binding domain, calmodulin-like binding, and thioredoxin. RNA-sequencing analysis of leaf tissue from ‘Alamo’, a rust-resistant switchgrass cultivar, and ‘Dacotah’, a rust-susceptible switchgrass cultivar, identified 2634 high quality variants in the RGHs between the two cultivars. RNA-sequencing data from field-grown cultivar ‘Summer’ plants indicated that the expression of some of these RGHs was developmentally regulated. Our results provide useful insight into the molecular structure, distribution, and expression patterns of members of the NB-LRR gene family in switchgrass. These results also provide a foundation for future work aimed at elucidating the molecular mechanisms underlying disease resistance in this important bioenergy crop.« less

Identification, characterization, and gene expression analysis of nucleotide binding site (NB)-type resistance gene homologues in switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Frazier, Taylor P.; Palmer, Nathan A.; Xie, Fuliang

Switchgrass ( Panicum virgatum L.) is a warm-season perennial grass that can be used as a second generation bioenergy crop. However, foliar fungal pathogens, like switchgrass rust, have the potential to significantly reduce switchgrass biomass yield. Despite its importance as a prominent bioenergy crop, a genome-wide comprehensive analysis of NB-LRR disease resistance genes has yet to be performed in switchgrass. In this study, we used a homology-based computational approach to identify 1011 potential NB-LRR resistance gene homologs (RGHs) in the switchgrass genome (v 1.1). In addition, we identified 40 RGHs that potentially contain unique domains including major sperm protein domain,more » jacalin-like binding domain, calmodulin-like binding, and thioredoxin. RNA-sequencing analysis of leaf tissue from ‘Alamo’, a rust-resistant switchgrass cultivar, and ‘Dacotah’, a rust-susceptible switchgrass cultivar, identified 2634 high quality variants in the RGHs between the two cultivars. RNA-sequencing data from field-grown cultivar ‘Summer’ plants indicated that the expression of some of these RGHs was developmentally regulated. Our results provide useful insight into the molecular structure, distribution, and expression patterns of members of the NB-LRR gene family in switchgrass. These results also provide a foundation for future work aimed at elucidating the molecular mechanisms underlying disease resistance in this important bioenergy crop.« less

Allelopathic Potential of Switchgrass ( Panicum virgatum L.) on Perennial Ryegrass ( Lolium perenne L.) and Alfalfa ( Medicago sativa L.)

NASA Astrophysics Data System (ADS)

Shui, Junfeng; An, Yu; Ma, Yongqing; Ichizen, Nobumasa

2010-10-01

This study investigated allelopathy and its chemical basis in nine switchgrass ( Panicum virgatum L.) accessions. Perennial ryegrass ( Lolium perenne L.) and alfalfa ( Medicago sativa L.) were used as test species. Undiluted aqueous extracts (5 g plant tissue in 50 ml water) from the shoots and roots of most of the switchgrass accessions inhibited the germination and growth of the test species. However, the allelopathic effect of switchgrass declined when extracts were diluted 5- or 50-fold. Seedling growth was more sensitive than seed germination as an indicator of allelopathic effect. Allelopathic effect was related to switchgrass ecotype but not related to ploidy level. Upland accessions displayed stronger allelopathic potential than lowland accessions. The aqueous extract from one switchgrass accession was separated into phenols, organic acids, neutral chemicals, and alkaloids, and then these fractions were bioassayed to test for allelopathic potential. Alkaloids had the strongest allelopathic effect among the four chemical fractions. In summary, the results indicated that switchgrass has allelopathic potential; however, there is not enough evidence to conclude that allelopathic advantage is the main factor that has contributed to the successful establishment of switchgrass on China’s Loess Plateau.

Allelopathic potential of switchgrass (Panicum virgatum L.) on perennial ryegrass (Lolium perenne L.) and alfalfa (Medicago sativa L.).

PubMed

Shui, Junfeng; An, Yu; Ma, Yongqing; Ichizen, Nobumasa

2010-10-01

This study investigated allelopathy and its chemical basis in nine switchgrass (Panicum virgatum L.) accessions. Perennial ryegrass (Lolium perenne L.) and alfalfa (Medicago sativa L.) were used as test species. Undiluted aqueous extracts (5 g plant tissue in 50 ml water) from the shoots and roots of most of the switchgrass accessions inhibited the germination and growth of the test species. However, the allelopathic effect of switchgrass declined when extracts were diluted 5- or 50-fold. Seedling growth was more sensitive than seed germination as an indicator of allelopathic effect. Allelopathic effect was related to switchgrass ecotype but not related to ploidy level. Upland accessions displayed stronger allelopathic potential than lowland accessions. The aqueous extract from one switchgrass accession was separated into phenols, organic acids, neutral chemicals, and alkaloids, and then these fractions were bioassayed to test for allelopathic potential. Alkaloids had the strongest allelopathic effect among the four chemical fractions. In summary, the results indicated that switchgrass has allelopathic potential; however, there is not enough evidence to conclude that allelopathic advantage is the main factor that has contributed to the successful establishment of switchgrass on China's Loess Plateau.

Plant community and white-tailed deer nutritional carrying capacity response to intercropping switchgrass in loblolly pine plantations

NASA Astrophysics Data System (ADS)

Greene, Ethan Jacob

Switchgrass (Panicum virgatum L.) is a cellulosic feedstock for alternative energy production that could grow well between planted pines (Pinus spp.). Southeastern planted pine occupies 15.8 million hectares and thus, switchgrass intercropping could affect biodiversity if broadly implemented. Therefore, I evaluated effects of intercropping switchgrass in loblolly pine (P. taeda L.) plantations on plant community diversity, plant biomass production, and white-tailed deer (Odocoileus virginianus Zimmerman) nutritional carrying capacity. In a randomized complete block design, I assigned three treatments (switchgrass intercropped, switchgrass monoculture, and a "control" of traditional pine management) to 4 replicates of 10-ha experimental units in Kemper County, Mississippi during 2014-2015. I detected 246 different plant species. Switchgrass intercropping reduced plant species richness and diversity but maintained evenness. I observed reduced forb and high-use deer forage biomass but only in intercropped alleys (interbeds). Soil micronutrient interactions affected forage protein of deer plants. White-tailed deer nutritional carrying capacity remained unaffected.

Kinetics of methane production from the codigestion of switchgrass and Spirulina platensis algae.

PubMed

El-Mashad, Hamed M

2013-03-01

Anaerobic batch digestion of four feedstocks was conducted at 35 and 50 °C: switchgrass; Spirulina platensis algae; and two mixtures of both switchgrass and S. platensis. Mixture 1 was composed of 87% switchgrass (based on volatile solids) and 13% S. platensis. Mixture 2 was composed of 67% switchgrass and 33% S. platensis. The kinetics of methane production from these feedstocks was studied using four first order models: exponential, Gompertz, Fitzhugh, and Cone. The methane yields after 40days of digestion at 35 °C were 355, 127, 143 and 198 ml/g VS, respectively for S. platensis, switchgrass, and Mixtures 1 and 2, while the yields at 50 °C were 358, 167, 198, and 236 ml/g VS, respectively. Based on Akaike's information criterion, the Cone model best described the experimental data. The Cone model was validated with experimental data collected from the digestion of a third mixture that was composed of 83% switchgrass and 17% S. platensis. Published by Elsevier Ltd.

Sulfite (SPORL) pretreatment of switchgrass for enzymatic saccharification

Treesearch

D.S. Zhang; Q. Yang; J.Y. Zhu; X.J. Pan

2013-01-01

SPORL (Sulfite Pretreatment to Overcome Recalcitrance of Lignocellulose) pretreatment was applied to switchgrass and optimized through an experimental design using Response Surface Methodology within the range of temperature (163–197 °C), time (3–37 min), sulfuric acid dosage (0.8–4.2% on switchgrass), and sodium sulfite dosage (0.6–7.4% on switchgrass)....

Integrating future scenario‐based crop expansion and crop conditions to map switchgrass biofuel potential in eastern Nebraska, USA

USGS Publications Warehouse

Gu, Yingxin; Wylie, Bruce K.

2018-01-01

Switchgrass (Panicum virgatum) has been evaluated as one potential source for cellulosic biofuel feedstocks. Planting switchgrass in marginal croplands and waterway buffers can reduce soil erosion, improve water quality, and improve regional ecosystem services (i.e. it serves as a potential carbon sink). In previous studies, we mapped high risk marginal croplands and highly erodible cropland buffers that are potentially suitable for switchgrass development, which would improve ecosystem services and minimally impact food production. In this study, we advance our previous study results and integrate future crop expansion information to develop a switchgrass biofuel potential ensemble map for current and future croplands in eastern Nebraska. The switchgrass biomass productivity and carbon benefits (i.e. NEP: net ecosystem production) for the identified biofuel potential ensemble areas were quantified. The future scenario‐based (‘A1B’) land use and land cover map for 2050, the US Geological Survey crop type and Compound Topographic Index (CTI) maps, and long‐term (1981–2010) averaged annual precipitation data were used to identify future crop expansion regions that are suitable for switchgrass development. Results show that 2528 km2 of future crop expansion regions (~3.6% of the study area) are potentially suitable for switchgrass development. The total estimated biofuel potential ensemble area (including cropland buffers, marginal croplands, and future crop expansion regions) is 4232 km2 (~6% of the study area), potentially producing 3.52 million metric tons of switchgrass biomass per year. Converting biofuel ensemble regions to switchgrass leads to potential carbon sinks (the total NEP for biofuel potential areas is 0.45 million metric tons C) and is environmentally sustainable. Results from this study improve our understanding of environmental conditions and ecosystem services of current and future cropland systems in eastern Nebraska and provide useful information to land managers to make land use decisions regarding switchgrass development.

Understanding changes in cellulose crystalline structure of lignocellulosic biomass during ionic liquid pretreatment by XRD.

PubMed

Zhang, Jiafu; Wang, Yixun; Zhang, Liye; Zhang, Ruihong; Liu, Guangqing; Cheng, Gang

2014-01-01

X-ray diffraction (XRD) was used to understand the interactions of cellulose in lignocellulosic biomass with ionic liquids (ILs). The experiment was designed in such a way that the process of swelling and solubilization of crystalline cellulose in plant cell walls was followed by XRD. Three different feedstocks, switchgrass, corn stover and rice husk, were pretreated using 1-butyl-3-methylimidazolium acetate ([C4mim][OAc]) at temperatures of 50-130°C for 6h. At a 5 wt.% biomass loading, increasing pretreatment temperature led to a drop in biomass crystallinity index (CrI), which was due to swelling of crystalline cellulose. After most of the crystalline cellulose was swollen with IL molecules, a low-order structure was found in the pretreated samples. Upon further increasing temperature, cellulose II structure started to form in the pretreated biomass samples as a result of solubilization of cellulose in [C4mim][OAc] and subsequent regeneration. Copyright © 2013 Elsevier Ltd. All rights reserved.

AmeriFlux US-AR2 ARM USDA UNL OSU Woodward Switchgrass 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Billesbach, Dave; Bradford, James

This is the AmeriFlux version of the carbon flux data for the site US-AR2 ARM USDA UNL OSU Woodward Switchgrass 2. Site Description - The ARM USDA UNL OSU Woodward Switchgrass 2 tower is located on public land owned by the USDA-ARS Southern Plains Range Research Station in Woodward, Oklahoma. The site is on a former wheat field that is in the process of changing to switchgrass. A companion site (ARM USDA UNL OSU Woodward Switchgrass 1) is on a former native prairie. Previous wheat was planted in Fall 2008. In Spring 2009, herbicide was applied to kill the wheatmore » prior to switchgrass planting. Later in the year, the site was sprayed with post-emergence herbicide. In 2010, fertilization occurred before herbicide was sprayed for broadleaf control.« less

Economics of intercropping loblolly pine and switchgrass for bioenergy markets in the southeastern United States

Treesearch

Andres Susaeta; Janaki Alavalapati Pankaj Lal; D. Evan Mercer; Douglas Carter

2012-01-01

Abstract The main objective of this study was to assess the economics of alley cropping of loblolly pine (Pinus taeda L.) and switchgrass (Panicum virgatum) in the southern United States. Assuming a price range of switchgrass between $15 and $50 Mg-1 and yield of 12 Mg ha-1 year-1, we investigated the effect of switchgrass production on the optimal forest management...

Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass

PubMed Central

Fu, Chunxiang; Mielenz, Jonathan R.; Xiao, Xirong; Ge, Yaxin; Hamilton, Choo Y.; Rodriguez, Miguel; Chen, Fang; Foston, Marcus; Ragauskas, Arthur; Bouton, Joseph; Dixon, Richard A.; Wang, Zeng-Yu

2011-01-01

Switchgrass is a leading dedicated bioenergy feedstock in the United States because it is a native, high-yielding, perennial prairie grass with a broad cultivation range and low agronomic input requirements. Biomass conversion research has developed processes for production of ethanol and other biofuels, but they remain costly primarily because of the intrinsic recalcitrance of biomass. We show here that genetic modification of switchgrass can produce phenotypically normal plants that have reduced thermal-chemical (≤180 °C), enzymatic, and microbial recalcitrance. Down-regulation of the switchgrass caffeic acid O-methyltransferase gene decreases lignin content modestly, reduces the syringyl:guaiacyl lignin monomer ratio, improves forage quality, and, most importantly, increases the ethanol yield by up to 38% using conventional biomass fermentation processes. The down-regulated lines require less severe pretreatment and 300–400% lower cellulase dosages for equivalent product yields using simultaneous saccharification and fermentation with yeast. Furthermore, fermentation of diluted acid-pretreated transgenic switchgrass using Clostridium thermocellum with no added enzymes showed better product yields than obtained with unmodified switchgrass. Therefore, this apparent reduction in the recalcitrance of transgenic switchgrass has the potential to lower processing costs for biomass fermentation-derived fuels and chemicals significantly. Alternatively, such modified transgenic switchgrass lines should yield significantly more fermentation chemicals per hectare under identical process conditions. PMID:21321194

Anthracnose disease of switchgrass caused by the novel fungal species Colletotrichum navitas.

PubMed

Crouch, Jo Anne; Beirn, Lisa A; Cortese, Laura M; Bonos, Stacy A; Clarke, Bruce B

2009-12-01

In recent years perennial grasses such as the native tallgrass prairie plant Panicum virgatum (switchgrass) have taken on a new role in the North American landscape as a plant-based source of renewable energy. Because switchgrass is a native plant, it has been suggested that disease problems will be minimal, but little research in this area has been conducted. Recently, outbreaks of switchgrass anthracnose disease have been reported from the northeastern United States. Incidences of switchgrass anthracnose are known in North America since 1886 through herbarium specimens and disease reports, but the causal agent of this disease has never been experimentally determined or taxonomically evaluated. In the present work, we evaluate the causal agent of switchgrass anthracnose, a new species we describe as Colletotrichum navitas (navitas=Latin for energy). Multilocus molecular phylogenetics and morphological characters show C. navitas is a novel species in the falcate-spored graminicolous group of the genus Colletotrichum; it is most closely related to the corn anthracnose pathogen Colletotrichum graminicola. We present a formal description and illustrations for C. navitas and provide experimental confirmation that this organism is responsible for switchgrass anthracnose disease.

Environmental and economic analysis of switchgrass production for water quality improvement in northeast Kansas.

PubMed

Nelson, Richard G; Ascough, James C; Langemeier, Michael R

2006-06-01

The primary objectives of this research were to determine SWAT model predicted reductions in four water quality indicators (sediment yield, surface runoff, nitrate nitrogen (NO(3)-N) in surface runoff, and edge-of-field erosion) associated with producing switchgrass (Panicum virgatum) on cropland in the Delaware basin in northeast Kansas, and evaluate switchgrass break-even prices. The magnitude of potential switchgrass water quality payments based on using switchgrass as an alternative energy source was also estimated. SWAT model simulations showed that between 527,000 and 1.27 million metric tons (Mg) of switchgrass could be produced annually across the basin depending upon nitrogen (N) fertilizer application levels (0-224 kg N ha(-1)). The predicted reductions in sediment yield, surface runoff, NO(3)-N in surface runoff, and edge-of-field erosion as a result of switchgrass plantings were 99, 55, 34, and 98%, respectively. The average annual cost per hectare for switchgrass ranged from about 190 US dollars with no N applied to around 345 US dollars at 224 kg N ha(-1) applied. Edge-of-field break-even price per Mg ranged from around 41 US dollars with no N applied to slightly less than 25 US dollars at 224 kg N ha(-1) applied. A majority of the switchgrass produced had an edge-of-field break-even price of 30 Mg(-1) US dollars or less. Savings of at least 50% in each of the four water quality indicators could be attained for an edge-of-field break-even price of 22-27.49 US dollars Mg(-1).

New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits

PubMed Central

2011-01-01

Background Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18) to duodecaploid (12X = 108). Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. Results A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective). Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. Conclusions The construction of the first switchgrass BAC library and comparative analysis of homoeologous harboring OsBRI1 orthologs present a glimpse into the switchgrass genome structure and complexity. Data obtained demonstrate the feasibility of using HICF fingerprinting to resolve the homoeologous chromosomes of the two distinct genomes in switchgrass, providing a robust and accurate BAC-based physical platform for this species. The genomic resources and sequence data generated will lay the foundation for deciphering the switchgrass genome and lead the way for an accurate genome sequencing strategy. PMID:21767393

New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits.

PubMed

Saski, Christopher A; Li, Zhigang; Feltus, Frank A; Luo, Hong

2011-07-18

Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18) to duodecaploid (12X = 108). Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective). Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. The construction of the first switchgrass BAC library and comparative analysis of homoeologous harboring OsBRI1 orthologs present a glimpse into the switchgrass genome structure and complexity. Data obtained demonstrate the feasibility of using HICF fingerprinting to resolve the homoeologous chromosomes of the two distinct genomes in switchgrass, providing a robust and accurate BAC-based physical platform for this species. The genomic resources and sequence data generated will lay the foundation for deciphering the switchgrass genome and lead the way for an accurate genome sequencing strategy.

Economic and Environmental Assessment of Switchgrass Production on High-Fertility Soil and an Assessment of Anaerobic Digests as an Intermediate Market for Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rhykerd, Robert; Bierma, Thomas; Jin, Guang

2014-03-31

This project had two parts. Part 1 was an economic and environmental assessment of switchgrass production on high-fertility soil, and included an assessment of the effects of field irrigation with treated municipal wastewater (Studies A,C, and E in Final Agreement). Part 2 was an assessment of methods to enhance anaerobic digestion of switchgrass, and included evaluation of several other potential biomass feedstocks (Studies B and D in Final Agreement). Results from Part 1 demonstrated that switchgrass does not compete economically against a corn and soybean rotation on highly productive soils. All four varieties of switchgrass lost money while corn andmore » soybeans were profitable in all four years of this study. Breakeven prices for the four switchgrass varieties were calculated using production costs. The installation of a center pivot irrigation system had minimal impact on crop production and corn and soybean production remained profitable in the year the irrigator was installed. Because of drought and delays in installing the wastewater treatment plant, the irrigation system was not used until year 4 of this study. Therefore, longer term studies evaluating multiple year studies on the impact of irrigation on switchgrass are warranted. Results from irrigating with treated municipal wastewater showed no negative impact on soil quality. Results from Part 2 demonstrated that anaerobic digestion (AD) of switchgrass could be significantly enhanced using low heat (100oC) and mild caustic pretreatment without fine-grinding. Heat for pretreatment could be available from biogas-based combined heat and power (CHP) systems. In bench-top digesters simulating municipal wastewater treatment AD, methane production of coarse-ground switchgrass increased over 20-fold with pretreatment compared to untreated switchgrass. Bench-top studies simulating dairy-based AD also found high specific methane yield, but even untreated switchgrass digested reasonably well, indicating the value of AD micro-organisms acclimated to lignocellulosic feedstock. Similar results were found for corn stover. However, oak and maple leaves – representing common urban yard waste – were more resistant to digestion even with pretreatment. More pretreatment research is required before yard waste could become a significant feedstock for the low/medium-solids digesters common to municipal wastewater or livestock operations.« less

The effect of switchgrass loadings on feedstock solubilization and biofuel production by Clostridium thermocellum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Verbeke, Tobin J.; Garcia, Gabriela M.; Elkins, James G.

High solids loading fermentations are necessary for the industrialization of lignocellulosic ethanol. To date, only a few studies have investigated the effect of solids loadings on microorganisms of interest for consolidated bioprocessing (CBP). Here, the effect that various switchgrass loadings have on Clostridium thermocellum solubilization and bioconversion are investigated. C. thermocellum was grown for ten days on 10, 25 or 50 g/L switchgrass or Avicel at equivalent glucan loadings. Avicel was completely consumed at all loadings, but total cellulose solubilization decreased from 63% to 37% as switchgrass loadings increased from 10 g/L to 50 g/L. Washed, spent switchgrass could bemore » additionally hydrolyzed and fermented in second-round fermentations suggesting access to fermentable substrates was not the limiting factor at higher feedstock loadings. Fermentations of Avicel or cellobiose using culture medium supplemented with 50% spent fermentation broth identified that compounds present in the samples collected from the 25 or 50 g/L switchgrass loadings were the most inhibitory to continued fermentation. Finally, recalcitrance alone cannot fully account for differences in solubilization and end-production formation between switchgrass and Avicel at increased substrate loadings. Effort to decouple metabolic inhibition from inhibition of hydrolysis suggest that C. thermocellum’s hydrolytic machinery is more vulnerable to inhibition from switchgrass-derived inhibitors than is the bacterium’s metabolism.« less

The effect of switchgrass loadings on feedstock solubilization and biofuel production by Clostridium thermocellum

DOE PAGES

Verbeke, Tobin J.; Garcia, Gabriela M.; Elkins, James G.

2017-11-30

High solids loading fermentations are necessary for the industrialization of lignocellulosic ethanol. To date, only a few studies have investigated the effect of solids loadings on microorganisms of interest for consolidated bioprocessing (CBP). Here, the effect that various switchgrass loadings have on Clostridium thermocellum solubilization and bioconversion are investigated. C. thermocellum was grown for ten days on 10, 25 or 50 g/L switchgrass or Avicel at equivalent glucan loadings. Avicel was completely consumed at all loadings, but total cellulose solubilization decreased from 63% to 37% as switchgrass loadings increased from 10 g/L to 50 g/L. Washed, spent switchgrass could bemore » additionally hydrolyzed and fermented in second-round fermentations suggesting access to fermentable substrates was not the limiting factor at higher feedstock loadings. Fermentations of Avicel or cellobiose using culture medium supplemented with 50% spent fermentation broth identified that compounds present in the samples collected from the 25 or 50 g/L switchgrass loadings were the most inhibitory to continued fermentation. Finally, recalcitrance alone cannot fully account for differences in solubilization and end-production formation between switchgrass and Avicel at increased substrate loadings. Effort to decouple metabolic inhibition from inhibition of hydrolysis suggest that C. thermocellum’s hydrolytic machinery is more vulnerable to inhibition from switchgrass-derived inhibitors than is the bacterium’s metabolism.« less

A model for deploying switchgrass for bioenergy in an intensive agricultural landscape

USDA-ARS?s Scientific Manuscript database

Switchgrass bioenergy research has been conducted in Nebraska since 1990. In that time, significant progress has been made in switchgrass breeding and genetics, molecular genetics, establishment, fertility management, production economics, production energetics, harvest and storage management, ecos...

Comparison of solid-state and submerged-state fermentation for the bioprocessing of switchgrass to ethanol and acetate by Clostridium phytofermentans.

PubMed

Jain, Abhiney; Morlok, Charles K; Henson, J Michael

2013-01-01

The conversion of sustainable energy crops using microbiological fermentation to biofuels and bioproducts typically uses submerged-state processes. Alternatively, solid-state fermentation processes have several advantages when compared to the typical submerged-state processes. This study compares the use of solid-state versus submerged-state fermentation using the mesophilic anaerobic bacterium Clostridium phytofermentans in the conversion of switchgrass to the end products of ethanol, acetate, and hydrogen. A shift in the ratio of metabolic products towards more acetate and hydrogen production than ethanol production was observed when C. phytofermentans was grown under solid-state conditions as compared to submerged-state conditions. Results indicated that the end product concentrations (in millimolar) obtained using solid-state fermentation were higher than using submerged-state fermentation. In contrast, the total fermentation products (in weight of product per weight of carbohydrates consumed) and switchgrass conversion were higher for submerged-state fermentation. The conversion of xylan was greater than glucan conversion under both fermentation conditions. An initial pH of 7 and moisture content of 80 % resulted in maximum end products formation. Scanning electron microscopy study showed the presence of biofilm formed by C. phytofermentans growing on switchgrass under submerged-state fermentation whereas bacterial cells attached to surface and no apparent biofilm was observed when grown under solid-state fermentation. To our knowledge, this is the first study reporting consolidated bioprocessing of a lignocellulosic substrate by a mesophilic anaerobic bacterium under solid-state fermentation conditions.

Extensive genetic diversity present within North American switchgrass germplasm

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a perennial, native North American grass currently grown for ecological restoration and forage purposes that has potential as a biofuel feedstock crop. Understanding the genetic diversity of switchgrass can provide insight into allelic variants important in devel...

Switchgrass cultivars alter microbial contribution to deep soil C

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a perennial, cellulosic biofuel feedstock capable of growing under a wide variety of climatic conditions on land marginally suited to cultivated crops. Due to its perennial nature and deep rooting characteristics, switchgrass contributes to soil C sequestration ...

Efficient Methods of Estimating Switchgrass Biomass Supplies

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is being developed as a biofuel feedstock for the United States. Efficient and accurate methods to estimate switchgrass biomass feedstock supply within a production area will be required by biorefineries. Our main objective was to determine the effectiveness of in...

The application of ultrasound in the enzymatic hydrolysis of switchgrass

USDA-ARS?s Scientific Manuscript database

In a series of experiments, untreated and ammonium hydroxide pretreated Klenow lowland variety switchgrasses are converted to reducing sugars using low frequency (20 kHz) ultrasound and commercially-available cellulase enzyme. Results from experiments using untreated and pretreated switchgrasses wit...

Gas exchange and stand-level estimates of water use and gross primary productivity in an experimental pine and switchgrass intercrop forestry system on the Lower Coastal Plain of North Carolina, U.S.A

Treesearch

Janine M. Albaugha; Jean-Christophe Domeca; Chris A. Maier; Eric B. Sucre; Zakiya H. Leggett; John S. King

2014-01-01

Despite growing interest in using switchgrass (Panicum virgatum L.) as a biofuel, there are limiteddata on the physiology of this species and its effect on stand water use and carbon (C) assimilationwhen grown as a forest intercrop for bioenergy. Therefore, we quantified gas exchange rates of switch-grass within intercropped plots and in pure switchgrass plots during...

Model evaluation of plant metal content and biomass yield for the phytoextraction of heavy metals by switchgrass.

PubMed

Chen, Bo-Ching; Lai, Hung-Yu; Juang, Kai-Wei

2012-06-01

To better understand the ability of switchgrass (Panicum virgatum L.), a perennial grass often relegated to marginal agricultural areas with minimal inputs, to remove cadmium, chromium, and zinc by phytoextraction from contaminated sites, the relationship between plant metal content and biomass yield is expressed in different models to predict the amount of metals switchgrass can extract. These models are reliable in assessing the use of switchgrass for phytoremediation of heavy-metal-contaminated sites. In the present study, linear and exponential decay models are more suitable for presenting the relationship between plant cadmium and dry weight. The maximum extractions of cadmium using switchgrass, as predicted by the linear and exponential decay models, approached 40 and 34 μg pot(-1), respectively. The log normal model was superior in predicting the relationship between plant chromium and dry weight. The predicted maximum extraction of chromium by switchgrass was about 56 μg pot(-1). In addition, the exponential decay and log normal models were better than the linear model in predicting the relationship between plant zinc and dry weight. The maximum extractions of zinc by switchgrass, as predicted by the exponential decay and log normal models, were about 358 and 254 μg pot(-1), respectively. To meet the maximum removal of Cd, Cr, and Zn, one can adopt the optimal timing of harvest as plant Cd, Cr, and Zn approach 450 and 526 mg kg(-1), 266 mg kg(-1), and 3022 and 5000 mg kg(-1), respectively. Due to the well-known agronomic characteristics of cultivation and the high biomass production of switchgrass, it is practicable to use switchgrass for the phytoextraction of heavy metals in situ. Copyright © 2012 Elsevier Inc. All rights reserved.

Improving water quality in the Chesapeake Bay using payments for ecosystem services for perennial biomass for bioenergy and biofuel production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Woodbury, Peter B.; Kemanian, Armen R.; Jacobson, Michael

Replacing row crops with perennial bioenergy crops may reduce nitrogen (N) loading to surface waters. We estimated the benefits, costs, and potential for replacing maize with switchgrass to meet required N loading reduction targets for the Chesapeake Bay (CB) of 26.9 Gg -1. After subtracting the potential reduction in N loading due to improved N fertilizer practices for maize, a further 22.8 Gg reduction is required. Replacing maize with fertilized switchgrass could reduce N loading to the CB by 18 kg ha -1 y -1, meeting 31% of the N reduction target. The break-even price of fertilized switchgrass to providemore » the same profit as maize in the CB is 111 $Mg -1 (oven-dry basis throughout). Growers replacing maize with switchgrass could receive an ecosystem service payment of 148 ha -1 based on the price paid in Maryland for planting a rye cover crop. For our estimated average switchgrass yield of 9.9 Mg ha -1, and the greater N loading reduction of switchgrass compared to a cover crop, this equates to 24 dollars Mg -1. The annual cost of this ecosystem service payment to induce switchgrass planting is 13.29 dollars kg -1 of N. Using the POLYSYS model to account for competition among food, feed, and biomass markets, we found that with the ecosystem service payment for switchgrass of 25 $ Mg -1 added to a farm-gate price of 111 dollars Mg -1, 11% of the N loading reduction target could be met while also producing 1.3 Tg of switchgrass, potentially yielding 420 dam 3 y -1 of ethanol.« less

Improving water quality in the Chesapeake Bay using payments for ecosystem services for perennial biomass for bioenergy and biofuel production

DOE PAGES

Woodbury, Peter B.; Kemanian, Armen R.; Jacobson, Michael; ...

2017-02-03

Replacing row crops with perennial bioenergy crops may reduce nitrogen (N) loading to surface waters. We estimated the benefits, costs, and potential for replacing maize with switchgrass to meet required N loading reduction targets for the Chesapeake Bay (CB) of 26.9 Gg -1. After subtracting the potential reduction in N loading due to improved N fertilizer practices for maize, a further 22.8 Gg reduction is required. Replacing maize with fertilized switchgrass could reduce N loading to the CB by 18 kg ha -1 y -1, meeting 31% of the N reduction target. The break-even price of fertilized switchgrass to providemore » the same profit as maize in the CB is 111 $Mg -1 (oven-dry basis throughout). Growers replacing maize with switchgrass could receive an ecosystem service payment of 148 ha -1 based on the price paid in Maryland for planting a rye cover crop. For our estimated average switchgrass yield of 9.9 Mg ha -1, and the greater N loading reduction of switchgrass compared to a cover crop, this equates to 24 dollars Mg -1. The annual cost of this ecosystem service payment to induce switchgrass planting is 13.29 dollars kg -1 of N. Using the POLYSYS model to account for competition among food, feed, and biomass markets, we found that with the ecosystem service payment for switchgrass of 25 $ Mg -1 added to a farm-gate price of 111 dollars Mg -1, 11% of the N loading reduction target could be met while also producing 1.3 Tg of switchgrass, potentially yielding 420 dam 3 y -1 of ethanol.« less

Mapping marginal croplands suitable for cellulosic feedstock crops in the Great Plains, United States

USGS Publications Warehouse

Gu, Yingxin; Wylie, Bruce K.

2016-01-01

Growing cellulosic feedstock crops (e.g., switchgrass) for biofuel is more environmentally sustainable than corn-based ethanol. Specifically, this practice can reduce soil erosion and water quality impairment from pesticides and fertilizer, improve ecosystem services and sustainability (e.g., serve as carbon sinks), and minimize impacts on global food supplies. The main goal of this study was to identify high-risk marginal croplands that are potentially suitable for growing cellulosic feedstock crops (e.g., switchgrass) in the US Great Plains (GP). Satellite-derived growing season Normalized Difference Vegetation Index, a switchgrass biomass productivity map obtained from a previous study, US Geological Survey (USGS) irrigation and crop masks, and US Department of Agriculture (USDA) crop indemnity maps for the GP were used in this study. Our hypothesis was that croplands with relatively low crop yield but high productivity potential for switchgrass may be suitable for converting to switchgrass. Areas with relatively low crop indemnity (crop indemnity <$2 157 068) were excluded from the suitable areas based on low probability of crop failures. Results show that approximately 650 000 ha of marginal croplands in the GP are potentially suitable for switchgrass development. The total estimated switchgrass biomass productivity gain from these suitable areas is about 5.9 million metric tons. Switchgrass can be cultivated in either lowland or upland regions in the GP depending on the local soil and environmental conditions. This study improves our understanding of ecosystem services and the sustainability of cropland systems in the GP. Results from this study provide useful information to land managers for making informed decisions regarding switchgrass development in the GP.

Switchgrass Genetics: Status, Future Directions, and Implications for Simulations

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a C4 polymorphic species with two ploidy levels, two major and numerous minor ecotypes adapted to different plant hardiness zones and ecoregions in its range. Switchgrasses are determinate, photoperiod sensitive, and require short days to induce flowering. Photo...

AmeriFlux US-AR1 ARM USDA UNL OSU Woodward Switchgrass 1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Billesbach, Dave; Bradford, James

This is the AmeriFlux version of the carbon flux data for the site US-AR1 ARM USDA UNL OSU Woodward Switchgrass 1. Site Description - The ARM USDA UNL OSU Woodward Switchgrass 1 tower is located on public land owned by the USDA-ARS Southern Plains Range Research Station in Woodward, Oklahoma. The site is on a former native prairie that is in the process of changing to switchgrass. A second companion site (ARM USDA UNL OSU Woodward Switchgrass 2) is on a former wheat field. In Spring 2009, the former native prairie site was burned, cattle were put on the pasturemore » to graze down emergent grass, and broadleaf herbicide was sprayed. In Summer 2009, the cattle were removed from the pasture, and the site was sprayed with herbicide to kill all grass. In Spring 2010, prior to the planting of switchgrass, final herbicide was sprayed to kill cheat grass and to control broadleaf plants.« less

Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus

DOE PAGES

Henske, John K.; Gilmore, Sean P.; Knop, Doriv; ...

2017-12-20

Anaerobic gut fungi are the primary colonizers of plant material in the rumen microbiome, but are poorly studied due to a lack of characterized isolates. While most genera of gut fungi form extensive rhizoidal networks, which likely participate in mechanical disruption of plant cell walls, fungi within the Caecomyces genus do not possess these rhizoids. Here, we describe a novel fungal isolate, Caecomyces churrovis, which forms spherical sporangia with a limited rhizoidal network yet secretes a diverse set of carbohydrate active enzymes (CAZymes) for plant cell wall hydrolysis. Despite lacking an extensive rhizoidal system, C. churrovis is capable of growthmore » on fibrous substrates like switchgrass, reed canary grass, and corn stover, although faster growth is observed on soluble sugars. Gut fungi have been shown to use enzyme complexes (fungal cellulosomes) in which CAZymes bind to non-catalytic scaffoldins to improve biomass degradation efficiency. However, transcriptomic analysis and enzyme activity assays reveal that C. churrovis relies more on free enzymes compared to other gut fungal isolates. Only 15% of CAZyme transcripts contain non-catalytic dockerin domains in C. churrovis, compared to 30% in rhizoid-forming fungi. Furthermore, C. churrovis is enriched in GH43 enzymes that provide complementary hemicellulose degrading activities, suggesting that a wider variety of these activities are required to degrade plant biomass in the absence of an extensive fungal rhizoid network. In conclusion, molecular characterization of a non-rhizoid-forming anaerobic fungus fills a gap in understanding the roles of CAZyme abundance and associated degradation mechanisms during lignocellulose breakdown within the rumen microbiome.« less

Manipulating microRNAs for improved biomass and biofuels from plant feedstocks.

PubMed

Trumbo, Jennifer Lynn; Zhang, Baohong; Stewart, Charles Neal

2015-04-01

Petroleum-based fuels are nonrenewable and unsustainable. Renewable sources of energy, such as lignocellulosic biofuels and plant metabolite-based drop-in fuels, can offset fossil fuel use and reverse environmental degradation through carbon sequestration. Despite these benefits, the lignocellulosic biofuels industry still faces many challenges, including the availability of economically viable crop plants. Cell wall recalcitrance is a major economic barrier for lignocellulosic biofuels production from biomass crops. Sustainability and biomass yield are two additional, yet interrelated, foci for biomass crop improvement. Many scientists are searching for solutions to these problems within biomass crop genomes. MicroRNAs (miRNAs) are involved in almost all biological and metabolic process in plants including plant development, cell wall biosynthesis and plant stress responses. Because of the broad functions of their targets (e.g. auxin response factors), the alteration of plant miRNA expression often results in pleiotropic effects. A specific miRNA usually regulates a biologically relevant bioenergy trait. For example, relatively low miR156 overexpression leads to a transgenic feedstock with enhanced biomass and decreased recalcitrance. miRNAs have been overexpressed in dedicated bioenergy feedstocks such as poplar and switchgrass yielding promising results for lignin reduction, increased plant biomass, the timing of flowering and response to harsh environments. In this review, we present the status of miRNA-related research in several major biofuel crops and relevant model plants. We critically assess published research and suggest next steps for miRNA manipulation in feedstocks for increased biomass and sustainability for biofuels and bioproducts. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Henske, John K.; Gilmore, Sean P.; Knop, Doriv

Anaerobic gut fungi are the primary colonizers of plant material in the rumen microbiome, but are poorly studied due to a lack of characterized isolates. While most genera of gut fungi form extensive rhizoidal networks, which likely participate in mechanical disruption of plant cell walls, fungi within the Caecomyces genus do not possess these rhizoids. Here, we describe a novel fungal isolate, Caecomyces churrovis, which forms spherical sporangia with a limited rhizoidal network yet secretes a diverse set of carbohydrate active enzymes (CAZymes) for plant cell wall hydrolysis. Despite lacking an extensive rhizoidal system, C. churrovis is capable of growthmore » on fibrous substrates like switchgrass, reed canary grass, and corn stover, although faster growth is observed on soluble sugars. Gut fungi have been shown to use enzyme complexes (fungal cellulosomes) in which CAZymes bind to non-catalytic scaffoldins to improve biomass degradation efficiency. However, transcriptomic analysis and enzyme activity assays reveal that C. churrovis relies more on free enzymes compared to other gut fungal isolates. Only 15% of CAZyme transcripts contain non-catalytic dockerin domains in C. churrovis, compared to 30% in rhizoid-forming fungi. Furthermore, C. churrovis is enriched in GH43 enzymes that provide complementary hemicellulose degrading activities, suggesting that a wider variety of these activities are required to degrade plant biomass in the absence of an extensive fungal rhizoid network. In conclusion, molecular characterization of a non-rhizoid-forming anaerobic fungus fills a gap in understanding the roles of CAZyme abundance and associated degradation mechanisms during lignocellulose breakdown within the rumen microbiome.« less

Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus.

PubMed

Henske, John K; Gilmore, Sean P; Knop, Doriv; Cunningham, Francis J; Sexton, Jessica A; Smallwood, Chuck R; Shutthanandan, Vaithiyalingam; Evans, James E; Theodorou, Michael K; O'Malley, Michelle A

2017-01-01

Anaerobic gut fungi are the primary colonizers of plant material in the rumen microbiome, but are poorly studied due to a lack of characterized isolates. While most genera of gut fungi form extensive rhizoidal networks, which likely participate in mechanical disruption of plant cell walls, fungi within the Caecomyces genus do not possess these rhizoids. Here, we describe a novel fungal isolate, Caecomyces churrovis , which forms spherical sporangia with a limited rhizoidal network yet secretes a diverse set of carbohydrate active enzymes (CAZymes) for plant cell wall hydrolysis. Despite lacking an extensive rhizoidal system, C. churrovis is capable of growth on fibrous substrates like switchgrass, reed canary grass, and corn stover, although faster growth is observed on soluble sugars. Gut fungi have been shown to use enzyme complexes (fungal cellulosomes) in which CAZymes bind to non-catalytic scaffoldins to improve biomass degradation efficiency. However, transcriptomic analysis and enzyme activity assays reveal that C. churrovis relies more on free enzymes compared to other gut fungal isolates. Only 15% of CAZyme transcripts contain non-catalytic dockerin domains in C. churrovis , compared to 30% in rhizoid-forming fungi. Furthermore, C. churrovis is enriched in GH43 enzymes that provide complementary hemicellulose degrading activities, suggesting that a wider variety of these activities are required to degrade plant biomass in the absence of an extensive fungal rhizoid network. Overall, molecular characterization of a non-rhizoid-forming anaerobic fungus fills a gap in understanding the roles of CAZyme abundance and associated degradation mechanisms during lignocellulose breakdown within the rumen microbiome.

Natural hybrids and gene flow between upland and lowland switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a perennial grass native to the North American tallgrass prairie and savanna habitats and is broadly adapted to the central and eastern USA. Upland and lowland ecotypes represent the two major taxa within switchgrass, which have distinct, but overlapping distribu...

Genetic variation of flowering time and biomass in switchgrass

USDA-ARS?s Scientific Manuscript database

The timing of phase change from juvenile (vegetative) to adult with reproductive competence is a key factor influencing biomass yield of switchgrass. A decline in biomass yield is typically observed in switchgrass immediately following completion of flowering. In temperate regions of the USA, if flo...

Switchgrass ecotypes alter microbial contribution to deep-soil C

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a C4, perennial grass that is being developed as a bioenergy crop for the United States. While aboveground biomass production is well documented for switchgrass ecotypes (lowland, upland), little is known about the impact of plant belowground productivity on mic...

Genome-size variation in switchgrass (Panicum virgatum): flow cytometry and cytology reveal rampant aneuploidy

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.), a native perennial dominant of the prairies of North America, has been targeted as a model herbaceous species for biofeedstock development. A flow-cytometric survey of a core set of 11 primarily upland polyploid switchgrass accessions indicated that there was con...

Genetic control of flowering and biomass in switchgrass

USDA-ARS?s Scientific Manuscript database

Early flowering can negatively affect biomass yield of switchgrass. In temperate regions of the USA, flowering occurs in switchgrass around the time of peak biomass yield (about 5 to 8 weeks prior to killing frost), effectively reducing the length of the growing season. The use of late-flowering swi...

Isolation, Characterization, and Quantification of Steroidal Saponins in Switchgrass (Panicum virgatum L.)

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) has been identified for development into an efficient and environment friendly biomass energy crop. A recent five-year study demonstrated that switchgrass grown for biofuel production produced 540 percent more energy than what is needed to grow, harvest and process...

Diversity and population structure of northern switchgrass as revealed though exome capture sequencing

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a polyploid, perennial grass species that is native to North America, and is being developed as a future biofuels feedstock crop. Switchgrass is present primarily in two ecotypes: a northern upland ecotype composed of tetraploid and octoploid accessions, and a so...

Transcriptional analysis of flowering time in switchgrass

USDA-ARS?s Scientific Manuscript database

Over the past two decades, switchgrass (Panicum virgatum) has emerged as a viable biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; a longer vegetative stage leads to an increase in overall biomass yield. The goal of this study was to gain insight into the...

Preference by horses for bedding pellets made from switchgrass (Panicum virgatum) straw

USDA-ARS?s Scientific Manuscript database

The bedding system used for stalled horses can impact their health and well-being. This study examined the saponin concentration in switchgrass (Panicum virgatum) straw, and bedding pellets made from switchgrass straw. Further, this study determined the palatability of bedding pellets made from sw...

Transmission of Switchgrass mosaic virus by Graminella aureovitatta

USDA-ARS?s Scientific Manuscript database

Switchgrass mosaic virus (SwMV) was identified in switchgrass (Panicum virgatum) and was proposed as a new marafivirus based on its genome sequence and comparison with its closest relative, Maize rayado fino virus (MRFV), a type member of the genus, Marafivirus. MRFV only infects maize (Zea mays) an...

Switchgrass and pecan biochar amendments to a sandy coastal soil

USDA-ARS?s Scientific Manuscript database

Sandy soils of the wet, warm SE Coastal Plain have poor physical characteristics and low carbon contents. To improve soil properties, we added switchgrass (Panicum virgatum) and non-activated pecan (Carya illinoinensis) biochar. Switchgrass was ground to a fine powder and added to soil at rates of 0...

Switchgrass cultivars alter microbial contribution to deep soil C in Nebraska

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a C4, perennial grass that is being developed as a bioenergy crop for the United States. While aboveground biomass production is well documented for switchgrass ecotypes (lowland, upland) and cultivars, there has been little focus on the impact of plant belowgro...

Comparative study of switchgrass cultivars using RNA sequencing technology

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a C4 perennial grass, identified as a promising bioenergy crop. Switchgrass exists in two ecotypes, upland and lowland, which are heterotic, or genetically complementary to each other. The objectives of this study are to assess the potential of SNP markers as a b...

Modeling differential growth in switchgrass cultivars across the Central and Southern Great Plains

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) has been recognized as a potential biofuel crop, because it is adapted to a wide range of environmental and climatic conditions. Zones of adaptation for many switchgrass cultivars are well documented and attributed to local adaptation to the temperature and photope...

Impact of harvest time and switchgrass cultivar on sugar release through enzymatic hydrolysis

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a native North American prairie grass being developed for bioenergy production in the central and eastern USA. The objective of this study was to identify the impacts harvest time and switchgrass cultivar had on sugar release variables determined through enzymat...

Co-processing of agricultural plastic waste and switchgrass via tail gas reactive pyrolysis

USDA-ARS?s Scientific Manuscript database

Mixtures of agricultural plastic waste in the form of polyethylene hay bale covers (PE) (4-37%) and switchgrass were investigated using the US Department of Agriculture’s tail gas reactive pyrolysis (TGRP) at different temperatures (400-570 deg C). TGRP of switchgrass and plastic mixtures significan...

Accuracy of genomic prediction in switchgrass (Panicum virgatum L.) improved by accounting for linkage disequilibrium

USDA-ARS?s Scientific Manuscript database

Switchgrass is a relatively high-yielding and environmentally sustainable biomass crop, but further genetic gains in biomass yield must be achieved to make it an economically viable bioenergy feedstock. Genomic selection is an attractive technology to generate rapid genetic gains in switchgrass and ...

Switchgrass harvest time management can impact biomass yield and nutrient content

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a dedicated energy crop native to much of North America. While high-biomass yield is of significant importance for the development of switchgrass as a bioenergy crop, nutrient content in the biomass as it relates to biofuel conversion efficiency is also critical...

Breeding and Selection of New Switchgrass Varieties for Increased Biomass Production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taliaferro, C.M.

2003-05-27

Switchgrass breeding and genetics research was conducted from 1992-2002 at the Oklahoma State University as part of the national DOE-Bioenergy Feedstock Development Program (BFDP) effort to develop the species as a bioenergy feedstock crop. The fundamental objective of the program was to implement and conduct a breeding program to increase biomass yield capability in switchgrass and develop cultivars for the central and southern United States. Supporting research objectives included: (1) switchgrass germplasm collection, characterization, and enhancement; (2) elucidation of cytogenetic and breeding behavior; and (3) identification of best breeding procedures.

Synergistic and Antagonistic Effects of Salinity and pH on Germination in Switchgrass (Panicum virgatum L.)

PubMed Central

Liu, Yuan; Wang, Quanzhen; Zhang, Yunwei; Cui, Jian; Chen, Guo; Xie, Bao; Wu, Chunhui; Liu, Haitao

2014-01-01

The effects of salt-alkaline mixed stress on switchgrass were investigated by evaluating seed germination and the proline, malondialdehyde (MDA) and soluble sugar contents in three switchgrass (Panicum virgatum L.) cultivars in order to identify which can be successfully produced on marginal lands affected by salt-alkaline mixed stress. The experimental conditions consisted of four levels of salinity (10, 60, 110 and 160 mM) and four pH levels (7.1, 8.3, 9.5 and 10.7). The effects of salt-alkaline mixed stress with equivalent coupling of the salinity and pH level on the switchgrass were explored via model analyses. Switchgrass was capable of germinating and surviving well in all treatments under low-alkaline pH (pH≤8.3), regardless of the salinity. However, seed germination and seedling growth were sharply reduced at higher pH values in conjunction with salinity. The salinity and pH had synergetic effects on the germination percentage, germination index, plumular length and the soluble sugar and proline contents in switchgrass. However, these two factors exhibited antagonistic effects on the radicular length of switchgrass. The combined effects of salinity and pH and the interactions between them should be considered when evaluating the strength of salt-alkaline mixed stress. PMID:24454834

Development of a system for characterizing biomass quality of lignocellulosic feedstocks for biochemical conversion

NASA Astrophysics Data System (ADS)

Murphy, Patrick Thomas

The purpose of this research was twofold: (i) to develop a system for screening lignocellulosic biomass feedstocks for biochemical conversion to biofuels and (ii) to evaluate brown midrib corn stover as feedstock for ethanol production. In the first study (Chapter 2), we investigated the potential of corn stover from bm1-4 hybrids for increased ethanol production and reduced pretreatment intensity compared to corn stover from the isogenic normal hybrid. Corn stover from hybrid W64A X A619 and respective isogenic bm hybrids was pretreated by aqueous ammonia steeping using ammonium hydroxide concentrations from 0 to 30%, by weight, and the resulting residues underwent simultaneous saccharification and cofermentation (SSCF) to ethanol. Dry matter (DM) digested by SSCF increased with increasing ammonium hydroxide concentration across all genotypes (P>0.0001) from 277 g kg-1 DM in the control to 439 g kg-1 DM in the 30% ammonium hydroxide pretreatment. The bm corn stover materials averaged 373 g kg-1 DM of DM digested by SSCF compared with 335 g kg-1 DM for the normal corn stover (P<0.0001). Of the bm mutations, bm3 had (i) the greatest effect on cell-wall carbohydrate hydrolysis of corn stover, (ii) the lowest initial cell-wall carbohydrate concentration, (iii) the lowest dry matter remaining after pretreatment, and (iv) the highest amount of monosaccharides released during enzymatic hydrolysis. However, bm corn stover did not reduce the severity of aqueous ammonia steeping pretreatment needed to maximize DM hydrolysis during SSCF compared with normal corn stover. In the remaining studies (Chapters 3 thru 5), a system for analyzing the quality of lignocellulosic biomass feedstocks for biochemical conversion to biofuels (i.e., pretreatment, enzymatic hydrolysis, and fermentation) was developed. To accomplish this, a carbohydrate availability model was developed to characterize feedstock quality. The model partitions carbohydrates within a feedstock material into fractions based on their availability to be converted to fermentable sugars, including non-structural carbohydrates (CN) (monosaccharides, starches, oligosaccharides), biochemically available carbohydrates (CB) (structural carbohydrates susceptible to enzymatic hydrolysis) with an associated 1st-order availability rate constant (k B) and unavailable carbohydrates (CU) (hemicellulose and cellulose in close association with lignin). The model partitions the noncarbohydrate dry matter into extractives, lignin, and ash. Quality parameters were determined using a biomass quality assay that combined established wet-chemistry analyses techniques, including total non-structural carbohydrates (TNC), alcohol insoluble residue (AIR), simultaneous saccharification and fermentation (SSCF), and Klason lignin. The next study evaluated multiple high-throughput (HTP) modifications to the original assay methods, including (i) using filter bags with batch sample processing, (ii) replacement of AIR with neutral detergent fiber (NDF) as a cell-wall isolation procedure, and (iii) elimination of the fermentation organism in the SSCF procedures used to determine biochemically available carbohydrates. The original and the HTP assay methods were compared using corn cobs, hybrid poplar, kenaf, and switchgrass. Biochemically available carbohydrates increased with the HTP methods in the corn cobs, hybrid poplar, and switchgrass, but remained the same in the kenaf. Total available carbohydrates increased and unavailable carbohydrates decreased with the HTP methods in the corn cobs and switchgrass and remained the same in the hybrid poplar and kenaf. There were no differences in total carbohydrates (CT) between the two methods. The final study evaluated the variability of biomass quality parameters in a set of corn stover samples, and developed calibration equations for determining parameter values using near infrared reflectance spectroscopy (NIRS). Fifty-two corn stover samples harvested in Iowa and Wisconsin in 2005 and 2006 were analyzed using the HTP assay for determining feedstock quality for biochemical conversion. Non-structural carbohydrates ranged from 84 to 155 g kg -1 DM, CB ranged from 354 to 557 g kg-1 DM, kB ranged from 0.20 to 0.33 h-1, CA ranged from 463 to 699 g kg-1 DM, and neutral detergent lignin (NDL) ranged from 32 to 74 g kg-1 DM. Significant differences (P<0.0001) among samples were observed for all parameters, except for the availability rate constant of CB. (Abstract shortened by UMI.)

Growth promotion and colonization of switchgrass (Panicum virgatum) cv. Alamo by bacterial endophyte Burkholderia phytofirmans strain PsJN

PubMed Central

2012-01-01

Background Switchgrass is one of the most promising bioenergy crop candidates for the US. It gives relatively high biomass yield and can grow on marginal lands. However, its yields vary from year to year and from location to location. Thus it is imperative to develop a low input and sustainable switchgrass feedstock production system. One of the most feasible ways to increase biomass yields is to harness benefits of microbial endophytes. Results We demonstrate that one of the most studied plant growth promoting bacterial endophytes, Burkholderia phytofirmans strain PsJN, is able to colonize and significantly promote growth of switchgrass cv. Alamo under in vitro, growth chamber, and greenhouse conditions. In several in vitro experiments, the average fresh weight of PsJN-inoculated plants was approximately 50% higher than non-inoculated plants. When one-month-old seedlings were grown in a growth chamber for 30 days, the PsJN-inoculated Alamo plants had significantly higher shoot and root biomass compared to controls. Biomass yield (dry weight) averaged from five experiments was 54.1% higher in the inoculated treatment compared to non-inoculated control. Similar results were obtained in greenhouse experiments with transplants grown in 4-gallon pots for two months. The inoculated plants exhibited more early tillers and persistent growth vigor with 48.6% higher biomass than controls. We also found that PsJN could significantly promote growth of switchgrass cv. Alamo under sub-optimal conditions. However, PsJN-mediated growth promotion in switchgrass is genotype specific. Conclusions Our results show B. phytofirmans strain PsJN significantly promotes growth of switchgrass cv. Alamo under different conditions, especially in the early growth stages leading to enhanced production of tillers. This phenomenon may benefit switchgrass establishment in the first year. Moreover, PsJN significantly stimulated growth of switchgrass cv. Alamo under sub-optimal conditions, indicating that the use of the beneficial bacterial endophytes may boost switchgrass growth on marginal lands and significantly contribute to the development of a low input and sustainable feedstock production system. PMID:22647367

Transcriptome Profiling of Rust Resistance in Switchgrass Using RNA-Seq Analysis

DOE PAGES

Serba, Desalegn D.; Uppalapati, Srinivasa Rao; Mukherjee, Shreyartha; ...

2015-03-16

Switchgrass rust caused by Puccinia emaculata is a major limiting factor for switchgrass (Panicum virgatum L.) production, especially in monoculture. Natural populations of switchgrass displayed diverse reactions to P. emaculata when evaluated in an Ardmore, OK, field. In order to identify the differentially expressed genes during the rust infection process and the mechanisms of switchgrass rust resistance, transcriptome analysis using RNA-Seq was conducted in two pseudo-F 1 parents ('PV281' and 'NFGA472'), and three moderately resistant and three susceptible progenies selected from a three-generation, four-founder switchgrass population (K5 x A4) x (AP13 x VS16). On average, 23.5 million reads per samplemore » (leaf tissue was collected at 0, 24, and 60 h post-inoculation (hpi)) were obtained from paired-end (2 x 100 bp) sequencing on the Illumina HiSeq2000 platform. Furthermore, mapping of the RNA-Seq reads to the switchgrass reference genome (AP13 ver. 1.1 assembly) constructed a total of 84,209 transcripts from 98,007 gene loci among all of the samples. Further analysis revealed that host defense- related genes, including the nucleotide binding site-leucinerich repeat domain containing disease resistance gene analogs, play an important role in resistance to rust infection. Rust-induced gene (RIG) transcripts inherited across generations were identified. The rust-resistant gene transcripts can be a valuable resource for developing molecular markers for rust resistance. Finally we identified the rust-resistant genotypes and gene transcripts which can expedite rust-resistant cultivar development in switchgrass.« less

Transcriptome Profiling of Rust Resistance in Switchgrass Using RNA-Seq Analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Serba, Desalegn D.; Uppalapati, Srinivasa Rao; Mukherjee, Shreyartha

Switchgrass rust caused by Puccinia emaculata is a major limiting factor for switchgrass (Panicum virgatum L.) production, especially in monoculture. Natural populations of switchgrass displayed diverse reactions to P. emaculata when evaluated in an Ardmore, OK, field. In order to identify the differentially expressed genes during the rust infection process and the mechanisms of switchgrass rust resistance, transcriptome analysis using RNA-Seq was conducted in two pseudo-F 1 parents ('PV281' and 'NFGA472'), and three moderately resistant and three susceptible progenies selected from a three-generation, four-founder switchgrass population (K5 x A4) x (AP13 x VS16). On average, 23.5 million reads per samplemore » (leaf tissue was collected at 0, 24, and 60 h post-inoculation (hpi)) were obtained from paired-end (2 x 100 bp) sequencing on the Illumina HiSeq2000 platform. Furthermore, mapping of the RNA-Seq reads to the switchgrass reference genome (AP13 ver. 1.1 assembly) constructed a total of 84,209 transcripts from 98,007 gene loci among all of the samples. Further analysis revealed that host defense- related genes, including the nucleotide binding site-leucinerich repeat domain containing disease resistance gene analogs, play an important role in resistance to rust infection. Rust-induced gene (RIG) transcripts inherited across generations were identified. The rust-resistant gene transcripts can be a valuable resource for developing molecular markers for rust resistance. Finally we identified the rust-resistant genotypes and gene transcripts which can expedite rust-resistant cultivar development in switchgrass.« less

Early lignin pathway enzymes and routes to chlorogenic acid in switchgrass (Panicum virgatum L.).

PubMed

Escamilla-Treviño, Luis L; Shen, Hui; Hernandez, Timothy; Yin, Yanbin; Xu, Ying; Dixon, Richard A

2014-03-01

Studying lignin biosynthesis in Panicum virgatum (switchgrass) has provided a basis for generating plants with reduced lignin content and increased saccharification efficiency. Chlorogenic acid (CGA, caffeoyl quinate) is the major soluble phenolic compound in switchgrass, and the lignin and CGA biosynthetic pathways potentially share intermediates and enzymes. The enzyme hydroxycinnamoyl-CoA: quinate hydroxycinnamoyltransferase (HQT) is responsible for CGA biosynthesis in tobacco, tomato and globe artichoke, but there are no close orthologs of HQT in switchgrass or in other monocotyledonous plants with complete genome sequences. We examined available transcriptomic databases for genes encoding enzymes potentially involved in CGA biosynthesis in switchgrass. The protein products of two hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT) genes (PvHCT1a and PvHCT2a), closely related to lignin pathway HCTs from other species, were characterized biochemically and exhibited the expected HCT activity, preferring shikimic acid as acyl acceptor. We also characterized two switchgrass coumaroyl shikimate 3'-hydroxylase (C3'H) enzymes (PvC3'H1 and PvC3'H2); both of these cytochrome P450s had the capacity to hydroxylate 4-coumaroyl shikimate or 4-coumaroyl quinate to generate caffeoyl shikimate or CGA. Another switchgrass hydroxycinnamoyl transferase, PvHCT-Like1, is phylogenetically distant from HCTs or HQTs, but exhibits HQT activity, preferring quinic acid as acyl acceptor, and could therefore function in CGA biosynthesis. The biochemical features of the recombinant enzymes, the presence of the corresponding activities in plant protein extracts, and the expression patterns of the corresponding genes, suggest preferred routes to CGA in switchgrass.

Transcriptomic analysis reveals vacuolar Na+ (K+)/H+ antiporter gene contributing to growth, development, and defense in switchgrass (Panicum virgatum L.).

PubMed

Huang, Yanhua; Cui, Xin; Cen, Huifang; Wang, Kehua; Zhang, Yunwei

2018-04-10

Intracellular Na + (K + )/H + antiporters (NHXs) have pivotal functions in regulating plant growth, development, and resistance to a range of stresses. To gain insight into the molecular events underlying their actions in switchgrass (Panicum virgatum L.), we analyzed transcriptomic changes between PvNHX1-overexpression transgenic lines and wild-type (WT) plants using RNA sequencing (RNA-seq) technology. The comparison of transcriptomic data from the WT and transgenic plants revealed a large number of differentially expressed genes (DEGs) in the latter. Gene ontology (GO) and KEGG pathway analyses showed that these DEGs were associated with a wide range of functions, and participated in many biological processes. For example, we found that PvNHX1 had an important role in plant growth through its regulation of photosynthetic activity and cell expansion. In addition, PvNHX1 regulated K + homeostasis, cell expansion and pollen development, indicating that it has unique and specific roles in flower development. We also found that transgenic switchgrass exhibited a higher level of transcription of defense-related genes, especially those involved in disease resistance. We showed that PvNHX1 had an important role in plant growth and development through its regulation of photosynthetic activity, cell expansion, K + homeostasis, and pollen development. Additionally, PvNHX1 overexpression activated a complex signal transduction network in response to various biotic and abiotic stresses. In relation to plant growth, development, and defense responses, PvNHX1 also had a vital regulatory role in the formation of a series of plant hormones and transcription factors (TFs). The reliability of the RNA-seq data was confirmed by quantitative real-time PCR. Our data provide a valuable foundation for further research into the molecular mechanisms and physiological roles of NHXs in plants.

Transcriptional Analysis of Flowering Time in Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tornqvist, Carl-Erik; Vaillancourt, Brieanne; Kim, Jeongwoon

Over the past two decades, switchgrass (Panicum virgatum) has emerged as a priority biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; therefore, increasing the length of vegetative growth will lead to an increase in overall biomass yield. The goal of this study was to gain insight into the control of flowering time in switchgrass that would assist in development of cultivars with longer vegetative phases through delayed flowering. RNA sequencing was used to assess genome-wide expression profiles across a developmental series between switchgrass genotypes belonging to the two main ecotypes: upland, typically earlymore » flowering, and lowland, typically late flowering. Leaf blades and tissues enriched for the shoot apical meristem (SAM) were collected in a developmental series from emergence through anthesis for RNA extraction. RNA from samples that flanked the SAM transition stage was sequenced for expression analyses. The analyses revealed differential expression patterns between early- and late-flowering genotypes for known flowering time orthologs. Namely, genes shown to play roles in photoperiod response and the circadian clock in other species were identified as potential candidates for regulating flowering time in the switchgrass genotypes analyzed. Based on their expression patterns, many of the differentially expressed genes could also be classified as putative promoters or repressors of flowering. The candidate genes presented here may be used to guide switchgrass improvement through marker-assisted breeding and/or transgenic or gene editing approaches.Over the past two decades, switchgrass (Panicum virgatum) has emerged as a priority biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; therefore, increasing the length of vegetative growth will lead to an increase in overall biomass yield. The goal of this study was to gain insight into the control of flowering time in switchgrass that would assist in development of cultivars with longer vegetative phases through delayed flowering. RNA sequencing was used to assess genome-wide expression profiles across a developmental series between switchgrass genotypes belonging to the two main ecotypes: upland, typically early flowering, and lowland, typically late flowering. Leaf blades and tissues enriched for the shoot apical meristem (SAM) were collected in a developmental series from emergence through anthesis for RNA extraction. RNA from samples that flanked the SAM transition stage was sequenced for expression analyses. The analyses revealed differential expression patterns between early- and late-flowering genotypes for known flowering time orthologs. Namely, genes shown to play roles in photoperiod response and the circadian clock in other species were identified as potential candidates for regulating flowering time in the switchgrass genotypes analyzed. Based on their expression patterns, many of the differentially expressed genes could also be classified as putative promoters or repressors of flowering. The candidate genes presented here may then be used to guide switchgrass improvement through marker-assisted breeding and/or transgenic or gene editing approaches.« less

Transcriptional Analysis of Flowering Time in Switchgrass

DOE PAGES

Tornqvist, Carl-Erik; Vaillancourt, Brieanne; Kim, Jeongwoon; ...

2017-04-27

Over the past two decades, switchgrass (Panicum virgatum) has emerged as a priority biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; therefore, increasing the length of vegetative growth will lead to an increase in overall biomass yield. The goal of this study was to gain insight into the control of flowering time in switchgrass that would assist in development of cultivars with longer vegetative phases through delayed flowering. RNA sequencing was used to assess genome-wide expression profiles across a developmental series between switchgrass genotypes belonging to the two main ecotypes: upland, typically earlymore » flowering, and lowland, typically late flowering. Leaf blades and tissues enriched for the shoot apical meristem (SAM) were collected in a developmental series from emergence through anthesis for RNA extraction. RNA from samples that flanked the SAM transition stage was sequenced for expression analyses. The analyses revealed differential expression patterns between early- and late-flowering genotypes for known flowering time orthologs. Namely, genes shown to play roles in photoperiod response and the circadian clock in other species were identified as potential candidates for regulating flowering time in the switchgrass genotypes analyzed. Based on their expression patterns, many of the differentially expressed genes could also be classified as putative promoters or repressors of flowering. The candidate genes presented here may be used to guide switchgrass improvement through marker-assisted breeding and/or transgenic or gene editing approaches.Over the past two decades, switchgrass (Panicum virgatum) has emerged as a priority biofuel feedstock. The bulk of switchgrass biomass is in the vegetative portion of the plant; therefore, increasing the length of vegetative growth will lead to an increase in overall biomass yield. The goal of this study was to gain insight into the control of flowering time in switchgrass that would assist in development of cultivars with longer vegetative phases through delayed flowering. RNA sequencing was used to assess genome-wide expression profiles across a developmental series between switchgrass genotypes belonging to the two main ecotypes: upland, typically early flowering, and lowland, typically late flowering. Leaf blades and tissues enriched for the shoot apical meristem (SAM) were collected in a developmental series from emergence through anthesis for RNA extraction. RNA from samples that flanked the SAM transition stage was sequenced for expression analyses. The analyses revealed differential expression patterns between early- and late-flowering genotypes for known flowering time orthologs. Namely, genes shown to play roles in photoperiod response and the circadian clock in other species were identified as potential candidates for regulating flowering time in the switchgrass genotypes analyzed. Based on their expression patterns, many of the differentially expressed genes could also be classified as putative promoters or repressors of flowering. The candidate genes presented here may then be used to guide switchgrass improvement through marker-assisted breeding and/or transgenic or gene editing approaches.« less

Response of Alamo switchgrass tissue chemistry and biomass to nitrogen fertilization in West Tennessee, USA

USDA-ARS?s Scientific Manuscript database

The purpose of this research was to examine above- and belowground responses to nitrogen fertilization in 5-year old “Alamo” switchgrass (Panicum virgatum). A fertilizer experiment included spring and fall sampling of switchgrass grown under annual applications of 0, 67, and 202 kg N ha-1. Nitrogen ...

Switchgrass (Panicum virgatum L.) genotypes differ between coastal sites and inland road corridors in the Northeastern US

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a North American grass that exhibits vast genetic diversity across its geographic range. In the Northeast, switchgrass was restricted to a narrow zone adjacent to the coastal salt marsh, but current populations inhabit inland road verges raising questions about t...

Impact of harvest time and cultivar on conversion of switchgrass to bio-oils via fast pyrolysis

USDA-ARS?s Scientific Manuscript database

The study of the effects of harvest time on switchgrass (Panicum virgatum L.) biomass and bioenergy production reported herein is the final part complementing two prior studies reporting on the harvest of six switchgrass cultivars grown at three northern United States locations over three years, har...

Switchgrass growth and pine-switchgrass interactions in established intercropping systems

DOE PAGES

Tian, Shiying; Cacho, Julian F.; Youssef, Mohamed A.; ...

2016-06-22

Intercropping switchgrass ( Panicum virgatum L.) with loblolly pine ( Pinus taeda L.) has been proposed for producing bioenergy feedstock in the southeastern United States. This study investigated switchgrass growth and pine–switchgrass interactions at two established experimental fields (7-year-old Lenoir site and 5-year-old Carteret site) located on the coastal plain of eastern United States. Position effects (edge and center of switchgrass alley in intercropping plots) and treatment effects (intercropping vs. grass-only) on above ground switchgrass growth were evaluated. Interspecific interactions with respect to capturing resources (light, soil water, and nitrogen) were investigated by measuring photosynthetically active radiation (PAR) above grassmore » canopy, soil moisture, and soil mineral nitrogen contents. Switchgrass growth was significantly (P = 0.001) affected by treatments in Lenoir and by position (P < 0.0001) in both study sites. Relative to the center, PAR above grass canopy at edge in both sites was about 48% less during the growing season. Soil water content during the growing season at the edge of grass alley was significantly (P = 0.0001) lower by 23% than at the center in Lenoir, while no significant (P = 0.42) difference was observed in Carteret, in spite of more grass growth at center at both sites. Soil mineral nitrogen content at the center of intercropping plots in Lenoir (no fertilization during 2015) was significantly (P < 0.07) lower than at the edge during the peak of growing season (June, July, and August), but not during early and late parts of growing season (May, September, and November). Position effects on soil water and mineral nitrogen were less evident under conditions with higher external inputs (rainfall and fertilization) and lower plant uptake during nongrowing seasons. Here, results from this study contributed to a better understanding of above- and belowground pine–switchgrass interactions which is necessary to properly manage this new cultivation system for bioenergy production in the southeastern United States.« less

Genetic Transformation of Switchgrass

NASA Astrophysics Data System (ADS)

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Genetic transformation of switchgrass.

PubMed

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

2009-01-01

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Soil carbon dynamics beneath switchgrass as indicated by stable isotope analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garten, C.T. Jr.; Wullschleger, S.D.

2000-04-01

Surface (0--40 cm) soil organic carbon (SOC) dynamics were studied beneath four switchgrass (Panicum virgatum L.) field trails in the southeastern US. Soil organic carbon was partitioned into particulate organic matter (POM) and mineral-associated organic matter (MOM). Most (75--90%) of the SOC at each study site was affiliated with MOM (<0.053 mm). Changes in stable carbon isotope ratios were used to derive carbon inputs to and losses from POM and MOM at each site. Inventories of existing SOC and new C{sub 4}-derived SOC beneath switchgrass decreased with increasing soil depth. Approximately 5 yr after establishment, 19 to 31% of themore » existing SOC inventories beneath switchgrass had been derived from new C{sub 4}-carbon inputs. Calculated turnover times of POM and MOM ranged from 2.4 to 4.3 yr and 26 to 40 yr, respectively. The turnover time of SOC in the POM fraction increased with decreasing mean annual temperature. A simple, two-compartment model was parameterized to predict the potential for soil carbon sequestration under switchgrass. An example calculation with the model indicated a measurable and verifiable recovery of soil carbon (=12% increase) on degraded lands through one decade of switchgrass production. The potential to sequester carbon through switchgrass cultivation will depend on initial soil carbon inventories, prevailing climate, soil types and site management.« less

Soil carbon dynamics beneath switchgrass as indicated by stable isotope analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garten Jr, Charles T; Wullschleger, Stan D

2000-04-01

Surface (0-40 cm) soil organic carbon (SOC) dynamics were studied beneath four switchgrass (Panicum virgatum L.) field trials in the southeastern United States. Soil organic carbon was partitioned into particulate organic matter (POM) and mineral-associated organic matter (MOM). Most (75-90%) of the SOC at each study site was affiliated with MOM (<0.053 mm). Changes in stable carbon isotope ratios were used to derive carbon inputs to and losses from POM and MOM at each site. Inventories of existing SOC and new C4-derived SOC beneath switchgrass decreased with increasing soil depth. Approximately 5 yr after establishment, 19 to 31% of themore » existing SOC inventories beneath switchgrass had been derived from new C{sub 4}-carbon inputs. Calculated turnover times of POM and MOM ranged from 2.4 to 4.3 yr and 26 to 40 yr, respectively. The turnover time of SOC in the POM fraction increased with decreasing mean annual temperature. A simple, two-compartment model was parameterized to predict the potential for soil carbon sequestration under switchgrass. An example calculation with the model indicated a measurable and verifiable recovery of soil carbon ({approx}12% increase) on degraded lands through one decade of switchgrass production. The potential to sequester carbon through switchgrass cultivation will depend on initial soil carbon inventories, prevailing climate, soil type, and site management.« less

Switchgrass cultivar

DOEpatents

Wu, Yanqi; Taliaferro, Charles M.

2012-10-02

A new cultivar of switchgrass `Cimarron` (SL93 2001-1) having increased biomass yield is provided. The switchgrass comprises all the morphological and physiological properties of the cultivar grown from a seed deposited under American Type Culture Collection (ATCC) No. PTA-10116. The invention also provides seeds, progeny, parts and methods of use of Cimarron, such as for the production of biofuels.

Legacies in switchgrass resistance to and recovery from drought suggest that good years can sustain plants through bad years

USDA-ARS?s Scientific Manuscript database

The warm-season perennial switchgrass (Panicum virgatum) is a candidate bioenergy crop. To be successful, switchgrass production must be maintained on low-quality landscapes with minimal inputs while facing future climates that are expected to be more extreme and more variable. We propose that ant...

Impact of harvest time and switchgrass cultivar on conversion to sugars and pyrolysis oils using biochemical and thermochemical routes

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.), a perennial grass native to much of North America, is undergoing development as a dedicated energy crop. While high-biomass yield is necessary for the development of switchgrass as a bioenergy crop, composition of the biomass and nutrient content as they relate to...

Silicon cantilever functionalization for cellulose-specific chemical force imaging of switchgrass

DOE PAGES

Lee, Ida; Evans, Barbara R.; Foston, Marcus B.; ...

2015-05-08

A method for direct functionalization of silicon and silicon nitride cantilevers with bifunctional silanes was tested with model surfaces to determine adhesive forces for different hydrogen-bonding chemistries. Application for biomass surface characterization was tested by mapping switchgrass and isolated switchgrass cellulose in topographic and force-volume mode using a cellulose-specific cantilever.

The WRKY transcription factor family and senescence in switchgrass.

PubMed

Rinerson, Charles I; Scully, Erin D; Palmer, Nathan A; Donze-Reiner, Teresa; Rabara, Roel C; Tripathi, Prateek; Shen, Qingxi J; Sattler, Scott E; Rohila, Jai S; Sarath, Gautam; Rushton, Paul J

2015-11-09

Early aerial senescence in switchgrass (Panicum virgatum) can significantly limit biomass yields. WRKY transcription factors that can regulate senescence could be used to reprogram senescence and enhance biomass yields. All potential WRKY genes present in the version 1.0 of the switchgrass genome were identified and curated using manual and bioinformatic methods. Expression profiles of WRKY genes in switchgrass flag leaf RNA-Seq datasets were analyzed using clustering and network analyses tools to identify both WRKY and WRKY-associated gene co-expression networks during leaf development and senescence onset. We identified 240 switchgrass WRKY genes including members of the RW5 and RW6 families of resistance proteins. Weighted gene co-expression network analysis of the flag leaf transcriptomes across development readily separated clusters of co-expressed genes into thirteen modules. A visualization highlighted separation of modules associated with the early and senescence-onset phases of flag leaf growth. The senescence-associated module contained 3000 genes including 23 WRKYs. Putative promoter regions of senescence-associated WRKY genes contained several cis-element-like sequences suggestive of responsiveness to both senescence and stress signaling pathways. A phylogenetic comparison of senescence-associated WRKY genes from switchgrass flag leaf with senescence-associated WRKY genes from other plants revealed notable hotspots in Group I, IIb, and IIe of the phylogenetic tree. We have identified and named 240 WRKY genes in the switchgrass genome. Twenty three of these genes show elevated mRNA levels during the onset of flag leaf senescence. Eleven of the WRKY genes were found in hotspots of related senescence-associated genes from multiple species and thus represent promising targets for future switchgrass genetic improvement. Overall, individual WRKY gene expression profiles could be readily linked to developmental stages of flag leaves.

The culturable endophytic fungal communities of switchgrass grown on a coal-mining site and their effects on plant growth

PubMed Central

Amna, Amna; Opiyo, Stephen Obol

2018-01-01

Plants have a diverse endophytic microbiome that is functionally important for their growth, development, and health. In this study, the diversity and specificity of culturable endophytic fungal communities were explored in one of the most important biofuel crops, switchgrass plants (Panicum virgatum L.), which have been cultivated on a reclaimed coal-mining site for more than 20 years. The endophytic fungi were isolated from the surface-sterilized shoot (leaf and stem), root, and seed tissues of switchgrass plants and then cultured for identification. A total of 1339 fungal isolates were found and 22 operational taxonomic units (OTUs) were sequence identified by internal transcribed spacer (ITS) primers and grouped into 7 orders and 4 classes. Although a diverse range of endophytic fungi associated with switchgrass were documented, the most abundant class, order, and species were Sordariomycetes, Hypocreales, and Fusarium spp. respectively. About 86% of the isolated endophytic fungi were able to enhance the heights of the shoots; 69% could increase the shoot fresh weights; and 62% could improve the shoot dry weights after being reintroduced back into the switchgrass plants, which illustrated their functional importance. Through the Shannon Diversity Index analysis, we observed a gradation of species diversity, with shoots and roots having the similar values and seeds having a lesser value. It was observed that the switchgrass plants showing better growth performance displayed higher endophytic fungal species diversity and abundance. It was also discovered that the rhizosphere soil organic matter content was positively correlated with the fungal species diversity. All these data demonstrate the functional association of these beneficial endophytic fungi with switchgrass and their great potential in improving the switchgrass growth and biomass to benefit the biofuel industry by reducing chemical inputs and burden to the environment. PMID:29902231

The culturable endophytic fungal communities of switchgrass grown on a coal-mining site and their effects on plant growth.

PubMed

Xia, Ye; Amna, Amna; Opiyo, Stephen Obol

2018-01-01

Plants have a diverse endophytic microbiome that is functionally important for their growth, development, and health. In this study, the diversity and specificity of culturable endophytic fungal communities were explored in one of the most important biofuel crops, switchgrass plants (Panicum virgatum L.), which have been cultivated on a reclaimed coal-mining site for more than 20 years. The endophytic fungi were isolated from the surface-sterilized shoot (leaf and stem), root, and seed tissues of switchgrass plants and then cultured for identification. A total of 1339 fungal isolates were found and 22 operational taxonomic units (OTUs) were sequence identified by internal transcribed spacer (ITS) primers and grouped into 7 orders and 4 classes. Although a diverse range of endophytic fungi associated with switchgrass were documented, the most abundant class, order, and species were Sordariomycetes, Hypocreales, and Fusarium spp. respectively. About 86% of the isolated endophytic fungi were able to enhance the heights of the shoots; 69% could increase the shoot fresh weights; and 62% could improve the shoot dry weights after being reintroduced back into the switchgrass plants, which illustrated their functional importance. Through the Shannon Diversity Index analysis, we observed a gradation of species diversity, with shoots and roots having the similar values and seeds having a lesser value. It was observed that the switchgrass plants showing better growth performance displayed higher endophytic fungal species diversity and abundance. It was also discovered that the rhizosphere soil organic matter content was positively correlated with the fungal species diversity. All these data demonstrate the functional association of these beneficial endophytic fungi with switchgrass and their great potential in improving the switchgrass growth and biomass to benefit the biofuel industry by reducing chemical inputs and burden to the environment.

Evaluation of Switchgrass as a co-firing fuel in the Southeast

DOE Office of Scientific and Technical Information (OSTI.GOV)

Southern Research Institute

2001-11-01

The ''Evaluation of Switchgrass as a Co-Firing Fuel in the Southeast'' is a comprehensive project incorporating the highest yielding variety of switchgrass, unique harvesting methods, detailed parametric evaluations in a state-of-the-art combustion research facility, and a full-scale demonstration in a tangentially-fired Alabama Power Company power boiler. These features were incorporated into the project to reduce the technical and economic risk of yielding a practical renewable energy option for the southeastern US. There are particular incentives for proving the feasibility of switchgrass as a biomass fuel in the southeastern US. Even though agriculture is a predominant industry much of the landmore » in this region is under-utilized, marginal farmland. As a result, some of the poorest counties in the nation are located in this region. The yields of switchgrass are substantially higher in the southeastern US than in other regions. Yield, or productivity, is a critical factor in determining the feasibility of biomass fuel. Yields in small research plots in the region averaged 25.8 Mg/ha (11.5 tons/acre) over the period 1990-1994. Achievable commercial yield in the southeastern US will likely be about 15.7 Mg/ha (7 tons/acre) with currently available varieties. Use of switchgrass as a supplemental fuel for coal-fired utility boilers could create an enormous market for growers. The Southern Company has 23,000 MW of coal-fired capacity in the southeast. If only 1% of this capacity was provided by switchgrass instead of coal, 74,500 ha (184,000 acres) of production would be needed. This would generate 1,288,000 tons of switchgrass which, if valued at $35/ton, would amount to over $45 million.« less

Effects of torrefaction and densification on switchgrass pyrolysis products

DOE PAGES

Yang, Zixu; Sarkar, Madhura; Kumar, Ajay; ...

2014-12-01

Abstract The pyrolysis behaviors of four types of pretreated switchgrass (torrefied at 230 and 270 °C, densification, and torrefaction at 270 ºC followed by densification) were studied at three temperatures (500, 600, 700 ºC) using a pyroprobe attached to a gas chromatogram mass spectroscopy (Py-GC/MS). The torrefaction of switchgrass improved its oxygen to carbon ratio and energy content. Contents of anhydrous sugars and phenols in pyrolysis products of torrefied switchgrass were higher than those in pyrolysis products of raw switchgrass. As the torrefaction temperature increased from 230 to 270 °C, the contents of anhydrous sugars and phenols in pyrolysis productsmore » increased whereas content of guaiacols decreased. High pyrolysis temperature (600 and 700 °C as compared to 500 °C) enhanced decomposition of lignin and anhydrous sugars, leading to increase in phenols, aromatics and furans. Densification enhanced depolymerization of cellulose and hemicellulose during pyrolysis.« less

Intra-Annual Changes in Biomass, Carbon, and Nitrogen Dynamics at 4-Year Old Switchgrass Field Trials in West Tennessee, USA

USDA-ARS?s Scientific Manuscript database

Switchgrass is a potential bioenergy crop that could promote soil C sequestration in some environments. We compared four switchgrass cultivars on a well-drained Alfisol to test for differences in biomass, C, and N dynamics during the fourth growing season. There was no difference (P >0.05) among cul...

Utility of remote sensing-based surface energy balance models to track water stress in rain-fed switchgrass under dry and wet conditions

USDA-ARS?s Scientific Manuscript database

The ability of remote sensing-based surface energy balance (SEB) models to track water stress in rain-fed switchgrass has not been explored yet. In this paper, the theoretical framework of crop water stress index (CWSI) was utilized to estimate CWSI in rain-fed switchgrass (Panicum virgatum L.) usin...

Early competitive effects on growth of loblolly pine grown in co-culture with switchgrass

Treesearch

Kurt J. Krapfl; Scott D. Roberts; Randall J. Rosseau; Jeff A. Hatten

2015-01-01

This study: (1) examined competitive interactions between switchgrass and loblolly pine grown in co-culture, and (2) assessed early growth rates of loblolly pine as affected by differing switchgrass competition treatments. Co-cultures were established and monitored on two Upper Coastal Plain sites for 2 years. The Pontotoc site has a history of agricultural use with...

Microbial nitrogen cycling response to forest-based bioenergy production.

PubMed

Minick, Kevan J; Strahm, Brian D; Fox, Thomas R; Sucre, Eric B; Leggett, Zakiya H

2015-12-01

Concern over rising atmospheric CO2 and other greenhouse gases due to fossil fuel combustion has intensified research into carbon-neutral energy production. Approximately 15.8 million ha of pine plantations exist across the southeastern United States, representing a vast land area advantageous for bioenergy production without significant landuse change or diversion of agricultural resources from food production. Furthermore, intercropping of pine with bioenergy grasses could provide annually harvestable, lignocellulosic biomass feedstocks along with production of traditional wood products. Viability of such a system hinges in part on soil nitrogen (N) availability and effects of N competition between pines and grasses on ecosystem productivity. We investigated effects of intercropping loblolly pine (Pinus taeda) with switchgrass (Panicum virgatum) on microbial N cycling processes in the Lower Coastal Plain of North Carolina, USA. Soil samples were collected from bedded rows of pine and interbed space of two treatments, composed of either volunteer native woody and herbaceous vegetation (pine-native) or pure switchgrass (pine-switchgrass) in interbeds. An in vitro 15N pool-dilution technique was employed to quantify gross N transformations at two soil depths (0-5 and 5-15 cm) on four dates in 2012-2013. At the 0-5 cm depth in beds of the pine-switchgrass treatment, gross N mineralization was two to three times higher in November and February compared to the pine-native treatment, resulting in increased NH4(+) availability. Gross and net nitrification were also significantly higher in February in the same pine beds. In interbeds of the pine-switchgrass treatment, gross N mineralization was lower from April to November, but higher in February, potentially reflecting positive effects of switchgrass root-derived C inputs during dormancy on microbial activity. These findings indicate soil N cycling and availability has increased in pine beds of the pine-switchgrass treatment compared to those of the pine-native treatment, potentially alleviating any negative effects of N competition between pine and switchgrass. We expect that reduced soil C in the pine-switchgrass treatment, effects of pine and switchgrass rooting on soil C availability, and plant N demand are major factors influencing soil N transformations. Future research should examine rooting architecture in-intercropped systems and the effects on soil microbial communities and function.

Survival of Salmonella, Escherichia coli 0157:H7, non-0157 shiga toxin producing E.coli, and potential surrogate bacteria in crop soil as affected by the addition of fast pyrolysis-generated switchgrass biochar

USDA-ARS?s Scientific Manuscript database

Fast pyrolysis of switchgrass (and resultant biochar) can be used for bio-fuel production, soil amendments for fertilizing crops, binding heavy metals, and sequestering environmental biocarbon. To determine the influence of fast pyrolysis-generated switchgrass biochar on survival of foodborne path...

Watershed-scale Hydrology and Water Quality Impact of Switchgrass Intercropping in Southern Managed Pine Forests

NASA Astrophysics Data System (ADS)

Chescheir, G. M.; Birgand, F.; Allen, E.; Bennett, E.; Carter, T.; Dobbs, N.; Muwamba, A.; Amatya, D. M.; Youssef, M.; Nettles, J. E.

2016-12-01

The use of marginal land for cellulosic biofuel crop production is an attractive solution to preserve agricultural land for food production. The space available between rows of young loblolly pine (Pinus taeda) trees offers enough light to support growth of biofuel crops for several years. A five year field study was conducted to assess the hydrology and water quality impacts of switchgrass (Panicum virgatum) intercropping with pine trees in watersheds of the southeastern US. Paired-watershed studies were replicated in Mississippi and Alabama on upland sites, and in North Carolina on a flat lowland site. In each state, the impact of switchgrass intercropping was assessed from differences in water and nutrient yields from contiguous 20-40 ha watersheds established as: conventional young pine plantation, switchgrass intercropped in young pine plantation, switchgrass only, and mid-rotation mature pine plantation. A total of 14 watersheds were equipped with continuous flow monitoring stations, flow proportional water samplers, groundwater wells, soil moisture sensors and weather stations. Data collection continued through a two year pre-treatment period, a one year treatment period when field operations were conducted to establish switchgrass, and a two year post-treatment period when the established switchgrass was fertilized and harvested annually. Our results showed that significant increases in total suspended solids (TSS) and nitrogen (N) loading occurred during the treatment periods at the upland sites in MS and AL. During the post treatment periods, TSS and N loading decreased to levels near those observed in pretreatment. At the lowland site, only nitrogen loading was increased during the treatment period. Concentrations of TSS at the lowland site were two orders of magnitude lower than those observed at the upland sites and were not significantly affected by the treatment. Inherent flow variability between watersheds within sites made detection of subtle differences in hydrology and water quality difficult to detect. Increases in N loading were not significant in response to annual fertilization of switchgrass. Our results suggest that intercropping switchgrass in managed pine forests does not significantly change the typically benign hydrology and water quality of the managed forests.

Genetic Improvement of Switchgrass and Other Herbaceous Plants for Use as Biomass Fuel Feedstock

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vogel, K.P.

2001-01-11

It should be highly feasible to genetically modify the feedstock quality of switchgrass and other herbaceous plants using both conventional and molecular breeding techniques. Effectiveness of breeding to modify herbages of switchgrass and other perennial and annual herbaceous species has already been demonstrated. The use of molecular markers and transformation technology will greatly enhance the capability of breeders to modify the plant structure and cell walls of herbaceous plants. It will be necessary to monitor gene flow to remnant wild populations of plants and have strategies available to curtail gene flow if it becomes a potential problem. It also willmore » be necessary to monitor plant survival and long-term productivity as affected by genetic changes that improve forage quality. Information on the conversion processes that will be used and the biomass characteristics that affect conversion efficiency and rate is absolutely essential as well as information on the relative economic value of specific traits. Because most forage or biomass quality characteristics are highly affected by plant maturity, it is suggested that plant material of specific maturity stages be used in research to determining desirable feedstock quality characteristics. Plant material could be collected at various stages of development from an array of environments and storage conditions that could be used in conversion research. The same plant material could be used to develop NIRS calibrations that could be used by breeders in their selection programs and also to develop criteria for a feedstock quality assessment program. Breeding for improved feedstock quality will likely affect the rate of improvement of biomass production per acre. If the same level of resources are used, multi-trait breeding simply reduces the selection pressure and hence the breeding progress that can be made for a single trait unless all the traits are highly correlated. Since desirable feedstock traits are likely to be similar to IVDMD, it is likely that they will not be highly positively correlated with yield. Hence to achieve target yields and improve specific quality traits, it will likely be necessary to increase the resources available to plant breeders. Marker assisted selection will be extremely useful in breeding for quality traits, particularly for traits that can be affected by modifying a few genes. Genetic markers are going to be needed for monitoring gene flow to wild populations. Transformation will be a very useful tool for determining the affects of specific genes on biomass feedstock quality.« less

Flowability parameters for chopped switchgrass, wheat straw and corn stover

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chevanan, Nehru; Womac, A.R.; Bitra, V.S.P.

2009-02-01

A direct shear cell to measure the shear strength and flow properties of chopped switchgrass, wheat straw, and corn stover was designed, fabricated, and tested. Yield loci (r2=0.99) determined at pre-consolidation pressures of 3.80 kPa and 5.02 kPa indicated that chopped biomass followed Mohr-Coulomb failure. Normal stress significantly affected the displacement required for shear failure, as well as the friction coefficient values for all three chopped biomass types. Displacement at shear failure ranged from 30 to 80 mm, and depended on pre-consolidation pressure, normal stress, and particle size. Friction coefficient was inversely related to normal stress, and was highest formore » chopped corn stover. Also, chopped corn stover exhibited the highest angle of internal friction, unconfined yield strength, major consolidation strength, and cohesive strength, all of which indicated increased challenges in handling chopped corn stover. The measured angle of internal friction and cohesive strength indicated that chopped biomass cannot be handled by gravity alone. The measured angle of internal friction and cohesive strength were 43 and 0.75 kPa for chopped switchgrass; 44 and 0.49 kPa for chopped wheat straw; and 48 and 0.82 kPa for chopped corn stover. Unconfined yield strength and major consolidation strength used for characterization of bulk flow materials and design of hopper dimensions were 3.4 and 10.4 kPa for chopped switchgrass; 2.3 and 9.6 kPa for chopped wheat straw and 4.2 and 11.8 kPa for chopped corn stover. These results are useful for development of efficient handling, storage, and transportation systems for biomass in biorefineries.« less

Switchgrass potential on reclaimed surface mines for biofuel production in West Virginia

NASA Astrophysics Data System (ADS)

Marra, Michael A.

The high cost and environmental risks associated with non-renewable energy sources has caused an increased interest in, and development of renewable biofuels. Switchgrass (Panicum virgatum L.), a warm season perennial grass, has been investigated as a source of biofuel feedstock due to its high biomass production on marginal soils, its tolerance of harsh growing conditions, and its ability to provide habitat for wildlife and soil conservation cover. West Virginia contains vast expanses of reclaimed surface mine lands and could potentially benefit from the production of switchgrass as a biofuel feedstock. Furthermore, switchgrass production could satisfy Surface Mining Reclamation and Control Act of 1977 (SMCRA) requirements for reclamation bond release to mine operators. Three separate studies will be discussed in this thesis to determine if switchgrass grown on reclaimed surface mines can produce yields similar to yields from stands grown under normal agronomic conditions and what common surface mining reclamation practices may be most appropriate for growing switchgrass. The first study examined yield production of three commercially-available, upland switchgrass varieties grown on two reclaimed surface mines in production years two, three and four. The Hampshire Hill mine site, which was reclaimed in the late 1990s using top soil and treated municipal sludge, averaged 5,800 kg (ha-yr)-1 of switchgrass compared to 803 kg (ha-yr)-1 at the Hobet 21 site which was reclaimed with crushed, unweathered rock over compacted overburden. Site and variety interacted with Cave-in-Rock as the top performer at the more fertile Hampshire Hill site and Shawnee produced the highest yields at Hobet 21 (7,853 kg ha-1 and 1,086 kg ha-1 averaged across years, respectively). Switchgrass yields increased from 2009 to 2010, but declined from 2010 to 2011. Switchgrass yields from farmlands in this region averaged about 15000 kg (ha-yr)-1 in the research literature, so switchgrass grown on reclaimed lands appears to be about 50% lower. A second study to determine optimal nitrogen and mulch rates for switchgrass establishment began in June 2011 on two newly-reclaimed surface mines. Both sites were seeded at a rate of 11.2 kg pure live seed (PLS) ha-1 of Cave-in-Rock on replicated treatments of 0, 33.6 and 67.0 kg N ha-1, and high and low mulch rates of mulch applied as hydromulch. Switchgrass cover, frequency and yield improved with the addition of any amount of N fertilizer compared to no N application. There was no significant difference in yield associated with high and low levels of N. We also observed that yields were not affected by application of additional mulch. The final study compared a one- and two-harvest system in the fourth year of production at the Hampshire Hill and Hobet 21 sites. There was no increase in yield production utilizing a two-harvest system (2922 kg (ha-yr)-1, averaged across site) compared to a one-harvest system (3029 kg (ha-yr)-1). The data also showed that re-growth collected from July to October in the two-harvest system added negligible yield and that yield collected in July was comparable in one- and two-harvest systems.

Advances in biotechnology and genomics of switchgrass

PubMed Central

2013-01-01

Switchgrass (Panicum virgatum L.) is a C4 perennial warm season grass indigenous to the North American tallgrass prairie. A number of its natural and agronomic traits, including adaptation to a wide geographical distribution, low nutrient requirements and production costs, high water use efficiency, high biomass potential, ease of harvesting, and potential for carbon storage, make it an attractive dedicated biomass crop for biofuel production. We believe that genetic improvements using biotechnology will be important to realize the potential of the biomass and biofuel-related uses of switchgrass. Tissue culture techniques aimed at rapid propagation of switchgrass and genetic transformation protocols have been developed. Rapid progress in genome sequencing and bioinformatics has provided efficient strategies to identify, tag, clone and manipulate many economically-important genes, including those related to higher biomass, saccharification efficiency, and lignin biosynthesis. Application of the best genetic tools should render improved switchgrass that will be more economically and environmentally sustainable as a lignocellulosic bioenergy feedstock. PMID:23663491

Effect of ensiling and silage additives on biogas production and microbial community dynamics during anaerobic digestion of switchgrass.

PubMed

Zhao, Xiaoling; Liu, Jinhuan; Liu, Jingjing; Yang, Fuyu; Zhu, Wanbin; Yuan, Xufeng; Hu, Yuegao; Cui, Zongjun; Wang, Xiaofen

2017-10-01

Silage processing has a crucial positive impact on the methane yield of anaerobic treated substrates. Changes in the characteristics of switchgrass after ensiling with different additives and their effects on methane production and microbial community changes during anaerobic digestion were investigated. After ensiling (CK), methane yield was increased by 33.59% relative to that of fresh switchgrass (FS). In comparison with the CK treatment, methane production was improved by 17.41%, 13.08% and 8.72% in response to ensiling with LBr+X, LBr and X, respectively. A modified Gompertz model predicted that the optimum treatment was LBr+X, with a potential cumulative methane yield of 178.31mL/g total solids (TS) and a maximum biogas production rate of 44.39mL/g TS·d. Firmicutes and Bacteroidetes were the predominant bacteria in FS and silage switchgrass; however, the switchgrass treated with LBr+X was rich in Synergistetes, which was crucial for methane production. Copyright © 2017. Published by Elsevier Ltd.

Response of “Alamo” switchgrass tissue chemistry and biomass to nitrogen fertilization in West Tennessee, USA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garten, Charles T.; Brice, Deanne J.; Castro, Hector F.

2011-01-01

Switchgrass (Panicum virgatum) is a perennial, warm-season grass that has been identified as a potential biofuel feedstock over a large part of North America. We examined above- and belowground responses to nitrogen fertilization in “Alamo” switchgrass grown in West Tennessee, USA. The fertilizer study included a spring and fall sampling of 5-year old switchgrass grown under annual applications of 0, 67, and 202 kg N ha -1 (as ammonium nitrate). Fertilization changed switchgrass biomass allocation as indicated by root:shoot ratios. End-of-growing season root:shoot ratios (mean ± SE) declined significantly (P ≤ 0.05) at the highest fertilizer nitrogen treatment (2.16 ±more » 0.08, 2.02 ± 0.18, and 0.88 ± 0.14, respectively, at 0, 67, and 202 kg N ha -1). Fertilization also significantly increased above- and belowground nitrogen concentrations and decreased plant C:N ratios. Data are presented for coarse live roots, fine live roots, coarse dead roots, fine dead roots, and rhizomes. At the end of the growing season, there was more carbon and nitrogen stored in belowground biomass than aboveground biomass. Finally, fertilization impacted switchgrass tissue chemistry and biomass allocation in ways that potentially impact soil carbon cycle processes and soil carbon storage.« less

Short-term bird response to harvesting switchgrass for biomass in Iowa

USGS Publications Warehouse

Murray, L.D.; Best, Louis B.

2003-01-01

The Conservation Reserve Program (CRP) provides habitat for grassland birds, but as contracts expire, some CRP fields might be returned to rowcrop production. One alternative to returning CRP fields to rowcrops is to produce switchgrass (Panicum virgatum) for use as a biomass fuel. Because the biomass is harvested during the fall and winter, breeding birds would not be directly affected by mowing the fields bur might be influenced by changes in vegetation structure resulting from the harvest. We evaluated bird abundances and nest success in totally harvested, partially harvested (alternating cut and uncut strips), and nonharvested CRP switchgrass fields in southern Iowa, USA, in 1999 and 2000. Species richness did not differ among harvest treatments. Abundances of most species (16 of 18) were not affected by the harvesting of switchgrass fields, and strip width did not affect bird numbers in strip-harvested fields. Grasshopper sparrows (Ammodramus savannarum) were more abundant in harvested portions of fields, and more sedge wrens (Cistothorus platensis) were recorded in nonharvested areas. The residual vegetation in nonharvested areas provided nest cover for species that begin nesting early in the season (e.g., northern harrier [Circus cyaneus] and ring-necked pheasant [Phasianus colchicus]). Nest success rates of grasshopper sparrows and common yellowthroats (Geothlypis trichas) were similar to those reported by other studies in switchgrass fields and might be sufficient to maintain stable populations. In general, switchgrass biomass fields create breeding habitat for some grassland birds, and a mixture of harvested and nonharvested fields would be more beneficial to grassland birds than totally harvesting or partially harvesting all switchgrass fields.

Transgene autoexcision in switchgrass pollen mediated by the Bxb1 recombinase

PubMed Central

2014-01-01

Background Switchgrass (Panicum virgatum L.) has a great potential as a platform for the production of biobased plastics, chemicals and energy mainly because of its high biomass yield on marginal land and low agricultural inputs. During the last decade, there has been increased interest in the genetic improvement of this crop through transgenic approaches. Since switchgrass, like most perennial grasses, is exclusively cross pollinating and poorly domesticated, preventing the dispersal of transgenic pollen into the environment is a critical requisite for the commercial deployment of this important biomass crop. In this study, the feasibility of controlling pollen-mediated gene flow in transgenic switchgrass using the large serine site-specific recombinase Bxb1 has been investigated. Results A novel approach utilizing co-transformation of two separate vectors was used to test the functionality of the Bxb1/att recombination system in switchgrass. In addition, two promoters with high pollen-specific activity were identified and thoroughly characterized prior to their introduction into a test vector explicitly designed for both autoexcision and quantitative analyses of recombination events. Our strategy for developmentally programmed precise excision of the recombinase and marker genes in switchgrass pollen resulted in the generation of transgene-excised progeny. The autoexcision efficiencies were in the range of 22-42% depending on the transformation event and assay used. Conclusion The results presented here mark an important milestone towards the establishment of a reliable biocontainment system for switchgrass which will facilitate the development of this crop as a biorefinery feedstock through advanced biotechnological approaches. PMID:25148894

Genotypic diversity effects on biomass production in native perennial bioenergy cropping systems

DOE PAGES

Morris, Geoffrey P.; Hu, Zhenbin; Grabowski, Paul P.; ...

2015-10-03

The perennial grass species that are being developed as biomass feedstock crops harbor extensive genotypic diversity, but the effects of this diversity on biomass production are not well understood. We investigated the effects of genotypic diversity in switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii) on perennial biomass cropping systems in two experiments conducted over 2008–2014 at a 5.4-ha fertile field site in northeastern Illinois, USA. We varied levels of switchgrass and big bluestem genotypic diversity using various local and nonlocal cultivars – under low or high species diversity, with or without nitrogen inputs – and quantified establishment, biomass yield,more » and biomass composition. In one experiment (‘agronomic trial’), we compared three switchgrass cultivars in monoculture to a switchgrass cultivar mixture and three different species mixtures, with or without N fertilization. In another experiment (‘diversity gradient’), we varied diversity levels in switchgrass and big bluestem (1, 2, 4, or 6 cultivars per plot), with one or two species per plot. In both experiments, cultivar mixtures produced yields equivalent to or greater than the best cultivars. In the agronomic trial, the three switchgrass mixture showed the highest production overall, though not significantly different than best cultivar monoculture. In the diversity gradient, genotypic mixtures had one-third higher biomass production than the average monoculture, and none of the monocultures were significantly higher yielding than the average mixture. Year-to-year variation in yields was lowest in the three-cultivar switchgrass mixtures and Cave-In-Rock (the southern Illinois cultivar) and also reduced in the mixture of switchgrass and big bluestem relative to the species monocultures. The effects of genotypic diversity on biomass composition were modest relative to the differences among species and genotypes. Our findings suggest that local genotypes can be included in biomass cropping systems without compromising yields and that genotypic mixtures could help provide high, stable yields of high-quality biomass feedstocks.« less

Model optimization of cadmium and accumulation in switchgrass (Panicum virgatum L.): potential use for ecological phytoremediation in Cd-contaminated soils.

PubMed

Wang, Quanzhen; Gu, Muyu; Ma, Xiaomin; Zhang, Hongjuan; Wang, Yafang; Cui, Jian; Gao, Wei; Gui, Jing

2015-11-01

Soil pollution with heavy metals is an increasingly serious threat to the environment, food security, and human health. Therefore, it is urgent to develop economic and highly efficient soil restoration technology for environmental improvement; phytoremediation is an option that is safe, has low cost, and is environmentally friendly. However, in selecting hyperaccumulators or tolerant plants, theories and operation technologies for optimal restoration should be satisfied. In this study, the switchgrass growth response and performance of phytoextraction under the coupling effect of Cd and pH were investigated by evaluating seed germination, seedling growth, and the Cd content in the plant to evaluate the potential use of switchgrass as a phytoremediation plant in cadmium contaminated soil. This study conducted three sets of independent experiments with five levels of Cd concentrations, including two orthogonal matrix designs of combining Cd with pH values. The results showed that switchgrass was germinated well under all treatments (Cd concentration of 0-500 μM), but the seedling growth was significantly affected by Cd and pH, as shown by multivariate regression analyses. Hormesis was found during the growth of switchgrass plants exposed to low Cd concentrations under hydroponic conditions, and switchgrass plants were capable of developing with a Cd concentration of 100-175 μM and pH of 4.1-5.9. Mild acidic conditions can enhance the ability of Cd to accumulate in switchgrass. Switchgrass was moderately tolerant to Cd and may be used as a phytoremediation plant for Cd-contaminated soils in the future. Our results also suggest that hormetic effects should be taken into consideration in the phytoremediation of Cd-contaminated soils. We discuss the physiological and biochemical mechanisms contributing to the effective application of the plant for the phytoremediation of Cd-contaminated soils.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Adkins, Jaron; Jastrow, Julie D.; Morris, Geoffrey P.

Switchgrass (Panicum virgatum L), a cellulosic biofuel feedstock, may promote soil C 21 accumulation compared to annual cropping systems by increasing the amount and retention of 22 root-derived soil C inputs. The aim of this study was to assess how different switchgrass 23 cultivars impact soil C inputs and retention, whether these impacts vary with depth, and whether 24 specific root length (SRL) explains these impacts. We collected soil to a depth of 30 cm from six 25 switchgrass cultivars with root systems ranging from high to low SRL. The cultivars (C4 species) 26 were grown for 27 months onmore » soils previously dominated by C3 plants, allowing us to use the 27 natural difference in 13C isotopic signatures between C3 soils and C4 plants to quantify 28 switchgrass-derived C accumulation. The soil was fractionated into coarse particulate organic 29 matter (CPOM), fine particulate organic matter (FPOM), silt, and clay-sized fractions. We 30 measured total C and plant-derived C in all soil fractions across all depths. The study led to two main results: (1) bulk soil C concentrations beneath switchgrass cultivars varied by 40% in the 0-32 10 cm soil depth and by 70% in the 10-20 cm soil depth, and cultivars with high bulk soil C 33 concentrations tended to have relatively high C concentrations in the mineral soil fractions and 34 relatively low C concentrations in the POM fractions; (2) there were significant differences in 35 switchgrass-derived soil C between cultivars at the 0-10 cm depth, where soil C inputs ranged 36 from 1.2 to 3.2 mg C g-1 dry soil. There was also evidence of a positive correlation between SRL 37 and switchgrass-derived C inputs when one outlier data point was removed. These results 38 indicate that switchgrass cultivars differentially impact mechanisms contributing to soil C accumulation.« less

Genotypic diversity effects on biomass production in native perennial bioenergy cropping systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morris, Geoffrey P.; Hu, Zhenbin; Grabowski, Paul P.

The perennial grass species that are being developed as biomass feedstock crops harbor extensive genotypic diversity, but the effects of this diversity on biomass production are not well understood. We investigated the effects of genotypic diversity in switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii) on perennial biomass cropping systems in two experiments conducted over 2008–2014 at a 5.4-ha fertile field site in northeastern Illinois, USA. We varied levels of switchgrass and big bluestem genotypic diversity using various local and nonlocal cultivars – under low or high species diversity, with or without nitrogen inputs – and quantified establishment, biomass yield,more » and biomass composition. In one experiment (‘agronomic trial’), we compared three switchgrass cultivars in monoculture to a switchgrass cultivar mixture and three different species mixtures, with or without N fertilization. In another experiment (‘diversity gradient’), we varied diversity levels in switchgrass and big bluestem (1, 2, 4, or 6 cultivars per plot), with one or two species per plot. In both experiments, cultivar mixtures produced yields equivalent to or greater than the best cultivars. In the agronomic trial, the three switchgrass mixture showed the highest production overall, though not significantly different than best cultivar monoculture. In the diversity gradient, genotypic mixtures had one-third higher biomass production than the average monoculture, and none of the monocultures were significantly higher yielding than the average mixture. Year-to-year variation in yields was lowest in the three-cultivar switchgrass mixtures and Cave-In-Rock (the southern Illinois cultivar) and also reduced in the mixture of switchgrass and big bluestem relative to the species monocultures. The effects of genotypic diversity on biomass composition were modest relative to the differences among species and genotypes. Our findings suggest that local genotypes can be included in biomass cropping systems without compromising yields and that genotypic mixtures could help provide high, stable yields of high-quality biomass feedstocks.« less

Evaluation of intercropped switchgrass establishment under a range of experimental site preparation treatments in a forested setting on the Lower Coastal Plain of North Carolina, U.S.A

Treesearch

Janine M. Albaugh; Eric B. Sucre; Zakiya H. Leggett; Jean-Christophe Domec; John S. King

2012-01-01

There is growing interest in using switchgrass (Panicum virgatum L.) as a biofuel crop and for its potential to sequester carbon. However, there are limited data on the establishment success of this species when grown as a forest intercrop in coastal plain settings of the U.S. Southeast. Therefore, we studied establishment success of switchgrass...

National-level infrastructure and economic effects of switchgrass cofiring with coal in existing power plants for carbon mitigation.

PubMed

Morrow, William R; Griffin, W Michael; Matthews, H Scott

2008-05-15

We update a previously presented Linear Programming (LP) methodology for estimating state level costs for reducing CO2 emissions from existing coal-fired power plants by cofiring switchgrass, a biomass energy crop, and coal. This paper presents national level results of applying the methodology to the entire portion of the United States in which switchgrass could be grown without irrigation. We present incremental switchgrass and coal cofiring carbon cost of mitigation curves along with a presentation of regionally specific cofiring economics and policy issues. The results show that cofiring 189 million dry short tons of switchgrass with coal in the existing U.S. coal-fired electricity generation fleet can mitigate approximately 256 million short tons of carbon-dioxide (CO2) per year, representing a 9% reduction of 2005 electricity sector CO2 emissions. Total marginal costs, including capital, labor, feedstock, and transportation, range from $20 to $86/ton CO2 mitigated,with average costs ranging from $20 to $45/ton. If some existing power plants upgrade to boilers designed for combusting switchgrass, an additional 54 million tons of switchgrass can be cofired. In this case, total marginal costs range from $26 to $100/ton CO2 mitigated, with average costs ranging from $20 to $60/ton. Costs for states east of the Mississippi River are largely unaffected by boiler replacement; Atlantic seaboard states represent the lowest cofiring cost of carbon mitigation. The central plains states west of the Mississippi River are most affected by the boiler replacement option and, in general, go from one of the lowest cofiring cost of carbon mitigation regions to the highest. We explain the variation in transportation expenses and highlight regional cost of mitigation variations as transportation overwhelms other cofiring costs.

Transcriptional and physiological data reveal the dehydration memory behavior in switchgrass (Panicum virgatum L.).

PubMed

Zhang, Chao; Peng, Xi; Guo, Xiaofeng; Tang, Gaijuan; Sun, Fengli; Liu, Shudong; Xi, Yajun

2018-01-01

Switchgrass ( Panicum virgatum L.) is a model biofuel plant because of its high biomass, cellulose-richness, easy degradation to ethanol, and the availability of extensive genomic information. However, a little is currently known about the molecular responses of switchgrass plants to dehydration stress, especially multiple dehydration stresses. Studies on the transcriptional profiles of 35-day-old tissue culture plants revealed 741 dehydration memory genes. Gene Ontology and pathway analysis showed that these genes were enriched in phenylpropanoid biosynthesis, starch and sucrose metabolism, and plant hormone signal transduction. Further analysis of specific pathways combined with physiological data suggested that switchgrass improved its dehydration resistance by changing various aspects of its responses to secondary dehydration stress (D2), including the regulation of abscisic acid (ABA) and jasmonic acid (JA) biosynthesis and signal transduction, the biosynthesis of osmolytes (l-proline, stachyose and trehalose), energy metabolism (i.e., metabolic process relating to photosynthetic systems, glycolysis, and the TCA cycle), and lignin biosynthesis. The transcriptional data and chemical substance assays showed that ABA was significantly accumulated during both primary (D1) and secondary (D2) dehydration stresses, whereas JA accumulated during D1 but became significantly less abundant during D2. This suggests the existence of a complicated signaling network of plant hormones in response to repeated dehydration stresses. A homology analysis focusing on switchgrass, maize, and Arabidopsis revealed the conservation and species-specific distribution of dehydration memory genes. The molecular responses of switchgrass plants to successive dehydration stresses have been systematically characterized, revealing a previously unknown transcriptional memory behavior. These results provide new insights into the mechanisms of dehydration stress responses in plants. The genes and pathways identified in this study will be useful for the genetic improvement of switchgrass and other crops.

Reduction of ethanol yield from switchgrass infected with rust caused by Puccinia emaculata

DOE PAGES

Sykes, Virginia R.; Allen, Fred L.; Mielenz, Jonathan R.; ...

2015-10-16

Switchgrass ( Panicum virgatum) is an important biofuel crop candidate thought to have low disease susceptibility. As switchgrass production becomes more prevalent, monoculture and production fields in close proximity to one another may increase the spread and severity of diseases such as switchgrass rust caused by the pathogen Puccinia emaculata. The objective of this research was to examine the impact of rust on ethanol yield in switchgrass. In 2010 and 2012, naturally infected leaves from field-grown Alamo and Kanlow in Knoxville, TN (2010, 2012) and Crossville, TN (2012) were visually categorized as exhibiting low, medium, or high disease based onmore » the degree of chlorosis and sporulation. P. emaculata was isolated from each disease range to confirm infection. Samples from 2010 were acid/heat pretreated and subjected to two runs of simultaneous saccharification and fermentation (SSF) with Saccharomyces cerevisiae D 5A to measure ethanol yield. Near-infrared spectroscopy (NIRS) was used to estimate ethanol yield for 2012 samples. SSF and NIRS data were analyzed separately using ANOVA. Disease level effects were significant within both models (P < 0.05) and both models explained a large amount of variation in ETOH (SSF: R 2 = 0.99, NIRS: R 2 = 0.99). In the SSF dataset, ethanol was reduced by 35 % in samples exhibiting medium disease symptoms and by 55 % in samples exhibiting high disease symptoms. In the NIRS dataset, estimated ethanol was reduced by 10 % in samples exhibiting medium disease symptoms and by 21 % in samples exhibiting high disease symptoms. Lastly, results indicate that switchgrass rust will likely have a negative impact on ethanol yield in switchgrass grown as a biofuel crop.« less

Reduction of ethanol yield from switchgrass infected with rust caused by Puccinia emaculata

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sykes, Virginia R.; Allen, Fred L.; Mielenz, Jonathan R.

Switchgrass ( Panicum virgatum) is an important biofuel crop candidate thought to have low disease susceptibility. As switchgrass production becomes more prevalent, monoculture and production fields in close proximity to one another may increase the spread and severity of diseases such as switchgrass rust caused by the pathogen Puccinia emaculata. The objective of this research was to examine the impact of rust on ethanol yield in switchgrass. In 2010 and 2012, naturally infected leaves from field-grown Alamo and Kanlow in Knoxville, TN (2010, 2012) and Crossville, TN (2012) were visually categorized as exhibiting low, medium, or high disease based onmore » the degree of chlorosis and sporulation. P. emaculata was isolated from each disease range to confirm infection. Samples from 2010 were acid/heat pretreated and subjected to two runs of simultaneous saccharification and fermentation (SSF) with Saccharomyces cerevisiae D 5A to measure ethanol yield. Near-infrared spectroscopy (NIRS) was used to estimate ethanol yield for 2012 samples. SSF and NIRS data were analyzed separately using ANOVA. Disease level effects were significant within both models (P < 0.05) and both models explained a large amount of variation in ETOH (SSF: R 2 = 0.99, NIRS: R 2 = 0.99). In the SSF dataset, ethanol was reduced by 35 % in samples exhibiting medium disease symptoms and by 55 % in samples exhibiting high disease symptoms. In the NIRS dataset, estimated ethanol was reduced by 10 % in samples exhibiting medium disease symptoms and by 21 % in samples exhibiting high disease symptoms. Lastly, results indicate that switchgrass rust will likely have a negative impact on ethanol yield in switchgrass grown as a biofuel crop.« less

Investigation of Pleurotus ostreatus pretreatment on switchgrass for ethanol production

NASA Astrophysics Data System (ADS)

Slavens, Shelyn Gehle

Fungal pretreatment using the white-rot fungus Pleurotus ostreatus on switchgrass for ethanol production was studied. In a small-scale storage study, small switchgrass bales were inoculated with fungal spawn and automatically watered to maintain moisture. Sampled at 25, 53, and 81 d, the switchgrass composition was determined and liquid hot water (LHW) pretreatment was conducted. Fungal pretreatment significantly decreased the xylan and lignin content; glucan was not significantly affected by fungal loading. The glucan, xylan, and lignin contents significantly decreased with increased fungal pretreatment time. The effects of the fungal pretreatment were not highly evident after the LHW pretreatment, showing only changes based on sampling time. Although other biological activity within the bales increased cellulose degradation, the fungal pretreatment successfully reduced the switchgrass lignin and hemicellulose contents. In a laboratory-scale nutrient supplementation study, copper, manganese, glucose, or water was added to switchgrass to induce production of ligninolytic enzymes by P. ostreatus. After 40 d, ligninolytic enzyme activities and biomass composition were determined and simultaneous saccharification and fermentation (SSF) was conducted to determine ethanol yield. Laccase activity was similar for all supplements and manganese peroxidase (MnP) activity was significantly less in copper-treated samples than in the other fungal-inoculated samples. The fungal pretreatment reduced glucan, xylan, and lignin content, while increasing extractable sugars content. The lowest lignin contents occurred in the water-fungal treated samples and produced the greatest ethanol yields. The greatest lignin contents occurred in the copper-fungal treated samples and produced the lowest ethanol yields. Manganese-fungal and glucose-fungal treated samples had similar, intermediate lignin contents and produced similar, intermediate ethanol yields. Ethanol yields from switchgrass were increased significantly by fungal pretreatment.

Site preparation for switchgrass intercropping in loblolly pine plantations reduces retained trees and snags, but maintains downed woody debris

USGS Publications Warehouse

Loman, Zachary G.; Riffell, Samuel K.; Miller, Darrin A.; Martin, James A.; Vilella, Francisco

2013-01-01

Within young pine (Pinus spp.) plantations, coarse woody debris (CWD) and green trees are important habitat structures that may be impacted by the production of biofuel feedstock. Therefore, we compared site preparation procedures associated with switchgrass (Panicum virgatum L.) intercropping to determine effects on CWD and green trees in stands (n = 24) site-prepared for intercropping, with switchgrass only, or pine plantation in Mississippi, USA. Following site preparation, CWD dispersal or volume did not differ between intercropped and control stands. Intercropped stands had significantly fewer retained trees and snags. Switchgrass monocultures had no retained trees or piles and significantly fewer pieces and less volume of CWD than the other treatments. Our results suggest switchgrass intercropping may provide similar habitat quality to traditional pine plantations for wildlife species using these areas in the year following disturbance, but may provide a less suitable habitat for species that require snags. However, the relationship between snag reduction and wildlife population response in an intercropped setting is not clear and should be further investigated. Regardless, if retaining snags is a desired outcome, site preparation for switchgrass should be restricted to the interbed area where it will be cultivated as opposed to extensive debris removal from the entire site.                   

Grassland bird response to harvesting switchgrass as a biomass energy crop

USGS Publications Warehouse

Roth, A.M.; Sample, D.W.; Ribic, C.A.; Paine, L.; Undersander, D.J.; Bartelt, G.A.

2005-01-01

The combustion of perennial grass biomass to generate electricity may be a promising renewable energy option. Switchgrass (Panicum virgatum) grown as a biofuel has the potential to provide a cash crop for farmers and quality nesting cover for grassland birds. In southwestern Wisconsin (near lat. 42??52???, long. 90??08???), we investigated the impact of an August harvest of switchgrass for bioenergy on community composition and abundance of Wisconsin grassland bird species of management concern. Harvesting the switchgrass in August resulted in changes in vegetation structure and bird species composition the following nesting season. In harvested transects, residual vegetation was shorter and the litter layer was reduced in the year following harvest. Grassland bird species that preferred vegetation of short to moderate height and low to moderate density were found in harvested areas. Unharvested areas provided tall, dense vegetation structure that was especially attractive to tall-grass bird species, such as sedge wren (Cistothorus platensis) and Henslow's sparrow (Ammodramus henslowii). When considering wildlife habitat value in harvest management of switchgrass for biofuel, leaving some fields unharvested each year would be a good compromise, providing some habitat for a larger number of grassland bird species of management concern than if all fields were harvested annually. In areas where most idle grassland habitat present on the landscape is tallgrass, harvest of switchgrass for biofuel has the potential to increase the local diversity of grassland birds.

Extraction and characterization of triglycerides from coffeeweed and switchgrass seeds as potential feedstocks for biodiesel production.

PubMed

Armah-Agyeman, Grace; Gyamerah, Michael; Biney, Paul O; Woldesenbet, Selamawit

2016-10-01

Although switchgrass has been developed as a biofuel feedstock and its potential for bioethanol and bio-oil from fast pyrolysis reported in the literature, the use of the seeds of switchgrass as a source of triglycerides for biodiesel production has not been reported. Similarly, the potential for extracting triglycerides from coffeeweed (an invasive plant of no current economic value) needs to be investigated to ascertain its potential economic use for biodiesel production. The results show that coffeeweed and switchgrass seeds contain known triglycerides which are 983 and 1000 g kg(-1) respectively of the fatty acids found in edible vegetable oils such as sunflower, corn and soybean oils. In addition, the triglyceride yields of 53-67 g kg(-1) of the seed samples are in the range of commercial oil-producing seeds such as corn (42 g kg(-1) ). The results also indicate that the two non-edible oils could be used as substitutes for edible oil for biodiesel production. In addition, the use of seeds of switchgrass for non-edible oil production (as a feedstock for the production of biodiesel) further increases the total biofuel yield when switchgrass is cultivated for use as energy feedstock for pyrolysis oil and biodiesel production. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Using satellite vegetation and compound topographic indices to map highly erodible cropland buffers for cellulosic biofuel crop developments in eastern Nebraska, USA

USGS Publications Warehouse

Gu, Yingxin; Wylie, Bruce K.

2015-01-01

Cultivating annual row crops in high topographic relief waterway buffers has negative environmental effects and can be environmentally unsustainable. Growing perennial grasses such as switchgrass (Panicum virgatum L.) for biomass (e.g., cellulosic biofuel feedstocks) instead of annual row crops in these high relief waterway buffers can improve local environmental conditions (e.g., reduce soil erosion and improve water quality through lower use of fertilizers and pesticides) and ecosystem services (e.g., minimize drought and flood impacts on production; improve wildlife habitat, plant vigor, and nitrogen retention due to post-senescence harvest for cellulosic biofuels; and serve as carbon sinks). The main objectives of this study are to: (1) identify cropland areas with high topographic relief (high runoff potentials) and high switchgrass productivity potential in eastern Nebraska that may be suitable for growing switchgrass, and (2) estimate the total switchgrass production gain from the potential biofuel areas. Results indicate that about 140,000 hectares of waterway buffers in eastern Nebraska are suitable for switchgrass development and the total annual estimated switchgrass biomass production for these suitable areas is approximately 1.2 million metric tons. The resulting map delineates high topographic relief croplands and provides useful information to land managers and biofuel plant investors to make optimal land use decisions regarding biofuel crop development and ecosystem service optimization in eastern Nebraska.

Improving Processing and Performance of Pure Lignin Carbon Fibers through Hardwood and Herbaceous Lignin Blends.

PubMed

Hosseinaei, Omid; Harper, David P; Bozell, Joseph J; Rials, Timothy G

2017-07-01

Lignin/lignin blends were used to improve fiber spinning, stabilization rates, and properties of lignin-based carbon fibers. Organosolv lignin from Alamo switchgrass ( Panicum virgatum ) and yellow poplar ( Liriodendron tulipifera ) were used as blends for making lignin-based carbon fibers. Different ratios of yellow poplar:switchgrass lignin blends were prepared (50:50, 75:25, and 85:15 w/w ). Chemical composition and thermal properties of lignin samples were determined. Thermal properties of lignins were analyzed using thermogravimetric analysis and differential scanning calorimetry. Thermal analysis confirmed switchgrass and yellow poplar lignin form miscible blends, as a single glass transition was observed. Lignin fibers were produced via melt-spinning by twin-screw extrusion. Lignin fibers were thermostabilized at different rates and subsequently carbonized. Spinnability of switchgrass lignin markedly improved by blending with yellow poplar lignin. On the other hand, switchgrass lignin significantly improved thermostabilization performance of yellow poplar fibers, preventing fusion of fibers during fast stabilization and improving mechanical properties of fibers. These results suggest a route towards a 100% renewable carbon fiber with significant decrease in production time and improved mechanical performance.

Improving Processing and Performance of Pure Lignin Carbon Fibers through Hardwood and Herbaceous Lignin Blends

PubMed Central

Hosseinaei, Omid; Bozell, Joseph J.; Rials, Timothy G.

2017-01-01

Lignin/lignin blends were used to improve fiber spinning, stabilization rates, and properties of lignin-based carbon fibers. Organosolv lignin from Alamo switchgrass (Panicum virgatum) and yellow poplar (Liriodendron tulipifera) were used as blends for making lignin-based carbon fibers. Different ratios of yellow poplar:switchgrass lignin blends were prepared (50:50, 75:25, and 85:15 w/w). Chemical composition and thermal properties of lignin samples were determined. Thermal properties of lignins were analyzed using thermogravimetric analysis and differential scanning calorimetry. Thermal analysis confirmed switchgrass and yellow poplar lignin form miscible blends, as a single glass transition was observed. Lignin fibers were produced via melt-spinning by twin-screw extrusion. Lignin fibers were thermostabilized at different rates and subsequently carbonized. Spinnability of switchgrass lignin markedly improved by blending with yellow poplar lignin. On the other hand, switchgrass lignin significantly improved thermostabilization performance of yellow poplar fibers, preventing fusion of fibers during fast stabilization and improving mechanical properties of fibers. These results suggest a route towards a 100% renewable carbon fiber with significant decrease in production time and improved mechanical performance. PMID:28671571

Identification and Characterization of microRNA319a and Its Putative Target Gene, PvPCF5, in the Bioenergy Grass Switchgrass (Panicum virgatum).

PubMed

Xie, Qi; Liu, Xue; Zhang, Yinbing; Tang, Jinfu; Yin, Dedong; Fan, Bo; Zhu, Lihuang; Han, Liebao; Song, Guilong; Li, Dayong

2017-01-01

Due to its high biomass yield, low environmental impact, and widespread adaptability to poor soils and harsh conditions, switchgrass ( Panicum virgatum L.), a warm-region perennial herbaceous plant, has attracted much attention in recent years. However, little is known about microRNAs (miRNAs) and their functions in this bioenergy grass. Here, we identified and characterized a miRNA gene, Pvi-MIR319a , encoding microRNA319a in switchgrass. Transgenic rice lines generated by overexpressing the Pvi-MIR319a precursor gene exhibited broader leaves and delayed flowering compared with the control. Gene expression analysis indicated at least four putative target genes were downregulated. Additionally, we cloned a putative target gene ( PvPCF5 ) of Pvi-MIR319a from switchgrass. PvPCF5, a TCP transcription factor, is a nuclear-localized protein with transactivation activity and control the development of leaf. Our results suggest that Pvi-MIR319a and its target genes may be used as potential genetic regulators for future switchgrass genetic improvement.

Hydrogen production from switchgrass via a hybrid pyrolysis-microbial electrolysis process

DOE PAGES

Lewis, Alex J.; Ren, Shoujie; Ye, Philip; ...

2015-06-30

A new approach to hydrogen production using a hybrid pyrolysis-microbial electrolysis process is described. The aqueous stream generated during pyrolysis of switchgrass was used as a substrate for hydrogen production in a microbial electrolysis cell, achieving a maximum hydrogen production rate of 4.3 L H2/L-day at a loading of 10 g COD/L-anode-day. Hydrogen yields ranged from 50 3.2% to76 0.5% while anode coulombic efficiency ranged from 54 6.5% to 96 0.21%, respectively. Significant conversion of furfural, organic acids and phenolic molecules was observed under both batch and continuous conditions. The electrical and overall energy efficiency ranged from 149-175% and 48-63%,more » respectively. The results demonstrate the potential of the pyrolysis-microbial electrolysis process as a sustainable and efficient route for production of renewable hydrogen with significant implications for hydrocarbon production from biomass.« less

Direct measures of mechanical energy for knife mill size reduction of switchgrass, wheat straw, and corn stover.

PubMed

Bitra, Venkata S P; Womac, Alvin R; Igathinathane, C; Miu, Petre I; Yang, Yuechuan T; Smith, David R; Chevanan, Nehru; Sokhansanj, Shahab

2009-12-01

Lengthy straw/stalk of biomass may not be directly fed into grinders such as hammer mills and disc refiners. Hence, biomass needs to be preprocessed using coarse grinders like a knife mill to allow for efficient feeding in refiner mills without bridging and choking. Size reduction mechanical energy was directly measured for switchgrass (Panicum virgatum L.), wheat straw (Triticum aestivum L.), and corn stover (Zea mays L.) in an instrumented knife mill. Direct power inputs were determined for different knife mill screen openings from 12.7 to 50.8 mm, rotor speeds between 250 and 500 rpm, and mass feed rates from 1 to 11 kg/min. Overall accuracy of power measurement was calculated to be +/-0.003 kW. Total specific energy (kWh/Mg) was defined as size reduction energy to operate mill with biomass. Effective specific energy was defined as the energy that can be assumed to reach the biomass. The difference is parasitic or no-load energy of mill. Total specific energy for switchgrass, wheat straw, and corn stover chopping increased with knife mill speed, whereas, effective specific energy decreased marginally for switchgrass and increased for wheat straw and corn stover. Total and effective specific energy decreased with an increase in screen size for all the crops studied. Total specific energy decreased with increase in mass feed rate, but effective specific energy increased for switchgrass and wheat straw, and decreased for corn stover at increased feed rate. For knife mill screen size of 25.4 mm and optimum speed of 250 rpm, optimum feed rates were 7.6, 5.8, and 4.5 kg/min for switchgrass, wheat straw, and corn stover, respectively, and the corresponding total specific energies were 7.57, 10.53, and 8.87 kWh/Mg and effective specific energies were 1.27, 1.50, and 0.24 kWh/Mg for switchgrass, wheat straw, and corn stover, respectively. Energy utilization ratios were calculated as 16.8%, 14.3%, and 2.8% for switchgrass, wheat straw, and corn stover, respectively. These data will be useful for preparing the feed material for subsequent fine grinding operations and designing new mills.

High Temperature Carbonized Grass as a High Performance Sodium Ion Battery Anode.

PubMed

Zhang, Fang; Yao, Yonggang; Wan, Jiayu; Henderson, Doug; Zhang, Xiaogang; Hu, Liangbing

2017-01-11

Hard carbon is currently considered the most promising anode candidate for room temperature sodium ion batteries because of its relatively high capacity, low cost, and good scalability. In this work, switchgrass as a biomass example was carbonized under an ultrahigh temperature, 2050 °C, induced by Joule heating to create hard carbon anodes for sodium ion batteries. Switchgrass derived carbon materials intrinsically inherit its three-dimensional porous hierarchical architecture, with an average interlayer spacing of 0.376 nm. The larger interlayer spacing than that of graphite allows for the significant Na ion storage performance. Compared to the sample carbonized under 1000 °C, switchgrass derived carbon at 2050 °C induced an improved initial Coulombic efficiency. Additionally, excellent rate capability and superior cycling performance are demonstrated for the switchgrass derived carbon due to the unique high temperature treatment.

Low-heat, mild alkaline pretreatment of switchgrass for anaerobic digestion.

PubMed

Jin, Guang; Bierma, Tom; Walker, Paul M

2014-01-01

This study examines the effectiveness of alkaline pretreatment under mild heat conditions (100°C or 212°F) on the anaerobic co-digestion of switchgrass. The effects of alkaline concentration, types of alkaline, heating time and rinsing were evaluated. In addition to batch studies, continuous-feed studies were performed in triplicate to identify potential digester operational problems caused by switchgrass co-digestion while accounting for uncertainty due to digester variability. Few studies have examined anaerobic digestion of switchgrass or the effects of mild heating to enhance alkaline pretreatment prior to biomass digestion. Results indicate that pretreatment can significantly enhance digestion of coarse-ground (≤ 0.78 cm particle size) switchgrass. Energy conversion efficiency as high as 63% was observed, and was comparable or superior to fine-grinding as a pretreatment method. The optimal NaOH concentration was found to be 5.5% (wt/wt alkaline/biomass) with a 91.7% moisture level. No evidence of operational problems such as solids build-up, poor mixing, or floating materials were observed. These results suggest the use of waste heat from a generator could reduce the concentration of alkaline required to adequately pretreat lignocellulosic feedstock prior to anaerobic digestion.

A geospatial suitability model for drought-tolerant switchgrass

NASA Astrophysics Data System (ADS)

Lewis, S. M.; Kelly, M.

2011-12-01

A perennial grass native to the North America, switchgrass (Panicum virgatum) has been targeted by the USDA as a model mass bioenergy crop to replace petroleum energy products and meet policy demands. Although highly water use efficient, as a warm-season crop, switchgrass requires a significant amount of water during the growing season (April -September). However, locations that have highly reliable water availability are also ideal for profitable food crops (e.g. corn and soy growing regions) and food competition is a significant concern in regards to biofuel crops being grown on productive agricultural lands. Drier, marginal lands (lands on which normal agricultural crops are difficult to cultivate) are therefore potentially ideal locations to grow biofuel crops to ensure that food competition is not an issue. Genetics scientists at UC Davis are in the process of developing a modified variety of switchgrass that can withstand extended periods of drought while not substantially affecting overall yield. As this product is being developed, it is important to identify the potential geographical niche for this new drought-tolerant variety of switchgrass. This project introduces a geospatial approach that utilizes both physical and economic variables to identify ideal geographic locations for this innovative crop.

Nitrogen Fertilization Effects on Productivity and Nitrogen Loss in Three Grass-Based Perennial Bioenergy Cropping Systems

PubMed Central

Duran, Brianna E. L.; Duncan, David S.; Oates, Lawrence G.; Kucharik, Christopher J.; Jackson, Randall D.

2016-01-01

Nitrogen (N) fertilization can greatly improve plant productivity but needs to be carefully managed to avoid harmful environmental impacts. Nutrient management guidelines aimed at reducing harmful forms of N loss such as nitrous oxide (N2O) emissions and nitrate (NO3-) leaching have been tailored for many cropping systems. The developing bioenergy industry is likely to make use of novel cropping systems, such as polycultures of perennial species, for which we have limited nutrient management experience. We studied how a switchgrass (Panicum virgatum) monoculture, a 5-species native grass mixture and an 18-species restored prairie responded to annual fertilizer applications of 56 kg N ha-1 in a field-scale agronomic trial in south-central Wisconsin over a 2-year period. We observed greater fertilizer-induced N2O emissions and sub-rooting zone NO3- concentrations in the switchgrass monoculture than in either polyculture. Fertilization increased aboveground net primary productivity in the polycultures, but not in the switchgrass monoculture. Switchgrass was generally more productive, while the two polycultures did not differ from each other in productivity or N loss. Our results highlight differences between polycultures and a switchgrass monoculture in responding to N fertilization. PMID:26991790

Bulk density and compaction behavior of knife mill chopped switchgrass,wheat straw, and corn stover

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chevanan, Nehru; Womac, A.R.; Bitra, V.S.P.

2009-08-01

Bulk density of comminuted biomass significantly increased by vibration during handling and transportation, and by normal pressure during storage. Compaction characteristics affecting the bulk density of switchgrass, wheat straw, and corn stover chopped in a knife mill at different operating conditions and using four different classifying screens were studied. Mean loose-filled bulk densities were 67.5 18.4 kg/m3 for switchgrass, 36.1 8.6 kg/m3 for wheat straw, and 52.1 10.8 kg/m3 for corn stover. Mean tapped bulk densities were 81.8 26.2 kg/m3 for switchgrass, 42.8 11.7 kg/m3 for wheat straw, and 58.9 13.4 kg/m3 for corn stover. Percentage changes in compressibility duemore » to variation in particle size obtained from a knife mill ranged from 64.3 to 173.6 for chopped switchgrass, 22.2 51.5 for chopped wheat straw and 42.1 117.7 for chopped corn stover within the tested consolidation pressure range of 5 120 kPa. Pressure and volume relationship of chopped biomass during compression with application of normal pressure can be characterized by the Walker model and Kawakita and Ludde model. Parameter of Walker model was correlated to the compressibility with Pearson correlation coefficient greater than 0.9. Relationship between volume reduction in chopped biomass with respect to number of tappings studied using Sone s model indicated that infinite compressibility was highest for chopped switchgrass followed by chopped wheat straw and corn stover. Degree of difficulty in packing measured using the parameters of Sone s model indicated that the chopped wheat straw particles compacted very rapidly by tapping compared to chopped switchgrass and corn stover. These results are very useful for solving obstacles in handling bulk biomass supply logistics issues for a biorefinery.« less

Evaluating the impacts of landscape positions and nitrogen fertilizer rates on dissolved organic carbon on switchgrass land seeded on marginally yielding cropland.

PubMed

Lai, Liming; Kumar, Sandeep; Mbonimpa, Eric G; Hong, Chang Oh; Owens, Vance N; Neupane, Ram P

2016-04-15

Dissolved organic carbon (DOC) through leaching into the soils is another mechanism of net C loss. It plays an important role in impacting the environment and impacted by soil and crop management practices. However, little is known about the impacts of landscape positions and nitrogen (N) fertilizer rates on DOC leaching in switchgrass (Panicum virgatum L.). This experimental design included three N fertilizer rates [0 (low); 56 (medium); 112 (high) kg N ha(-1)] and three landscape positions (shoulder, backslope and footslope). Daily average DOC contents at backslope were significantly lower than that at shoulder and footslope. The DOC contents from the plots that received medium N rate were also significantly lower than the plots that received low N rates. The interactions of landscape and N rates on DOC contents were different in every year from 2009 to 2014, however, no significant consistent trend of DOC contents was observed over time. Annual average DOC contents from the plots managed with low N rate were higher than those with high N rate. These contents at the footslope were higher than that at the shoulder position. Data show that there is a moderate positive relationship between the total average DOC contents and the total average switchgrass biomass yields. Overall, the DOC contents from leachate in the switchgrass land were significantly influenced by landscape positions and N rates. The N fertilization reduced DOC leaching contents in switchgrass field. The switchgrass could retain soil and environment sustainability to some extent. These findings will assist in understanding the mechanism of changes in DOC contents with various parameters in the natural environment and crop management systems. However, use of long-term data might help to better assess the effects of above factors on DOC leaching contents and loss in the switchgrass field in the future. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bulk density and compaction behavior of knife mill chopped switchgrass, wheat straw, and corn stover.

PubMed

Chevanan, Nehru; Womac, Alvin R; Bitra, Venkata S P; Igathinathane, C; Yang, Yuechuan T; Miu, Petre I; Sokhansanj, Shahab

2010-01-01

Bulk density of comminuted biomass significantly increased by vibration during handling and transportation, and by normal pressure during storage. Compaction characteristics affecting the bulk density of switchgrass, wheat straw, and corn stover chopped in a knife mill at different operating conditions and using four different classifying screens were studied. Mean loose-filled bulk densities were 67.5+/-18.4 kg/m(3) for switchgrass, 36.1+/-8.6 kg/m(3) for wheat straw, and 52.1+/-10.8 kg/m(3) for corn stover. Mean tapped bulk densities were 81.8+/-26.2 kg/m(3) for switchgrass, 42.8+/-11.7 kg/m(3) for wheat straw, and 58.9+/-13.4 kg/m(3) for corn stover. Percentage changes in compressibility due to variation in particle size obtained from a knife mill ranged from 64.3 to 173.6 for chopped switchgrass, 22.2-51.5 for chopped wheat straw and 42.1-117.7 for chopped corn stover within the tested consolidation pressure range of 5-120 kPa. Pressure and volume relationship of chopped biomass during compression with application of normal pressure can be characterized by the Walker model and Kawakita and Ludde model. Parameter of Walker model was correlated to the compressibility with Pearson correlation coefficient greater than 0.9. Relationship between volume reduction in chopped biomass with respect to number of tappings studied using Sone's model indicated that infinite compressibility was highest for chopped switchgrass followed by chopped wheat straw and corn stover. Degree of difficulty in packing measured using the parameters of Sone's model indicated that the chopped wheat straw particles compacted very rapidly by tapping compared to chopped switchgrass and corn stover. These results are very useful for solving obstacles in handling bulk biomass supply logistics issues for a biorefinery.

Generation of Neo Octaploid Switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) exists as multiple cytotypes with octaploid and tetraploid populations occupying distinct, overlapping ranges. These cytotypes tend to show differences in adaptation, yield potential, and other characters, but the specific result of whole genome duplication is not ...

Comparative study of organosolv lignin extracted from prairie cordgrass, switchgrass and corn stover.

PubMed

Cybulska, Iwona; Brudecki, Grzegorz; Rosentrater, Kurt; Julson, James L; Lei, Hanwu

2012-08-01

Lignin extracted from prairie cordgrass, switchgrass, and corn stover (using ethyl acetate-ethanol-water organosolv pretreatment) was analyzed and characterized using several methods. These methods included analysis of purity (by determination of Klason lignin, carbohydrate, and ash contents), solubility (with several organic solvents), phenolic group analysis (ultraviolet ionization difference spectra, and nitrobenzene oxidation), and general functional group analysis (by (1)H NMR). Results showed that all the examined lignin samples were relatively pure (contained over 50% Klason lignin, less than 5% carbohydrate contamination, and less than 3% ash), but switchgrass-derived lignin was observed to be the purest. All the lignins were found to contain high amounts of phenolic groups, while switchgrass-derived lignin was the most phenolic, according to the ionization difference spectra. Nitrobenzene oxidation revealed that all the lignin samples contained available guaiacyl units in high amounts. Copyright © 2012 Elsevier Ltd. All rights reserved.

Predicted avian responses to bioenergy development scenarios in an intensive agricultural landscape

USGS Publications Warehouse

Uden, Daniel R.; Allen, Craig R.; Mitchell, Rob B.; McCoy, Tim D.; Guan, Qingfeng

2015-01-01

Conversion of native prairie to agriculture has increased food and bioenergy production but decreased wildlife habitat. However, enrollment of highly erodible cropland in conservation programs has compensated for some grassland loss. In the future, climate change and production of second-generation perennial biofuel crops could further transform agricultural landscapes and increase or decrease grassland area. Switchgrass (Panicum virgatum) is an alternative biofuel feedstock that may be economically and environmentally superior to maize (Zea mays) grain for ethanol production on marginally productive lands. Switchgrass could benefit farmers economically and increase grassland area, but there is uncertainty as to how conversions between rowcrops, switchgrass monocultures and conservation grasslands might occur and affect wildlife. To explore potential impacts on grassland birds, we developed four agricultural land-use change scenarios for an intensively cultivated landscape, each driven by potential future climatic changes and ensuing irrigation limitations, ethanol demand, commodity prices, and continuation of a conservation program. For each scenario, we calculated changes in area for landcover classes and predicted changes in grassland bird abundances. Overall, birds responded positively to the replacement of rowcrops with switchgrass and negatively to the conversion of conservation grasslands to switchgrass or rowcrops. Landscape context and interactions between climate, crop water use, and irrigation availability could influence future land-use, and subsequently, avian habitat quality and quantity. Switchgrass is likely to provide higher quality avian habitat than rowcrops but lower quality habitat than conservation grasslands, and therefore, may most benefit birds in heavily cultivated, irrigation dependent landscapes under warmer and drier conditions, where economic profitability may also encourage conversions to drought tolerant bioenergy feedstocks.

Modeling analysis on germination and seedling growth using ultrasound seed pretreatment in switchgrass.

PubMed

Wang, Quanzhen; Chen, Guo; Yersaiyiti, Hayixia; Liu, Yuan; Cui, Jian; Wu, Chunhui; Zhang, Yunwei; He, Xueqing

2012-01-01

Switchgrass is a perennial C4 plant with great potential as a bioenergy source and, thus, a high demand for establishment from seed. This research investigated the effects of ultrasound treatment on germination and seedling growth in switchgrass. Using an orthogonal matrix design, conditions for the ultrasound pretreatment in switchgrass seed, including sonication time (factor A), sonication temperature (factor B) and ultrasound output power (factor C), were optimized for germinating and stimulating seedling growth (indicated as plumular and radicular lengths) through modeling analysis. The results indicate that sonication temperature (B) was the most effective factor for germination, whereas output power (C) had the largest effect on seedling growth when ultrasound treatment was used. Combined with the analyses of range, variance and models, the final optimal ultrasonic treatment conditions were sonication for 22.5 min at 39.7°C and at an output power of 348 W, which provided the greatest germination percentage and best seedling growth. For this study, the orthogonal matrix design was an efficient method for optimizing the conditions of ultrasound seed treatment on switchgrass. The electrical conductivity of seed leachates in three experimental groups (control, soaked in water only, and ultrasound treatment) was determined to investigate the effects of ultrasound on seeds and eliminate the effect of water in the ultrasound treatments. The results showed that the electrical conductivity of seed leachates during either ultrasound treatment or water bath treatment was significantly higher than that of the control, and that the ultrasound treatment had positive effects on switchgrass seeds.

Breeding bird community response to establishing intercropped switchgrass in intensively-managed pine stands

USGS Publications Warehouse

Loman, Zachary G.; Riffell, Samuel K.; Wheat, Bradley R.; Miller, Darrin A.; Martin, James A.; Vilella, Francisco

2014-01-01

Intercropping switchgrass (Panicum virgatum L.) between tree rows within young pine (Pinus spp.) plantations is a potential method to generate lignocellulosic biofuel feedstocks within intensively managed forests. Intensively managed pine supports a diverse avian assemblage potentially affected by establishment and maintenance of an annual biomass feedstock via changes in plant communities, dead wood resources, and habitat structure. We sought to understand how establishing switchgrass on an operational scale affects bird communities within intercropped plantations as compared to typical intensively managed loblolly pine (Pinus taeda L.) forest. We conducted breeding bird point counts using distance sampling for three years (2011–2013) following establishment of intercropped switchgrass stands (6 replicates), traditionally-managed pine plantations, and switchgrass-only plots (0.1 km2 minimum) in Kemper Co., MS. We detected 59 breeding bird species from 11,195 detections. Neotropical migrants and forest-edge associated species were less abundant in intercropped plots than controls the first two years after establishment and more abundant in year three. Short distance migrants and residents were scarce in intercropped and control plots initially, and did not differ between these two treatments in any year. Species associated with pine-grass habitat structure were less abundant initially in intercropped plots, but converged with pine controls in subsequent years. Switchgrass monocultures provided minimal resources for birds. If songbird conservation is a management priority, managers should consider potential reductions of some breeding birds for one to two years following intercropping. It is unclear how these relationships may change outside the breeding season and as stands age.

Comparative transcriptome profiling of upland (VS16) and lowland (AP13) ecotypes of switchgrass.

PubMed

Ayyappan, Vasudevan; Saha, Malay C; Thimmapuram, Jyothi; Sripathi, Venkateswara R; Bhide, Ketaki P; Fiedler, Elizabeth; Hayford, Rita K; Kalavacharla, Venu Kal

2017-01-01

Transcriptomes of two switchgrass genotypes representing the upland and lowland ecotypes will be key tools in switchgrass genome annotation and biotic and abiotic stress functional genomics. Switchgrass (Panicum virgatum L.) is an important bioenergy feedstock for cellulosic ethanol production. We report genome-wide transcriptome profiling of two contrasting tetraploid switchgrass genotypes, VS16 and AP13, representing the upland and lowland ecotypes, respectively. A total of 268 million Illumina short reads (50 nt) were generated, of which, 133 million were obtained in AP13 and the rest 135 million in VS16. More than 90% of these reads were mapped to the switchgrass reference genome (V1.1). We identified 6619 and 5369 differentially expressed genes in VS16 and AP13, respectively. Gene ontology and KEGG pathway analysis identified key genes that regulate important pathways including C4 photosynthesis, photorespiration and phenylpropanoid metabolism. A series of genes (33) involved in photosynthetic pathway were up-regulated in AP13 but only two genes showed higher expression in VS16. We identified three dicarboxylate transporter homologs that were highly expressed in AP13. Additionally, genes that mediate drought, heat, and salinity tolerance were also identified. Vesicular transport proteins, syntaxin and signal recognition particles were seen to be up-regulated in VS16. Analyses of selected genes involved in biosynthesis of secondary metabolites, plant-pathogen interaction, membrane transporters, heat, drought and salinity stress responses confirmed significant variation in the relative expression reflected in RNA-Seq data between VS16 and AP13 genotypes. The phenylpropanoid pathway genes identified here are potential targets for biofuel conversion.

Responses of switchgrass to precipitation changes: Nonlinear and asymmetric?

USDA-ARS?s Scientific Manuscript database

Background/Question/Methods: Climate changes, including chronic changes in precipitation amounts, will influence plant physiology, biomass and productivity, and soil respiration. However, such precipitation effects on switchgrass, a major bioenergy crop, have not been well investigated. Two preci...

Sodium Hydroxide Pretreatment of Switchgrass for Ethanol Production

USDA-ARS?s Scientific Manuscript database

Lignocellulose-to-ethanol conversion is a promising technology to supplement corn-based ethanol production. However, the recalcitrant structure of lignocellulosic material is a major obstacle to the efficient conversion. To improve the enzymatic digestibility of switchgrass for the fermentable sugar...

Do yield and quality of big bluestem and switchgrass feedstock decline over winter?

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) and big bluestem (Andropogon gerdardii Vitman) are potential bioenergy feedstocks for thermochemical platforms. Feedstock storage, fall harvest constraints, and environmental benefits provided by perennials are rationales for developing localized perennial feedstock...

Selecting, Establishing, and Managing Switchgrass (Panicum virgatum) for Biofuels

NASA Astrophysics Data System (ADS)

Parrish, David J.; Fike, John H.

Switchgrass is being widely considered as a feedstock for biofuel production. Much remains to be learned about ideal feedstock characteristics, but switchgrass offers many advantages already and can perhaps be manipulated to offer more. When planning to grow switchgrass, select a cultivar that is well adapted to the location - generally a lowland cultivar for the southern United States and an upland cultivar at higher latitudes. Plant non-dormant seed after soils are well warmed, preferably with no-till methods and always with good weed control. Except for weeds, few pests appear to be widespread; but disease and insect pests could become more important as acreages increase. Fertilization requirements are relatively low, with 50 kg N/ha/year being a good “generic” recommendation where a single harvest is taken after plants have senesced; more will be needed if biomass is harvested while still green. Switchgrass should be harvested no more than twice per year and may generally be expected to produce 12 to ≥20 mg/ha/year across its usual range of distribution. A single harvest may provide for maximum sustainable yields - especially if the harvest is taken after tops die back at the end of the season. Several harvesting technologies are available, but the preferred technology may depend on logistics and economics associated with the local processing point, or biorefinery.

Nitrogen Fertilization Effects on Productivity and Nitrogen Loss in Three Grass-Based Perennial Bioenergy Cropping Systems

DOE PAGES

Duran, Brianna E. L.; Duncan, David S.; Oates, Lawrence G.; ...

2016-03-18

Nitrogen (N) fertilization can greatly improve plant productivity but needs to be carefully managed to avoid harmful environmental impacts. Nutrient management guidelines aimed at reducing harmful forms of N loss such as nitrous oxide (N 2O) emissions and nitrate (NO 3 -) leaching have been tailored for many cropping systems. The developing bioenergy industry is likely to make use of novel cropping systems, such as polycultures of perennial species, for which we have limited nutrient management experience. We studied how a switchgrass (Panicum virgatum) monoculture, a 5-species native grass mixture and an 18- species restored prairie responded to annual fertilizermore » applications of 56 kg N ha -1 in a fieldscale agronomic trial in south-central Wisconsin over a 2-year period.We observed greater fertilizer-induced N 2O emissions and sub-rooting zone NO 3 - concentrations in the switchgrass monoculture than in either polyculture. Fertilization increased aboveground net primary productivity in the polycultures, but not in the switchgrass monoculture. Switchgrass was generally more productive, while the two polycultures did not differ from each other in productivity or N loss. In conclusion, our results highlight differences between polycultures and a switchgrass monoculture in responding to N fertilization.« less

Genetic variation, population structure and linkage disequilibrium in Switchgrass with ISSR, SCoT and EST-SSR markers.

PubMed

Zhang, Yu; Yan, Haidong; Jiang, Xiaomei; Wang, Xiaoli; Huang, Linkai; Xu, Bin; Zhang, Xinquan; Zhang, Lexin

2016-01-01

To evaluate genetic variation, population structure, and the extent of linkage disequilibrium (LD), 134 switchgrass ( Panicum virgatum L.) samples were analyzed with 51 markers, including 16 ISSRs, 20 SCoTs, and 15 EST-SSRs. In this study, a high level of genetic variation was observed in the switchgrass samples and they had an average Nei's gene diversity index (H) of 0.311. A total of 793 bands were obtained, of which 708 (89.28 %) were polymorphic. Using a parameter marker index (MI), the efficiency of the three types of markers (ISSR, SCoT, and EST-SSR) in the study were compared and we found that SCoT had a higher marker efficiency than the other two markers. The 134 switchgrass samples could be divided into two sub-populations based on STRUCTURE, UPGMA clustering, and principal coordinate analyses (PCA), and upland and lowland ecotypes could be separated by UPGMA clustering and PCA analyses. Linkage disequilibrium analysis revealed an average r 2 of 0.035 across all 51 markers, indicating a trend of higher LD in sub-population 2 than that in sub-population 1 ( P  < 0.01). The population structure revealed in this study will guide the design of future association studies using these switchgrass samples.

Selecting, establishing, and managing switchgrass (Panicum virgatum) for biofuels.

PubMed

Parrish, David J; Fike, John H

2009-01-01

Switchgrass is being widely considered as a feedstock for biofuel production. Much remains to be learned about ideal feedstock characteristics, but switchgrass offers many advantages already and can perhaps be manipulated to offer more. When planning to grow switchgrass, select a cultivar that is well adapted to the location - generally a lowland cultivar for the southern United States and an upland cultivar at higher latitudes. Plant non-dormant seed after soils are well warmed, preferably with no-till methods and always with good weed control. Except for weeds, few pests appear to be widespread; but disease and insect pests could become more important as acreages increase. Fertilization requirements are relatively low, with 50 kg N/ha/year being a good "generic" recommendation where a single harvest is taken after plants have senesced; more will be needed if biomass is harvested while still green. Switchgrass should be harvested no more than twice per year and may generally be expected to produce 12 to >or=20 mg/ha/year across its usual range of distribution. A single harvest may provide for maximum sustainable yields - especially if the harvest is taken after tops die back at the end of the season. Several harvesting technologies are available, but the preferred technology may depend on logistics and economics associated with the local processing point, or biorefinery.

Big bluestem and switchgrass feedstock harvest timing: Nitrous oxide response to feedstock harvest timing

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) and big bluestem (Andropogon gerdardii Vitman) are potential bioenergy feedstocks. Feedstock storage limitations, labor constraints for harvest, and environmental benefits provided by perennials are rationales for developing localized perennial feedstock as an alter...

Switchgrass nitrogen response and estimated production costs on diverse sites

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) has been the principal perennial herbaceous crop investigated for bioenergy production in North America given its high production potential, relatively low input requirements, and potential suitability for use on marginal lands. Few large trials have determined swit...

Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax.

PubMed

Pigna, Gaia; Dhillon, Taniya; Dlugosz, Elizabeth M; Yuan, Joshua S; Gorman, Connor; Morandini, Piero; Lenaghan, Scott C; Stewart, C Neal

2016-12-01

Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 µM 2,4-D and 4.4 µM BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 µM 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 ± 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hydrologic cost-effectiveness ratio favors switchgrass production on marginal croplands over existing grasslands

PubMed Central

Yimam, Yohannes Tadesse; Ochsner, Tyson E.; Fox, Garey A.

2017-01-01

Switchgrass (Panicum virgatum L.) has attracted attention as a promising second generation biofuel feedstock. Both existing grasslands and marginal croplands have been suggested as targets for conversion to switchgrass, but the resulting production potentials and hydrologic impacts are not clear. The objectives of this study were to model switchgrass production on existing grasslands (scenario-I) and on marginal croplands that have severe to very severe limitations for crop production (scenario-II) and to evaluate the effects on evapotranspiration (ET) and streamflow. The Soil and Water Assessment Tool (SWAT) was applied to the 1063 km2 Skeleton Creek watershed in north-central Oklahoma, a watershed dominated by grasslands (35%) and winter wheat cropland (47%). The simulated average annual yield (2002–2011) for rainfed Alamo switchgrass for both scenarios was 12 Mg ha-1. Yield varied spatially under scenario-I from 6.1 to 15.3 Mg ha-1, while under scenario-II the range was from 8.2 to 13.8 Mg ha-1. Comparison of average annual ET and streamflow between the baseline simulation (existing land use) and scenario-I showed that scenario-I had 5.6% (37 mm) higher average annual ET and 27.7% lower streamflow, representing a 40.7 million m3 yr-1 streamflow reduction. Compared to the baseline, scenario-II had only 0.5% higher ET and 3.2% lower streamflow, but some monthly impacts were larger. In this watershed, the water yield reduction per ton of biomass production (i.e. hydrologic cost-effectiveness ratio) was more than 5X greater under scenario-I than under scenario-II. These results suggest that, from a hydrologic perspective, it may be preferable to convert marginal cropland to switchgrass production rather than converting existing grasslands. PMID:28792541

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kenkel, Philip; Holcomb, Rodney B.

In order for the biofuel industry to meet the RFS benchmarks for biofuels, new feedstock sources and production systems will have to be identified and evaluated. The Southern Plains has the potential to produce over a billion gallons of biofuels from regionally produced alternative crops, agricultural residues, and animal fats. While information on biofuel conversion processes is available, it is difficult for entrepreneurs, community planners and other interested individuals to determine the feasibility of biofuel processes or to match production alternatives with feed stock availability and community infrastructure. This project facilitates the development of biofuel production from these regionally availablemore » feed stocks. Project activities are concentrated in five major areas. The first component focused on demonstrating the supply of biofuel feedstocks. This involves modeling the yield and cost of production of dedicated energy crops at the county level. In 1991 the DOE selected switchgrass as a renewable source to produce transportation fuel after extensive evaluations of many plant species in multiple location (Caddel et al,. 2010). However, data on the yield and cost of production of switchgrass are limited. This deficiency in demonstrating the supply of biofuel feedstocks was addressed by modeling the potential supply and geographic variability of switchgrass yields based on relationship of available switchgrass yields to the yields of other forage crops. This model made it possible to create a database of projected switchgrass yields for five different soil types at the county level. A major advantage of this methodology is that the supply projections can be easily updated as improved varieties of switchgrass are developed and additional yield data becomes available. The modeling techniques are illustrated using the geographic area of Oklahoma. A summary of the regional supply is then provided.« less

Hydrologic cost-effectiveness ratio favors switchgrass production on marginal croplands over existing grasslands.

PubMed

Yimam, Yohannes Tadesse; Ochsner, Tyson E; Fox, Garey A

2017-01-01

Switchgrass (Panicum virgatum L.) has attracted attention as a promising second generation biofuel feedstock. Both existing grasslands and marginal croplands have been suggested as targets for conversion to switchgrass, but the resulting production potentials and hydrologic impacts are not clear. The objectives of this study were to model switchgrass production on existing grasslands (scenario-I) and on marginal croplands that have severe to very severe limitations for crop production (scenario-II) and to evaluate the effects on evapotranspiration (ET) and streamflow. The Soil and Water Assessment Tool (SWAT) was applied to the 1063 km2 Skeleton Creek watershed in north-central Oklahoma, a watershed dominated by grasslands (35%) and winter wheat cropland (47%). The simulated average annual yield (2002-2011) for rainfed Alamo switchgrass for both scenarios was 12 Mg ha-1. Yield varied spatially under scenario-I from 6.1 to 15.3 Mg ha-1, while under scenario-II the range was from 8.2 to 13.8 Mg ha-1. Comparison of average annual ET and streamflow between the baseline simulation (existing land use) and scenario-I showed that scenario-I had 5.6% (37 mm) higher average annual ET and 27.7% lower streamflow, representing a 40.7 million m3 yr-1 streamflow reduction. Compared to the baseline, scenario-II had only 0.5% higher ET and 3.2% lower streamflow, but some monthly impacts were larger. In this watershed, the water yield reduction per ton of biomass production (i.e. hydrologic cost-effectiveness ratio) was more than 5X greater under scenario-I than under scenario-II. These results suggest that, from a hydrologic perspective, it may be preferable to convert marginal cropland to switchgrass production rather than converting existing grasslands.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sarath, Gautam

This grant funded work was undertaken to develop fundamental biological knowledge of the factors affecting the complex plant trait “fitness” in switchgrass (Panicum virgatum L.), a plant being developed as a biomass crop. Using a diverse range of latitudinally-adapted switchgrass plants, genomic, molecular and physiological studies were performed to track a number of different aspects of plant genetics and physiology over the course of the growing season. Work was performed on both genetically unrelated and genetically related plants. Plants were established in the field from seedlings raised in a greenhouse, or from clones present in other field nurseries. Field grownmore » plants were used as the source of all tissues. The three objectives of this proposal were:(1) Transcript Profiling, Metabolomics, and C and N Partitioning and Recycling in Crowns and Rhizomes of Switchgrass over two growing seasons; (2) Gene Profiling During Regreening and Dormancy of Bulked Segregants; (3) Extent of Linkage Disequilibrium in Populations for Adaptation and Fitness Traits Being Developed for Central and Northern USA, that Show Significant Heterosis. Objective 1 results: Plants were labeled using 13CO2 (a stable isotope) using an acrylic chamber constructed specifically for this purpose. Plants became labeled with 13C and label decayed in aerial tissues over the course of the growing season. Varying amounts of 13C were recovered in the rhizomes. These data are being analyzed. Plants were also labeled with 15N-urea. Plants absorbed significant amounts of label that was remobilized to the growing shoots. N-dynamics would suggest that a portion of the 15N absorbed into the crowns and rhizomes is sequestered below ground. Variable amounts of 15N were translocated from the shoots to the roots over the course of the growing season. Polar metabolites extracted from a diverse array of rhizomes were analyzed using GCMS. Data indicated that there was a significant shift in metabolite pools over the course of the growing season, and differences in the levels of specific metabolites could be linked to the progression of dormancy. Several metabolites that accumulate in dormant rhizomes were identified. Some of these metabolites could be potentially linked to winter-survival of switchgrass. Extensive high-throughput sequencing was conducted on crown and rhizome samples collected from field grown plants. Initial work was performed on a Roche 454 system. All later work was performed on an Illumina sequencing-by-synthesis system. Some of these datasets have been published as peer-reviewed papers, other data are currently being analyzed and being readied for publication. Objective 2 results: Genetically related but phenotypically divergent plants from an octaploid switchgrass population were grown in a replicated field nursery. Rhizomes were harvested at four different times over the course of the growing season from plants with high winter survival and those with lower winter survival. RNA-Seq was performed on harvested materials. Initial analysis suggests that plants with lowered winter survival experience a greater level of cellular stress in dormant tissues. This aspect of plant function is being probed in greater depth. Objective 3 results: A total of 592 individual clones with three clonal replications in a randomized complete block design from each of five populations used in Objective 1 studies were rated for heading date in 2012 and 2014, green-up day of year in 2013, anthesis date in 2012, and yield in 2012, they were also subjected to NIR spectroscopy to derive cell wall composition estimates based on prior NIR calibrations. Plants were genotyped via a genotyping by sequencing (GBS) approach from reduced representation libraries constructed with adaptors that identified each individual. Libraries generated with the restriction enzyme PstI and called SNPs using Samtools after alignment to version 1.1 of the switchgrass genome sequence. A total of approximately 40,000 SNPs were found. These were then further filtered to eliminate markers with a minor allele frequency of < 0.05. The results of population analysis using STRUCTURE with expected population sizes or cluster numbers (K), clearly shows the hybrid composition of the KxS population and discriminated easily between upland (Summer) and lowland (Kanlow) populations. Under an assumption of 5 distinct populations there were detectable differences in allele frequencies between subpopulations within the three Kanlow populations particularly with respect to Kanlow EM and Kanlow base. We detected 110 SNPs with an allele frequency difference of ≥ 0.2 between Kanlow EM and Kanlow base populations, while 120 SNPs showed an allele frequency difference of ≥ 0.15 between Kanlow N1 and Kanlow base populations. These data are being readied for publication.« less

Hydrogen production from switchgrass via an integrated pyrolysis-microbial electrolysis process.

PubMed

Lewis, A J; Ren, S; Ye, X; Kim, P; Labbe, N; Borole, A P

2015-11-01

A new approach to hydrogen production using an integrated pyrolysis-microbial electrolysis process is described. The aqueous stream generated during pyrolysis of switchgrass was used as a substrate for hydrogen production in a microbial electrolysis cell, achieving a maximum hydrogen production rate of 4.3 L H2/L anode-day at a loading of 10 g COD/L-anode-day. Hydrogen yields ranged from 50±3.2% to 76±0.5% while anode Coulombic efficiency ranged from 54±6.5% to 96±0.21%, respectively. Significant conversion of furfural, organic acids and phenolic molecules was observed under both batch and continuous conditions. The electrical and overall energy efficiency ranged from 149-175% and 48-63%, respectively. The results demonstrate the potential of the pyrolysis-microbial electrolysis process as a sustainable and efficient route for production of renewable hydrogen with significant implications for hydrocarbon production from biomass. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of precipitation changes on switchgrass photosynthesis, growth, and biomass: A mesocosm experiment

USDA-ARS?s Scientific Manuscript database

Climate changes, including chronic changes in precipitation amounts, will influence plant physiology and growth. However, such precipitation effects on switchgrass, a major bioenergy crop, have not been well investigated. We conducted a two-year precipitation simulation experiment using large pots...

The WRKY transcription factor family and senescence in switchgrass

USDA-ARS?s Scientific Manuscript database

Background: Early aerial senescence in switchgrass (Panicum virgatum) can significantly limit biomass yields. WRKY transcription factors that can regulate senescence could be used to reprogram senescence and enhance biomass yields. Methods: All potential WRKY genes present in the version 1.0 of the...

Evaluation of switchgrass for improved biomass yield on marginal land

USDA-ARS?s Scientific Manuscript database

The national strategy is to produce bioenergy crops on marginal cropland where there will be no competition with food production. The characteristics of switchgrass (Panicum virgatum) make it an excellent candidate for sustainable biomass production on marginal land. However, few studies have evalu...

Genomic prediction for winter survival in lowland switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a North American native perennial warm season grass and a promising cellulosic bioenergy feedstock. It has two ecotypes – lowland and upland. The lowland ecotype has generated considerable interest because of its higher biomass compared to the upland ecotype. How...

Topsoil thickness influences nitrogen management of switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is an attractive bioenergy crop option for eroded portions of claypan landscapes where grain crop production is marginally profitable. Topsoil thickness above the claypan or depth to claypan (DTC) can vary widely within fields and little information exists on its im...

Switchgrass composition and yield response to alternative soil amendments under intensified heat and drought conditions

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) and guinea grass (Panicum maximum Jacq.) have been proposed as sustainable alternatives to fossil fuels in temperate and tropical environments, respectively; although still requiring non-renewable inputs, notably, fertilizer-nitrogen (N). Furthermore, climate change...

Depolymerization and hydrodeoxygenation of switchgrass lignin with formic acid.

PubMed

Xu, Weiyin; Miller, Stephen J; Agrawal, Pradeep K; Jones, Christopher W

2012-04-01

Organosolv switchgrass lignin is depolymerized and hydrodeoxygenated with a formic acid hydrogen source, 20 wt % Pt/C catalyst, and ethanol solvent. The combination of formic acid and Pt/C is found to promote production of higher fractions of lower molecular weight compounds in the liquid products. After 4 h of reaction, all of the switchgrass lignin is solubilized and 21 wt % of the biomass is shown to be converted into seven prominent molecular species that are identified and quantified. Reaction time is shown to be an important variable in affecting changes in product distributions and bulk liquid product properties. At 20 h of reaction, the lignin is significantly depolymerized to form liquid products with a 76 % reduction in the weighted average molecular weight. Elemental analysis also shows that the resultant liquid products have a 50 % reduction in O/C and 10 % increase in H/C molar ratios compared to the switchgrass lignin after 20 h. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Biomass logistics analysis for large scale biofuel production: case study of loblolly pine and switchgrass.

PubMed

Lu, Xiaoming; Withers, Mitch R; Seifkar, Navid; Field, Randall P; Barrett, Steven R H; Herzog, Howard J

2015-05-01

The objective of this study was to assess the costs, energy consumption and greenhouse gas (GHG) emissions throughout the biomass supply chain for large scale biofuel production. Two types of energy crop were considered, switchgrass and loblolly pine, as representative of herbaceous and woody biomass. A biomass logistics model has been developed to estimate the feedstock supply system from biomass production through transportation. Biomass in the form of woodchip, bale and pellet was investigated with road, railway and waterway transportation options. Our analysis indicated that the farm or forest gate cost is lowest for loblolly pine whole tree woodchip at $39.7/dry tonne and highest for switchgrass round bale at $72.3/dry tonne. Switchgrass farm gate GHG emissions is approximately 146kgCO2e/dry tonne, about 4 times higher than loblolly pine. The optimum biomass transportation mode and delivered form are determined by the tradeoff between fixed and variable costs for feedstock shipment. Copyright © 2015 Elsevier Ltd. All rights reserved.

Natural variation in genes potentially involved in plant architecture and adaptation in switchgrass (Panicum virgatum L.).

PubMed

Bahri, Bochra A; Daverdin, Guillaume; Xu, Xiangyang; Cheng, Jan-Fang; Barry, Kerrie W; Brummer, E Charles; Devos, Katrien M

2018-06-14

Advances in genomic technologies have expanded our ability to accurately and exhaustively detect natural genomic variants that can be applied in crop improvement and to increase our knowledge of plant evolution and adaptation. Switchgrass (Panicum virgatum L.), an allotetraploid (2n = 4× = 36) perennial C4 grass (Poaceae family) native to North America and a feedstock crop for cellulosic biofuel production, has a large potential for genetic improvement due to its high genotypic and phenotypic variation. In this study, we analyzed single nucleotide polymorphism (SNP) variation in 372 switchgrass genotypes belonging to 36 accessions for 12 genes putatively involved in biomass production to investigate signatures of selection that could have led to ecotype differentiation and to population adaptation to geographic zones. A total of 11,682 SNPs were mined from ~ 15 Gb of sequence data, out of which 251 SNPs were retained after filtering. Population structure analysis largely grouped upland accessions into one subpopulation and lowland accessions into two additional subpopulations. The most frequent SNPs were in homozygous state within accessions. Sixty percent of the exonic SNPs were non-synonymous and, of these, 45% led to non-conservative amino acid changes. The non-conservative SNPs were largely in linkage disequilibrium with one haplotype being predominantly present in upland accessions while the other haplotype was commonly present in lowland accessions. Tajima's test of neutrality indicated that PHYB, a gene involved in photoperiod response, was under positive selection in the switchgrass population. PHYB carried a SNP leading to a non-conservative amino acid change in the PAS domain, a region that acts as a sensor for light and oxygen in signal transduction. Several non-conservative SNPs in genes potentially involved in plant architecture and adaptation have been identified and led to population structure and genetic differentiation of ecotypes in switchgrass. We suggest here that PHYB is a key gene involved in switchgrass natural selection. Further analyses are needed to determine whether any of the non-conservative SNPs identified play a role in the differential adaptation of upland and lowland switchgrass.

Influence of endophytic root bacteria on the growth, cadmium tolerance and uptake of switchgrass (Panicum virgatum L.).

PubMed

Afzal, S; Begum, N; Zhao, H; Fang, Z; Lou, L; Cai, Q

2017-08-01

This study aimed to evaluate the effect of five cadmium-tolerant endophytic plant growth-promoting bacteria (PGPB) strains on the biomass and cadmium tolerance, and accumulation of the bioenergy plant switchgrass (Panicum virgatum L.) under cadmium (Cd)-contaminated conditions. Five bacterial strains (Bj05, Le14, Ps14, So02 and Bo03) that could tolerate up to 2000 μmol l -1 CdCl 2 with a high Cd-binding capacity were isolated from plants grown in Cd-contaminated soil. These strains could promote the growth of switchgrass in the presence of 20 μmol l -1 CdCl 2 . Under Cd stress, both the root and shoot length and biomass in two switchgrass cultivars, Alamo and Cave-in-Rock (CIR), increased significantly after treatment with PGPB (P ˂ 0·05). Of the five PGPB strains, Bj05 and Le14 most effectively alleviated the Cd-induced growth inhibition of Alamo plants. Interestingly, the five PGPB strains increased Cd tolerance and decreased the accumulation and translocation factor (TF) of Cd in switchgrass when grown in the presence of 20 μmol l -1 CdCl 2 . The Cd concentrations in roots and shoots of Alamo and CIR plants were significantly reduced (P ˂ 0·05) compared with noninoculated plants. By 16S rRNA sequencing, these PGPB strains were classified as Pseudomonas putida Bj05, Pseudomonas fluorescens Ps14, and Enterobacter spp. Le14, So02 and Bo03. Inoculation with PGPB protects plants from the inhibitory effects of Cd, improves plant growth and decreases Cd concentration. These observations suggest that these strains could be used to promote growth and lower the bioavailability of Cd in switchgrass. These strains are potential candidates for the development of low Cd-accumulating switchgrass, particularly in areas of Cd contamination and pollution, and could be used efficiently for the bioremediation of contaminated soil. © 2017 The Society for Applied Microbiology.

evaluation of the bioconversion of genetically modified switchgrass using simultaneous saccharification and fermentation ans a consolidated bioprocessing approach

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yee, Kelsey L; Rodriguez, Jr., Miguel; Tschaplinski, Timothy J

2012-01-01

Abstract Background: The inherent recalcitrance of lignocellulosic biomass is one of the major economic hurdles for the production of fuels and chemicals from biomass. Additionally, lignin is recognized as having a negative impact on enzymatic hydrolysis of biomass, and as a result much interest has been placed on modifying the lignin pathway to improve bioconversion of lignocellulosic feedstocks. Results: Previous results showed down-regulation of the caffeic acid 3-O-methyl transferase (COMT) gene in the lignin pathway yielded switchgrass (Panicum virgatum) that was more susceptible to bioconversion after dilute acid pretreatment. Here we examined the response of these plant lines to mildermore » pretreatment conditions with yeast-based SSF, CBP with Clostridium thermocellum, and fermentations with the cellulolytic extreme thermophiles, Caldicellulosiruptor bescii and Caldicellulosiruptor obsidiansis. Unlike the S. cerevisiae SSF conversions, fermentations of pretreated down-regulated COMT transgenic switchgrass with C. thermocellum showed an apparent inhibition of fermentation not observed in the wild-type switchgrass. This inhibition can be eliminated by hot water extraction of the pretreated biomass which resulted in superior conversion yield with transgenic versus wild-type switchgrass for C. thermocellum, also exceeding the yeast-based SSF yield. Further fermentation evaluation of the transgenic switchgrass indicated differential inhibition for the Caldicellulosiruptor strains, which could not be rectified by additional processing conditions. Gas chromatography-mass spectrometry metabolite profiling was used to examine the fermentation broth to elucidate the relative abundance of lignin derived aromatic compounds. The types and abundance of fermentation-derived lignin constituents varied between C. thermocellum and each of the Caldicellulosiruptor strains. Conclusions: The down-regulation of the COMT gene improves the bioconversion of switchgrass relative to the wild-type regardless of the pretreatment condition or fermentation microorganism. However, bacterial fermentations demonstrated strain-dependent sensitivity to the COMT transgenic biomass, likely due to additional soluble lignin pathway-derived constituents resulting from the COMT gene disruption. Removal of these inhibitory constituents permitted completion of fermentation by C. thermocellum, but not by the Caldicellulosiruptor strains. The reason for this difference needs to be explored further.« less

Using electrical resistance probes for moisture determination in switchgrass windrows

USDA-ARS?s Scientific Manuscript database

Determining moisture levels in windrowed biomass is important for both forage producers and researchers. Energy crops such as switchgrass have been troublesome when using the standard methods set for electrical resistance meters. The objectives of this study were to i) develop the methodologies need...

Seasonal below-ground metabolism in switchgrass

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum) a perennial, polyploid, C4 warm-season grass is one of the foremost herbaceous species being advanced as a source of biomass for biofuel end uses. At the end of every growing season, the aerial tissues senesce, and the below-ground rhizomes become dormant. Future growt...

Determining switchgrass biomass supplies for cellulosic biorefineries

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is being developed into a bioenergy crop for use in temperate regions of the USA. Information on spatial and temporial variation for stands and biomass yield among and within fields in large agroecoregions is not available. A reliable feedstock supply will be essent...

Life cycle analysis of switchgrass converted via pyrolysis, gasification, and fermentation

USDA-ARS?s Scientific Manuscript database

The US is promoting and developing low carbon fuel sources. Perennial bioenergy crops such as switchgrass (Panicum virgatum L.) are one viable source for low carbon transportation fuels. The objective is to determine the net greenhouse gas (GHG) emissions from different conversion methods (pyrolysi...

Switchgrass growth and effects on biomass accumulation, moisture content, and nutrient removal

USDA-ARS?s Scientific Manuscript database

Temporal patterns of plant growth, composition, and nutrient removal impact development of models for predicting optimal harvest times of switchgrass (Panicum virgatum L.) for bioenergy. Objectives were to characterize seasonal trends in yield, tissue moisture, ash content, leaf area index (LAI), in...

Detection and characterization of the first North American mastrevirus in Switchgrass

USDA-ARS?s Scientific Manuscript database

Virus infections have the potential to reduce biomass yields in energy crops, including Panicum virgatum (switchgrass). As a first step towards managing virus-induced biomass reductions, deep sequencing was used to identify viruses associated with mosaic symptoms in switch grass, which detected thre...

Impact of switchgrass harvest time on biomass yield and conversion

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a perennial grass native to much of North America being developed as a dedicated energy crop for conversion to biofuels. Breeding efforts are focused on producing high-yielding cultivars that can maintain high yield across multiple environments, including poor so...

Grass invasion into switchgrass managed for biomass energy

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum) is a warm-season perennial grass and is the model herbaceous perennial bioenergy feedstock. Although it is indigenous to North American grasslands east of the Rocky Mountains and has been planted for forage and conservation purposes for more than 75 years, there is con...

Nitrogen management of switchgrass and miscanthus on marginal soils

USDA-ARS?s Scientific Manuscript database

Miscanthus × giganteus and switchgrass yield and fertilizer N requirements have been well studied in Europe and parts of the United States, but few reports have investigated their production on eroded claypan soils economically marginal for grain crops. This study was conducted to evaluate yield pot...

Topsoil depth influences switchgrass nitrogen management on claypan soils

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is an attractive forage or bioenergy crop option for eroded portions of claypan landscapes where grain crop production is marginally profitable. Topsoil depth to the claypan can vary widely within fields and little information exists on the impacts of the topsoil de...

Structural analysis of pyrolytic lignins isolated from switchgrass fast pyrolysis oil

USDA-ARS?s Scientific Manuscript database

Structural characterization of lignin extracted from the bio-oil produced by fast pyrolysis of switchgrass (Panicum virgatum) is reported. This new information is important to understanding the utility of lignin as a chemical feedstock in a pyrolysis based biorefinery. Pyrolysis induces a variety of...

Comparing corn stover and switchgrass biochar: characterization and sorption properties

USDA-ARS?s Scientific Manuscript database

A switchgrass biochar (SB) produced by fast pyrolysis and a corn stover biochar (CSB) from a slow pyrolysis process were mechanically milled and characterized. Both of these biochars are very cost-effective and originate as residues from bioenergy production and the corn industry, respectively. Thes...

Genome-Wide Identification, Phylogeny, and Expression Analysis of ARF Genes Involved in Vegetative Organs Development in Switchgrass.

PubMed

Wang, Jianli; Wu, Zhenying; Shen, Zhongbao; Bai, Zetao; Zhong, Peng; Ma, Lichao; Pan, Duofeng; Zhang, Ruibo; Li, Daoming; Zhang, Hailing; Fu, Chunxiang; Han, Guiqing; Guo, Changhong

2018-01-01

Auxin response factors (ARFs) have been reported to play vital roles during plant growth and development. In order to reveal specific functions related to vegetative organs in grasses, an in-depth study of the ARF gene family was carried out in switchgrass ( Panicum virgatum L.), a warm-season C4 perennial grass that is mostly used as bioenergy and animal feedstock. A total of 47 putative ARF genes ( PvARFs ) were identified in the switchgrass genome (2n = 4x = 36), 42 of which were anchored to the seven pairs of chromosomes and found to be unevenly distributed. Sixteen PvARFs were predicted to be potential targets of small RNAs (microRNA160 and 167). Phylogenetically speaking, PvARFs were divided into seven distinct subgroups based on the phylogeny, exon/intron arrangement, and conserved motif distribution. Moreover, 15 pairs of PvARFs have different temporal-spatial expression profiles in vegetative organs (2nd, 3rd, and 4th internode and leaves), which implies that different PvARFs have specific functions in switchgrass growth and development. In addition, at least 14 pairs of PvARFs respond to naphthylacetic acid (NAA) treatment, which might be helpful for us to study on auxin response in switchgrass. The comprehensive analysis, described here, will facilitate the future functional analysis of ARF genes in grasses.

Switchgrass Biomass Quality as Affected by Nitrogen Rate, Harvest Time, and Storage

DOE PAGES

Ibrahim, Mostafa; Hong, Chang Oh.; Singh, Shikha; ...

2017-01-25

The purpose of this study was to assess the changes in switchgrass (Panicum virgatum L.) biomass quality as affected by N rate, harvest time, and storage. This research was conducted near Bristol, SD, in 2010 and 2011. Treatments included three N rates (0, 56, and 112 kg N ha –1) applied annually and each N rate replicated four times. After a killing frost, all of the plots were harvested and baled in large round bales in October 2010 and November 2011. An area of about 30 m 2 from each plot was left unharvested to represent storage of standing switchgrassmore » over the winter and to determine dry matter yields. Switchgrass was analyzed for hemicellulose, cellulose, lignin, mineral elements, N, and C. In the first season, storage of the fall harvested switchgrass bales numerically increased the concentrations of hemicellulose, lignin, and N. In the second season, they increased significantly. Mineral elements significantly increased in both sampling seasons. Delaying harvest until spring decreased lignin, N, and mineral elements concentration, and increased cellulose and hemicellulose concentrations, but also reduced biomass yield. Results from this study suggest that delaying the switchgrass harvest until spring increased the overall feedstock quality for ethanol production, but yield reductions must be considered to determine the overall economic impact of a delayed harvest.« less

Tassel Removal Positively Affects Biomass Production Coupled with Significantly Increasing Stem Digestibility in Switchgrass

PubMed Central

Zhao, Chunqiao; Fan, Xifeng; Hou, Xincun; Zhu, Yi; Yue, Yuesen; Zhang, Shuang; Wu, Juying

2015-01-01

In this study, tassels of Cave-in-Rock (upland) and Alamo (lowland) were removed at or near tassel emergence to explore its effects on biomass production and quality. Tassel-removed (TR) Cave-in-Rock and Alamo both exhibited a significant (P<0.05) increase in plant heights (not including tassel length), tiller number, and aboveground biomass dry weight (10% and 12%, 30% and 13%, 13% and 18%, respectively by variety) compared to a control (CK) treatment. Notably, total sugar yields of TR Cave-in-Rock and Alamo stems increased significantly (P<0.05 or 0.01) by 19% and 19%, 21% and 14%, 52% and 18%, respectively by variety, compared to those of control switchgrass under 3 treatments by direct enzymatic hydrolysis (DEH), enzymatic hydrolysis after 1% NaOH pretreatment (EHAL) and enzymatic hydrolysis after 1% H2SO4 pretreatment (EHAC). These differences were mainly due to significantly (P<0.05 or 0.01) higher cellulose content, lower cellulose crystallinity indexes (CrI) caused by higher arabinose (Ara) substitution in xylans, and lower S/G ratio in lignin. However, the increases of nitrogen (N) and sulphur (S) concentration negatively affects the combustion quality of switchgrass aboveground biomass. This work provides information for increasing biomass production and quality in switchgrass and also facilitates the inhibition of gene dispersal of switchgrass in China. PMID:25849123

Switchgrass Biomass Quality as Affected by Nitrogen Rate, Harvest Time, and Storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ibrahim, Mostafa; Hong, Chang Oh.; Singh, Shikha

The purpose of this study was to assess the changes in switchgrass (Panicum virgatum L.) biomass quality as affected by N rate, harvest time, and storage. This research was conducted near Bristol, SD, in 2010 and 2011. Treatments included three N rates (0, 56, and 112 kg N ha –1) applied annually and each N rate replicated four times. After a killing frost, all of the plots were harvested and baled in large round bales in October 2010 and November 2011. An area of about 30 m 2 from each plot was left unharvested to represent storage of standing switchgrassmore » over the winter and to determine dry matter yields. Switchgrass was analyzed for hemicellulose, cellulose, lignin, mineral elements, N, and C. In the first season, storage of the fall harvested switchgrass bales numerically increased the concentrations of hemicellulose, lignin, and N. In the second season, they increased significantly. Mineral elements significantly increased in both sampling seasons. Delaying harvest until spring decreased lignin, N, and mineral elements concentration, and increased cellulose and hemicellulose concentrations, but also reduced biomass yield. Results from this study suggest that delaying the switchgrass harvest until spring increased the overall feedstock quality for ethanol production, but yield reductions must be considered to determine the overall economic impact of a delayed harvest.« less

[Effect of ensilage on bioconversion of switchgrass to ethanol based on liquid hot water pretreatment].

PubMed

Wu, Wentao; Ju, Meiting; Liu, Jinpeng; Liu, Boqun

2016-04-25

Ensilage is a traditional way of preserving fresh biomass. However, in order to apply ensilage to the ethanol biorefinery, two parameters need to be evaluated: quantity and quality changes of the biomass; and its effects on bioconversion process. To study these two aspects, switchgrass harvested on three different time points (Early, mid and late fall) were used as feedstock. The early fall harvested biomass was ensiled at 5 moisture levels ranging from 30% to 70%. Silage of 40% moisture and 3 other raw switchgrass were pretreated with liquid hot water, followed by enzymatic hydrolysis as well as simultaneous saccharification and fermentation. After 21 days storage pH values of all silages decreased below 4.0 and the dry matter losses were less than 2.0%, and structural sugars contents did not change dramatically. Liquid hot water caused more hemicellulose dissolution in the silage than in unensiled switchgrass. However, ensilage also increased the risk of releasing more sugar degradation products; After enzymatic hydrolysis, silage obtained higher total glucose, xylose and galactose yields than raw materials; After simultaneous saccharification and fermentation, ethanol concentration in silage was 12.1 g/L, higher than the unensiled switchgrass (10.3 g/L, 9.7 g/L and 10.6 g/L for early, mid and late fall respectively). Our results suggest that ensilage helps increase pretreatment efficiency and sugar yield, which increases final ethanol production.

Quantitative trait loci mapping for flowering time in a switchgrass pseudo-F2 population

USDA-ARS?s Scientific Manuscript database

Flowering is an important developmental event in switchgrass (Panicum virgatum) because the onset of flowering causes the cessation of vegetative growth and biomass accumulation. The objective of this study was to generate a linkage map using single nucleotide polymorphism (SNP) markers to identify ...

Sodium hydroxide pretreatment of genetically modified switchgrass for improved enzymatic release of sugars

USDA-ARS?s Scientific Manuscript database

Overcoming biomass recalcitrance to biological conversion has been the focus of enormous research efforts in the cellulosic biofuel area in the past decades. In this study, Alamo switchgrass was genetically transformed to suppress the expression of 4-coumarate-CoA ligase (4CL). The transgenic plants...

Nitrogen and harvest impact on biomass yield of established switchgrass (Panicum virgatum L.)

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) has been identified as the model herbaceous biomass energy crop by the United States Department of Energy as it is capable of being a viable bioenergy feedstock while providing multiple environmental benefits when grown on marginal soil landscapes. Nitrogen (N) fert...

Nitrogen uptake by corn and switchgrass plants in soils of varying depths in Central Missouri

USDA-ARS?s Scientific Manuscript database

Sustainable biomass feedstock production systems involve biomass generation from non-agricultural or marginal lands with minimal external inputs. Switchgrass based alley cropping systems have been proposed as biomass feedstock crop systems in marginal lands. In many areas in the Midwest United State...

First report of Puccinia emaculata on switchgrass (Panicum virgatum L.) in Tennessee

USDA-ARS?s Scientific Manuscript database

In the spring of 2007, switchgrass accessions and cultivars Alamo, Kanlow, SL-93-2001, and NSL 2001-1 (lowland), Blackwell (upland), Grenville, Falcon, and Miami (unknown ploidy levels) were sown at the East Tennessee Research and Extension Center in Knoxville, Tennessee for evaluation and controlle...

N2 fixation of common and hairy vetches when intercropped into switchgrass

USDA-ARS?s Scientific Manuscript database

Interest in alternatives to synthetic nitrogen (N) fertilizer for switchgrass (Panicum virgatum L.) forage and bioenergy production continues to increase, and interseeding legumes into swards may be one such prospect. Common vetch (Vicia sativa L.) occurs naturally throughout the U.S. and has fewer ...

Switchgrass biomass composition traits and their effects on its digestion by ruminants and bioconversion to ethanol

USDA-ARS?s Scientific Manuscript database

Six generations of divergent breeding in switchgrass (Panicum virgatum L.) for forage in vitro digestibility (IVDMD) resulted in significant changes in 20 biomass composition traits. Stepwise multi-regression was used to determine which of the 20 composition traits had largest significant effects on...

Field productivities of Switchgrass for conversion to sugars and ethanol

USDA-ARS?s Scientific Manuscript database

Switchgrass (SG, Panicum virgatum) is a warm season perennial grass being developed as a dedicated bioenergy crop. The Agricultural Research Service has recently released a new cultivar of SG named Liberty, which is a cross between Summer and Kanlow cultivars. In prior studies Liberty grown on 4 sit...

Comparison of Dilute Acid and Ionic Liquid Pretreatment of Switchgrass: Biomass Recalcitrance, Delignification and Enzymatic Saccharification

USDA-ARS?s Scientific Manuscript database

The efficiency of two biomass pretreatment technologies, dilute acid hydrolysis and dissolution in an ionic liquid, are compared in terms of delignification, saccharification efficiency and saccharide yields with switchgrass serving as a model bioenergy crop. When subject to ionic liquid pretreatme...

QTL x environment interactions and latitudinal adaptation in switchgrass (Panicum virgatum)

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum) is a polyploid C4 perennial grass that is native to North America and has been championed as a promising biofuel feedstock. It is a common member of most native prairie communities and exhibits extensive phenotypic variability and adaptation across its range, especial...

Water use efficiency by switchgrass compared to a native grass or a native grass alfalfa mixture

USDA-ARS?s Scientific Manuscript database

Development of sustainable cellulosic biofuel systems requires knowing the water use efficiency (WUE) of potential bioenergy crops. Impact of early and late season droughts on WUE and soil water deficits were evaluated in switchgrass (Panicum virgatum L.), western wheatgrass (Pascopyrum smithii (Ry...

Swithchgrass biomass quality as affected by nitrogen rate, harvest time and storage

USDA-ARS?s Scientific Manuscript database

The main purpose of this study was to assess the sustainability of switchgrass biomass quality as affected by storage after harvesting, delaying the harvest time, and applying different rates of nitrogen (N). The present study was conducted at Bristol, South Dakota under switchgrass land previously ...

Responses of switchgrass soil respiration and its components to precipitation gradient in a mescocosm study

USDA-ARS?s Scientific Manuscript database

The objectives of this study were to investigate the effects of the precipitation changes on soil, microbial and root respirations of switchgrass soils, and the relationships between soil respiration and plant growth, soil moisture and temperature. A mesocosm experiment was conducted with five prec...

Topsoil thickness and harvest management influence switchgrass production and profitability

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is an attractive dual use forage and/or biomass crop option for eroded or marginal soils where corn (Zea mays L.) grain production often is not profitable. Topsoil thickness, especially above soils with a claypan, relates to crop productivity and nutrient removal an...

Effects of site preparation for pine forest/switchgrass Intercropping on water quality

Treesearch

A. Muwamba; D. M. Amatya; H. Ssegane; G.M. Chescheir; T. Appelboom; E.W. Tollner; J. E. Nettles; M. A. Youssef; F. Birgand; R. W. Skaggs; S. Tian

2015-01-01

A study was initiated to investigate the sustainability effects of intercropping switchgrass (Panicum virgatum L.) in a loblolly pine (Pinus taeda L.) plantation. This forest-based biofuel system could possibly provide biomass from the perennial energy grass while maintaining the economics and environmental benefits of a forest...

Switchgrass Compositional Variations Arising from Spatial Distribution and Legume Intercropping

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum) is a high–yielding, second-generation feedstock that can be grown on marginal land with minimal inputs. Due to the high genetic diversity within and among cultivars of this species, there may be a great amount of genotype x environment-induced differences among seconda...

Biomass production and nutrient removal by switchgrass under irrigation

USDA-ARS?s Scientific Manuscript database

Switchgrass has been identified to supply a major portion of U.S. energy needs when used as a fuel. Assessments of the export of essential plant nutrients are needed to determine impacts on soil fertility that will influence fertilizer recommendations since the nutrients contained in the above groun...

Intercropping with switchgrass improves net greenhouse gas balance in hybrid poplar plantations on a sand soil

USDA-ARS?s Scientific Manuscript database

Highly productive, commercial hybrid poplar plantations are being managed in the Pacific Northwest for high-value timber production at relatively low stocking densities under irrigation. The open understory was used to produce switchgrass (Panicum virgatum) prior to canopy closure. The objectives ...

Hydraulic properties affected by topsoil thickness in switchgrass and corn-soybean cropping systems

USDA-ARS?s Scientific Manuscript database

Loss of productive topsoil by soil erosion over time can reduce the productive capacity of soil and can significantly affect soil hydraulic properties. This study evaluated the effects of reduced topsoil thickness and perennial switchgrass (Panicum virgatum L.) versus corn (Zea mays L.)/soybean [Gly...

Microwave pretreatment of switchgrass for bioethanol production

NASA Astrophysics Data System (ADS)

Keshwani, Deepak Radhakrishin

Lignocellulosic materials are promising alternative feedstocks for bioethanol production. These materials include agricultural residues, cellulosic waste such as newsprint and office paper, logging residues, and herbaceous and woody crops. However, the recalcitrant nature of lignocellulosic biomass necessitates a pretreatment step to improve the yield of fermentable sugars. The overall goal of this dissertation is to expand the current state of knowledge on microwave-based pretreatment of lignocellulosic biomass. Existing research on bioenergy and value-added applications of switchgrass is reviewed in Chapter 2. Switchgrass is an herbaceous energy crop native to North America and has high biomass productivity, potentially low requirements for agricultural inputs and positive environmental impacts. Based on results from test plots, yields in excess of 20 Mg/ha have been reported. Environmental benefits associated with switchgrass include the potential for carbon sequestration, nutrient recovery from run-off, soil remediation and provision of habitats for grassland birds. Published research on pretreatment of switchgrass reported glucose yields ranging from 70-90% and xylose yields ranging from 70-100% after hydrolysis and ethanol yields ranging from 72-92% after fermentation. Other potential value-added uses of switchgrass include gasification, bio-oil production, newsprint production and fiber reinforcement in thermoplastic composites. Research on microwave-based pretreatment of switchgrass and coastal bermudagrass is presented in Chapter 3. Pretreatments were carried out by immersing the biomass in dilute chemical reagents and exposing the slurry to microwave radiation at 250 watts for residence times ranging from 5 to 20 minutes. Preliminary experiments identified alkalis as suitable chemical reagents for microwave-based pretreatment. An evaluation of different alkalis identified sodium hydroxide as the most effective alkali reagent. Under optimum pretreatment conditions, 82% glucose and 63% xylose yields were achieved for switchgrass, and 87% glucose and 59% xylose yields were achieved for coastal bermudagrass following enzymatic hydrolysis of the pretreated biomass. The optimum enzyme loadings were 15 FPU/g and 20 CBU/g for switchgrass and 10 FPU/g and 20 CBU/g for coastal bermudagrass. Dielectric properties for dilute sodium hydroxide solutions were measured and compared to solid loss, lignin reduction and reducing sugar levels in hydrolyzates. Results indicate that the dielectric loss tangent of alkali solutions is a potential indicator of the severity of microwave-based pretreatments. Modeling of pretreatment processes can be a valuable tool in process simulations of bioethanol production from lignocellulosic biomass. Chapter 4 discusses three different approaches that were used to model delignification and carbohydrate loss during microwave-based pretreatment of switchgrass: statistical linear regression modeling, kinetic modeling using a time-dependent rate coefficient, and a Mamdani-type fuzzy inference system. The dielectric loss tangent of the alkali reagent and pretreatment time were used as predictors in all models. The statistical linear regression model for delignification gave comparable root mean square error (RMSE) values for training and testing data and predictions were approximately within 1% of experimental values. The kinetic model for delignification and xylan loss gave comparable RMSE values for training and testing data sets and predictions were approximately within 2% of experimental values. The kinetic model for cellulose loss was not as effective and predictions were only within 5-7% of experimental values. The time-dependent rate coefficients of the kinetic models calculated from experimental data were consistent with the heterogeneity (or lack thereof) of individual biomass components. The Mamdani-type fuzzy inference system was shown to be an effective means to model pretreatment processes and gave the most accurate predictions (<3%) for cellulose loss.

Label-free quantitative proteomics for the extremely thermophilic bacterium Caldicellulosiruptor obsidiansis reveal distinct abundance patterns upon growth on cellobiose, crystalline cellulose, and switchgrass.

PubMed

Lochner, Adriane; Giannone, Richard J; Keller, Martin; Antranikian, Garabed; Graham, David E; Hettich, Robert L

2011-12-02

Mass spectrometric analysis of Caldicellulosiruptor obsidiansis cultures grown on four different carbon sources identified 65% of the cells' predicted proteins in cell lysates and supernatants. Biological and technical replication together with sophisticated statistical analysis were used to reliably quantify protein abundances and their changes as a function of carbon source. Extracellular, multifunctional glycosidases were significantly more abundant on cellobiose than on the crystalline cellulose substrates Avicel and filter paper, indicating either disaccharide induction or constitutive protein expression. Highly abundant flagellar, chemotaxis, and pilus proteins were detected during growth on insoluble substrates, suggesting motility or specific substrate attachment. The highly abundant extracellular binding protein COB47_0549 together with the COB47_1616 ATPase might comprise the primary ABC-transport system for cellooligosaccharides, while COB47_0096 and COB47_0097 could facilitate monosaccharide uptake. Oligosaccharide degradation can occur either via extracellular hydrolysis by a GH1 β-glycosidase or by intracellular phosphorolysis using two GH94 enzymes. When C. obsidiansis was grown on switchgrass, the abundance of hemicellulases (including GH3, GH5, GH51, and GH67 enzymes) and certain sugar transporters increased significantly. Cultivation on biomass also caused a concerted increase in cytosolic enzymes for xylose and arabinose fermentation.

Caldicellulosiruptor Core and Pangenomes Reveal Determinants for Noncellulosomal Thermophilic Deconstruction of Plant Biomass

PubMed Central

Blumer-Schuette, Sara E.; Giannone, Richard J.; Zurawski, Jeffrey V.; Ozdemir, Inci; Ma, Qin; Yin, Yanbin; Xu, Ying; Kataeva, Irina; Poole, Farris L.; Adams, Michael W. W.; Hamilton-Brehm, Scott D.; Elkins, James G.; Larimer, Frank W.; Land, Miriam L.; Hauser, Loren J.; Cottingham, Robert W.; Hettich, Robert L.

2012-01-01

Extremely thermophilic bacteria of the genus Caldicellulosiruptor utilize carbohydrate components of plant cell walls, including cellulose and hemicellulose, facilitated by a diverse set of glycoside hydrolases (GHs). From a biofuel perspective, this capability is crucial for deconstruction of plant biomass into fermentable sugars. While all species from the genus grow on xylan and acid-pretreated switchgrass, growth on crystalline cellulose is variable. The basis for this variability was examined using microbiological, genomic, and proteomic analyses of eight globally diverse Caldicellulosiruptor species. The open Caldicellulosiruptor pangenome (4,009 open reading frames [ORFs]) encodes 106 GHs, representing 43 GH families, but only 26 GHs from 17 families are included in the core (noncellulosic) genome (1,543 ORFs). Differentiating the strongly cellulolytic Caldicellulosiruptor species from the others is a specific genomic locus that encodes multidomain cellulases from GH families 9 and 48, which are associated with cellulose-binding modules. This locus also encodes a novel adhesin associated with type IV pili, which was identified in the exoproteome bound to crystalline cellulose. Taking into account the core genomes, pangenomes, and individual genomes, the ancestral Caldicellulosiruptor was likely cellulolytic and evolved, in some cases, into species that lost the ability to degrade crystalline cellulose while maintaining the capacity to hydrolyze amorphous cellulose and hemicellulose. PMID:22636774

Predicted harvest time effects on switchgrass moisture content, nutrient concentration, yield, and profitability

USDA-ARS?s Scientific Manuscript database

Production costs change with harvest date of switchgrass (Panicum virgatum L.) as a result of nutrient recycling and changes in yield of this perennial crop. This study examines the range of cost of production from an early, yield-maximizing harvest date to a late winter harvest date at low moisture...

Effect of alfalfa (medicago sativa) on fermentation profile and nutritive value of switchgrass (panicum virgatum) and bermudagrass (cynodon dactylon) silages

USDA-ARS?s Scientific Manuscript database

An experiment was conducted at the University of Kentucky Spindletop Farm in Lexington, Kentucky between October and November, 2009 to evaluate the effect of different percentages of alfalfa (Medicago sativa) as mixtures in switchgrass (Panicum virgatus) and bermudagrass (Cynodon dactylon) silages. ...

Fertilizer recommendations for switchgrass: Quantifying economic effects on quality and yield

USDA-ARS?s Scientific Manuscript database

Switchgrass (Panicum virgatum L.) is a native, perennial warm season grass that is suited for biomass production for conversion to renewable fuels as well as feed production on marginal soils. Yield responses to macro nutrients of N, P and K, have shown N to be the major driver for capturing yield p...

First report of Tilletia pulcherrima on switchgrass (Panicum virgatum L.) in Texas

USDA-ARS?s Scientific Manuscript database

Switchgrass seed samples of cultivars ‘Blackwell’ and ‘Alamo’ from Bamert Seed Company, Muleshoe, Texas, and grown in Texas were examined microscopically for bunt fungi. Fourteen completely bunted seeds of ‘Blackwell’ and four in ‘Alamo’ were detected in 100 g samples of each. Teliospores were globo...

Switchgrass response to nitrogen fertilizer across diverse environments in the USA: a regional feedstock partnership report

USDA-ARS?s Scientific Manuscript database

The Regional Feedstock Partnership is a collaborative effort between the Sun Grant Initiative (through Land Grant Universities), the US Department of Energy, and the US Department of Agriculture. One segment of this partnership is the field-scale evaluation of switchgrass (Panicum virgatum L.) in di...

Effect of topsoil thickness on soil water infiltration in corn-soybean rotation and switchgrass production systems

USDA-ARS?s Scientific Manuscript database

Switchgrass and corn are sometimes used as a resource for biofuel production. The effect of production management systems on water infiltration is very critical in claypan landscape to increase production as well as minimize economic and environmental risks. The objective of this study was to evalua...

Effects of precipitation changes on aboveground net primary production and soil respiration in a switchgrass field

USDA-ARS?s Scientific Manuscript database

This study attempted to test whether switchgrass aboveground net primary production (ANPP) responds to precipitation (PPT) changes in a double asymmetry pattern as framed by Knapp et al. (2016), and whether it is held true for other ecosystem processes such as soil respiration (SR). Data were colle...

Environmental sustainability of intercropping switchgrass in a loblolly pine forest

Treesearch

George Chescheir; Francois Birgand; Mohamed Youssef; Jami Nettles; Devendra Amatya

2016-01-01

A multi-institutional watershed study has been conducted since 2010 to quantify the environmental sustainability of planting switchgrass (Panicum virgatum L.) between wide rows of loblolly pine (Pinus taeda L.). The hypothesized advantage of this intercropping system is the production of biofuel feedstock to provide additional...

Insect and plant-derived miRNAs in greenbug (schizaphis graminum) and yellow sugarcane aphid (sipha flava) revealed by deep sequencing

USDA-ARS?s Scientific Manuscript database

Schizaphis graminum (green bug; GB) and Sipha flava yellow sugarcane aphid; YSA) are two cereal aphid species with broad host ranges capable of establishing on sorghum (Sorghum bicolor) and several switchgrass (Panicum virgatum) cultivars. Switchgrass and sorghum are staple renewable bioenergy crops...

Karyotype variation is indicative of subgenomic and ecotypic differentiation in switchgrass

USDA-ARS?s Scientific Manuscript database

A cytogenetic study was conducted on a dihaploid individual (2n'='2X'='18) of switchgrass to establish a chromosome karyotype. Size differences, condensation patterns, and arm-length ratios were used as identifying features and fluorescence in-situ hybridization (FISH) assigned 5S and 45S rDNA loci...

30 years of progress toward increased biomass yield of switchgrass and big bluestem

USDA-ARS?s Scientific Manuscript database

Breeding to improved biomass production of switchgrass and big bluestem for conversion to bioenergy began in 1992. The purpose of this study was (1) to develop a platform for uniform regional testing of cultivars and experimental populations for these species and (2) to estimate the gains made by br...

Genetic parameters and prediction of breeding values in switchgrass bred for bioenergy

USDA-ARS?s Scientific Manuscript database

Estimating genetic parameters is an essential step in breeding by recurrent selection to maximize genetic gains over time. This study evaluated the effects of selection on genetic variation across two successive cycles (C1 and C2) of a ‘Summer’x‘Kanlow’ switchgrass (Panicum virgatum L.) population. ...

Greenhouse gas fluxes and root productivity in a switchgrass and loblolly pine intercropping system for bioenergy production

Treesearch

Paliza Shrestha; John R. Seiler; Brian D. Strahm; Eric B. Sucre; Zakiya H. Leggett

2015-01-01

This study is part of a larger collaborative effort to determine the overall environmental sustainability of intercropping pine (Pinus taeda L.) and switchgrass (Panicum virgatum L.), both of which are promising feedstock for bioenergy production in the Lower Coastal Plain in North Carolina.

Topsoil thickness effects on corn, soybean, and switchgrass production on claypan soils

USDA-ARS?s Scientific Manuscript database

Diminished topsoil thickness or depth to claypan (DTC) is a major cause of yield and profit depression in corn (Zea mays L.) and to a lesser extent in soybean (Glycine max [L.]) production on claypan soils. Perennial grasses such as switchgrass (Panicum virgatum L.) may be more resilient to reduced ...

Association of proteomics changes with Al-sensitive root zones in switchgrass

USDA-ARS?s Scientific Manuscript database

In this paper, we report on aluminum (Al)-induced root proteomic changes in switchgrass. After growth in a hydroponic culture system supplemented with 400 uM of Al, plants began to show signs of physiological stress such as a reduction in photosynthetic rate. At this time, the basal 2-cmlong root ti...

Evaluation of chopped switchgrass and chopped bermudagrass as litter materials over multiple heavy broiler flocks

USDA-ARS?s Scientific Manuscript database

Chopped switchgrass (SG) and chopped bermudagrass (BG) were evaluated as alternatives to pine shavings (PS) for broiler litter over 3 flocks. Twenty-four pens were filled with the 3 litter types. Live performance parameters included mortality, BW, BW gain, feed consumption, and feed conversion. Mort...

Switchgrass (Panicum virgatum) Intercropping within Managed Loblolly Pine (Pinus taeda) Does Not Affect Wild Bee Communities.

PubMed

Campbell, Joshua W; Miller, Darren A; Martin, James A

2016-11-04

Intensively-managed pine ( Pinus spp.) have been shown to support diverse vertebrate communities, but their ability to support invertebrate communities, such as wild bees, has not been well-studied. Recently, researchers have examined intercropping switchgrass ( Panicum virgatum ), a native perennial, within intensively managed loblolly pine ( P. taeda ) plantations as a potential source for cellulosic biofuels. To better understand potential effects of intercropping on bee communities, we investigated visitation of bees within three replicates of four treatments of loblolly pine in Mississippi, U.S.A.: 3-4 year old pine plantations and 9-10 year old pine plantations with and without intercropped switchgrass. We used colored pan traps to capture bees during the growing seasons of 2013 and 2014. We captured 2507 bees comprised of 18 different genera during the two-year study, with Lasioglossum and Ceratina being the most common genera captured. Overall, bee abundances were dependent on plantation age and not presence of intercropping. Our data suggests that switchgrass does not negatively impact or promote bee communities within intensively-managed loblolly pine plantations.

Microbial community analysis of switchgrass planted and unplanted soil microcosms displaying PCB dechlorination

PubMed Central

Liang, Yi; Meggo, Richard; Hu, Dingfei; Schnoor, Jerald L.; Mattes, Timothy E.

2015-01-01

Polychlorinated biphenyls (PCBs) pose potential risks to human and environmental health because they are carcinogenic, persistent and bioaccumulative. In this study we investigated bacterial communities in soil microcosms spiked with PCB 52, 77 and 153. Switchgrass (Panicum virgatum) was employed to improve overall PCB removal and redox cycling (i.e. sequential periods of flooding followed by periods of no flooding) was performed in an effort to promote PCB dechlorination. Lesser chlorinated PCB transformation products were detected in all microcosms, indicating the occurrence of PCB dechlorination. Terminal restriction fragment length polymorphism (T-RFLP) and clone library analysis showed that PCB spiking, switchgrass planting and redox cycling affected the microbial community structure. Putative organohalide-respiring Chloroflexi populations, which were not found in unflooded microcosms, were enriched after two weeks of flooding in the redox-cycled microcosms. Sequences classified as Geobacter sp. were detected in all microcosms, and were most abundant in the switchgrass-planted microcosm spiked with PCB congeners. The presence of possible organohalide-respiring bacteria in these soil microcosms suggests they play a role in PCB dechlorination therein. PMID:25820643

Microbial community analysis of switchgrass planted and unplanted soil microcosms displaying PCB dechlorination.

PubMed

Liang, Yi; Meggo, Richard; Hu, Dingfei; Schnoor, Jerald L; Mattes, Timothy E

2015-08-01

Polychlorinated biphenyls (PCBs) pose potential risks to human and environmental health because they are carcinogenic, persistent, and bioaccumulative. In this study, we investigated bacterial communities in soil microcosms spiked with PCB 52, 77, and 153. Switchgrass (Panicum virgatum) was employed to improve overall PCB removal, and redox cycling (i.e., sequential periods of flooding followed by periods of no flooding) was performed in an effort to promote PCB dechlorination. Lesser chlorinated PCB transformation products were detected in all microcosms, indicating the occurrence of PCB dechlorination. Terminal restriction fragment length polymorphism (T-RFLP) and clone library analysis showed that PCB spiking, switchgrass planting, and redox cycling affected the microbial community structure. Putative organohalide-respiring Chloroflexi populations, which were not found in unflooded microcosms, were enriched after 2 weeks of flooding in the redox-cycled microcosms. Sequences classified as Geobacter sp. were detected in all microcosms and were most abundant in the switchgrass-planted microcosm spiked with PCB congeners. The presence of possible organohalide-respiring bacteria in these soil microcosms suggests that they play a role in PCB dechlorination therein.

Impact of AFEX™ Pretreatment and Extrusion Pelleting on Pellet Physical Properties and Sugar Recovery from Corn Stover, Prairie Cord Grass, and Switchgrass.

PubMed

Sundaram, Vijay; Muthukumarappan, Kasiviswanathan

2016-05-01

The effects of AFEX™ pretreatment, feedstock moisture content (5,10, and 15 % wb), particle size (screen sizes of 2, 4, and 8 mm), and extrusion temperature (75, 100, and 125 °C) on pellet bulk density, pellet hardness, and sugar recovery from corn stover, prairie cord grass, and switchgrass were investigated. Pellets were produced from untreated and AFEX™ pretreated feedstocks using a laboratory-scale extruder. AFEX™ pretreatment increased subsequent pellet bulk density from 453.0 to 650.6 kg m(-3) for corn stover from 463.2 to 680.1 kg m(-3) for prairie cord grass, and from 433.9 to 627.7 kg m(-3) for switchgrass. Maximum pellet hardness of 2342.8, 2424.3, and 1298.6 N was recorded for AFEX™ pretreated corn stover, prairie cord grass, and switchgrass, respectively. Glucose yields of AFEX™ corn stover pellets, prairie cord grass, and switchgrass pellets varied from 88.9 to 94.9 %, 90.1 to 94.9 %, and 87.0 to 92.9 %, respectively. Glucose and xylose yields of AFEX™ pellets were not affected by the extruder barrel temperature and the hammer mill screen size. The results obtained showed that low temperature and large particle size during the extrusion pelleting process can be employed for AFEX™-treated biomass without compromising sugar yields.

Soil physical and hydrological properties under three biofuel crops in Ohio

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bonin, Catherine; Lal, Dr. Rattan; Schmitz, Matthias

While biofuel crops are widely studied and compared for their energy and carbon footprints, less is known about their effects on other soil properties, particularly hydrologic characteristics. Soils under three biofuel crops, corn (Zea mays), switchgrass (Panicum virgatum), and willow (Salix spp.), were analyzed seven years after establishment to assess the effects on soil bulk density ({rho}{sub b}), penetration resistance (PR), water-holding capacity, and infiltration characteristics. The PR was the highest under corn, along with the lowest associated water content, while PR was 50-60% lower under switchgrass. In accordance with PR data, surface (0-10 cm) bulk density also tended tomore » be lower under switchgrass. Both water infiltration rates and cumulative infiltration amounts varied widely among and within the three crops. Because the Philip model did not fit the data, results were analyzed using the Kostiakov model instead. Switchgrass plots had an average cumulative infiltration of 69 cm over 3 hours with a constant infiltration rate of 0.28 cm min{sup -1}, compared with 37 cm and 0.11 cm min{sup -1} for corn, and 26 cm and 0.06 cm min{sup -1} for willow, respectively. Results suggest that significant changes in soil physical and hydrologic properties may require more time to develop. Soils under switchgrass may have lower surface bulk density, higher field water capacity, and a more rapid water infiltration rate than those under corn or willow.« less

Accuracy of Genomic Prediction in Switchgrass (Panicum virgatum L.) Improved by Accounting for Linkage Disequilibrium

PubMed Central

Ramstein, Guillaume P.; Evans, Joseph; Kaeppler, Shawn M.; Mitchell, Robert B.; Vogel, Kenneth P.; Buell, C. Robin; Casler, Michael D.

2016-01-01

Switchgrass is a relatively high-yielding and environmentally sustainable biomass crop, but further genetic gains in biomass yield must be achieved to make it an economically viable bioenergy feedstock. Genomic selection (GS) is an attractive technology to generate rapid genetic gains in switchgrass, and meet the goals of a substantial displacement of petroleum use with biofuels in the near future. In this study, we empirically assessed prediction procedures for genomic selection in two different populations, consisting of 137 and 110 half-sib families of switchgrass, tested in two locations in the United States for three agronomic traits: dry matter yield, plant height, and heading date. Marker data were produced for the families’ parents by exome capture sequencing, generating up to 141,030 polymorphic markers with available genomic-location and annotation information. We evaluated prediction procedures that varied not only by learning schemes and prediction models, but also by the way the data were preprocessed to account for redundancy in marker information. More complex genomic prediction procedures were generally not significantly more accurate than the simplest procedure, likely due to limited population sizes. Nevertheless, a highly significant gain in prediction accuracy was achieved by transforming the marker data through a marker correlation matrix. Our results suggest that marker-data transformations and, more generally, the account of linkage disequilibrium among markers, offer valuable opportunities for improving prediction procedures in GS. Some of the achieved prediction accuracies should motivate implementation of GS in switchgrass breeding programs. PMID:26869619

Nitrogen rate and landscape impacts on life cycle energy use and emissions from switchgrass-derived ethanol

DOE PAGES

Mbonimpa, Eric G.; Kumar, Sandeep; Owens, Vance N.; ...

2015-08-24

Switchgrass-derived ethanol has been proposed as an alternative to fossil fuels to improve sustainability of the US energy sector. In this study, life cycle analysis (LCA) was used to estimate the environmental benefits of this fuel. To better define the LCA environmental impacts associated with fertilization rates and farm-landscape topography, results from a controlled experiment were analyzed. Data from switchgrass plots planted in 2008, consistently managed with three nitrogen rates (0, 56, and 112 kg N ha –1), two landscape positions (shoulder and footslope), and harvested annually (starting in 2009, the year after planting) through 2014 were used as inputmore » into the Greenhouse gases, Regulated Emissions and Energy use in transportation (GREET) model. Simulations determined nitrogen (N) rate and landscape impacts on the life cycle energy and emissions from switchgrass ethanol used in a passenger car as ethanol–gasoline blends (10% ethanol:E10, 85% ethanol:E85s). Results indicated that E85s may lead to lower fossil fuels use (58 to 77%), greenhouse gas (GHG) emissions (33 to 82%), and particulate matter (PM2.5) emissions (15 to 54%) in comparison with gasoline. However, volatile organic compounds (VOCs) and other criteria pollutants such as nitrogen oxides (NOx), particulate matter (PM10), and sulfur dioxides (SO x) were higher for E85s than those from gasoline. Nitrogen rate above 56 kg N ha –1 yielded no increased biomass production benefits; but did increase (up to twofold) GHG, VOCs, and criteria pollutants. Lower blend (E10) results were closely similar to those from gasoline. The landscape topography also influenced life cycle impacts. Biomass grown at the footslope of fertilized plots led to higher switchgrass biomass yield, lower GHG, VOCs, and criteria pollutants in comparison with those at the shoulder position. Lastly, results also showed that replacing switchgrass before maximum stand life (10–20 years.) can further reduce the energy and emissions reduction benefits.« less

Assessing the Impacts of Land Use Change from Cotton to Perennial Bioenergy Grasses on Hydrological Fluxes and Water Quality in a Semi-Arid Agricultural Watershed Using the APEX Model

NASA Astrophysics Data System (ADS)

Chen, Y.; Ale, S.; Rajan, N.

2015-12-01

The semi-arid Texas High Plains (THP) region, where cotton (Gossypium hirsutum L.) is grown in vast acreage, has the potential to grow perennial bioenergy grasses. A change in land use from cotton cropping systems to perennial grasses such as Alamo switchgrass (Panicum virgatum L.) and Miscanthus giganteus (Miscanthus sinensis Anderss. [Poaceae]) can significantly affect regional hydrologic cycle and water quality. Assessing the impacts of this potential land use change on hydrology and water quality enables the environmental assessment of feasibility to grow perennial grasses in this region to meet the U.S. national bioenergy target of 2022. The Agricultural Policy/Environmental eXtender (APEX) model was used in this study to assess the impacts of replacing cotton with switchgrass and Miscanthus on water and nitrogen balances in the upstream subwatershed of the Double Mountain Fork Brazos watershed in the THP, which contains 52% cotton land use. The APEX model was initially calibrated against observed streamflow and crop yield data. Since observed data on nitrogen loads in streamflow was not available for this subwatershed, we calibrated the APEX model against the SWAT-simulated nitrogen loads at the outlet of this subwatershed, which were obtained in a parallel study. The calibrated APEX model was used to simulate the impacts of land use change from cotton to Miscanthus and switchgrass on surface and subsurface water and nitrogen balances. Preliminary results revealed that the average (1994-2009) annual surface runoff decreased by 84% and 66% under the irrigated and dryland switchgrass scenarios compared to the baseline scenarios. Average annual percolation increased by 106% and 57% under the irrigated and dryland switchgrass scenarios relative to the baseline scenarios. Preliminary results also indicated Miscanthus and switchgrass appeared to be superior to cotton in terms of better water conservation and water quality, and minimum crop management requirements.

Modifying a Cow-Calf Biophysical Simulation Model for Analyses of Alternative Enterprises

NASA Astrophysics Data System (ADS)

Lutes, Jennifer L.

Cow-calf producers in the United States, tasked with providing beef calves for the beef industry, have had a multitude of difficulties to overcome in recent years. Producers in northwest Arkansas were negatively impacted by high hay prices coupled with low beef cattle market prices due to severe drought experienced in portions of 2010, 2011, and 2012. During this time they also faced high grain prices, due to a record low harvest, combined with portions of the corn harvest diverted from human and animal feed to ethanol production. Tight lending policies of this time, reminiscent of the housing market crash in 2008, along with the negative public attention associated with high levels of greenhouse gas emissions associated with beef production, lead to a tough situation for cattle producers faced with increasing input costs, decreased revenue, and lack of access to loans. With these issues in mind, this research aimed to determine if incorporating switchgrass (Panicum virgatum) production on a cow-calf farm could serve to increase net returns, decrease income volatility, lower net greenhouse gas (GHG) emissions without decreasing beef output, and provide a viable source of feedstock for a potential bio-refinery. The study determined that switchgrass is a potential solution to these problems and thus aimed to discover differences in switchgrass supply under different government policies in four northwestern counties in Arkansas to an as-yet, non-existent bio-refinery. It was determined that growing switchgrass on pastureland, once devoted to cow-calf production, is a viable enterprise diversification tool that under the right conditions could be used to improve producer financial and environmental outcomes. However, bioenergy production is slow to gain traction in the US due to adverse market conditions from low fossil fuel prices. Thus, in the US, there are only a few bio-refineries currently online and accepting lignocellulosic biomass, however none of them are close enough to northwest Arkansas to incentivize biomass production in this region. With this in mind, the results from an individual farm with switchgrass were extrapolated to a four county region to determine potential biomass supply for a hypothetical biorefinery. In conjunction with this analysis, two potential policies aimed at increasing biomass supply and lowering carbon emissions, were analyzed for their implications on the financial and environmental wellbeing of farms. It turns out, each of the two policies, the Biomass Crop Assistance Program (BCAP) and a Carbon Offset Program (CO), encourage the production of switchgrass and policy outcomes are most favorable when land of adequate quality is chosen to support higher switchgrass yield. At lower yield levels, the inclusion of switchgrass on pastures leads to less positive environmental outcomes and increased producer income variance.

The use of marker-data transformations to account for linkage disequilibrium in genomic selection: a case study in switchgrass (Panicum virgatum L.)

USDA-ARS?s Scientific Manuscript database

Genomic selection (GS) is an attractive technology to generate rapid genetic gains, particularly in perennial grass species like switchgrass, where phenotyping generally requires at least two years of field trial. In this study, we empirically assessed prediction procedures for GS in two different p...

An exergy based assessment of the production and conversion of switchgrass, equine waste and forest residue to bio-oil using fast pyrolysis

USDA-ARS?s Scientific Manuscript database

The resource efficiency of biofuel production via biomass pyrolysis is evaluated using exergy as an assessment metric. Three feedstocks, important to various sectors of US agriculture, switchgrass, forest residue and equine waste are considered for conversion to bio-oil (pyrolysis oil) via fast pyro...

Characterization of biobased polyurethane foams employing lignin fractionated from microwave liquefied switchgrass

Treesearch

Xingyan Huang; Cornelis F. De Hoop; Jiulong Xie; Chung-Yun Hse; Jinqiu Qi; Tingxing Hu

2017-01-01

Lignin samples fractionated from microwave liquefied switchgrass were applied in the preparation of semirigid polyurethane (PU) foams without purification.The objective of this study was to elucidate the influence of lignin in the PU matrix on themorphological, chemical, mechanical, and thermal properties of thePU foams.The scanning electron microscopy (SEM) images...

Complete genome sequence of switchgrass mosaic virus, a member of a proposed new species in the genus Marafivirus

USDA-ARS?s Scientific Manuscript database

The complete genome sequence of a virus recently detected in switchgrass (Panicum virgatum) was determined and was found to be closely related to Maize rayado fino virus (MRFV), genus Marafivirus, family Tymoviridae. The genomic RNA is 6408 nucleotides long, excluding the poly (A) tail, and encodes...

Economic potential for switchgrass production in the U.S. Northern Plains: A minimum-data analysis

USDA-ARS?s Scientific Manuscript database

There is a demand for timely information to support policy decision making. There is also interest in the potential for alternative crops such as switchgrass to be used for ethanol production and which would have a positive impact on net greenhouse gas emission. This paper uses a new minimum-data mo...

Water quality effects of switchgrass intercropping on pine forest in Coastal North Carolina.

Treesearch

Augustine Muwamba; Devendra Amatya; George M Chescheir; Jamie Nettles; Timothy Appelboom; Herbert Ssegane; Ernest Tollner; Mohamed Youssef; Francois Birgand; R. Wayne Skaggs; Shiying Tian

2017-01-01

Interplanting a cellulosic bioenergy crop (switchgrass, Panicum virgatum L.) between loblolly pine (Pinus taeda L.) rows could potentially provide a sustainable source of bio-feedstock without competing for land currently in food production. The objectives of this study were to: (1) quantify the concentrations and loads of drainage water nitrogen (N) and phosphorus (...

Comparative relationships and chromosome evolution in switchgrass (Panicum virgatum) and its genomic model, foxtail millet (Setaria italica)

USDA-ARS?s Scientific Manuscript database

A switchgrass (Panicum virgatum L.) consensus map was developed that combined data from two mapping populations and integrated recombination data from both parents of this largely obligate outcrossing species. The consensus map consisted of 1,321 loci and spanned 2,122 cM. An analysis of the distri...

Different Effects of Corn Ethanol and Switchgrass-Based Biofuels on Soil Erosion and Nutrients Loads in the Iowa River Basin

NASA Astrophysics Data System (ADS)

Wu, Y.; Liu, S.

2010-12-01

Biofuels have become important alternative energy resources and their use is likely to expand in the foreseeable future. The expansion of corn-based ethanol production has resulted in a tightening of the global corn supply-and-demand balance. Perennial grasses such as switchgrass (Panicum virgatum) are also being considered as candidates for biofuel feedstocks. Expansion of biofuel production will generate diverse impacts on the economy and environment. How to optimize land use activities to address the need for biofuel production while protecting the environment is still a grand challenge. Unfortunately, little is known about the effects of biofuel-oriented alternative land uses on water quality. Can we produce the same amount or more biofuels by converting some cornfields to switchgrass, for example, while reducing environmental pressure? The objective of this study was to evaluate the potential impacts of land cover change from corn to switchgrass (e.g., on marginal lands) and related management activities on water quality in the Iowa River Basin (drainage area of 32,360 km2) using the Soil and Water Assessment Tool (SWAT). The model was calibrated and validated under baseline conditions using daily streamflow and sediment, and monthly nutrients at Wapello, Iowa (near the outlet of the watershed). The preliminary results show that both the annual average sediment yield and nitrate nitrogen load would decrease when marginal corn areas are converted to switchgrass. However, the magnitude of change depends greatly on the detailed management practices, such as techniques and amount of fertilization, harvesting practices, and residue management.

Farm-level feasibility of bioenergy depends on variations across multiple sectors

NASA Astrophysics Data System (ADS)

Myhre, Mitchell; Barford, Carol

2013-03-01

The potential supply of bioenergy from farm-grown biomass is uncertain due to several poorly understood or volatile factors, including land availability, yield variability, and energy prices. Although biomass production for liquid fuel has received more attention, here we present a case study of biomass production for renewable heat and power in the state of Wisconsin (US), where heating constitutes at least 30% of total energy demand. Using three bioenergy systems (50 kW, 8.8 MW and 50 MW) and Wisconsin farm-level data, we determined the net farm income effect of producing switchgrass (Panicum virgatum) as a feedstock, either for on-farm use (50 kW system) or for sale to an off-farm energy system operator (8.8 and 50 MW systems). In southern counties, where switchgrass yields approach 10 Mg ha-1 yr-1, the main determinants of economic feasibility were the available land area per farm, the ability to utilize bioheat, and opportunity cost assumptions. Switchgrass yield temporal variability was less important. For the state median farm size and switchgrass yield, at least 25% (50 kW system) or 50% (8.8 MW system) bioheat utilization was required to economically offset propane or natural gas heat, respectively, and purchased electricity. Offsetting electricity only (50 MW system) did not generate enough revenue to meet switchgrass production expenses. Although the opportunity cost of small-scale (50 kW) on-farm bioenergy generation was higher, it also held greater opportunity for increasing farm net income, especially by replacing propane-based heat.

Accuracy of genomic prediction in switchgrass ( Panicum virgatum L.) improved by accounting for linkage disequilibrium

DOE PAGES

Ramstein, Guillaume P.; Evans, Joseph; Kaeppler, Shawn M.; ...

2016-02-11

Switchgrass is a relatively high-yielding and environmentally sustainable biomass crop, but further genetic gains in biomass yield must be achieved to make it an economically viable bioenergy feedstock. Genomic selection (GS) is an attractive technology to generate rapid genetic gains in switchgrass, and meet the goals of a substantial displacement of petroleum use with biofuels in the near future. In this study, we empirically assessed prediction procedures for genomic selection in two different populations, consisting of 137 and 110 half-sib families of switchgrass, tested in two locations in the United States for three agronomic traits: dry matter yield, plant height,more » and heading date. Marker data were produced for the families’ parents by exome capture sequencing, generating up to 141,030 polymorphic markers with available genomic-location and annotation information. We evaluated prediction procedures that varied not only by learning schemes and prediction models, but also by the way the data were preprocessed to account for redundancy in marker information. More complex genomic prediction procedures were generally not significantly more accurate than the simplest procedure, likely due to limited population sizes. Nevertheless, a highly significant gain in prediction accuracy was achieved by transforming the marker data through a marker correlation matrix. Our results suggest that marker-data transformations and, more generally, the account of linkage disequilibrium among markers, offer valuable opportunities for improving prediction procedures in GS. Furthermore, some of the achieved prediction accuracies should motivate implementation of GS in switchgrass breeding programs.« less

Accuracy of genomic prediction in switchgrass ( Panicum virgatum L.) improved by accounting for linkage disequilibrium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ramstein, Guillaume P.; Evans, Joseph; Kaeppler, Shawn M.

Switchgrass is a relatively high-yielding and environmentally sustainable biomass crop, but further genetic gains in biomass yield must be achieved to make it an economically viable bioenergy feedstock. Genomic selection (GS) is an attractive technology to generate rapid genetic gains in switchgrass, and meet the goals of a substantial displacement of petroleum use with biofuels in the near future. In this study, we empirically assessed prediction procedures for genomic selection in two different populations, consisting of 137 and 110 half-sib families of switchgrass, tested in two locations in the United States for three agronomic traits: dry matter yield, plant height,more » and heading date. Marker data were produced for the families’ parents by exome capture sequencing, generating up to 141,030 polymorphic markers with available genomic-location and annotation information. We evaluated prediction procedures that varied not only by learning schemes and prediction models, but also by the way the data were preprocessed to account for redundancy in marker information. More complex genomic prediction procedures were generally not significantly more accurate than the simplest procedure, likely due to limited population sizes. Nevertheless, a highly significant gain in prediction accuracy was achieved by transforming the marker data through a marker correlation matrix. Our results suggest that marker-data transformations and, more generally, the account of linkage disequilibrium among markers, offer valuable opportunities for improving prediction procedures in GS. Furthermore, some of the achieved prediction accuracies should motivate implementation of GS in switchgrass breeding programs.« less

Solar-induced direct biomass-to-electricity hybrid fuel cell using polyoxometalates as photocatalyst and charge carrier.

PubMed

Liu, Wei; Mu, Wei; Liu, Mengjie; Zhang, Xiaodan; Cai, Hongli; Deng, Yulin

2014-01-01

The current polymer-exchange membrane fuel cell technology cannot directly use biomass as fuel. Here we present a solar-induced hybrid fuel cell that is directly powered with natural polymeric biomasses, such as starch, cellulose, lignin, and even switchgrass and wood powders. The fuel cell uses polyoxometalates as the photocatalyst and charge carrier to generate electricity at low temperature. This solar-induced hybrid fuel cell combines some features of solar cells, fuel cells and redox flow batteries. The power density of the solar-induced hybrid fuel cell powered by cellulose reaches 0.72 mW cm(-2), which is almost 100 times higher than cellulose-based microbial fuel cells and is close to that of the best microbial fuel cells reported in literature. Unlike most cell technologies that are sensitive to impurities, the cell reported in this study is inert to most organic and inorganic contaminants present in the fuels.

Solar-induced direct biomass-to-electricity hybrid fuel cell using polyoxometalates as photocatalyst and charge carrier

NASA Astrophysics Data System (ADS)

Liu, Wei; Mu, Wei; Liu, Mengjie; Zhang, Xiaodan; Cai, Hongli; Deng, Yulin

2014-02-01

The current polymer-exchange membrane fuel cell technology cannot directly use biomass as fuel. Here we present a solar-induced hybrid fuel cell that is directly powered with natural polymeric biomasses, such as starch, cellulose, lignin, and even switchgrass and wood powders. The fuel cell uses polyoxometalates as the photocatalyst and charge carrier to generate electricity at low temperature. This solar-induced hybrid fuel cell combines some features of solar cells, fuel cells and redox flow batteries. The power density of the solar-induced hybrid fuel cell powered by cellulose reaches 0.72 mW cm-2, which is almost 100 times higher than cellulose-based microbial fuel cells and is close to that of the best microbial fuel cells reported in literature. Unlike most cell technologies that are sensitive to impurities, the cell reported in this study is inert to most organic and inorganic contaminants present in the fuels.

Soil nitrogen dynamics in switchgrass seeded to a marginally yielding cropland of South Dakota

USDA-ARS?s Scientific Manuscript database

Soil nitrate (NO3-), nitrate leaching, and nitrous oxide (N2O) emissions for 2009 through 2015 were monitored to explore N dynamics in switchgrass (Panicum virgatum L.) seeded to a marginally yielding cropland. Our findings indicated that N rate impacted soil NO3- (0-5 cm depth) and soil surface N2O...

Impacts of switchgrass intercropping in traditional pine forests on hydrology and water quality in the southeastern United States.

Treesearch

Devendra Amatya; G.M. Chescheir; J.E. Nettles

2016-01-01

Preliminary results indicate that switchgrass (Panicum virgatum L.), grown as a cellulosic biofuel between managed loblolly pine (Pinus taeda L.) beds on the Atlantic Coastal Plain forests has no significant effect on shallow ground water table and stream outflows. Although management operations (e.g. harvesting, shearing between pine rows, raking, and bedding)...

Production of Butanol (a Biofuel) from Agricultural Residues: Part II - Use of Corn Stover and Switchgrass Hydrolysates

USDA-ARS?s Scientific Manuscript database

Acetone butanol ethanol (ABE or AB, or solvent) was produced from hydrolyzed corn stover and switchgrass using Clostridium beijerinckii P260. A control experiment using glucose resulted in the production of 21.06 gL**-1 total ABE. In this experiment, an AB yield and productivity of 0.41 and 0.31 g...

Research and Technology Development for Genetic Improvement of Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kausch, Albert; Rhodes, Richard

This research adds to the understanding of switchgrass genetics and the increasing of biomass relevant to production of bioenergy. Switchgrass, Panicum virgatum L., and its related species are well known as potential bioenergy crops since the early 1990s. There are global economic, political, US national security and environmental pressures to increase renewable biofuel production and utilization to offset gasoline and diesel fuel use and climate change, especially in the liquid fuel transportation sector. To realize the potential of bioenergy crops, rapid genetic improvement of the most promising perennial grass feedstocks, such as switchgrass, are anticipated by current genomics, association genetics,more » marker assisted breeding, hybrid plant development, advanced tissue culture, conventional genetics and other approaches to increase yield, processability, and regional adaptation. The technical effectiveness and economic feasibility of the methods or techniques investigated are demonstrated by several publications, presentations and patents produced as an outcome and deliverable of this research. This project is of a broad benefit to the public not only through the dissemination of this information but also to the development of new methods which will be applied to future bioenergy crop improvement as well as other crops.« less

Glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass.

PubMed

Gladden, John M; Allgaier, Martin; Miller, Christopher S; Hazen, Terry C; VanderGheynst, Jean S; Hugenholtz, Philip; Simmons, Blake A; Singer, Steven W

2011-08-15

Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60°C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80°C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

Fermentation characteristics and nutritive value of low moisture silage made from mature bermudagrass (C. dactylon) and switchgrass (P. virgatum) in mixture with alfalfa (M. sativa) or treated with urea and plantain (Musa AAB

USDA-ARS?s Scientific Manuscript database

Two experiments were conducted at the University of Kentucky Spindletop Farm in Lexington, Kentucky between October and November, 2009 to evaluate the effect of different percentages of alfalfa (Medicago sativa) as mixtures in switchgrass (Panicum virgatus) and bermudagrass (Cynodon dactylon) silage...

Comparison of ethanol production from corn cobs and switchgrass following a pyrolysis-based biorefinery approach.

PubMed

Luque, Luis; Oudenhoven, Stijn; Westerhof, Roel; van Rossum, Guus; Berruti, Franco; Kersten, Sascha; Rehmann, Lars

2016-01-01

One of the main obstacles in lignocellulosic ethanol production is the necessity of pretreatment and fractionation of the biomass feedstocks to produce sufficiently pure fermentable carbohydrates. In addition, the by-products (hemicellulose and lignin fraction) are of low value, when compared to dried distillers grains (DDG), the main by-product of corn ethanol. Fast pyrolysis is an alternative thermal conversion technology for processing biomass. It has recently been optimized to produce a stream rich in levoglucosan, a fermentable glucose precursor for biofuel production. Additional product streams might be of value to the petrochemical industry. However, biomass heterogeneity is known to impact the composition of pyrolytic product streams, as a complex mixture of aromatic compounds is recovered with the sugars, interfering with subsequent fermentation. The present study investigates the feasibility of fast pyrolysis to produce fermentable pyrolytic glucose from two abundant lignocellulosic biomass sources in Ontario, switchgrass (potential energy crop) and corn cobs (by-product of corn industry). Demineralization of biomass removes catalytic centers and increases the levoglucosan yield during pyrolysis. The ash content of biomass was significantly decreased by 82-90% in corn cobs when demineralized with acetic or nitric acid, respectively. In switchgrass, a reduction of only 50% for both acids could be achieved. Conversely, levoglucosan production increased 9- and 14-fold in corn cobs when rinsed with acetic and nitric acid, respectively, and increased 11-fold in switchgrass regardless of the acid used. After pyrolysis, different configurations for upgrading the pyrolytic sugars were assessed and the presence of potentially inhibitory compounds was approximated at each step as double integral of the UV spectrum signal of an HPLC assay. The results showed that water extraction followed by acid hydrolysis and solvent extraction was the best upgrading strategy. Ethanol yields achieved based on initial cellulose fraction were 27.8% in switchgrass and 27.0% in corn cobs. This study demonstrates that ethanol production from switchgrass and corn cobs is possible following a combined thermochemical and fermentative biorefinery approach, with ethanol yields comparable to results in conventional pretreatments and fermentation processes. The feedstock-independent fermentation ability can easily be assessed with a simple assay.

Deciphering Natural Allelic Variation in Switchgrass for Biomass Yield and Quality Using a Nested Association Mapping Population

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saha, Malay C.; Brummer, E. Charles; Kaeppler, Shawn

Switchgrass (Panicum virgatum L.) is a C4 grass with high biomass yield potential and a model species for bioenergy feedstock development. Understanding the genetic basis of quantitative traits is essential to facilitate genome-enabled breeding programs. The nested association mapping (NAM) analysis combines the best features of both bi-parental and association analyses and can provide high power and high resolution in QTL detection and will ensure significant improvements in biomass yield and quality. To develop a NAM population of switchgrass, 15 highly diverse genotypes with specific characteristics were selected from a diversity panel and crossed to a recurrent parent, AP13, amore » genotype selected for whole genome sequencing and parent of a mapping population. Ten genotypes from each of the 15 F1 families were then chain crossed. Progenies form each family were randomly selected to develop the NAM population. The switchgrass NAM population consists of a total of 2000 genotypes from 15 families. All the progenies, founder parents, F1 parents (n=2350) were evaluated in replicated field trials at Ardmore, OK and Knoxville, TN. Phenotypic data on plant height, tillering ability, regrowth, flowering time, and biomass yield were collected. Dried biomass samples were also analyzed using prediction equations of NIRS at the Noble Foundation and for lignin content, S/G ratio, and sugar release characteristics at the NREL. Genomic shotgun sequencing of 15 switchgrass NAM founder parental genomes at JGI produced 28-66 Gb high-quality sequence data. Alignment of these sequences with the reference genome, AP13 (v3.0), revealed that up to 99% of the genomic sequences mapped to the reference genome. A total of 2,149 individuals from NAM populations were sequenced by exome capture and two sets of 15 SNP matrices (one for each family) were generated. QTL associated with important traits have been identified and verified in breeding populations. The QTL detected and their associated markers can be used in molecular breeding programs to facilitate development of improved switchgrass cultivars for biofuel production.« less

Seedling Biomass Partition and Water Use Efficiency of Switchgrass and Milkvetch in Monocultures and Mixtures in Response to Various Water Availabilities

NASA Astrophysics Data System (ADS)

Xu, Bingcheng; Deng, Xiping; Zhang, Suiqi; Shan, Lun

2010-10-01

Seedling biomass and allocation, transpiration water use efficiency (TWUE), and species competition between switchgrass ( Panicum virgatum L.) and milkvetch ( Astragalus adsurgens Pall.) were investigated in a pot-cultivated experiment under different levels of water availability. The experiment was conducted using a simple replacement design in which switchgrass and milkvetch were grown in growth chamber with ten seedlings per pot, in three combinations of the two species (0:10, 5:5 and 10:0). Five water treatments included sufficient water supply (HW), gradual soil drying from HW (DHW), moderate water stress (LW), gradual soil drying from LW (DLW), and re-establishment of LW conditions after 12 days of drying from LW (RLW). Water treatments were applied over a 15-day period. Biomass production and its partitioning, and TWUE were determined at the end of the experiment. Species competitive indices (competitive ratio (CR), aggressivity (A) and relative yield total (RYT)) were calculated from the biomass dry weight data for shoots, roots and total biomass. Water stress significantly reduced seedling biomass production but increased root:shoot ratios in both monocultures and mixtures. In the RLW treatment, only switchgrass monocultures displayed compensatory biomass production and TWUE, while both species demonstrated compensatory growth in the mixture. Switchgrass was the dominant species and much more aggressive than milkvetch in the LW treatment, while in the other four treatments milkvetch was the dominant species as measured by the positive value of aggressivity and higher values of CR. The total biomass RYT values of the two species were higher than 1.0, indicating some degree of resource complimentarity. In the two-species mixture, although the biomass production was lower than that of milkvetch in the monoculture, there was better TWUE, especially under low and fluctuating water availability.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu, Chaowei; Reddy, Amitha P.; Simmons, Christopher W.

Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methodsmore » included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under thermophilic and high-solid conditions. The results suggest that the presence of microorganisms may be more important than their relative abundance in retaining an active microbial community.« less

Impacts of root traits and genotypic diversity in switchgrass cropping systems on biogeochemical cycling of soil carbon and nitrogen

NASA Astrophysics Data System (ADS)

De Graaff, M. A.; Jastrow, J. D.; Adkins, J.; Johns, A. C.; Morris, G.; Six, J.

2016-12-01

Land-use change for bioenergy production can create greenhouse gas (GHG) emissions through disturbance of soil carbon (C) pools, but native species with extensive root systems may rapidly repay the GHG debt by enhancing soil C sequestration upon land-use change, particularly when grown in diverse mixtures. Here we investigated how root traits and genotypic diversity in switchgrass (Panicum virgatum) impacts yield, nitrogen (N) cycling and soil C stabilization. Owing to extensive within-species variation in root morphology and architecture among the switchgrass cultivars, we hypothesized that increasing cultivar diversity would enhance belowground niche differentiation, thereby increasing N use efficiency, yield, and ultimately soil C stabilization. Our experiment was conducted at the Fermilab National Environmental Research Park, in northeastern Illinois, USA, where we varied the level of switchgrass genotypic diversity using various local and non-local cultivars (1, 2, 4, or 6 cultivars per plot) in a replicated field trial. We found that genotypic mixtures had one-third higher biomass production than the average monoculture, and no monoculture was significantly higher yielding than the average mixture. Further, year-to-year variation in yields was reduced in the mixture of switchgrass relative to the species monocultures. Despite positive impacts of increased intraspecific diversity on biomass production, we found no effect on N use efficiency, or soil C sequestration. However there were differences among cultivars in soil C input and soil C stabilization. These differences were related to specific root length (SRL), where greater SRL was accompanied by more root-derived soil C. Our findings suggest SRL is a root trait that affects soil C input, and that genotypic mixtures could help provide high, stable yields of high-quality biomass feedstocks.

Toxicity of coal fly ash (CFA) and toxicological response of switchgrass in mycorrhiza-mediated CFA-soil admixtures.

PubMed

Awoyemi, Olushola M; Dzantor, E Kudjo

2017-10-01

Increasing support for the use of Coal fly ash (CFA) in agriculture has necessitated a better understanding of the effects of the CFA in various cropping schemes. Experiments were conducted to assess mutagenic response of a mutant strain of Salmonella enterica serovar Typhimurium (TA100) to varying concentrations of CFA-water extracts, determine oxidative stress in switchgrass (Panicum virgatum L.) at varying levels of CFA-soil admixtures, and evaluate mycorrhiza-mediated modulation of oxidative stress responses of CFA-grown switchgrass. The TA100 exposed to 0%, 5%, 10%, 15%, 20% and 25% (w/v) CFA-water extracts elicited significant (p < 0.05) mutagenic responses at 20% and 25% extract levels but not below the 15% level. In greenhouse pot experiment, CFA-soil admixtures at 7.5% and 15% (w/w) significantly (p < 0.05) decreased the activities of superoxide dismutase (SOD) by 19.1% and 28.3% respectively, compared to control soil (0% w/w CFA/soil). Under the same conditions, activities of glutathione peroxidase (GPx) decreased by 75.9% and 66.9%. In contrast to the antioxidant enzyme activities, levels of malondialdehyde (MDA) an indicator of lipid peroxidation increased significantly (p < 0.05) by 30.49% and 38.38%. Inoculation of 7.5% and 15% CFA-soil admixtures with arbuscular mycorrhizal fungi (AMF), Rhizophaga clarus enhanced the activities of both SOD and GPx in the switchgrass, while it significantly (p < 0.05) reduced the levels of MDA. The study demonstrated that incorporation of CFA (at concentrations considered to be non-mutagenic against TA100) as soil amendment produced concentration-dependent oxidative stress responses in switchgrass; however, inoculation of the CFA-soil admixtures with AMF significantly modulated the oxidative stress responses. Copyright © 2017 Elsevier Inc. All rights reserved.

Soil and variety effects on energy use and carbon emissions associated with switchgrass-based ethanol production in Mississippi

DOE Office of Scientific and Technical Information (OSTI.GOV)

Woli, Prem; Paz, Joel O.; Baldwin, Brian S.

2012-06-29

High biomass production potential, wide adaptability, low input requirement, and low environmental risk make switchgrass an economically and ecologically viable energy crop.The inherent variablity in switchgrass productivity due to variations in soil and variety could affect the sustainability and eco-friendliness of switchgrass-based ethanol production. This study examined the soil and variety effects on these variables. Three locations in Mississippi were selected based on latitude and potential acreage. Using ALMANAC, switchgrass biomass yields were simulated for several scenarios of soils and varities. The simulated yields were fed to IBSAL to compute energy use and CO2 emissions in various operations in themore » biomass supply From the energy and emissions values, the sustainability and eco-friendliness of ethanol production were determined using net energy value (NEV) and carbon credit balance (CCB) as indicators, respectively. Soil and variety effects on NEV and CCB were analyzed using the Kruskal-Wallis test. Results showed significant differences in NEV and CCB across soils and varieties. Both NEV and CCB increased in the direction of heavier to lighter soils and on the order of north-upland , south-upland, north-lowland, and south-lowland varieties. Only north-upland and south-lowland varieties were significantly significantly different because they were different in both cytotype and ecotype. Gaps between lowland and upland varieties were smaller in a dry year than in a wet year. The NEV and CCB increased in the direction of dry to wet year. From south to north, they decreased for lowland cytotypes but increased for upland cytotypes. Thus, the differences among varieties decreased northwards.« less

Non-targeted Colonization by the Endomycorrhizal Fungus, Serendipita vermifera, in Three Weeds Typically Co-occurring with Switchgrass

PubMed Central

Ray, Prasun; Guo, Yingqing; Kolape, Jaydeep; Craven, Kelly D.

2018-01-01

Serendipita vermifera (=Sebacina vermifera; isolate MAFF305830) is a mycorrhizal fungus originally isolated from the roots of an Australian orchid that we have previously shown to be beneficial in enhancing biomass yield and drought tolerance in switchgrass, an important bioenergy crop for cellulosic ethanol production in the United States. However, almost nothing is known about how this root-associated fungus proliferates and grows through the soil matrix. Such information is critical to evaluate the possibility of non-target effects, such as unintended spread to weedy plants growing near a colonized switchgrass plant in a field environment. A microcosm experiment was conducted to study movement of vegetative mycelia of S. vermifera between intentionally inoculated switchgrass (Panicum virgatum L.) and nearby weeds. We constructed size-exclusion microcosms to test three different common weeds, large crabgrass (Digitaria sanguinalis L.), Texas panicum (Panicum texanum L.), and Broadleaf signalgrass (Brachiaria platyphylla L.), all species that typically co-occur in Southern Oklahoma and potentially compete with switchgrass. We report that such colonization of non-target plants by S. vermifera can indeed occur, seemingly via co-mingled root systems. As a consequence of colonization, significant enhancement of growth was noted in signalgrass, while a mild increase (albeit not significant) was evident in crabgrass. Migration of the fungus seems unlikely in root-free bulk soil, as we failed to see transmission when the roots were kept separate. This research is the first documentation of non-targeted colonization of this unique root symbiotic fungus and highlights the need for such assessments prior to deployment of biological organisms in the field. PMID:29375607

Development of Kinetics and Mathematical Models for High-Pressure Gasification of Lignite-Switchgrass Blends: Cooperative Research and Development Final Report, CRADA Number CRD-11-447

DOE Office of Scientific and Technical Information (OSTI.GOV)

Iisa, Kristiina

2016-04-06

NREL will work with Participant as a subtier partner under DE-FOA-0000240 titled "Co-Production of Power, Fuels, and Chemicals via Coal/Biomass Mixtures." The goal of the project is to determine the gasification characteristics of switchgrass and lignite mixtures and develop kinetic models. NREL will utilize a pressurized thermogravimetric analyzer to measure the reactivity of chars generated in a pressurized entrained-flow reactor at Participant's facilities and to determine the evolution of gaseous species during pyrolysis of switchgrass-lignite mixtures. Mass spectrometry and Fourier-transform infrared analysis will be used to identify and quantify the gaseous species. The results of the project will aid inmore » defining key reactive properties of mixed coal biomass fuels.« less

Carbon Calculator for Land Use Change from Biofuels Production (CCLUB). Users' Manual and Technical Documentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, Jennifer B.; Qin, Zhangcai; Mueller, Steffen

The Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) calculates carbon emissions from land use change (LUC) for four different ethanol production pathways including corn grain ethanol and cellulosic ethanol from corn stover, Miscanthus, and switchgrass. This document discusses the version of CCLUB released September 30, 2014 which includes corn and three cellulosic feedstocks: corn stover, Miscanthus, and switchgrass.

Phytoremediation of biosolids from an end-of-life municipal lagoon using cattail (Typha latifolia L.) and switchgrass (Panicum virgatum L.).

PubMed

Jeke, Nicholson N; Hassan, Adenike O; Zvomuya, Francis

2017-03-04

Land spreading of biosolids as a disposal option is expensive and can disperse pathogens and contaminants in the environment. This growth room study examined phytoremediation using switchgrass (Panicum virgatum L.) and cattail (Typha latifolia L.) as an alternative to land spreading of biosolids. Seedlings were transplanted into pots containing 3.9 kg of biosolids (dry wt.). Aboveground biomass (AGB) was harvested either once or twice during each 90-day growth period. Switchgrass AGB yield was greater with two harvests than with one harvest during the first 90-day growth period, whereas cattail yield was not affected by harvest frequency. In the second growth period, harvesting frequency did not affect the yield of either plant species. However, repeated harvesting significantly improved nitrogen (N) and phosphorus (P) uptake by both plants in the first period. Phytoextraction of P was significantly greater for switchgrass (3.9% of initial biosolids P content) than for cattail (2.8%), while plant species did not have a significant effect on N phytoextraction. The trace element accumulation in the AGB of both plant species was negligible. Phytoextraction rates attained in this study suggest that phytoremediation can effectively remove P from biosolids and offers a potentially viable alternative to the disposal of biosolids on agricultural land.

Effect of acid, steam explosion, and size reduction pretreatments on bio-oil production from sweetgum, switchgrass, and corn stover.

PubMed

Wang, Hui; Srinivasan, Radhakrishnan; Yu, Fei; Steele, Philip; Li, Qi; Mitchell, Brian; Samala, Aditya

2012-05-01

Bio-oil produced from biomass by fast pyrolysis has the potential to be a valuable substitute for fossil fuels. In a recent work on pinewood, we found that pretreatment alters the structure and chemical composition of biomass, which influence fast pyrolysis. In this study, we evaluated dilute acid, steam explosion, and size reduction pretreatments on sweetgum, switchgrass, and corn stover feedstocks. Bio-oils were produced from untreated and pretreated feedstocks in an auger reactor at 450 °C. The bio-oil's physical properties of pH, water content, acid value, density, and viscosity were measured. The chemical characteristics of the bio-oils were determined by gas chromatography-mass spectrometry. The results showed that bio-oil yield and composition were influenced by the pretreatment method and feedstock type. Bio-oil yields of 52, 33, and 35 wt% were obtained from medium-sized (0.68-1.532 mm) untreated sweetgum, switchgrass, and corn stover, respectively, which were higher than the yields from other sizes. Bio-oil yields of 56, 46, and 51 wt% were obtained from 1% H(2)SO(4)-treated medium-sized sweetgum, switchgrass, and corn stover, respectively, which were higher than the yields from untreated and steam explosion treatments.

Spatially-explicit life cycle assessment of sun-to-wheels transportation pathways in the U.S.

PubMed

Geyer, Roland; Stoms, David; Kallaos, James

2013-01-15

Growth in biofuel production, which is meant to reduce greenhouse gas (GHG) emissions and fossil energy demand, is increasingly seen as a threat to food supply and natural habitats. Using photovoltaics (PV) to directly convert solar radiation into electricity for battery electric vehicles (BEVs) is an alternative to photosynthesis, which suffers from a very low energy conversion efficiency. Assessments need to be spatially explicit, since solar insolation and crop yields vary widely between locations. This paper therefore compares direct land use, life cycle GHG emissions and fossil fuel requirements of five different sun-to-wheels conversion pathways for every county in the contiguous U.S.: Ethanol from corn or switchgrass for internal combustion vehicles (ICVs), electricity from corn or switchgrass for BEVs, and PV electricity for BEVs. Even the most land-use efficient biomass-based pathway (i.e., switchgrass bioelectricity in U.S. counties with hypothetical crop yields of over 24 tonnes/ha) requires 29 times more land than the PV-based alternative in the same locations. PV BEV systems also have the lowest life cycle GHG emissions throughout the U.S. and the lowest fossil fuel inputs, except for locations with hypothetical switchgrass yields of 16 or more tonnes/ha. Including indirect land use effects further strengthens the case for PV.

Impacts of variability in cellulosic biomass yields on energy security.

PubMed

Mullins, Kimberley A; Matthews, H Scott; Griffin, W Michael; Anex, Robert

2014-07-01

The practice of modeling biomass yields on the basis of deterministic point values aggregated over space and time obscures important risks associated with large-scale biofuel use, particularly risks related to drought-induced yield reductions that may become increasingly frequent under a changing climate. Using switchgrass as a case study, this work quantifies the variability in expected yields over time and space through switchgrass growth modeling under historical and simulated future weather. The predicted switchgrass yields across the United States range from about 12 to 19 Mg/ha, and the 80% confidence intervals range from 20 to 60% of the mean. Average yields are predicted to decrease with increased temperatures and weather variability induced by climate change. Feedstock yield variability needs to be a central part of modeling to ensure that policy makers acknowledge risks to energy supplies and develop strategies or contingency plans that mitigate those risks.

Utilization of lignocellulosic polysaccharides

NASA Astrophysics Data System (ADS)

Fenske, John James

Lignocellulosic biomass represents a vast supply of fermentable carbohydrates and functional aromatic compounds. Conversion of lignocellulosics to ethanol and other useful products would be of widespread economical and environmental benefit. Better understanding of the behavior of different lignocellulosic feedstocks in fermentation protocols as well as catalytic activities involved in lignocellulosic depolymerization will further enhance the commercial viability of biomass-to-ethanol conversion processes. The relative toxicity of the combined non-xylose components in prehydrolysates derived from three different lignocellulosic biomass feedstocks (poplar, corn stover and switchgrass, or Panicum virgatum L.) was determined using a Pichia stipits fermentation assay. The relative toxicity of the prehydrolysates, in decreasing order, was poplar-derived prehydrolysates > switchgrass-derived prehydrolysates > corn stover-derived prehydrolysates. Ethanol yields averaged 74%, 83% and 88% of control values for poplar, switchgrass and corn stover prehydrolysates, respectively. Volumetric ethanol productivities (g ethanol lsp{-1} hsp{-1}) averaged 32%, 70% and 102% of control values for poplar, switchgrass and corn stover prehydrolysates, respectively. Ethanol productivities correlated closely with acetate concentrations in the prehydrolysates; however, regression lines correlating acetate concentrations and ethanol productivities were found to be feedstock-dependent. Differences in the relative toxicity of xylose-rich prehydrolysates derived from woody and herbaceous feedstocks are likely due to the relative abundance of a variety of inhibitory compounds, e.g. acetate and aromatic compounds. Fourteen aromatic monomers present in prehydrolysates prepared from corn stover, switchgrass, and poplar were tentatively identified by comparison with published mass spectra. The concentrations of the aromatic monomers totaled 112, 141 and 247 mg(l)sp{-1} for corn stover, switchgrass and poplar prehydrolysates, respectively. The woody and herbaceous feedstocks differed in both amount and type of aromatic monomers. The cellulases of Trichoderma reesei are the most widely studied for use in the depolymerization of lignocellulosics. The Trichoderma cellobiohydrolases CBH1 and CBH2 are traditionally categorized as exo-acting cellulases. A simple individual-based model was created to explore the potential effects of native endo activity on substrate-velocity profiles. The model results indicate that an enzyme with a small amount of endo activity will show an apparent substrate inhibition as substrate levels are increased. Actual hydrolysis studies using affinity chromatography-purified CBH2 preparations from three laboratories indicate that CBH2 has native endo activity, while CBH1 does not.

Temporal dynamics of fibrolytic and methanogenic rumen microorganisms during in situ incubation of switchgrass determined by 16S rRNA gene profiling.

PubMed

Piao, Hailan; Lachman, Medora; Malfatti, Stephanie; Sczyrba, Alexander; Knierim, Bernhard; Auer, Manfred; Tringe, Susannah G; Mackie, Roderick I; Yeoman, Carl J; Hess, Matthias

2014-01-01

The rumen microbial ecosystem is known for its biomass-degrading and methane-producing phenotype. Fermentation of recalcitrant plant material, comprised of a multitude of interwoven fibers, necessitates the synergistic activity of diverse microbial taxonomic groups that inhabit the anaerobic rumen ecosystem. Although interspecies hydrogen (H2) transfer, a process during which bacterially generated H2 is transferred to methanogenic Archaea, has obtained significant attention over the last decades, the temporal variation of the different taxa involved in in situ biomass-degradation, H2 transfer and the methanogenesis process remains to be established. Here we investigated the temporal succession of microbial taxa and its effect on fiber composition during rumen incubation using 16S rRNA amplicon sequencing. Switchgrass filled nylon bags were placed in the rumen of a cannulated cow and collected at nine time points for DNA extraction and 16S pyrotag profiling. The microbial community colonizing the air-dried and non-incubated (0 h) switchgrass was dominated by members of the Bacilli (recruiting 63% of the pyrotag reads). During in situ incubation of the switchgrass, two major shifts in the community composition were observed: Bacilli were replaced within 30 min by members belonging to the Bacteroidia and Clostridia, which recruited 34 and 25% of the 16S rRNA reads generated, respectively. A second significant shift was observed after 16 h of rumen incubation, when members of the Spirochaetes and Fibrobacteria classes became more abundant in the fiber-adherent community. During the first 30 min of rumen incubation ~13% of the switchgrass dry matter was degraded, whereas little biomass degradation appeared to have occurred between 30 min and 4 h after the switchgrass was placed in the rumen. Interestingly, methanogenic members of the Euryarchaeota (i.e., Methanobacteria) increased up to 3-fold during this period of reduced biomass-degradation, with peak abundance just before rates of dry matter degradation increased again. We hypothesize that during this period microbial-mediated fibrolysis was temporarily inhibited until H2 was metabolized into CH4 by methanogens. Collectively, our results demonstrate the importance of inter-species interactions for the biomass-degrading and methane-producing phenotype of the rumen microbiome-both microbially facilitated processes with global significance.

Registration of NE Trailblazer C-1, NE Trailblazer C0, NE Trailblazer C2, NE Trailblazer C3, NE Trailblazer C4, and NE Trailblazer C5 Switchgrass Germplasms

USDA-ARS?s Scientific Manuscript database

NE Trailblazer C-1 (GP-101, PI 672015), NE Trailblazer C0 (GP-100, PI 672014), NE Trailblazer C2 (GP-102, PI 672016), NE Trailblazer C3 (GP-103, PI 672017), NE Trailblazer C4 (GP-104, PI 672018), and NE Trailblazer C5 (GP-105, PI 672019) switchgrass (Panicum virgatum L.) germplasms were released by ...

Life cycle assessment of switchgrass- and corn stover-derived ethanol-fueled automobiles.

PubMed

Spatari, Sabrina; Zhang, Yimin; MacLean, Heather L

2005-12-15

Utilizing domestically produced cellulose-derived ethanol for the light-duty vehicle fleet can potentially improve the environmental performance and sustainability of the transport and energy sectors of the economy. A life cycle assessment model was developed to examine environmental implications of the production and use of ethanol in automobiles in Ontario, Canada. The results were compared to those of low-sulfur reformulated gasoline (RFG) in a functionally equivalent automobile. Two time frames were evaluated, one near-term (2010), which examines converting a dedicated energy crop (switchgrass) and an agricultural residue (corn stover) to ethanol; and one midterm (2020), which assumes technological improvements in the switchgrass-derived ethanol life cycle. Near-term results show that, compared to a RFG automobile, life cycle greenhouse gas (GHG) emissions are 57% lower for an E85-fueled automobile derived from switchgrass and 65% lower for ethanol from corn stover, on a grams of CO2 equivalent per kilometer basis. Corn stover ethanol exhibits slightly lower life cycle GHG emissions, primarily due to sharing emissions with grain production. Through projected improvements in crop and ethanol yields, results for the mid-term scenario show that GHG emissions could be 25-35% lower than those in 2010 and that, even with anticipated improvements in RFG automobiles, E85 automobiles could still achieve up to 70% lower GHG emissions across the life cycle.

Switchgrass Cultivar/Ecotype Selection and Management for Biofuels in the Upper Southeast USA

PubMed Central

Parrish, David J.; Wolf, Dale D.

2014-01-01

Switchgrass (Panicum virgatum L.), a perennial warm-season grass indigenous to the eastern USA, has potential as a biofuels feedstock. The objective of this study was to investigate the performance of upland and lowland switchgrass cultivars under different environments and management treatments. Four cultivars of switchgrass were evaluated from 2000 to 2001 under two management regimes in plots established in 1992 at eight locations in the upper southeastern USA. Two management treatments included 1) a single annual harvest (in late October to early November) and a single application of 50 kg N/ha/yr and 2) two annual harvests (in midsummer and November) and a split application of 100 kg N/ha/yr. Biomass yields averaged 15 Mg/ha/yr and ranged from 10 to 22 Mg/ha/yr across cultivars, managements, locations, and years. There was no yield advantage in taking two harvests of the lowland cultivars (Alamo and Kanlow). When harvested twice, upland cultivars (Cave-in-Rock and Shelter) provided yields equivalent to the lowland ecotypes. Tiller density was 36% lower in stands cutting only once per year, but the stands appeared vigorous after nine years of such management. Lowland cultivars and a one-cutting management (after the tops have senesced) using low rates of applied N (50 kg/ha) are recommended. PMID:25105170

Soil carbon inventories under a bioenergy crop (switchgrass): Measurement limitations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garten, C.T. Jr.; Wullschleger, S.D.

Approximately 5 yr after planting, coarse root carbon (C) and soil organic C (SOC) inventories were compared under different types of plant cover at four switchgrass (Panicum virgatum L.) production field trials in the southeastern USA. There was significantly more coarse root C under switchgrass (Alamo variety) and forest cover than tall fescue (Festuca arundinacea Schreb.), corn (Zea mays L.), or native pastures of mixed grasses. Inventories of SOC under switchgrass were not significantly greater than SOC inventories under other plant covers. At some locations the statistical power associated with ANOVA of SOC inventories was low, which raised questions aboutmore » whether differences in SOC could be detected statistically. A minimum detectable difference (MDD) for SOC inventories was calculated. The MDD is the smallest detectable difference between treatment means once the variation, significance level, statistical power, and sample size are specified. The analysis indicated that a difference of {approx}50 mg SOC/cm{sup 2} or 5 Mg SOC/ha, which is {approx}10 to 15% of existing SOC, could be detected with reasonable sample sizes and good statistical power. The smallest difference in SOC inventories that can be detected, and only with exceedingly large sample sizes, is {approx}2 to 3%. These measurement limitations have implications for monitoring and verification of proposals to ameliorate increasing global atmospheric CO{sub 2} concentrations by sequestering C in soils.« less

Effects of precipitation changes on switchgrass photosynthesis, growth, and biomass: A mesocosm experiment.

PubMed

Hui, Dafeng; Yu, Chih-Li; Deng, Qi; Dzantor, E Kudjo; Zhou, Suping; Dennis, Sam; Sauve, Roger; Johnson, Terrance L; Fay, Philip A; Shen, Weijun; Luo, Yiqi

2018-01-01

Climate changes, including chronic changes in precipitation amounts, will influence plant physiology and growth. However, such precipitation effects on switchgrass, a major bioenergy crop, have not been well investigated. We conducted a two-year precipitation simulation experiment using large pots (95 L) in an environmentally controlled greenhouse in Nashville, TN. Five precipitation treatments (ambient precipitation, and -50%, -33%, +33%, and +50% of ambient) were applied in a randomized complete block design with lowland "Alamo" switchgrass plants one year after they were established from tillers. The growing season progression of leaf physiology, tiller number, height, and aboveground biomass were determined each growing season. Precipitation treatments significantly affected leaf physiology, growth, and aboveground biomass. The photosynthetic rates in the wet (+50% and +33%) treatments were significantly enhanced by 15.9% and 8.1%, respectively, than the ambient treatment. Both leaf biomass and plant height were largely increased, resulting in dramatically increases in aboveground biomass by 56.5% and 49.6% in the +50% and +33% treatments, respectively. Compared to the ambient treatment, the drought (-33% and -50%) treatments did not influence leaf physiology, but the -50% treatment significantly reduced leaf biomass by 37.8%, plant height by 16.3%, and aboveground biomass by 38.9%. This study demonstrated that while switchgrass in general is a drought tolerant grass, severe drought significantly reduces Alamo's growth and biomass, and that high precipitation stimulates its photosynthesis and growth.

Switchgrass Cultivar/Ecotype Selection and Management for Biofuels in the Upper Southeast USA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lemus, Rocky; Parrish, David J.; Wolf, Dale D.

Switchgrass ( Panicum virgatum L.), a perennial warm-season grass indigenous to the eastern USA, has potential as a biofuels feedstock. The objective of this study was to investigate the performance of upland and lowland switchgrass cultivars under different environments and management treatments. Four cultivars of switchgrass were evaluated from 2000 to 2001 under two management regimes in plots established in 1992 at eight locations in the upper southeastern USA. Two management treatments included 1) a single annual harvest (in late October to early November) and a single application of 50 kg N/ha/yr and 2) two annual harvests (in midsummer and November)more » and a split application of 100 kg N/ha/yr. Biomass yields averaged 15 Mg/ha/yr and ranged from 10 to 22 Mg/ha/yr across cultivars, managements, locations, and years. There was no yield advantage in taking two harvests of the lowland cultivars (Alamo and Kanlow). When harvested twice, upland cultivars (Cave-in-Rock and Shelter) provided yields equivalent to the lowland ecotypes. Tiller density was 36% lower in stands cutting only once per year, but the stands appeared vigorous after nine years of such management. Lowland cultivars and a one-cutting management (after the tops have senesced) using low rates of applied N (50 kg/ha) are recommended.« less

Switchgrass Cultivar/Ecotype Selection and Management for Biofuels in the Upper Southeast USA

DOE PAGES

Lemus, Rocky; Parrish, David J.; Wolf, Dale D.

2014-01-01

Switchgrass ( Panicum virgatum L.), a perennial warm-season grass indigenous to the eastern USA, has potential as a biofuels feedstock. The objective of this study was to investigate the performance of upland and lowland switchgrass cultivars under different environments and management treatments. Four cultivars of switchgrass were evaluated from 2000 to 2001 under two management regimes in plots established in 1992 at eight locations in the upper southeastern USA. Two management treatments included 1) a single annual harvest (in late October to early November) and a single application of 50 kg N/ha/yr and 2) two annual harvests (in midsummer and November)more » and a split application of 100 kg N/ha/yr. Biomass yields averaged 15 Mg/ha/yr and ranged from 10 to 22 Mg/ha/yr across cultivars, managements, locations, and years. There was no yield advantage in taking two harvests of the lowland cultivars (Alamo and Kanlow). When harvested twice, upland cultivars (Cave-in-Rock and Shelter) provided yields equivalent to the lowland ecotypes. Tiller density was 36% lower in stands cutting only once per year, but the stands appeared vigorous after nine years of such management. Lowland cultivars and a one-cutting management (after the tops have senesced) using low rates of applied N (50 kg/ha) are recommended.« less

Effects on Environmental and Socioeconomic Sustainability of Producing Ethanol from Perennial Grasses

NASA Astrophysics Data System (ADS)

Dale, V. H.; Parish, E. S.

2016-12-01

Using perennial grasses to produce ethanol can enhance progress toward sustainability. A suite of 35 environmental and socioeconomic sustainability indicators was considered in a holistic sustainability assessment of a five-year switchgrass-to-ethanol production experiment centered on a demonstration-scale biorefinery in Vonore, Tennessee. By combining field measurements, literature review and expert opinion, the team was able to rate 28 of the 35 recommended sustainability indicators. The team combined these ratings within a multi-attribute decision support system tool and used this information to compare the sustainability of producing 2118 hectares of no-till switchgrass relative to two alternative business-as-usual scenarios of unmanaged pasture and tilled corn production. The results suggest that East Tennessee switchgrass production improves environmental quality overall and can be beneficial to the counties surrounding the biorefinery in terms of dollars earned and jobs created. The timing of switchgrass production also provides an opportunity to use inactive equipment and laborers. By incorporating a landscape design approach, the opportunities, constraints and most reasonable paths forward for growing bioenergy feedstock in specific context can be assessed in a way that adapts and improves local practices. Lessons learned from this case study are being incorporated into sustainability assessments of corn stover in Iowa and a variety of bioenergy feedstocks in diverse settings. The overall goal is to develop sound management practices that can address the multiple and sometimes competing demands of stakeholders.

Energy Potential and Greenhouse Gas Emissions from Bioenergy Cropping Systems on Marginally Productive Cropland

PubMed Central

Schmer, Marty R.; Vogel, Kenneth P.; Varvel, Gary E.; Follett, Ronald F.; Mitchell, Robert B.; Jin, Virginia L.

2014-01-01

Low-carbon biofuel sources are being developed and evaluated in the United States and Europe to partially offset petroleum transport fuels. Current and potential biofuel production systems were evaluated from a long-term continuous no-tillage corn (Zea mays L.) and switchgrass (Panicum virgatum L.) field trial under differing harvest strategies and nitrogen (N) fertilizer intensities to determine overall environmental sustainability. Corn and switchgrass grown for bioenergy resulted in near-term net greenhouse gas (GHG) reductions of −29 to −396 grams of CO2 equivalent emissions per megajoule of ethanol per year as a result of direct soil carbon sequestration and from the adoption of integrated biofuel conversion pathways. Management practices in switchgrass and corn resulted in large variation in petroleum offset potential. Switchgrass, using best management practices produced 3919±117 liters of ethanol per hectare and had 74±2.2 gigajoules of petroleum offsets per hectare which was similar to intensified corn systems (grain and 50% residue harvest under optimal N rates). Co-locating and integrating cellulosic biorefineries with existing dry mill corn grain ethanol facilities improved net energy yields (GJ ha−1) of corn grain ethanol by >70%. A multi-feedstock, landscape approach coupled with an integrated biorefinery would be a viable option to meet growing renewable transportation fuel demands while improving the energy efficiency of first generation biofuels. PMID:24594783

Improved tissue culture conditions for the emerging C4 model Panicum hallii.

PubMed

Grant, Joshua N; Burris, Jason N; Stewart, C Neal; Lenaghan, Scott C

2017-04-27

Panicum hallii Vasey (Hall's panicgrass) is a compact, perennial C 4 grass in the family Poaceae, which has potential to enable bioenergy research for switchgrass (Panicum virgatum L.). Unlike P. hallii, switchgrass has a large genome, allopolyploidy, self-incompatibility, a long life cycle, and large stature-all suboptimal traits for rapid genetics research. Herein we improved tissue culture methodologies for two inbred P. hallii populations: FIL2 and HAL2, to enable further development of P. hallii as a model C 4 plant. The optimal seed-derived callus induction medium was determined to be Murashige and Skoog (MS) medium supplemented with 40 mg L -1 L-cysteine, 300 mg L -1 L-proline, 3% sucrose, 1 g L -1 casein hydrolysate, 3 mg L -1 2,4-dichlorophenoxyacetic acid (2,4-D), and 45 μg L -1 6-benzylaminopurine (BAP), which resulted in callus induction of 51 ± 29% for FIL2 and 81 ± 19% for HAL2. The optimal inflorescence-derived callus induction was observed on MP medium (MS medium supplemented with 2 g L -1 L-proline, 3% maltose, 5 mg L -1 2,4-D, and 500 μg L -1 BAP), resulting in callus induction of 100 ± 0.0% for FIL2 and 84 ± 2.4% for HAL2. Shoot regeneration rates of 11.5 ± 0.8 shoots/gram for FIL2 and 11.3 ± 0.6 shoots/gram for HAL2 were achieved using seed-induced callus, whereas shoot regeneration rates of 26.2 ± 2.6 shoots/gram for FIL2 and 29.3 ± 3.6 shoots/gram for HAL2 were achieved from inflorescence-induced callus. Further, cell suspension cultures of P. hallii were established from seed-derived callus, providing faster generation of callus tissue compared with culture using solidified media (1.41-fold increase for FIL2 and 3.00-fold increase for HAL2). Aside from abbreviated tissue culture times from callus induction to plant regeneration for HAL2, we noted no apparent differences between FIL2 and HAL2 populations in tissue culture performance. For both populations, the cell suspension cultures outperformed tissue cultures on solidified media. Using the methods developed in this work, P. hallii callus was induced from seeds immediately after harvest in a shorter time and with higher frequencies than switchgrass. For clonal propagation, P. hallii callus was established from R1 inflorescences, similar to switchgrass, which further strengthens the potential of this plant as a C 4 model for genetic studies. The rapid cycling (seed-to-seed time) and ease of culture, further demonstrate the potential utility of P. hallii as a C 4 model plant.

Identification of Cell Wall Synthesis Regulatory Genes Controlling Biomass Characteristics and Yield in Rice (Oryza Sativa)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peng, Zhaohua PEng; Ronald, Palmela; Wang, Guo-Liang

This project aims to identify the regulatory genes of rice cell wall synthesis pathways using a cell wall removal and regeneration system. We completed the gene expression profiling studies following the time course from cell wall removal to cell wall regeneration in rice suspension cells. We also completed, total proteome, nuclear subproteome and histone modification studies following the course from cell wall removal and cell wall regeneration process. A large number of differentially expressed regulatory genes and proteins were identified. Meanwhile, we generated RNAi and over-expression transgenic rice for 45 genes with at least 10 independent transgenic lines for eachmore » gene. In addition, we ordered T-DNA and transposon insertion mutants for 60 genes from Korea, Japan, and France and characterized the mutants. Overall, we have mutants and transgenic lines for over 90 genes, exceeded our proposed goal of generating mutants for 50 genes. Interesting Discoveries a) Cell wall re-synthesis in protoplasts may involve a novel cell wall synthesis mechanism. The synthesis of the primary cell wall is initiated in late cytokinesis with further modification during cell expansion. Phragmoplast plays an essential role in cell wall synthesis. It services as a scaffold for building the cell plate and formation of a new cell wall. Only one phragmoplast and one new cell wall is produced for each dividing cell. When the cell wall was removed enzymatically, we found that cell wall re-synthesis started from multiple locations simultaneously, suggesting that a novel mechanism is involved in cell wall re-synthesis. This observation raised many interesting questions, such as how the starting sites of cell wall synthesis are determined, whether phragmoplast and cell plate like structures are involved in cell wall re-synthesis, and more importantly whether the same set of enzymes and apparatus are used in cell wall re-synthesis as during cytokinesis. Given that many known cell wall synthesis pathway genes are induced by removal of cell wall, some cell wall synthesis apparatus must be shared in both cases. The cell wall re-synthesis mechanism may have broad application because our preliminary assay indicates that the cell wall characteristics are highly different from those produced during cytokinesis. A thorough understanding on the regulation of cell wall re-synthesis may lead to improvement of cell wall characteristics. b) Removal of cell wall results in chromatin decondensation Another interesting observation was that removal of cell wall was associated with substantial chromatin change. Our DNA DAPI stain, chromatin MNase digestion, histone modification proteomics, protein differential expression analysis, and DNA oligo array studies all supported that substantial chromatin change was associated with removal of cell wall treatment. It is still under investigation if the chromatin change is associated with activation of cell wall synthesis genes, in which chromatin remodeling is required. Another possibility is that the cell wall is required for stabilizing the chromatin structure in plant cells. Given that spindle fiber is directly connected with both chromatin structure and cell wall synthesis, it is possible that there is an intrinsic connection between cell wall and chromatin.« less

Architecture of dermatophyte cell Walls: Electron microscopic and biochemical analysis

NASA Technical Reports Server (NTRS)

Nozawa, Y.; Kitajima, Y.

1984-01-01

A review with 83 references on the cell wall structure of dermatophytes is presented. Topics discussed include separation and preparation of cell walls; microstructure of cell walls by electron microscopy; chemical composition of cell walls; structural model of cell walls; and morphological structure of cell walls.

Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions.

PubMed

Yu, Chaowei; Reddy, Amitha P; Simmons, Christopher W; Simmons, Blake A; Singer, Steven W; VanderGheynst, Jean S

2015-01-01

Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methods included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. A stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under thermophilic and high-solid conditions. The results suggest that the presence of microorganisms may be more important than their relative abundance in retaining an active microbial community.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mbonimpa, Eric G.; Kumar, Sandeep; Owens, Vance N.

Switchgrass-derived ethanol has been proposed as an alternative to fossil fuels to improve sustainability of the US energy sector. In this study, life cycle analysis (LCA) was used to estimate the environmental benefits of this fuel. To better define the LCA environmental impacts associated with fertilization rates and farm-landscape topography, results from a controlled experiment were analyzed. Data from switchgrass plots planted in 2008, consistently managed with three nitrogen rates (0, 56, and 112 kg N ha –1), two landscape positions (shoulder and footslope), and harvested annually (starting in 2009, the year after planting) through 2014 were used as inputmore » into the Greenhouse gases, Regulated Emissions and Energy use in transportation (GREET) model. Simulations determined nitrogen (N) rate and landscape impacts on the life cycle energy and emissions from switchgrass ethanol used in a passenger car as ethanol–gasoline blends (10% ethanol:E10, 85% ethanol:E85s). Results indicated that E85s may lead to lower fossil fuels use (58 to 77%), greenhouse gas (GHG) emissions (33 to 82%), and particulate matter (PM2.5) emissions (15 to 54%) in comparison with gasoline. However, volatile organic compounds (VOCs) and other criteria pollutants such as nitrogen oxides (NOx), particulate matter (PM10), and sulfur dioxides (SO x) were higher for E85s than those from gasoline. Nitrogen rate above 56 kg N ha –1 yielded no increased biomass production benefits; but did increase (up to twofold) GHG, VOCs, and criteria pollutants. Lower blend (E10) results were closely similar to those from gasoline. The landscape topography also influenced life cycle impacts. Biomass grown at the footslope of fertilized plots led to higher switchgrass biomass yield, lower GHG, VOCs, and criteria pollutants in comparison with those at the shoulder position. Lastly, results also showed that replacing switchgrass before maximum stand life (10–20 years.) can further reduce the energy and emissions reduction benefits.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chung, Daehwan; Cha, Minseok; Snyder, Elise N.

In this paper, we report that the C. bescii genome does not encode an acetaldehyde/alcohol dehydrogenase or an acetaldehyde dehydrogenase and no ethanol production is detected in this strain. The recent introduction of an NADH-dependent AdhE from C. thermocellum (Fig. 1a) in an ldh mutant of this strain resulted in production of ethanol from un-pretreated switchgrass, but the thermolability of the C. thermocellum AdhE at the optimum growth temperature of C. bescii (78 °C) meant that ethanol was not produced above 65 °C. The adhB and adhE genes from Thermoanaerobacter pseudethanolicus 39E, an anaerobic thermophile that produces ethanol as amore » major fermentation product at 70 °C, were cloned and expressed in an ldh deletion mutant of C. bescii. The engineered strains produced ethanol at 75 °C, near the ethanol boiling point. The AdhB expressing strain produced ethanol (1.4 mM on Avicel, 0.4 mM on switchgrass) as well as acetate (13.0 mM on Avicel, 15.7 mM on switchgrass). The AdhE expressing strain produced more ethanol (2.3 mM on Avicel, 1.6 mM on switchgrass) and reduced levels of acetate (12.3 mM on Avicel, 15.1 mM on switchgrass). These engineered strains produce cellulosic ethanol at the highest temperature of any microorganism to date. In addition, the addition of 40 mM MOPS to the growth medium increased the maximal growth yield of C. bescii by approximately twofold. In conclusion, the utilization of thermostable enzymes will be critical to achieving high temperature CBP in bacteria such as C. bescii. The ability to produce ethanol at 75 °C, near its boiling point, raises the possibility that process optimization could allow in situ product removal of this end product to mitigate ethanol toxicity.« less

ASSESSING OF HERBIVOROUS AND BENEFICIAL INSECTS ON SWITCHGRASS IN UKRAINE.

PubMed

Stefanovska, T; Kucherovska, S; Pisdlisnyuk, V

2014-01-01

A perennial switchgrass, (Panicum virgatum L.), (C4) that is native to North America has good potential for biomass production because of its wide geographic distribution and adaptability to diverse environmental conditions. Insects can significantly impact the yield and quality of biofuel crops. If switchgrass are to be grown on marginally arable land or in monoculture, it are likely to be plagued with herbivore pests and plant diseases at a rate that exceeds what would be expected if the plants were not stressed in this manner. This biofuel crop has been under evaluation for commercial growing in Ukraine for eight years. However, insect diversity and the potential impact of pests on biomass production of this feedstock have not been accessed yet. The objective of our study, started in 2011, is a survey of switch grass insects by trophic groups and determine species that have pest status at two sites in the Central part of Ukraine (Kiev and Poltava regions). In Poltava site we investigated the effect of nine varieties of switchgrass (lowland and upland) to insects' diversity. We assessed changes over time in the densities of major insects' trophic groups, identifying potential pests and natural enemies. Obtained results indicates that different life stages of herbivorous insects from Hymenoptera, Homoptera, Diptera and Coleoptera orders were present on switchgrass during the growing season. Our study results suggests that choice of variety has an impact on trophic groups' structure and number of insects from different orders on swicthgrass. Herbivores and beneficial insects were the only groups that showed significant differences across sampling dates. The highest population of herbivores insects we recorded on 'Alamo' variety for studied years, although herbivore diversity tended to increase on 'Shelter', 'Alamo' and 'Cave-in-Rock' during 2012 and 2013. 'Dacotah', 'Nebraska', 'Sunburst', 'Forestburg' and 'Carthage' showed the highest level of beneficial insects during our study.

Cellulosic ethanol production via consolidated bioprocessing at 75 °C by engineered Caldicellulosiruptor bescii

DOE PAGES

Chung, Daehwan; Cha, Minseok; Snyder, Elise N.; ...

2015-10-06

In this paper, we report that the C. bescii genome does not encode an acetaldehyde/alcohol dehydrogenase or an acetaldehyde dehydrogenase and no ethanol production is detected in this strain. The recent introduction of an NADH-dependent AdhE from C. thermocellum (Fig. 1a) in an ldh mutant of this strain resulted in production of ethanol from un-pretreated switchgrass, but the thermolability of the C. thermocellum AdhE at the optimum growth temperature of C. bescii (78 °C) meant that ethanol was not produced above 65 °C. The adhB and adhE genes from Thermoanaerobacter pseudethanolicus 39E, an anaerobic thermophile that produces ethanol as amore » major fermentation product at 70 °C, were cloned and expressed in an ldh deletion mutant of C. bescii. The engineered strains produced ethanol at 75 °C, near the ethanol boiling point. The AdhB expressing strain produced ethanol (1.4 mM on Avicel, 0.4 mM on switchgrass) as well as acetate (13.0 mM on Avicel, 15.7 mM on switchgrass). The AdhE expressing strain produced more ethanol (2.3 mM on Avicel, 1.6 mM on switchgrass) and reduced levels of acetate (12.3 mM on Avicel, 15.1 mM on switchgrass). These engineered strains produce cellulosic ethanol at the highest temperature of any microorganism to date. In addition, the addition of 40 mM MOPS to the growth medium increased the maximal growth yield of C. bescii by approximately twofold. In conclusion, the utilization of thermostable enzymes will be critical to achieving high temperature CBP in bacteria such as C. bescii. The ability to produce ethanol at 75 °C, near its boiling point, raises the possibility that process optimization could allow in situ product removal of this end product to mitigate ethanol toxicity.« less

Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions

DOE PAGES

Yu, Chaowei; Reddy, Amitha P.; Simmons, Christopher W.; ...

2015-12-02

Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methodsmore » included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under thermophilic and high-solid conditions. The results suggest that the presence of microorganisms may be more important than their relative abundance in retaining an active microbial community.« less

Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

PubMed Central

Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

2016-01-01

The development of a range of sub-proteomic approaches to the plant cell wall has identified many of the cell wall proteins. However, it remains difficult to elucidate the precise biological role of each protein and the cell wall dynamics driven by their actions. The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. It therefore offers a unique opportunity to investigate the de novo construction process of the cell wall. This review deals with sub-proteomic approaches to the plant cell wall through the use of protoplasts, a methodology that will provide the basis for further exploration of cell wall proteins and cell wall dynamics. PMID:28248244

Carbon Calculator for Land Use Change from Biofuels Production (CCLUB) Users’ Manual and Technical Documentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, Jennifer B.; Qin, Zhangcai; Mueller, Steffen

Themore » $$\\underline{C}$$arbon $$\\underline{C}$$alculator for $$\\underline{L}$$and $$\\underline{U}$$se Change from $$\\underline{B}$$iofuels Production (CCLUB) calculates carbon emissions from land use change (LUC) for four different ethanol production pathways including corn grain ethanol and cellulosic ethanol from corn stover, Miscanthus, and switchgrass. This document discusses the version of CCLUB released September 30, 2014 which includes corn and three cellulosic feedstocks: corn stover, Miscanthus, and switchgrass.« less

Switchgrass ubiquitin promoter (PVUBI2) and uses thereof

DOEpatents

Stewart, C. Neal; Mann, David George James

2013-12-10

The subject application provides polynucleotides, compositions thereof and methods for regulating gene expression in a plant. Polynucleotides disclosed herein comprise novel sequences for a promoter isolated from Panicum virgatum (switchgrass) that initiates transcription of an operably linked nucleotide sequence. Thus, various embodiments of the invention comprise the nucleotide sequence of SEQ ID NO: 2 or fragments thereof comprising nucleotides 1 to 692 of SEQ ID NO: 2 that are capable of driving the expression of an operably linked nucleic acid sequence.

Characterization of the Sclerotinia sclerotiorum cell wall proteome.

PubMed

Liu, Longzhou; Free, Stephen J

2016-08-01

We used a proteomic analysis to identify cell wall proteins released from Sclerotinia sclerotiorum hyphal and sclerotial cell walls via a trifluoromethanesulfonic acid (TFMS) digestion. Cell walls from hyphae grown in Vogel's glucose medium (a synthetic medium lacking plant materials), from hyphae grown in potato dextrose broth and from sclerotia produced on potato dextrose agar were used in the analysis. Under the conditions used, TFMS digests the glycosidic linkages in the cell walls to release intact cell wall proteins. The analysis identified 24 glycosylphosphatidylinositol (GPI)-anchored cell wall proteins and 30 non-GPI-anchored cell wall proteins. We found that the cell walls contained an array of cell wall biosynthetic enzymes similar to those found in the cell walls of other fungi. When comparing the proteins in hyphal cell walls grown in potato dextrose broth with those in hyphal cell walls grown in the absence of plant material, it was found that a core group of cell wall biosynthetic proteins and some proteins associated with pathogenicity (secreted cellulases, pectin lyases, glucosidases and proteases) were expressed in both types of hyphae. The hyphae grown in potato dextrose broth contained a number of additional proteins (laccases, oxalate decarboxylase, peroxidase, polysaccharide deacetylase and several proteins unique to Sclerotinia and Botrytis) that might facilitate growth on a plant host. A comparison of the proteins in the sclerotial cell wall with the proteins in the hyphal cell wall demonstrated that sclerotia formation is not marked by a major shift in the composition of cell wall protein. We found that the S. sclerotiorum cell walls contained 11 cell wall proteins that were encoded only in Sclerotinia and Botrytis genomes. © 2015 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.

Water quality and quantity in the context of large-scale cellulosic biofuel production in the Mississippi-Atchafalaya River Basin

NASA Astrophysics Data System (ADS)

VanLoocke, A.; Bernacchi, C. J.; Twine, T. E.; Kucharik, C. J.

2012-12-01

Numerous socio-economic and environmental pressures have driven the need to increase domestic renewable energy production in the Midwest. The primary attempt at addressing this need has been to use maize; however, the leaching of residual nitrate from maize fertilizer into runoff drives the formation of the Gulf of Mexico hypoxic or "Dead" zone which can have significant environmental impacts on the marine ecosystems. As a result of the threat to benthic organisms and fisheries in this region, The Mississippi Basin/Gulf of Mexico Task Force has set in place goals to reduce the size of the hypoxic zone from the current size of ~ 20,000 km2 to < 5000 km2 by the year. It is predicted that annual dissolved inorganic nitrate (DIN) export would have to decrease by 30 to 55% to meet this goal. An alternative option to meet the renewable energy needs while reducing the environmental impacts associated with DIN export is to produce high-yielding, low fertilizer input perennial grasses such as switchgrass and miscanthus. Miscanthus and switchgrass have been shown to greatly reduce nitrate leaching at the plot scale, even during the establishment phase. This reduction in leaching is attributed to the perennial nature and the efficient recycling of nutrients via nutrient translocation. While these feedstocks are able to achieve higher productivity than maize grain with fewer inputs, they require more water, presenting the potential for environmental impacts on regional hydrologic cycle, including reductions in streamflow. The goal of this research is to determine the change in streamflow in the Mississippi-Atchafalaya River Basin (MARB) and the export of nitrogen from fertilizer to the Gulf of Mexico. To address this goal, we adapted a vegetation model capable of simulating the biogeochemistry of current crops as well as miscanthus and switchgrass, the Integrated Biosphere Simulator - agricultural version (Agro-IBIS) and coupled it with a hydrology model capable of simulating streamflow and nitrogen export, the Terrestrial Hydrology Model with Biogeochemistry. Simulations were conducted at varying fertilizer application rates and fraction coverages of miscanthus and switchgrass across the MARB. Data analysis indicated that there were reductions in runoff and streamflow throughout the MARB, with the largest differences occurring in drier portions of the regions. However differences in streamflow were only statistically resolved when miscanthus production was above 25% coverage and switchgrass above 35%. Compared to streamflow, statistically significant reductions in nitrogen export occurred at lower percent coverage, with unfertilized miscanthus having significant reductions at 10% and switchgrass at 25% coverage respectively, however this effect was smaller at higher fertilizer application rates. These results indicate that, given targeted management strategies, there is potential for miscanthus and switchgrass to provide key ecosystem services by reducing the export of DIN, while avoiding hydrologic impacts of reduced streamflow.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mei, Chuansheng; Nowak, Jerzy; Seiler, John

Switchgrass represents a promising feedstock crop for US energy sustainability. However, its broad utilization for bioenergy requires improvements of biomass yields and stress tolerance. In this DOE funded project, we have been working on harnessing beneficial bacterial endophytes to enhance switchgrass performance and to develop a low input feedstock production system for marginal lands that do not compete with the production of food crops. We have demonstrated that one of most promising plant growth-promoting bacterial endophytes, Burkholderia phytofirmans strain PsJN, is able to colonize roots and significantly promote growth of switchgrass cv. Alamo under in vitro, growth chamber, greenhouse, asmore » well as field conditions. Furthermore, PsJN bacterization improved growth and development of switchgrass seedlings, significantly stimulated plant root and shoot growth, and tiller number in the field, and enhanced biomass accumulation on both poor (p<0.001) and rich (p<0.05) soils, with more effective stimulation of plant growth in low fertility soil. Plant physiology measurements showed that PsJN inoculated Alamo had consistently lower transpiration, lower stomatal conductance, and higher water use efficiency in greenhouse conditions. These physiological changes may significantly contribute to the recorded growth enhancement. PsJN inoculation rapidly results in an increase in photosynthetic rates which contributes to the advanced growth and development. Some evidence suggests that this initial growth advantage decreases with time when resources are not limited such as in greenhouse studies. Additionally, better drought resistance and drought hardening were observed in PsJN inoculated switchgrass. Using the DOE-funded switchgrass EST microarray, in a collaboration with the Genomics Core Facility at the Noble Foundation, we have determined gene expression profile changes in both responsive switchgrass cv. Alamo and non-responsive cv. Cave-in-Rock (CR) following PsJN bacterization. With the MapMan software to analyze microarray data, the number of up- and down-regulated probes was calculated. The number of up-regulated probes in Alamo was 26, 14, 14, and 12% at 0.5, 2, 4 and 8 days after inoculation (DAI) with PsJN, respectively while the corresponding number in CR was 24, 22, 21, and 19%, respectively. In both cultivars, the largest number of up-regulated probes occurred at 0.5 DAI. Noticeable differences throughout the timeframe between Alamo and CR were that the number was dramatically decreased to half (12%) in Alamo but remained high in CR (approximately 20%). The number of down regulated genes demonstrated different trends in Alamo and CR. Alamo had an increasing trend from 9% at 0.5 DAI to 11, 17, and 28% at 2, 4, and 8 DAI, respectively. However, CR had 13% at 0.5 and 2 DAI, and declined to 10% at 4 and 8 DAI. With the aid of MapMan and PageMan, we mapped the response of the ID probes to the observed major gene regulatory network and major biosynthetic pathway changes associated with the beneficial bacterial endophyte infection, colonization, and early growth promotion process. We found significant differences in gene expression patterns between responsive and non-responsive cultivars in many pathways, including redox state regulation, signaling, proteolysis, transcription factors, as well as hormone (SA and JA in particular)-associated pathways. Form microarray data, a total of 50 key genes have been verified using qPCR. Ten of these genes were chosen for further functional study via either overexpression and/or RNAi knockout technologies. These genes were calmodulin-related calcium sensor protein (CAM), glutathione S-transferase (GST), histidine-containing phosphotransfer protein (H-221), 3 different zinc finger proteins (ZF-371, ZF131 and ZF242), EF hand transcription factor (EF-622), peroxidase, cellulose synthase catalytic submit A2 (CESA2), and Aux/IAA family. A total of 8 overexpression and 5 RNAi transgenic plants have been regenerated, and their gene expression levels determined using qPCR. Consequently high, medium and low expression lines were propagated in vitro for gene function study. When adequate numbers of individual transgenic lines were obtained, they were challenged with PsJN to see if PsJN promotes or inhibits growth of transgenic plants. Our results demonstrated that EF-622 overexpression, ZF-371, GST, H-221 and CAM RNAi transgenic lines lost responses to PsJN, i.e. PsJN had no growth promotive effects on these transgenic plants. Further study needs to be done to characterize this loss of responsiveness to PsJN. During this funding period, we have done more work related to this funded project and established collaborations with other institutions and obtained some interesting results, building a foundation for further research projects. For example, we isolated a naturally-occurring bacterium from surface-sterilized switchgrass seeds, identified as a unique Panteoa agglomerans species, and named strain PaKM. PaKM has been proved to be an efficient growth promoter of switchgrass over a broad spectrum of genotypes and has potential in applications with low input and sustainable production systems on marginal lands. In collaboration with Dr. Shuijin Hu (North Carolina State University), we conducted experiments on how endophyte-inoculated switchgrass affects soil N and P availability and the number of AMF in roots. Our preliminary results showed that PsJN increased AMF infection of switchgrass roots, and enhanced soil N availability and soil N mineralization on a low nutrient field. Further study of this phenomenon on different soils, over longer time periods, is needed to assess its potential impact on the productivity and longevity of switchgrass stands.« less

The Role of Auxin in Cell Wall Expansion

PubMed Central

2018-01-01

Plant cells are surrounded by cell walls, which are dynamic structures displaying a strictly regulated balance between rigidity and flexibility. Walls are fairly rigid to provide support and protection, but also extensible, to allow cell growth, which is triggered by a high intracellular turgor pressure. Wall properties regulate the differential growth of the cell, resulting in a diversity of cell sizes and shapes. The plant hormone auxin is well known to stimulate cell elongation via increasing wall extensibility. Auxin participates in the regulation of cell wall properties by inducing wall loosening. Here, we review what is known on cell wall property regulation by auxin. We focus particularly on the auxin role during cell expansion linked directly to cell wall modifications. We also analyze downstream targets of transcriptional auxin signaling, which are related to the cell wall and could be linked to acid growth and the action of wall-loosening proteins. All together, this update elucidates the connection between hormonal signaling and cell wall synthesis and deposition. PMID:29565829

The Role of Auxin in Cell Wall Expansion.

PubMed

Majda, Mateusz; Robert, Stéphanie

2018-03-22

Plant cells are surrounded by cell walls, which are dynamic structures displaying a strictly regulated balance between rigidity and flexibility. Walls are fairly rigid to provide support and protection, but also extensible, to allow cell growth, which is triggered by a high intracellular turgor pressure. Wall properties regulate the differential growth of the cell, resulting in a diversity of cell sizes and shapes. The plant hormone auxin is well known to stimulate cell elongation via increasing wall extensibility. Auxin participates in the regulation of cell wall properties by inducing wall loosening. Here, we review what is known on cell wall property regulation by auxin. We focus particularly on the auxin role during cell expansion linked directly to cell wall modifications. We also analyze downstream targets of transcriptional auxin signaling, which are related to the cell wall and could be linked to acid growth and the action of wall-loosening proteins. All together, this update elucidates the connection between hormonal signaling and cell wall synthesis and deposition.

Localization of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4,6-trinitrotoluene (TNT) in poplar and switchgrass plants using phosphor imager autoradiography.

PubMed

Brentner, Laura B; Mukherji, Sachiyo T; Walsh, Susan A; Schnoor, Jerald L

2010-02-01

Phosphor imager autoradiography is a technique for rapid, sensitive analysis of the localization of xenobiotics in plant tissues. Use of this technique is relatively new to research in the field of plant science, and the potential for enhancing visualization and understanding of plant uptake and transport of xenobiotics remains largely untapped. Phosphor imager autoradiography is used to investigate the uptake and translocation of the explosives 1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4,6-trinitrotoluene within Populus deltoides x nigra DN34 (poplar) and Panicum vigratum Alamo (switchgrass). In both plant types, TNT and/or TNT-metabolites remain predominantly in root tissues while RDX and/or RDX-metabolites are readily translocated to leaf tissues. Phosphor imager autoradiography is further investigated for use in semi-quantitative analysis of uptake of TNT by switchgrass. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Starace, Anne K.; Evans, Robert J.; Lee, David D.

A variety of hardwood, softwood, and herbaceous feedstocks (oak, southern yellow pine mix, loblolly pine, pinyon-juniper mix, and switchgrass) were each torrefied at 200, 250, and 300 °C. Each of the feedstocks was pyrolyzed and the resulting vapors were analyzed with a molecular beam mass spectrometer (py-MBMS). Compositional analysis was used to measure the total lignin content of three of the feedstocks (southern yellow pine, softwood; oak, hardwood; and switchgrass, herbaceous) before and after torrefaction at 300 °C, and large differences in the fraction of lignin lost during torrefaction were found between feedstocks, with oak having the largest decrease inmore » lignin during torrefaction and switchgrass having the least. Finally, it is hypothesized that these differences in the thermal degradation are due to, in part, the different ratios of S, G, and H lignins in the feedstocks. Additionally, the torrefaction of kraft lignin was studied using thermogravimetric analysis coupled with Fourier transform infrared spectroscopy (TGA-FTIR) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR).« less

Industrial hemp as a potential bioenergy crop in comparison with kenaf, switchgrass and biomass sorghum.

PubMed

Das, Lalitendu; Liu, Enshi; Saeed, Areej; Williams, David W; Hu, Hongqiang; Li, Chenlin; Ray, Allison E; Shi, Jian

2017-11-01

This study takes combined field trial, lab experiment, and economic analysis approaches to evaluate the potential of industrial hemp in comparison with kenaf, switchgrass and biomass sorghum. Agronomy data suggest that the per hectare yield (5437kg) of industrial hemp stem alone was at a similar level with switchgrass and sorghum; while the hemp plants require reduced inputs. Field trial also showed that ∼1230kg/ha hemp grain can be harvested in addition to stems. Results show a predicted ethanol yield of ∼82gallons/dry ton hemp stems, which is comparable to the other three tested feedstocks. A comparative cost analysis indicates that industrial hemp could generate higher per hectare gross profit than the other crops if both hemp grains and biofuels from hemp stem were counted. These combined evaluation results demonstrate that industrial hemp has great potential to become a promising regional commodity crop for producing both biofuels and value-added products. Copyright © 2017 Elsevier Ltd. All rights reserved.

Influence of Weather on the Predicted Moisture Content of Field Chopped EnergySorghum and Switchgrass

DOE PAGES

Popp, Michael P.; Searcy, Stephen S.; Sokhansanj, Shahab; ...

2015-03-25

To determine the effects of weather on harvested moisture content (MC) of switchgrass (Panicum virgatum) and energy sorghum (Sorghum bicolor), tracking of harvest progress on individual fields in the Integrated Biomass Supply and Logistics (IBSAL) model was modified to allow: i) rewetting of swathed material in the drying formulae; and ii) field queuing rules based on equipment availability and weather. Estimated crop yield and initial MC by harvest date, as observed in field trials, along with the modeling of different delays between mowing and harvest allowed estimation of harvested MC, annual tonnage processed and associated processing cost differences by cropmore » and location over 10 years. Extending the hours of annual equipment use had minor implications on cost of production. Energy sorghum proved difficult to dry in the field. Its higher yield, leading to shorter supply distance to the plant, may justify harvesting of energy sorghum early in the season with drier weather. Lastly, later harvest for lower-yielding switchgrass offers MC advantages.« less

Influence of Weather on the Predicted Moisture Content of Field Chopped EnergySorghum and Switchgrass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Popp, Michael P.; Searcy, Stephen S.; Sokhansanj, Shahab

To determine the effects of weather on harvested moisture content (MC) of switchgrass (Panicum virgatum) and energy sorghum (Sorghum bicolor), tracking of harvest progress on individual fields in the Integrated Biomass Supply and Logistics (IBSAL) model was modified to allow: i) rewetting of swathed material in the drying formulae; and ii) field queuing rules based on equipment availability and weather. Estimated crop yield and initial MC by harvest date, as observed in field trials, along with the modeling of different delays between mowing and harvest allowed estimation of harvested MC, annual tonnage processed and associated processing cost differences by cropmore » and location over 10 years. Extending the hours of annual equipment use had minor implications on cost of production. Energy sorghum proved difficult to dry in the field. Its higher yield, leading to shorter supply distance to the plant, may justify harvesting of energy sorghum early in the season with drier weather. Lastly, later harvest for lower-yielding switchgrass offers MC advantages.« less

Cell Wall Remodeling Enzymes Modulate Fungal Cell Wall Elasticity and Osmotic Stress Resistance

PubMed Central

Ene, Iuliana V.; Walker, Louise A.; Schiavone, Marion; Lee, Keunsook K.; Martin-Yken, Hélène; Dague, Etienne; Gow, Neil A. R.; Munro, Carol A.

2015-01-01

ABSTRACT The fungal cell wall confers cell morphology and protection against environmental insults. For fungal pathogens, the cell wall is a key immunological modulator and an ideal therapeutic target. Yeast cell walls possess an inner matrix of interlinked β-glucan and chitin that is thought to provide tensile strength and rigidity. Yeast cells remodel their walls over time in response to environmental change, a process controlled by evolutionarily conserved stress (Hog1) and cell integrity (Mkc1, Cek1) signaling pathways. These mitogen-activated protein kinase (MAPK) pathways modulate cell wall gene expression, leading to the construction of a new, modified cell wall. We show that the cell wall is not rigid but elastic, displaying rapid structural realignments that impact survival following osmotic shock. Lactate-grown Candida albicans cells are more resistant to hyperosmotic shock than glucose-grown cells. We show that this elevated resistance is not dependent on Hog1 or Mkc1 signaling and that most cell death occurs within 10 min of osmotic shock. Sudden decreases in cell volume drive rapid increases in cell wall thickness. The elevated stress resistance of lactate-grown cells correlates with reduced cell wall elasticity, reflected in slower changes in cell volume following hyperosmotic shock. The cell wall elasticity of lactate-grown cells is increased by a triple mutation that inactivates the Crh family of cell wall cross-linking enzymes, leading to increased sensitivity to hyperosmotic shock. Overexpressing Crh family members in glucose-grown cells reduces cell wall elasticity, providing partial protection against hyperosmotic shock. These changes correlate with structural realignment of the cell wall and with the ability of cells to withstand osmotic shock. PMID:26220968

DOE Office of Scientific and Technical Information (OSTI.GOV)

Raz-Yaseef, Naama; Billesbach, Dave P.; Fischer, Marc L.

The Southern Great Plains are characterized by a fine-scale mixture of different land-cover types, predominantly winter-wheat and grazed pasture, with relatively small areas of other crops, native prairie, and switchgrass. Recent droughts and predictions of increased drought in the Southern Great Plains, especially during the summer months, raise concern for these ecosystems. We measured ecosystem carbon and water fluxes with eddy-covariance systems over cultivated cropland for 10 years, and over lightly grazed prairie and new switchgrass fields for 2 years each. Growing-season precipitation showed the strongest control over net carbon uptake for all ecosystems, but with a variable effect: grassesmore » (prairie and switchgrass) needed at least 350 mm of precipitation during the growing season to become net carbon sinks, while crops needed only 100 mm. In summer, high temperatures enhanced evaporation and led to higher likelihood of dry soil conditions. Therefore, summer-growing native prairie species and switchgrass experienced more seasonal droughts than spring-growing crops. For wheat, the net reduction in carbon uptake resulted mostly from a decrease in gross primary production rather than an increase in respiration. Flux measurements suggested that management practices for crops were effective in suppressing evapotranspiration and decomposition (by harvesting and removing secondary growth), and in increasing carbon uptake (by fertilizing and conserving summer soil water). In light of future projections for wetter springs and drier and warmer summers in the Southern Great Plains, our study indicates an increased vulnerability in native ecosystems and summer crops over time.« less

Regional Climate Implications of Large-scale Cultivation of Biofuel Crops

NASA Astrophysics Data System (ADS)

Rowe, C. M.; Oglesby, R. J.; Hays, C. J.; van Etten, A. R.

2008-12-01

Conversion from corn-based ethanol to cellulosic ethanol has the potential to dramatically alter the production of biofuels in the United States and could result in large-scale changes in the agricultural landscape of vast areas of the country. Regions currently dominated by corn production could see widespread planting of switchgrass and other fast-growing, water-efficient sources of cellulose biomass. An often overlooked side effect of these land-cover changes could be a significant alteration of the energy fluxes between the land surface and the atmosphere with profound local, regional, and continental impacts on the climate system. Changes in the surface energy balance result primarily from differences in the seasonality of transpiration from corn versus switchgrass and could be enhanced as a result of a reduced need for irrigation of switchgrass in areas where corn can be produced only under irrigation. Preliminary modeling results using a simple "bucket" land surface model coupled to the WRF mesoscale model have demonstrated increases in summertime average daily maximum temperature of up to 4° C, smaller increases of up to 2° C in nighttime minimum temperatures and reductions in precipitation by up to 25% when corn was changed to switchgrass over the central United States. Improved parameterization of biofuel crops in more sophisticated land surface models will allow us to refine these preliminary estimates and assess the impacts of large-scale conversion to cellulosic biofuel crops, relative to greenhouse gas induced regional climate change.

Nitrogen fertilization challenges the climate benefit of cellulosic biofuels

DOE PAGES

Ruan, Leilei; Bhardwaj, Ajay K.; Hamilton, Stephen K.; ...

2016-06-01

Cellulosic biofuels are intended to improve future energy and climate security. Nitrogen (N) fertilizer is commonly recommended to stimulate yields but can increase losses of the greenhouse gas nitrous oxide (N 2O) and other forms of reactive N, including nitrate. We measured soil N2O emissions and nitrate leaching along a switchgrass ( Panicum virgatum) high resolution N-fertilizer gradient for three years post-establishment. Results revealed an exponential increase in annual N2O emissions that each year became stronger (R 2 > 0.9, P < 0.001) and deviated further from the fixed percentage assumed for IPCC Tier 1 emission factors. Concomitantly, switchgrass yieldsmore » became less responsive each year to N fertilizer. Nitrate leaching (and calculated indirect N2O emissions) also increased exponentially in response to N inputs, but neither methane (CH4) uptake nor soil organic carbon changed detectably. Overall, N fertilizer inputs at rates greater than crop need curtailed the climate benefit of ethanol production almost two-fold, from a maximum mitigation capacity of–5.71 ± 0.22 Mg CO 2e ha –1 yr –1 in switchgrass fertilized at 56 kgNha –1 to only –2.97 ± 0.18 MgCO 2e ha –1 yr –1 in switchgrass fertilized at 196 kgNha –1. In conclusion, minimizing N fertilizer use will be an important strategy for fully realizing the climate benefits of cellulosic biofuel production.« less

Analysis on storage off-gas emissions from woody, herbaceous, and torrefied biomass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tumuluru, Jaya Shankar; Lim, C. Jim; Bi, Xiaotao T.

Wood chips, torrefied wood chips, ground switchgrass, and wood pellets were tested for off-gas emissions during storage. Storage canisters with gas-collection ports were used to conduct experiments at room temperature of 20 °C and in a laboratory oven set at 40 °C. Commercially-produced wood pellets yielded the highest carbon monoxide (CO) emissions at both 20 and 40 °C (1600 and 13,000 ppmv), whereas torrefied wood chips emitted the lowest of about <200 and <2000 ppmv. Carbon dioxide (CO₂) emissions from wood pellets were 3000 ppmv and 42,000 ppmv, whereas torrefied wood chips registered at about 2000 and 25,000 ppmv, atmore » 20 and 40 °C at the end of 11 days of storage. CO emission factors (milligrams per kilogram of biomass) calculated were lowest for ground switchgrass and torrefied wood chips (2.68 and 4.86 mg/kg) whereas wood pellets had the highest CO of about 10.60 mg/kg, respectively, at 40 °C after 11 days of storage. In the case of CO₂, wood pellets recorded the lowest value of 55.46 mg/kg, whereas switchgrass recorded the highest value of 318.72 mg/kg. This study concludes that CO emission factor is highest for wood pellets, CO₂ is highest for switchgrass and CH₄ is negligible for all feedstocks except for wood pellets, which is about 0.374 mg/kg at the end of 11-day storage at 40 °C.« less

Analysis on storage off-gas emissions from woody, herbaceous, and torrefied biomass

DOE PAGES

Tumuluru, Jaya Shankar; Lim, C. Jim; Bi, Xiaotao T.; ...

2015-03-02

Wood chips, torrefied wood chips, ground switchgrass, and wood pellets were tested for off-gas emissions during storage. Storage canisters with gas-collection ports were used to conduct experiments at room temperature of 20 °C and in a laboratory oven set at 40 °C. Commercially-produced wood pellets yielded the highest carbon monoxide (CO) emissions at both 20 and 40 °C (1600 and 13,000 ppmv), whereas torrefied wood chips emitted the lowest of about <200 and <2000 ppmv. Carbon dioxide (CO₂) emissions from wood pellets were 3000 ppmv and 42,000 ppmv, whereas torrefied wood chips registered at about 2000 and 25,000 ppmv, atmore » 20 and 40 °C at the end of 11 days of storage. CO emission factors (milligrams per kilogram of biomass) calculated were lowest for ground switchgrass and torrefied wood chips (2.68 and 4.86 mg/kg) whereas wood pellets had the highest CO of about 10.60 mg/kg, respectively, at 40 °C after 11 days of storage. In the case of CO₂, wood pellets recorded the lowest value of 55.46 mg/kg, whereas switchgrass recorded the highest value of 318.72 mg/kg. This study concludes that CO emission factor is highest for wood pellets, CO₂ is highest for switchgrass and CH₄ is negligible for all feedstocks except for wood pellets, which is about 0.374 mg/kg at the end of 11-day storage at 40 °C.« less

Nitrogen fertilization challenges the climate benefit of cellulosic biofuels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ruan, Leilei; Bhardwaj, Ajay K.; Hamilton, Stephen K.

Cellulosic biofuels are intended to improve future energy and climate security. Nitrogen (N) fertilizer is commonly recommended to stimulate yields but can increase losses of the greenhouse gas nitrous oxide (N 2O) and other forms of reactive N, including nitrate. We measured soil N2O emissions and nitrate leaching along a switchgrass ( Panicum virgatum) high resolution N-fertilizer gradient for three years post-establishment. Results revealed an exponential increase in annual N2O emissions that each year became stronger (R 2 > 0.9, P < 0.001) and deviated further from the fixed percentage assumed for IPCC Tier 1 emission factors. Concomitantly, switchgrass yieldsmore » became less responsive each year to N fertilizer. Nitrate leaching (and calculated indirect N2O emissions) also increased exponentially in response to N inputs, but neither methane (CH4) uptake nor soil organic carbon changed detectably. Overall, N fertilizer inputs at rates greater than crop need curtailed the climate benefit of ethanol production almost two-fold, from a maximum mitigation capacity of–5.71 ± 0.22 Mg CO 2e ha –1 yr –1 in switchgrass fertilized at 56 kgNha –1 to only –2.97 ± 0.18 MgCO 2e ha –1 yr –1 in switchgrass fertilized at 196 kgNha –1. In conclusion, minimizing N fertilizer use will be an important strategy for fully realizing the climate benefits of cellulosic biofuel production.« less

An Analysis of the Climate Change Mitigation Potential through Soil Organic Carbon Sequestration in a Corn Belt Watershed

NASA Astrophysics Data System (ADS)

Bhattarai, M. D.; Secchi, S.; Schoof, J. T.

2015-12-01

The sequestration of carbon constitutes one of major options in agricultural climate change land-based mitigation. We examined the carbon sequestration potential of alternative agricultural land uses in an intensively farmed Corn Belt watershed. We Used downscaled data from eight atmosphere-ocean general circulation models (AOGCMs) for a simulation period between 2015 and 2099 with three emission pathways reflecting low, medium and high greenhouse gas scenarios. The use of downscaled data, coupled with high resolution land use and soil data, can help policy makers and land managers better understand spatial and temporal impacts of climate change. We consider traditional practices such as no-till corn-soybean rotations and continuous corn and include also switchgrass, a bioenergy crop. Our results show that switching from conventional tillage continuous corn to no-till corn-soybean can sequester the equivalent of 156,000 MtCO2 of soil organic carbon with a sequestration rate of 2.38 MtCO2 ha-1 yr-1 for the simulated period. Our results also indicate that switchgrass can sequester the equivalent of 282,000 MtCO2 of soil organic carbon with a sequestration rate of 4.4 MtCO2 ha-1 yr-1 for the period. Our finding also suggests that while climate change impacts corn and soybean yields, it does not have a significant effect on switchgrass yields possibly due to carbon fertilization effect on switchgrass yields.

Effects of precipitation changes on switchgrass photosynthesis, growth, and biomass: A mesocosm experiment

PubMed Central

Yu, Chih-Li; Deng, Qi; Dzantor, E. Kudjo; Zhou, Suping; Dennis, Sam; Sauve, Roger; Johnson, Terrance L.; Fay, Philip A.; Shen, Weijun; Luo, Yiqi

2018-01-01

Climate changes, including chronic changes in precipitation amounts, will influence plant physiology and growth. However, such precipitation effects on switchgrass, a major bioenergy crop, have not been well investigated. We conducted a two-year precipitation simulation experiment using large pots (95 L) in an environmentally controlled greenhouse in Nashville, TN. Five precipitation treatments (ambient precipitation, and -50%, -33%, +33%, and +50% of ambient) were applied in a randomized complete block design with lowland "Alamo" switchgrass plants one year after they were established from tillers. The growing season progression of leaf physiology, tiller number, height, and aboveground biomass were determined each growing season. Precipitation treatments significantly affected leaf physiology, growth, and aboveground biomass. The photosynthetic rates in the wet (+50% and +33%) treatments were significantly enhanced by 15.9% and 8.1%, respectively, than the ambient treatment. Both leaf biomass and plant height were largely increased, resulting in dramatically increases in aboveground biomass by 56.5% and 49.6% in the +50% and +33% treatments, respectively. Compared to the ambient treatment, the drought (-33% and -50%) treatments did not influence leaf physiology, but the -50% treatment significantly reduced leaf biomass by 37.8%, plant height by 16.3%, and aboveground biomass by 38.9%. This study demonstrated that while switchgrass in general is a drought tolerant grass, severe drought significantly reduces Alamo’s growth and biomass, and that high precipitation stimulates its photosynthesis and growth. PMID:29420600

Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends.

PubMed

Cianchetta, Stefano; Bregoli, Luca; Galletti, Stefania

2017-11-01

Giant reed, miscanthus, and switchgrass are considered prominent lignocellulosic feedstocks to obtain fermentable sugars for biofuel production. The bioconversion into sugars requires a delignifying pre-treatment step followed by hydrolysis with cellulase and other accessory enzymes like xylanase, especially in the case of alkali pre-treatments, which retain the hemicellulose fraction. Blends richer in accessory enzymes than commercial mix can be obtained growing fungi on feedstock-based substrates, thus ten selected Trichoderma isolates, including the hypercellulolytic strain Trichoderma reesei Rut-C30, were grown on giant reed, miscanthus, or switchgrass-based substrates. The produced enzymes were used to saccharify the corresponding feedstocks, compared to a commercial enzymatic mix (6 FPU/g). Feedstocks were acid (H 2 SO 4 0.2-2%, w/v) or alkali (NaOH 0.02-0.2%, w/v) pre-treated. A microplate-based approach was chosen for most of the experimental steps due to the large number of samples. The highest bioconversion was generally obtained with Trichoderma harzianum Or4/99 enzymes (78, 89, and 94% final sugar yields at 48 h for giant reed, miscanthus, and switchgrass, respectively), with significant increases compared to the commercial mix, especially with alkaline pre-treatments. The differences in bioconversion yields were only partially caused by xylanases (maximum R 2  = 0.5), indicating a role for other accessory enzymes.

Cell Wall Remodeling Enzymes Modulate Fungal Cell Wall Elasticity and Osmotic Stress Resistance.

PubMed

Ene, Iuliana V; Walker, Louise A; Schiavone, Marion; Lee, Keunsook K; Martin-Yken, Hélène; Dague, Etienne; Gow, Neil A R; Munro, Carol A; Brown, Alistair J P

2015-07-28

The fungal cell wall confers cell morphology and protection against environmental insults. For fungal pathogens, the cell wall is a key immunological modulator and an ideal therapeutic target. Yeast cell walls possess an inner matrix of interlinked β-glucan and chitin that is thought to provide tensile strength and rigidity. Yeast cells remodel their walls over time in response to environmental change, a process controlled by evolutionarily conserved stress (Hog1) and cell integrity (Mkc1, Cek1) signaling pathways. These mitogen-activated protein kinase (MAPK) pathways modulate cell wall gene expression, leading to the construction of a new, modified cell wall. We show that the cell wall is not rigid but elastic, displaying rapid structural realignments that impact survival following osmotic shock. Lactate-grown Candida albicans cells are more resistant to hyperosmotic shock than glucose-grown cells. We show that this elevated resistance is not dependent on Hog1 or Mkc1 signaling and that most cell death occurs within 10 min of osmotic shock. Sudden decreases in cell volume drive rapid increases in cell wall thickness. The elevated stress resistance of lactate-grown cells correlates with reduced cell wall elasticity, reflected in slower changes in cell volume following hyperosmotic shock. The cell wall elasticity of lactate-grown cells is increased by a triple mutation that inactivates the Crh family of cell wall cross-linking enzymes, leading to increased sensitivity to hyperosmotic shock. Overexpressing Crh family members in glucose-grown cells reduces cell wall elasticity, providing partial protection against hyperosmotic shock. These changes correlate with structural realignment of the cell wall and with the ability of cells to withstand osmotic shock. The C. albicans cell wall is the first line of defense against external insults, the site of immune recognition by the host, and an attractive target for antifungal therapy. Its tensile strength is conferred by a network of cell wall polysaccharides, which are remodeled in response to growth conditions and environmental stress. However, little is known about how cell wall elasticity is regulated and how it affects adaptation to stresses such as sudden changes in osmolarity. We show that elasticity is critical for survival under conditions of osmotic shock, before stress signaling pathways have time to induce gene expression and drive glycerol accumulation. Critical cell wall remodeling enzymes control cell wall flexibility, and its regulation is strongly dependent on host nutritional inputs. We also demonstrate an entirely new level of cell wall dynamism, where significant architectural changes and structural realignment occur within seconds of an osmotic shock. Copyright © 2015 Ene et al.

A Thermophilic Ionic Liquid-Tolerant Cellulase Cocktail for the Production of Cellulosic Biofuels

PubMed Central

Park, Joshua I.; Steen, Eric J.; Burd, Helcio; Evans, Sophia S.; Redding-Johnson, Alyssa M.; Batth, Tanveer; Benke, Peter I.; D'haeseleer, Patrik; Sun, Ning; Sale, Kenneth L.; Keasling, Jay D.; Lee, Taek Soon; Petzold, Christopher J.; Mukhopadhyay, Aindrila; Singer, Steven W.; Simmons, Blake A.; Gladden, John M.

2012-01-01

Generation of biofuels from sugars in lignocellulosic biomass is a promising alternative to liquid fossil fuels, but efficient and inexpensive bioprocessing configurations must be developed to make this technology commercially viable. One of the major barriers to commercialization is the recalcitrance of plant cell wall polysaccharides to enzymatic hydrolysis. Biomass pretreatment with ionic liquids (ILs) enables efficient saccharification of biomass, but residual ILs inhibit both saccharification and microbial fuel production, requiring extensive washing after IL pretreatment. Pretreatment itself can also produce biomass-derived inhibitory compounds that reduce microbial fuel production. Therefore, there are multiple points in the process from biomass to biofuel production that must be interrogated and optimized to maximize fuel production. Here, we report the development of an IL-tolerant cellulase cocktail by combining thermophilic bacterial glycoside hydrolases produced by a mixed consortia with recombinant glycoside hydrolases. This enzymatic cocktail saccharifies IL-pretreated biomass at higher temperatures and in the presence of much higher IL concentrations than commercial fungal cocktails. Sugars obtained from saccharification of IL-pretreated switchgrass using this cocktail can be converted into biodiesel (fatty acid ethyl-esters or FAEEs) by a metabolically engineered strain of E. coli. During these studies, we found that this biodiesel-producing E. coli strain was sensitive to ILs and inhibitors released by saccharification. This cocktail will enable the development of novel biomass to biofuel bioprocessing configurations that may overcome some of the barriers to production of inexpensive cellulosic biofuels. PMID:22649505

Mathematical models of lignin biosynthesis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Faraji, Mojdeh; Fonseca, Luis; Escamilla-Trevino, Luis

Lignin is a natural polymer that is interwoven with cellulose and hemicellulose within plant cell walls. Due to this molecular arrangement, lignin is a major contributor to the recalcitrance of plant materials with respect to the extraction of sugars and their fermentation into ethanol, butanol, and other potential bioenergy crops. The lignin biosynthetic pathway is similar, but not identical in different plant species. It is in each case comprised of a moderate number of enzymatic steps, but its responses to manipulations, such as gene knock-downs, are complicated by the fact that several of the key enzymes are involved in severalmore » reaction steps. This feature poses a challenge to bioenergy production, as it renders it difficult to select the most promising combinations of genetic manipulations for the optimization of lignin composition and amount.Here, we present several computational models than can aid in the analysis of data characterizing lignin biosynthesis. While minimizing technical details, we focus on the questions of what types of data are particularly useful for modeling and what genuine benefits the biofuel researcher may gain from the resulting models. We demonstrate our analysis with mathematical models for black cottonwood (Populus trichocarpa), alfalfa (Medicago truncatula), switchgrass (Panicum virgatum) and the grass Brachypodium distachyon. Despite commonality in pathway structure, different plant species show different regulatory features and distinct spatial and topological characteristics. The putative lignin biosynthes pathway is not able to explain the plant specific laboratory data, and the necessity of plant specific modeling should be heeded.« less

Mathematical models of lignin biosynthesis

DOE PAGES

Faraji, Mojdeh; Fonseca, Luis; Escamilla-Trevino, Luis; ...

2018-02-09

Lignin is a natural polymer that is interwoven with cellulose and hemicellulose within plant cell walls. Due to this molecular arrangement, lignin is a major contributor to the recalcitrance of plant materials with respect to the extraction of sugars and their fermentation into ethanol, butanol, and other potential bioenergy crops. The lignin biosynthetic pathway is similar, but not identical in different plant species. It is in each case comprised of a moderate number of enzymatic steps, but its responses to manipulations, such as gene knock-downs, are complicated by the fact that several of the key enzymes are involved in severalmore » reaction steps. This feature poses a challenge to bioenergy production, as it renders it difficult to select the most promising combinations of genetic manipulations for the optimization of lignin composition and amount.Here, we present several computational models than can aid in the analysis of data characterizing lignin biosynthesis. While minimizing technical details, we focus on the questions of what types of data are particularly useful for modeling and what genuine benefits the biofuel researcher may gain from the resulting models. We demonstrate our analysis with mathematical models for black cottonwood (Populus trichocarpa), alfalfa (Medicago truncatula), switchgrass (Panicum virgatum) and the grass Brachypodium distachyon. Despite commonality in pathway structure, different plant species show different regulatory features and distinct spatial and topological characteristics. The putative lignin biosynthes pathway is not able to explain the plant specific laboratory data, and the necessity of plant specific modeling should be heeded.« less

A Transcriptomic Analysis of Xylan Mutants Does Not Support the Existence of a Secondary Cell Wall Integrity System in Arabidopsis

PubMed Central

Faria-Blanc, Nuno; Mortimer, Jenny C.; Dupree, Paul

2018-01-01

Yeast have long been known to possess a cell wall integrity (CWI) system, and recently an analogous system has been described for the primary walls of plants (PCWI) that leads to changes in plant growth and cell wall composition. A similar system has been proposed to exist for secondary cell walls (SCWI). However, there is little data to support this. Here, we analyzed the stem transcriptome of a set of cell wall biosynthetic mutants in order to investigate whether cell wall damage, in this case caused by aberrant xylan synthesis, activates a signaling cascade or changes in cell wall synthesis gene expression. Our data revealed remarkably few changes to the transcriptome. We hypothesize that this is because cells undergoing secondary cell wall thickening have entered a committed programme leading to cell death, and therefore a SCWI system would have limited impact. The absence of transcriptomic responses to secondary cell wall alterations may facilitate engineering of the secondary cell wall of plants. PMID:29636762

A Transcriptomic Analysis of Xylan Mutants Does Not Support the Existence of a Secondary Cell Wall Integrity System in Arabidopsis.

PubMed

Faria-Blanc, Nuno; Mortimer, Jenny C; Dupree, Paul

2018-01-01

Yeast have long been known to possess a cell wall integrity (CWI) system, and recently an analogous system has been described for the primary walls of plants (PCWI) that leads to changes in plant growth and cell wall composition. A similar system has been proposed to exist for secondary cell walls (SCWI). However, there is little data to support this. Here, we analyzed the stem transcriptome of a set of cell wall biosynthetic mutants in order to investigate whether cell wall damage, in this case caused by aberrant xylan synthesis, activates a signaling cascade or changes in cell wall synthesis gene expression. Our data revealed remarkably few changes to the transcriptome. We hypothesize that this is because cells undergoing secondary cell wall thickening have entered a committed programme leading to cell death, and therefore a SCWI system would have limited impact. The absence of transcriptomic responses to secondary cell wall alterations may facilitate engineering of the secondary cell wall of plants.

Differential histone modification and protein expression associated with cell wall removal and regeneration in rice (Oryza sativa).

PubMed

Tan, Feng; Zhang, Kangling; Mujahid, Hana; Verma, Desh Pal S; Peng, Zhaohua

2011-02-04

The cell wall is a critical extracellular structure that provides protection and structural support in plant cells. To study the biological function of the cell wall and the regulation of cell wall resynthesis, we examined cellular responses to enzymatic removal of the cell wall in rice (Oryza sativa) suspension cells using proteomic approaches. We find that removal of cell wall stimulates cell wall synthesis from multiple sites in protoplasts instead of from a single site as in cytokinesis. Nucleus DAPI stain and MNase digestion further show that removal of the cell wall is concomitant with substantial chromatin reorganization. Histone post-translational modification studies using both Western blots and isotope labeling assisted quantitative mass spectrometry analyses reveal that substantial histone modification changes, particularly H3K18(AC) and H3K23(AC), are associated with the removal and regeneration of the cell wall. Label-free quantitative proteome analyses further reveal that chromatin associated proteins undergo dramatic changes upon removal of the cell wall, along with cytoskeleton, cell wall metabolism, and stress-response proteins. This study demonstrates that cell wall removal is associated with substantial chromatin change and may lead to stimulation of cell wall synthesis using a novel mechanism.

Plant cell wall proteomics: the leadership of Arabidopsis thaliana

PubMed Central

Albenne, Cécile; Canut, Hervé; Jamet, Elisabeth

2013-01-01

Plant cell wall proteins (CWPs) progressively emerged as crucial components of cell walls although present in minor amounts. Cell wall polysaccharides such as pectins, hemicelluloses, and cellulose represent more than 90% of primary cell wall mass, whereas hemicelluloses, cellulose, and lignins are the main components of lignified secondary walls. All these polymers provide mechanical properties to cell walls, participate in cell shape and prevent water loss in aerial organs. However, cell walls need to be modified and customized during plant development and in response to environmental cues, thus contributing to plant adaptation. CWPs play essential roles in all these physiological processes and particularly in the dynamics of cell walls, which requires organization and rearrangements of polysaccharides as well as cell-to-cell communication. In the last 10 years, plant cell wall proteomics has greatly contributed to a wider knowledge of CWPs. This update will deal with (i) a survey of plant cell wall proteomics studies with a focus on Arabidopsis thaliana; (ii) the main protein families identified and the still missing peptides; (iii) the persistent issue of the non-canonical CWPs; (iv) the present challenges to overcome technological bottlenecks; and (v) the perspectives beyond cell wall proteomics to understand CWP functions. PMID:23641247

Cell Wall Structure of Coccoid Green Algae as an Important Trade-Off Between Biotic Interference Mechanisms and Multidimensional Cell Growth.

PubMed

Dunker, Susanne; Wilhelm, Christian

2018-01-01

Coccoid green algae can be divided in two groups based on their cell wall structure. One group has a highly chemical resistant cell wall (HR-cell wall) containing algaenan. The other group is more susceptible to chemicals (LR-cell wall - Low resistant cell wall). Algaenan is considered as important molecule to explain cell wall resistance. Interestingly, cell wall types (LR- and HR-cell wall) are not in accordance with the taxonomic classes Chlorophyceae and Trebouxiophyceae, which makes it even more interesting to consider the ecological function. It was already shown that algaenan helps to protect against virus, bacterial and fungal attack, but in this study we show for the first time that green algae with different cell wall properties show different sensitivity against interference competition with the cyanobacterium Microcystis aeruginosa . Based on previous work with co-cultures of M. aeruginosa and two green algae ( Acutodesmus obliquus and Oocystis marssonii ) differing in their cell wall structure, it was shown that M. aeruginosa could impair only the growth of the green algae if they belong to the LR-cell wall type. In this study it was shown that the sensitivity to biotic interference mechanism shows a more general pattern within coccoid green algae species depending on cell wall structure.

The N-Linked Outer Chain Mannans and the Dfg5p and Dcw1p Endo-α-1,6-Mannanases Are Needed for Incorporation of Candida albicans Glycoproteins into the Cell Wall

PubMed Central

Ao, Jie; Chinnici, Jennifer L.; Maddi, Abhiram

2015-01-01

A biochemical pathway for the incorporation of cell wall protein into the cell wall of Neurospora crassa was recently proposed. In this pathway, the DFG-5 and DCW-1 endo-α-1,6-mannanases function to covalently cross-link cell wall protein-associated N-linked galactomannans, which are structurally related to the yeast outer chain mannans, into the cell wall glucan-chitin matrix. In this report, we demonstrate that the mannosyltransferase enzyme Och1p, which is needed for the synthesis of the N-linked outer chain mannan, is essential for the incorporation of cell wall glycoproteins into the Candida albicans cell wall. Using endoglycosidases, we show that C. albicans cell wall proteins are cross-linked into the cell wall via their N-linked outer chain mannans. We further demonstrate that the Dfg5p and Dcw1p α-1,6-mannanases are needed for the incorporation of cell wall glycoproteins into the C. albicans cell wall. Our results support the hypothesis that the Dfg5p and Dcw1p α-1,6-mannanases incorporate cell wall glycoproteins into the C. albicans cell wall by cross-linking outer chain mannans into the cell wall glucan-chitin matrix. PMID:26048011

Regulation of Cell Wall Biogenesis in Saccharomyces cerevisiae: The Cell Wall Integrity Signaling Pathway

PubMed Central

Levin, David E.

2011-01-01

The yeast cell wall is a strong, but elastic, structure that is essential not only for the maintenance of cell shape and integrity, but also for progression through the cell cycle. During growth and morphogenesis, and in response to environmental challenges, the cell wall is remodeled in a highly regulated and polarized manner, a process that is principally under the control of the cell wall integrity (CWI) signaling pathway. This pathway transmits wall stress signals from the cell surface to the Rho1 GTPase, which mobilizes a physiologic response through a variety of effectors. Activation of CWI signaling regulates the production of various carbohydrate polymers of the cell wall, as well as their polarized delivery to the site of cell wall remodeling. This review article centers on CWI signaling in Saccharomyces cerevisiae through the cell cycle and in response to cell wall stress. The interface of this signaling pathway with other pathways that contribute to the maintenance of cell wall integrity is also discussed. PMID:22174182

Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatment/biomass feedstock combinations.

PubMed

Banerjee, Goutami; Car, Suzana; Scott-Craig, John S; Borrusch, Melissa S; Walton, Jonathan D

2010-10-12

Enzymes for plant cell wall deconstruction are a major cost in the production of ethanol from lignocellulosic biomass. The goal of this research was to develop optimized synthetic mixtures of enzymes for multiple pretreatment/substrate combinations using our high-throughput biomass digestion platform, GENPLAT, which combines robotic liquid handling, statistical experimental design and automated Glc and Xyl assays. Proportions of six core fungal enzymes (CBH1, CBH2, EG1, β-glucosidase, a GH10 endo-β1,4-xylanase, and β-xylosidase) were optimized at a fixed enzyme loading of 15 mg/g glucan for release of Glc and Xyl from all combinations of five biomass feedstocks (corn stover, switchgrass, Miscanthus, dried distillers' grains plus solubles [DDGS] and poplar) subjected to three alkaline pretreatments (AFEX, dilute base [0.25% NaOH] and alkaline peroxide [AP]). A 16-component mixture comprising the core set plus 10 accessory enzymes was optimized for three pretreatment/substrate combinations. Results were compared to the performance of two commercial enzymes (Accellerase 1000 and Spezyme CP) at the same protein loadings. When analyzed with GENPLAT, corn stover gave the highest yields of Glc with commercial enzymes and with the core set with all pretreatments, whereas corn stover, switchgrass and Miscanthus gave comparable Xyl yields. With commercial enzymes and with the core set, yields of Glc and Xyl were highest for grass stovers pretreated by AP compared to AFEX or dilute base. Corn stover, switchgrass and DDGS pretreated with AFEX and digested with the core set required a higher proportion of endo-β1,4-xylanase (EX3) and a lower proportion of endo-β1,4-glucanase (EG1) compared to the same materials pretreated with dilute base or AP. An optimized enzyme mixture containing 16 components (by addition of α-glucuronidase, a GH11 endoxylanase [EX2], Cel5A, Cel61A, Cip1, Cip2, β-mannanase, amyloglucosidase, α-arabinosidase, and Cel12A to the core set) was determined for AFEX-pretreated corn stover, DDGS, and AP-pretreated corn stover. The optimized mixture for AP-corn stover contained more exo-β1,4-glucanase (i.e., the sum of CBH1 + CBH2) and less endo-β1,4-glucanase (EG1 + Cel5A) than the optimal mixture for AFEX-corn stover. Amyloglucosidase and β-mannanase were the two most important enzymes for release of Glc from DDGS but were not required (i.e., 0% optimum) for corn stover subjected to AP or AFEX. As a function of enzyme loading over the range 0 to 30 mg/g glucan, Glc release from AP-corn stover reached a plateau of 60-70% Glc yield at a lower enzyme loading (5-10 mg/g glucan) than AFEX-corn stover. Accellerase 1000 was superior to Spezyme CP, the core set or the 16-component mixture for Glc yield at 12 h, but the 16-component set was as effective as the commercial enzyme mixtures at 48 h. The results in this paper demonstrate that GENPLAT can be used to rapidly produce enzyme cocktails for specific pretreatment/biomass combinations. Pretreatment conditions and feedstock source both influence the Glc and Xyl yields as well as optimal enzyme proportions. It is predicted that it will be possible to improve synthetic enzyme mixtures further by the addition of additional accessory enzymes.

A model for cell wall dissolution in mating yeast cells: polarized secretion and restricted diffusion of cell wall remodeling enzymes induces local dissolution.

PubMed

Huberman, Lori B; Murray, Andrew W

2014-01-01

Mating of the budding yeast, Saccharomyces cerevisiae, occurs when two haploid cells of opposite mating types signal using reciprocal pheromones and receptors, grow towards each other, and fuse to form a single diploid cell. To fuse, both cells dissolve their cell walls at the point of contact. This event must be carefully controlled because the osmotic pressure differential between the cytoplasm and extracellular environment causes cells with unprotected plasma membranes to lyse. If the cell wall-degrading enzymes diffuse through the cell wall, their concentration would rise when two cells touched each other, such as when two pheromone-stimulated cells adhere to each other via mating agglutinins. At the surfaces that touch, the enzymes must diffuse laterally through the wall before they can escape into the medium, increasing the time the enzymes spend in the cell wall, and thus raising their concentration at the point of attachment and restricting cell wall dissolution to points where cells touch each other. We tested this hypothesis by studying pheromone treated cells confined between two solid, impermeable surfaces. This confinement increases the frequency of pheromone-induced cell death, and this effect is diminished by reducing the osmotic pressure difference across the cell wall or by deleting putative cell wall glucanases and other genes necessary for efficient cell wall fusion. Our results support the model that pheromone-induced cell death is the result of a contact-driven increase in the local concentration of cell wall remodeling enzymes and suggest that this process plays an important role in regulating cell wall dissolution and fusion in mating cells.

A Model for Cell Wall Dissolution in Mating Yeast Cells: Polarized Secretion and Restricted Diffusion of Cell Wall Remodeling Enzymes Induces Local Dissolution

PubMed Central

Huberman, Lori B.; Murray, Andrew W.

2014-01-01

Mating of the budding yeast, Saccharomyces cerevisiae, occurs when two haploid cells of opposite mating types signal using reciprocal pheromones and receptors, grow towards each other, and fuse to form a single diploid cell. To fuse, both cells dissolve their cell walls at the point of contact. This event must be carefully controlled because the osmotic pressure differential between the cytoplasm and extracellular environment causes cells with unprotected plasma membranes to lyse. If the cell wall-degrading enzymes diffuse through the cell wall, their concentration would rise when two cells touched each other, such as when two pheromone-stimulated cells adhere to each other via mating agglutinins. At the surfaces that touch, the enzymes must diffuse laterally through the wall before they can escape into the medium, increasing the time the enzymes spend in the cell wall, and thus raising their concentration at the point of attachment and restricting cell wall dissolution to points where cells touch each other. We tested this hypothesis by studying pheromone treated cells confined between two solid, impermeable surfaces. This confinement increases the frequency of pheromone-induced cell death, and this effect is diminished by reducing the osmotic pressure difference across the cell wall or by deleting putative cell wall glucanases and other genes necessary for efficient cell wall fusion. Our results support the model that pheromone-induced cell death is the result of a contact-driven increase in the local concentration of cell wall remodeling enzymes and suggest that this process plays an important role in regulating cell wall dissolution and fusion in mating cells. PMID:25329559

Evaluation of the Relative Merits of Herbaceous and Woody Crops for Use in Tunable Thermochemical Processing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Park, Joon-Hyun; Martinalbo, Ilya

This report summarizes the work and findings of the grant work conducted from January 2009 until September 2011 under the collaboration between Ceres, Inc. and Choren USA, LLC. This DOE-funded project involves a head-to-head comparison of two types of dedicated energy crops in the context of a commercial gasification conversion process. The main goal of the project was to gain a better understanding of the differences in feedstock composition between herbaceous and woody species, and how these differences may impact a commercial gasification process. In this work, switchgrass was employed as a model herbaceous energy crop, and willow as amore » model short-rotation woody crop. Both crops are species native to the U.S. with significant potential to contribute to U.S. goals for renewable liquid fuel production, as outlined in the DOE Billion Ton Update (http://www1.eere.energy.gov/biomass/billion_ton_update.html, 2011). In some areas of the U.S., switching between woody and herbaceous feedstocks or blending of the two may be necessary to keep a large-scale gasifier operating near capacity year round. Based on laboratory tests and process simulations it has been successfully shown that suitable high yielding switchgrass and willow varieties exist that meet the feedstock specifications for large scale entrained flow biomass gasification. This data provides the foundation for better understanding how to use both materials in thermochemical processes. It has been shown that both switchgrass and willow varieties have comparable ranges of higher heating value, BTU content and indistinguishable hydrogen/carbon ratios. Benefits of switchgrass, and other herbaceous feedstocks, include its low moisture content, which reduce energy inputs and costs for drying feedstock. Compared to the typical feedstock currently being used in the Carbo-V® process, switchgrass has a higher ash content, combined with a lower ash melting temperature. Whether or not this may cause inefficiencies in the process, needs to be verified by long term test runs. Currently, there are not sufficient operational test data available for the Carbo-V® process for the utilization of higher ash content feedstocks. The application of currently evolving biomass pretreatment technologies, such as pelletization and torrefaction, will be able to expand the portfolio of biomass varieties and species acceptable in gasification processes. Tests showed that 6 mm diameter pellets of switchgrass were superior to 8 mm diameter pellets produced in a flat dye press, and that torrefaction of switchgrass produced an excellent (but currently costly) feedstock that could be handled, crushed, and combusted in a manner compatible with any coal-fed gasification facility. Ceres will use this information in the development of high yielding, dedicated energy crops specifically tailored for thermochemical conversion. CHOREN will make use of the information for improvement or development of low cost, highly efficient biomass gasification processes that convert a wide variety of biomass feedstocks to fuels, chemicals, heat and power via the production of tar free green syngas on an industrial scale.« less

High-performance liquid-catalyst fuel cell for direct biomass-into-electricity conversion.

PubMed

Liu, Wei; Mu, Wei; Deng, Yulin

2014-12-01

Herein, we report high-performance fuel cells that are catalyzed solely by polyoxometalate (POM) solution without any solid metal or metal oxide. The novel design of the liquid-catalyst fuel cells (LCFC) changes the traditional gas-solid-surface heterogeneous reactions to liquid-catalysis reactions. With this design, raw biomasses, such as cellulose, starch, and even grass or wood powders can be directly converted into electricity. The power densities of the fuel cell with switchgrass (dry powder) and bush allamanda (freshly collected) are 44 mW cm(-2) and 51 mW cm(-2) respectively. For the cellulose-based biomass fuel cell, the power density is almost 3000 times higher than that of cellulose-based microbial fuel cells. Unlike noble-metal catalysts, POMs are tolerant to most organic and inorganic contaminants. Therefore, almost any raw biomass can be used directly to produce electricity without prior purification. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A study of the native cell wall structures of the marine alga Ventricaria ventricosa (Siphonocladales, Chlorophyceae) using atomic force microscopy.

PubMed

Eslick, Enid M; Beilby, Mary J; Moon, Anthony R

2014-04-01

A substantial proportion of the architecture of the plant cell wall remains unknown with a few cell wall models being proposed. Moreover, even less is known about the green algal cell wall. Techniques that allow direct visualization of the cell wall in as near to its native state are of importance in unravelling the spatial arrangement of cell wall structures and hence in the development of cell wall models. Atomic force microscopy (AFM) was used to image the native cell wall of living cells of Ventricaria ventricosa (V. ventricosa) at high resolution under physiological conditions. The cell wall polymers were identified mainly qualitatively via their structural appearance. The cellulose microfibrils (CMFs) were easily recognizable and the imaging results indicate that the V. ventricosa cell wall has a cross-fibrillar structure throughout. We found the native wall to be abundant in matrix polysaccharides existing in different curing states. The soft phase matrix polysaccharides susceptible by the AFM scanning tip existed as a glutinous fibrillar meshwork, possibly incorporating both the pectic- and hemicellulosic-type substances. The hard phase matrix producing clearer images, revealed coiled fibrillar structures associated with CMFs, sometimes being resolved as globular structures by the AFM tip. The coiling fibrillar structures were also seen in the images of isolated cell wall fragments. The mucilaginous component of the wall was discernible from the gelatinous cell wall matrix as it formed microstructural domains over the surface. AFM has been successful in imaging the native cell wall and revealing novel findings such as the 'coiling fibrillar structures' and cell wall components which have previously not been seen, that is, the gelatinous matrix phase.

Altered Cell Wall Plasticity Can Restrict Plant Growth under Ammonium Nutrition.

PubMed

Podgórska, Anna; Burian, Maria; Gieczewska, Katarzyna; Ostaszewska-Bugajska, Monika; Zebrowski, Jacek; Solecka, Danuta; Szal, Bożena

2017-01-01

Plants mainly utilize inorganic forms of nitrogen (N), such as nitrate (NO 3 - ) and ammonium (NH 4 + ). However, the composition of the N source is important, because excess of NH 4 + promotes morphological disorders. Plants cultured on NH 4 + as the sole N source exhibit serious growth inhibition, commonly referred to as "ammonium toxicity syndrome." NH 4 + -mediated suppression of growth may be attributable to both repression of cell elongation and reduction of cell division. The precondition for cell enlargement is the expansion of the cell wall, which requires the loosening of the cell wall polymers. Therefore, to understand how NH 4 + nutrition may trigger growth retardation in plants, properties of their cell walls were analyzed. We found that Arabidopsis thaliana using NH 4 + as the sole N source has smaller cells with relatively thicker cell walls. Moreover, cellulose, which is the main load-bearing polysaccharide revealed a denser assembly of microfibrils. Consequently, the leaf blade tissue showed elevated tensile strength and indicated higher cell wall stiffness. These changes might be related to changes in polysaccharide and ion content of cell walls. Further, NH 4 + toxicity was associated with altered activities of cell wall modifying proteins. The lower activity and/or expression of pectin hydrolyzing enzymes and expansins might limit cell wall expansion. Additionally, the higher activity of cell wall peroxidases can lead to higher cross-linking of cell wall polymers. Overall, the NH 4 + -mediated inhibition of growth is related to a more rigid cell wall structure, which limits expansion of cells. The changes in cell wall composition were also indicated by decreased expression of Feronia , a receptor-like kinase involved in the control of cell wall extension.

Altered Cell Wall Plasticity Can Restrict Plant Growth under Ammonium Nutrition

PubMed Central

Podgórska, Anna; Burian, Maria; Gieczewska, Katarzyna; Ostaszewska-Bugajska, Monika; Zebrowski, Jacek; Solecka, Danuta; Szal, Bożena

2017-01-01

Plants mainly utilize inorganic forms of nitrogen (N), such as nitrate (NO3–) and ammonium (NH4+). However, the composition of the N source is important, because excess of NH4+ promotes morphological disorders. Plants cultured on NH4+ as the sole N source exhibit serious growth inhibition, commonly referred to as “ammonium toxicity syndrome.” NH4+-mediated suppression of growth may be attributable to both repression of cell elongation and reduction of cell division. The precondition for cell enlargement is the expansion of the cell wall, which requires the loosening of the cell wall polymers. Therefore, to understand how NH4+ nutrition may trigger growth retardation in plants, properties of their cell walls were analyzed. We found that Arabidopsis thaliana using NH4+ as the sole N source has smaller cells with relatively thicker cell walls. Moreover, cellulose, which is the main load-bearing polysaccharide revealed a denser assembly of microfibrils. Consequently, the leaf blade tissue showed elevated tensile strength and indicated higher cell wall stiffness. These changes might be related to changes in polysaccharide and ion content of cell walls. Further, NH4+ toxicity was associated with altered activities of cell wall modifying proteins. The lower activity and/or expression of pectin hydrolyzing enzymes and expansins might limit cell wall expansion. Additionally, the higher activity of cell wall peroxidases can lead to higher cross-linking of cell wall polymers. Overall, the NH4+-mediated inhibition of growth is related to a more rigid cell wall structure, which limits expansion of cells. The changes in cell wall composition were also indicated by decreased expression of Feronia, a receptor-like kinase involved in the control of cell wall extension. PMID:28848567

Nest survival modelling using a multi-species approach in forests managed for timber and biofuel feedstock

USGS Publications Warehouse

Loman, Zachary G.; Monroe, Adrian; Riffell, Samuel K.; Miller, Darren A.; Vilella, Francisco; Wheat, Bradley R.; Rush, Scott A.; Martin, James A.

2018-01-01

Switchgrass (Panicum virgatum) intercropping is a novel forest management practice for biomass production intended to generate cellulosic feedstocks within intensively managed loblolly pine‐dominated landscapes. These pine plantations are important for early‐successional bird species, as short rotation times continually maintain early‐successional habitat. We tested the efficacy of using community models compared to individual surrogate species models in understanding influences on nest survival. We analysed nest data to test for differences in habitat use for 14 bird species in plots managed for switchgrass intercropping and controls within loblolly pine (Pinus taeda) plantations in Mississippi, USA.We adapted hierarchical models using hyper‐parameters to incorporate information from both common and rare species to understand community‐level nest survival. This approach incorporates rare species that are often discarded due to low sample sizes, but can inform community‐level demographic parameter estimates. We illustrate use of this approach in generating both species‐level and community‐wide estimates of daily survival rates for songbird nests. We were able to include rare species with low sample size (minimum n = 5) to inform a hyper‐prior, allowing us to estimate effects of covariates on daily survival at the community level, then compare this with a single‐species approach using surrogate species. Using single‐species models, we were unable to generate estimates below a sample size of 21 nests per species.Community model species‐level survival and parameter estimates were similar to those generated by five single‐species models, with improved precision in community model parameters.Covariates of nest placement indicated that switchgrass at the nest site (<4 m) reduced daily nest survival, although intercropping at the forest stand level increased daily nest survival.Synthesis and applications. Community models represent a viable method for estimating community nest survival rates and effects of covariates while incorporating limited data for rarely detected species. Intercropping switchgrass in loblolly pine plantations slightly increased daily nest survival at the research plot scale (0.1 km2), although at a local scale (50 m2) switchgrass negatively influenced nest survival. A likely explanation is intercropping shifted community composition, favouring species with greater disturbance tolerance.

Switchgrass (Panicum virgatum L.) Intraspecific Variation and Thermotolerance Classification Using in Vitro Seed Germination Assay

DOE PAGES

Seepaul, Ramdeo; Macoon, Bisoondat; Reddy, K. Raja; ...

2011-01-01

Cardinal temperatures for plant processes have been used for thermotolerance screening of genotypes, geoclimatic adaptability determination and phenological prediction. Current simulation models for switchgrass (Panicum virgatum L.) utilize single cardinal temperatures across genotypes for both vegetative and reproductive processes although in-tra-specific variation exists among genotypes. An experiment was conducted to estimate the cardinal temperatures for seed germination of 14 diverse switchgrass genotypes and to classify genotypes for temperature tolerance. Stratified seeds of each genotype were germinated at eight constant temperatures from 10 °C to 45 °C under a constant light intensity of 35 μmol m -2s -1 for 12 hdmore » -1. Germination was recorded at 6-h intervals in all treatments. Maximum seed germination (MSG) and germination rate (GR), estimated by fitting Sigmoidal function to germination-time series data, varied among genotypes. Quadratic and bilinear models best described the MSG and GR responses to temperature, respectively. The mean cardinal temperatures, T min, T opt, and T max, were 8.1, 26.6, and 45.1 °C for MSG and 11.1, 33.1, and 46.0 °C for GR, respectively. Cardinal temperatures for MSG and GR; however, varied significantly among genotypes. Genotypes were classified as sensitive (Cave-in-Rock, Dacotah, Expresso, Forestburg, Kanlow, Sunburst, Trailblazer, and Tusca), intermediate (Alamo, Blackwell, Carthage, Shawnee, and Shelter) and tolerant (Summer) to high temperature based on cumulative temperature response index (CTRI) estimated by summing individual response indices estimated from the MSG and GR cardinal temperatures. Similarly, genotypes were also classified as sensitive (Alamo, Blackwell, Carthage, Dacotah, Shawnee, Shelter and Summer), moderately sensitive (Cave-in-rock, Forestburg, Kanlow, Sunburst, and Tusca), moderately tolerant (Trailblazer), and tolerant (Expresso) to low temperatures. The cardinal temperature estimates would be useful to improve switchgrass models for field applications. Additionally, the identified cold- and heat-tolerant genotypes can be selected for niche environments and in switchgrass breeding programs to develop new genotypes for low and high temperature environments.« less

Kinetic analyses and pyrolytic behavior of Para grass (Urochloa mutica) for its bioenergy potential.

PubMed

Ahmad, Muhammad Sajjad; Mehmood, Muhammad Aamer; Al Ayed, Omar S; Ye, Guangbin; Luo, Huibo; Ibrahim, Muhammad; Rashid, Umer; Arbi Nehdi, Imededdine; Qadir, Ghulam

2017-01-01

The biomass of Urochloa mutica was subjected to thermal degradation analyses to understand its pyrolytic behavior for bioenergy production. Thermal degradation experiments were performed at three different heating rates, 10, 30 and 50°Cmin -1 using simultaneous thermogravimetric-differential scanning calorimetric analyzer, under an inert environment. The kinetic analyses were performed using isoconversional models of Kissenger-Akahira-Sunose (KAS) and Flynn-Wall-Ozawa (FWO). The high heating value was calculated as 15.04MJmol -1 . The activation energy (E) values were shown to be ranging from 103 through 233 kJmol -1 . Pre-exponential factors (A) indicated the reaction to follow first order kinetics. Gibbs free energy (ΔG) was measured to be ranging from 169 to 173kJmol -1 and 168 to 172kJmol -1 , calculated by KAS and FWO methods, respectively. We have shown that Para grass biomass has considerable bioenergy potential comparable to established bioenergy crops such as switchgrass and miscanthus. Copyright © 2016 Elsevier Ltd. All rights reserved.

Vulnerability of crops and native grasses to summer drying in the U.S. Southern Great Plains

DOE PAGES

Raz-Yaseef, Naama; Billesbach, Dave P.; Fischer, Marc L.; ...

2015-08-31

The Southern Great Plains are characterized by a fine-scale mixture of different land-cover types, predominantly winter-wheat and grazed pasture, with relatively small areas of other crops, native prairie, and switchgrass. Recent droughts and predictions of increased drought in the Southern Great Plains, especially during the summer months, raise concern for these ecosystems. We measured ecosystem carbon and water fluxes with eddy-covariance systems over cultivated cropland for 10 years, and over lightly grazed prairie and new switchgrass fields for 2 years each. Growing-season precipitation showed the strongest control over net carbon uptake for all ecosystems, but with a variable effect: grassesmore » (prairie and switchgrass) needed at least 350 mm of precipitation during the growing season to become net carbon sinks, while crops needed only 100 mm. In summer, high temperatures enhanced evaporation and led to higher likelihood of dry soil conditions. Therefore, summer-growing native prairie species and switchgrass experienced more seasonal droughts than spring-growing crops. For wheat, the net reduction in carbon uptake resulted mostly from a decrease in gross primary production rather than an increase in respiration. Flux measurements suggested that management practices for crops were effective in suppressing evapotranspiration and decomposition (by harvesting and removing secondary growth), and in increasing carbon uptake (by fertilizing and conserving summer soil water). In light of future projections for wetter springs and drier and warmer summers in the Southern Great Plains, our study indicates an increased vulnerability in native ecosystems and summer crops over time.« less

The feasibility of producing adequate feedstock for year–round cellulosic ethanol production in an intensive agricultural fuelshed

USGS Publications Warehouse

Uden, Daniel R.; Mitchell, Rob B.; Allen, Craig R.; Guan, Qingfeng; McCoy, Tim D.

2013-01-01

To date, cellulosic ethanol production has not been commercialized in the United States. However, government mandates aimed at increasing second-generation biofuel production could spur exploratory development in the cellulosic ethanol industry. We conducted an in-depth analysis of the fuelshed surrounding a starch-based ethanol plant near York, Nebraska that has the potential for cellulosic ethanol production. To assess the feasibility of supplying adequate biomass for year-round cellulosic ethanol production from residual maize (Zea mays) stover and bioenergy switchgrass (Panicum virgatum) within a 40-km road network service area of the existing ethanol plant, we identified ∼14,000 ha of marginally productive cropland within the service area suitable for conversion from annual rowcrops to switchgrass and ∼132,000 ha of maize-enrolled cropland from which maize stover could be collected. Annual maize stover and switchgrass biomass supplies within the 40-km service area could range between 429,000 and 752,000 metric tons (mT). Approximately 140–250 million liters (l) of cellulosic ethanol could be produced, rivaling the current 208 million l annual starch-based ethanol production capacity of the plant. We conclude that sufficient quantities of biomass could be produced from maize stover and switchgrass near the plant to support year-round cellulosic ethanol production at current feedstock yields, sustainable removal rates and bioconversion efficiencies. Modifying existing starch-based ethanol plants in intensive agricultural fuelsheds could increase ethanol output, return marginally productive cropland to perennial vegetation, and remove maize stover from productive cropland to meet feedstock demand.

Switchgrass as a biofuels crop for the upper Southeast

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parrish, D.J.; Wolf, D.D.

1993-12-31

Switchgrass (Panicum virgatum) has been identified in DOE-sponsored studies as a widely adapted, productive herbaceous candidate for biofuels cropping. It is a perennial that has been planted using no-till procedures, and it appears to have positive effects on the soils in which it grows. We have been looking at this species as a potential fuelcrop (as well as a valuable forage) for several years. In this presentation, we note several {open_quotes}lessons learned{close_quotes} about switchgrass establishment and management as an energy crop. Data include results from recent plantings in the upper Southeast USA and from cutting management studies. Six varieties ofmore » switchgrass (Alamo, Cave-in-Rock, Kanlow, Shelter, and two breeder`s lines) varied markedly in the success of their no-till establishment at eight locations across the upper Southeast. Better weed control, which was achieved at later planting dates, seemed to be the key. Yields obtained in the establishment stands revealed that two harvests per season are more productive (by 2 to 3 Mg/ha) than one, but the date of first cutting is crucial. First cutting should be from late-June to mid-July. A two-cut system may not be economically advantageous, however. Another cutting-management study detected losses of standing biomass at the end of the growing season. As much as 15% of the above-ground biomass present in early-September was no longer harvestable in early-November. We think this loss results from translocation of dry matter to below-ground parts.« less

Impacts of biofuels production alternatives on water quantity and quality in the Iowa River Basin

USGS Publications Warehouse

Wu, Y.; Liu, S.

2012-01-01

Corn stover as well as perennial grasses like switchgrass (Panicum virgatum) and miscanthus are being considered as candidates for the second generation biofuel feedstocks. However, the challenges to biofuel development are its effects on the environment, especially water quality. This study evaluates the long-term impacts of biofuel production alternatives (e.g., elevated corn stover removal rates and the potential land cover change) on an ecosystem with a focus on biomass production, soil erosion, water quantity and quality, and soil nitrate nitrogen concentration at the watershed scale. The Soil and Water Assessment Tool (SWAT) was modified for setting land cover change scenarios and applied to the Iowa River Basin (a tributary of the Upper Mississippi River Basin). Results show that biomass production can be sustained with an increased stover removal rate as long as the crop demand for nutrients is met with appropriate fertilization. Although a drastic increase (4.7–70.6%) in sediment yield due to erosion and a slight decrease (1.2–3.2%) in water yield were estimated with the stover removal rate ranging between 40% and 100%, the nitrate nitrogen load declined about 6–10.1%. In comparison to growing corn, growing either switchgrass or miscanthus can reduce sediment erosion greatly. However, land cover changes from native grass to switchgrass or miscanthus would lead to a decrease in water yield and an increase in nitrate nitrogen load. In contrast to growing switchgrass, growing miscanthus is more productive in generating biomass, but its higher water demand may reduce water availability in the study area.

The Cell Wall of the Human Fungal Pathogen Aspergillus fumigatus: Biosynthesis, Organization, Immune Response, and Virulence.

PubMed

Latgé, Jean-Paul; Beauvais, Anne; Chamilos, Georgios

2017-09-08

More than 90% of the cell wall of the filamentous fungus Aspergillus fumigatus comprises polysaccharides. Biosynthesis of the cell wall polysaccharides is under the control of three types of enzymes: transmembrane synthases, which are anchored to the plasma membrane and use nucleotide sugars as substrates, and cell wall-associated transglycosidases and glycosyl hydrolases, which are responsible for remodeling the de novo synthesized polysaccharides and establishing the three-dimensional structure of the cell wall. For years, the cell wall was considered an inert exoskeleton of the fungal cell. The cell wall is now recognized as a living organelle, since the composition and cellular localization of the different constitutive cell wall components (especially of the outer layers) vary when the fungus senses changes in the external environment. The cell wall plays a major role during infection. The recognition of the fungal cell wall by the host is essential in the initiation of the immune response. The interactions between the different pattern-recognition receptors (PRRs) and cell wall pathogen-associated molecular patterns (PAMPs) orientate the host response toward either fungal death or growth, which would then lead to disease development. Understanding the molecular determinants of the interplay between the cell wall and host immunity is fundamental to combatting Aspergillus diseases.

Cell Wall Composition and Candidate Biosynthesis Gene Expression During Rice Development

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Fan; Manisseri, Chithra; Fagerström, Alexandra

Cell walls of grasses, including cereal crops and biofuel grasses, comprise the majority of plant biomass and intimately influence plant growth, development and physiology. However, the functions of many cell wall synthesis genes, and the relationships among and the functions of cell wall components remain obscure. To better understand the patterns of cell wall accumulation and identify genes that act in grass cell wall biosynthesis, we characterized 30 samples from aerial organs of rice (Oryza sativa cv. Kitaake) at 10 developmental time points, 3-100 d post-germination. Within these samples, we measured 15 cell wall chemical components, enzymatic digestibility and 18more » cell wall polysaccharide epitopes/ligands. We also used quantitative reverse transcription-PCR to measure expression of 50 glycosyltransferases, 15 acyltransferases and eight phenylpropanoid genes, many of which had previously been identified as being highly expressed in rice. Most cell wall components vary significantly during development, and correlations among them support current understanding of cell walls. We identified 92 significant correlations between cell wall components and gene expression and establish nine strong hypotheses for genes that synthesize xylans, mixed linkage glucan and pectin components. This work provides an extensive analysis of cell wall composition throughout rice development, identifies genes likely to synthesize grass cell walls, and provides a framework for development of genetically improved grasses for use in lignocellulosic biofuel production and agriculture.« less

The Impact of Microfibril Orientations on the Biomechanics of Plant Cell Walls and Tissues.

PubMed

Ptashnyk, Mariya; Seguin, Brian

2016-11-01

The microscopic structure and anisotropy of plant cell walls greatly influence the mechanical properties, morphogenesis, and growth of plant cells and tissues. The microscopic structure and properties of cell walls are determined by the orientation and mechanical properties of the cellulose microfibrils and the mechanical properties of the cell wall matrix. Viewing the shape of a plant cell as a square prism with the axis aligning with the primary direction of expansion and growth, the orientation of the microfibrils within the side walls, i.e. the parts of the cell walls on the sides of the cells, is known. However, not much is known about their orientation at the upper and lower ends of the cell. Here we investigate the impact of the orientation of cellulose microfibrils within the upper and lower parts of the plant cell walls by solving the equations of linear elasticity numerically. Three different scenarios for the orientation of the microfibrils are considered. We also distinguish between the microstructure in the side walls given by microfibrils perpendicular to the main direction of the expansion and the situation where the microfibrils are rotated through the wall thickness. The macroscopic elastic properties of the cell wall are obtained using homogenization theory from the microscopic description of the elastic properties of the cell wall microfibrils and wall matrix. It is found that the orientation of the microfibrils in the upper and lower parts of the cell walls affects the expansion of the cell in the lateral directions and is particularly important in the case of forces acting on plant cell walls and tissues.

High temperature induced disruption of the cell wall integrity and structure in Pleurotus ostreatus mycelia.

PubMed

Qiu, Zhiheng; Wu, Xiangli; Gao, Wei; Zhang, Jinxia; Huang, Chenyang

2018-05-30

Fungal cells are surrounded by a tight cell wall to protect them from harmful environmental conditions and to resist lysis. The synthesis and assembly determine the shape, structure, and integrity of the cell wall during the process of mycelial growth and development. High temperature is an important abiotic stress, which affects the synthesis and assembly of cell walls. In the present study, the chitin and β-1,3-glucan concentrations in the cell wall of Pleurotus ostreatus mycelia were changed after high-temperature treatment. Significantly higher chitin and β-1,3-glucan concentrations were detected at 36 °C than those incubated at 28 °C. With the increased temperature, many aberrant chitin deposition patches occurred, and the distribution of chitin in the cell wall was uneven. Moreover, high temperature disrupts the cell wall integrity, and P. ostreatus mycelia became hypersensitive to cell wall-perturbing agents at 36 °C. The cell wall structure tended to shrink or distorted after high temperature. The cell walls were observed to be thicker and looser by using transmission electron microscopy. High temperature can decrease the mannose content in the cell wall and increase the relative cell wall porosity. According to infrared absorption spectrum, high temperature broke or decreased the glycosidic linkages. Finally, P. ostreatus mycelial cell wall was easily degraded by lysing enzymes after high-temperature treatment. In other words, the cell wall destruction caused by high temperature may be a breakthrough for P. ostreatus to be easily infected by Trichoderma.

Endoplasmic reticulum-derived reactive oxygen species (ROS) is involved in toxicity of cell wall stress to Candida albicans.

PubMed

Yu, Qilin; Zhang, Bing; Li, Jianrong; Zhang, Biao; Wang, Honggang; Li, Mingchun

2016-10-01

The cell wall is an important cell structure in both fungi and bacteria, and hence becomes a common antimicrobial target. The cell wall-perturbing agents disrupt synthesis and function of cell wall components, leading to cell wall stress and consequent cell death. However, little is known about the detailed mechanisms by which cell wall stress renders fungal cell death. In this study, we found that ROS scavengers drastically attenuated the antifungal effect of cell wall-perturbing agents to the model fungal pathogen Candida albicans, and these agents caused remarkable ROS accumulation and activation of oxidative stress response (OSR) in this fungus. Interestingly, cell wall stress did not cause mitochondrial dysfunction and elevation of mitochondrial superoxide levels. Furthermore, the iron chelator 2,2'-bipyridyl (BIP) and the hydroxyl radical scavengers could not attenuate cell wall stress-caused growth inhibition and ROS accumulation. However, cell wall stress up-regulated expression of unfold protein response (UPR) genes, enhanced protein secretion and promoted protein folding-related oxidation of Ero1, an important source of ROS production. These results indicated that oxidation of Ero1 in the endoplasmic reticulum (ER), rather than mitochondrial electron transport and Fenton reaction, contributed to cell wall stress-related ROS accumulation and consequent growth inhibition. Our findings uncover a novel link between cell wall integrity (CWI), ER function and ROS production in fungal cells, and shed novel light on development of strategies promoting the antifungal efficacy of cell wall-perturbing agents against fungal infections. Copyright © 2016 Elsevier Inc. All rights reserved.

Regulation of cell wall biosynthesis.

PubMed

Zhong, Ruiqin; Ye, Zheng-Hua

2007-12-01

Plant cell walls differ in their amount and composition among various cell types and even in different microdomains of the wall of a given cell. Plants must have evolved regulatory mechanisms controlling biosynthesis, targeted secretion, and assembly of wall components to achieve the heterogeneity in cell walls. A number of factors, including hormones, the cytoskeleton, glycosylphosphatidylinositol-anchored proteins, phosphoinositides, and sugar nucleotide supply, have been implicated in the regulation of cell wall biosynthesis or deposition. In the past two years, there have been important discoveries in transcriptional regulation of secondary wall biosynthesis. Several transcription factors in the NAC and MYB families have been shown to be the key switches for activation of secondary wall biosynthesis. These studies suggest a transcriptional network comprised of a hierarchy of transcription factors is involved in regulating secondary wall biosynthesis. Further investigation and integration of the regulatory players participating in the making of cell walls will certainly lead to our understanding of how wall amounts and composition are controlled in a given cell type. This may eventually allow custom design of plant cell walls on the basis of our needs.

Production of polyhydroxybutyrate in switchgrass, a value-added co-product in an important lignocellulosic biomass crop.

PubMed

Somleva, Maria N; Snell, Kristi D; Beaulieu, Julie J; Peoples, Oliver P; Garrison, Bradley R; Patterson, Nii A

2008-09-01

Polyhydroxyalkanoate bio-based plastics made from renewable resources can reduce petroleum consumption and decrease plastic waste disposal issues as they are inherently biodegradable in soil, compost and marine environments. In this paper, the successful engineering of the biomass crop switchgrass (Panicum virgatum L.) for the synthesis of polyhydroxybutyrate (PHB) is reported. Polymer production was monitored in more than 400 primary transformants grown under in vitro and glasshouse conditions. Plants containing up to 3.72% dry weight of PHB in leaf tissues and 1.23% dry weight of PHB in whole tillers were obtained. Results from the analysis of the polymer distribution at the cellular and whole plant levels are presented, and target areas for the improvement of PHB production are highlighted. Polymer accumulation was also analysed in the T(1) generation obtained from controlled crosses of transgenic plants. This study presents the first successful expression of a functional multigene pathway in switchgrass, and demonstrates that this high-yielding biomass crop is amenable to the complex metabolic engineering strategies necessary to produce high-value biomaterials with lignocellulose-derived biofuels.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shahrukh, Hassan; Oyedun, Adetoyese Olajire; Kumar, Amit

Here, a process model was developed to determine the net energy ratio (NER) for production of pellets from steam pretreated agricultural residue (AR) and energy crop (i.e. switchgrass in this case). The NER is a ratio of the net energy output to the total net energy input from non-renewable energy sources into a system. Scenarios were developed to measure the effects of temperature and level of steam pretreatment on the NER of steam pretreated AR- and switch grass-based pellets. The NER for the base case at 6 kg h -1 is 1.76 and 1.37 for steam-pretreated AR- and switchgrass-based pellets,more » respectively. The reason behind the difference is that more energy is required to dry switchgrass pellets than AR pellets. The sensitivity analysis for the model shows that the optimum temperature for steam pretreatment is 160 C with 50% pretreatment (half the feedstock is pretreated, while the rest is undergoes regular pelletization). The uncertainty results for NER for steam pretreated AR and switch grass pellets are 1.62 ± 0.10 and 1.42 ± 0.11, respectively.« less

Characteristics of Lignin Fractions from Dilute Acid Pretreated Switchgrass and Their Effect on Cellobiohydrolase from Trichoderma longibrachiatum

DOE PAGES

Yao, Lan; Yang, Haitao; Yoo, Chang Geun; ...

2018-02-06

To investigate the interactions between acid pretreated switchgrass lignin and cellobiohydrolase (CBH), three different lignin fractions were isolated from dilute acid pretreated switchgrass by (i) ethanol extraction, followed by (ii) dioxane/H2O extraction, and (iii) cellulase treatment, respectively. Structural properties of each lignin fraction were elucidated by GPC, 13C-NMR, and 2D-HSQC NMR analyses. The adsorptions of CBH to the isolated lignin fractions were also studied by Langmuir adsorption isotherms. Ethanol-extractable lignin fraction, mainly composed of syringyl (S) and guaiacyl (G) units, had the lowest molecular weight, while dioxane/H2O-extracted lignin fraction had the lowest S/G ratio with higher content of p-coumaric acidmore » (pCA) unit. The residual lignin fraction after enzymatic treatment had the highest S/G ratio without hydroxyphenyl (H) unit. Strong associations were found between lignin properties such as lignin composition and S/G ratio and its non-productive enzyme adsorption factors including the maximum adsorption capacity and binding strength.« less

Effects of Torrefaction Temperature on Pyrolysis Vapor Products of Woody and Herbaceous Feedstocks

DOE PAGES

Starace, Anne K.; Evans, Robert J.; Lee, David D.; ...

2016-06-17

A variety of hardwood, softwood, and herbaceous feedstocks (oak, southern yellow pine mix, loblolly pine, pinyon-juniper mix, and switchgrass) were each torrefied at 200, 250, and 300 °C. Each of the feedstocks was pyrolyzed and the resulting vapors were analyzed with a molecular beam mass spectrometer (py-MBMS). Compositional analysis was used to measure the total lignin content of three of the feedstocks (southern yellow pine, softwood; oak, hardwood; and switchgrass, herbaceous) before and after torrefaction at 300 °C, and large differences in the fraction of lignin lost during torrefaction were found between feedstocks, with oak having the largest decrease inmore » lignin during torrefaction and switchgrass having the least. Finally, it is hypothesized that these differences in the thermal degradation are due to, in part, the different ratios of S, G, and H lignins in the feedstocks. Additionally, the torrefaction of kraft lignin was studied using thermogravimetric analysis coupled with Fourier transform infrared spectroscopy (TGA-FTIR) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR).« less

Characteristics of Lignin Fractions from Dilute Acid Pretreated Switchgrass and Their Effect on Cellobiohydrolase from Trichoderma longibrachiatum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yao, Lan; Yang, Haitao; Yoo, Chang Geun

To investigate the interactions between acid pretreated switchgrass lignin and cellobiohydrolase (CBH), three different lignin fractions were isolated from dilute acid pretreated switchgrass by (i) ethanol extraction, followed by (ii) dioxane/H2O extraction, and (iii) cellulase treatment, respectively. Structural properties of each lignin fraction were elucidated by GPC, 13C-NMR, and 2D-HSQC NMR analyses. The adsorptions of CBH to the isolated lignin fractions were also studied by Langmuir adsorption isotherms. Ethanol-extractable lignin fraction, mainly composed of syringyl (S) and guaiacyl (G) units, had the lowest molecular weight, while dioxane/H2O-extracted lignin fraction had the lowest S/G ratio with higher content of p-coumaric acidmore » (pCA) unit. The residual lignin fraction after enzymatic treatment had the highest S/G ratio without hydroxyphenyl (H) unit. Strong associations were found between lignin properties such as lignin composition and S/G ratio and its non-productive enzyme adsorption factors including the maximum adsorption capacity and binding strength.« less

Biomass pyrolysis and combustion integral and differential reaction heats with temperatures using thermogravimetric analysis/differential scanning calorimetry.

PubMed

Shen, Jiacheng; Igathinathane, C; Yu, Manlu; Pothula, Anand Kumar

2015-06-01

Integral reaction heats of switchgrass, big bluestem, and corn stalks were determined using thermogravimetric analysis/differential scanning calorimetry (TGA/DSC). Iso-conversion differential reaction heats using TGA/DSC pyrolysis and combustion of biomass were not available, despite reports available on heats required and released. A concept of iso-conversion differential reaction heats was used to determine the differential reaction heats of each thermal characteristics segment of these materials. Results showed that the integral reaction heats were endothermic from 30 to 700°C for pyrolysis of switchgrass and big bluestem, but they were exothermic for corn stalks prior to 587°C. However, the integral reaction heats for combustion of the materials followed an endothermic to exothermic transition. The differential reaction heats of switchgrass pyrolysis were predominantly endothermic in the fraction of mass loss (0.0536-0.975), and were exothermic for corn stalks (0.0885-0.850) and big bluestem (0.736-0.919). Study results provided better insight into biomass thermal mechanism. Published by Elsevier Ltd.

Relating Nanoscale Accessibility within Plant Cell Walls to Improved Enzyme Hydrolysis Yields in Corn Stover Subjected to Diverse Pretreatments.

PubMed

Crowe, Jacob D; Zarger, Rachael A; Hodge, David B

2017-10-04

Simultaneous chemical modification and physical reorganization of plant cell walls via alkaline hydrogen peroxide or liquid hot water pretreatment can alter cell wall structural properties impacting nanoscale porosity. Nanoscale porosity was characterized using solute exclusion to assess accessible pore volumes, water retention value as a proxy for accessible water-cell walls surface area, and solute-induced cell wall swelling to measure cell wall rigidity. Key findings concluded that delignification by alkaline hydrogen peroxide pretreatment decreased cell wall rigidity and that the subsequent cell wall swelling resulted increased nanoscale porosity and improved enzyme binding and hydrolysis compared to limited swelling and increased accessible surface areas observed in liquid hot water pretreated biomass. The volume accessible to a 90 Å dextran probe within the cell wall was found to be correlated to both enzyme binding and glucose hydrolysis yields, indicating cell wall porosity is a key contributor to effective hydrolysis yields.

Following the compositional changes of fresh grape skin cell walls during the fermentation process in the presence and absence of maceration enzymes.

PubMed

Zietsman, Anscha J J; Moore, John P; Fangel, Jonatan U; Willats, William G T; Trygg, Johan; Vivier, Melané A

2015-03-18

Cell wall profiling technologies were used to follow compositional changes that occurred in the skins of grape berries (from two different ripeness levels) during fermentation and enzyme maceration. Multivariate data analysis showed that the fermentation process yielded cell walls enriched in hemicellulose components because pectin was solubilized (and removed) with a reduction as well as exposure of cell wall proteins usually embedded within the cell wall structure. The addition of enzymes caused even more depectination, and the enzymes unravelled the cell walls enabling better access to, and extraction of, all cell wall polymers. Overripe grapes had cell walls that were extensively hydrolyzed and depolymerized, probably by natural grape-tissue-ripening enzymes, and this enhanced the impact that the maceration enzymes had on the cell wall monosaccharide profile. The combination of the techniques that were used is an effective direct measurement of the hydrolysis actions of maceration enzymes on the cell walls of grape berry skin.

Employing proteomic analysis to compare Paracoccidioides lutzii yeast and mycelium cell wall proteins.

PubMed

Araújo, Danielle Silva; de Sousa Lima, Patrícia; Baeza, Lilian Cristiane; Parente, Ana Flávia Alves; Melo Bailão, Alexandre; Borges, Clayton Luiz; de Almeida Soares, Célia Maria

2017-11-01

Paracoccidioidomycosis is an important systemic mycosis caused by thermodimorphic fungi of the Paracoccidioides genus. During the infective process, the cell wall acts at the interface between the fungus and the host. In this way, the cell wall has a key role in growth, environment sensing and interaction, as well as morphogenesis of the fungus. Since the cell wall is absent in mammals, it may present molecules that are described as target sites for new antifungal drugs. Despite its importance, up to now few studies have been conducted employing proteomics in for the identification of cell wall proteins in Paracoccidioides spp. Here, a detailed proteomic approach, including cell wall-fractionation coupled to NanoUPLC-MS E , was used to study and compare the cell wall fractions from Paracoccidioides lutzii mycelia and yeast cells. The analyzed samples consisted of cell wall proteins extracted by hot SDS followed by extraction by mild alkali. In summary, 512 proteins constituting different cell wall fractions were identified, including 7 predicted GPI-dependent cell wall proteins that are potentially involved in cell wall metabolism. Adhesins previously described in Paracoccidioides spp. such as enolase, glyceraldehyde-3-phosphate dehydrogenase were identified. Comparing the proteins in mycelium and yeast cells, we detected some that are common to both fungal phases, such as Ecm33, and some specific proteins, as glucanase Crf1. All of those proteins were described in the metabolism of cell wall. Our study provides an important elucidation of cell wall composition of fractions in Paracoccidioides, opening a way to understand the fungus cell wall architecture. Copyright © 2017 Elsevier B.V. All rights reserved.

Engineering cell wall synthesis mechanism for enhanced PHB accumulation in E. coli.

PubMed

Zhang, Xing-Chen; Guo, Yingying; Liu, Xu; Chen, Xin-Guang; Wu, Qiong; Chen, Guo-Qiang

2018-01-01

The rigidity of bacterial cell walls synthesized by a complicated pathway limit the cell shapes as coccus, bar or ellipse or even fibers. A less rigid bacterium could be beneficial for intracellular accumulation of poly-3-hydroxybutyrate (PHB) as granular inclusion bodies. To understand how cell rigidity affects PHB accumulation, E. coli cell wall synthesis pathway was reinforced and weakened, respectively. Cell rigidity was achieved by thickening the cell walls via insertion of a constitutive gltA (encoding citrate synthase) promoter in front of a series of cell wall synthesis genes on the chromosome of several E. coli derivatives, resulting in 1.32-1.60 folds increase of Young's modulus in mechanical strength for longer E. coli cells over-expressing fission ring FtsZ protein inhibiting gene sulA. Cell rigidity was weakened by down regulating expressions of ten genes in the cell wall synthesis pathway using CRISPRi, leading to elastic cells with more spaces for PHB accumulation. The regulation on cell wall synthesis changes the cell rigidity: E. coli with thickened cell walls accumulated only 25% PHB while cell wall weakened E. coli produced 93% PHB. Manipulation on cell wall synthesis mechanism adds another possibility to morphology engineering of microorganisms. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Salt stress causes cell wall damage in yeast cells lacking mitochondrial DNA.

PubMed

Gao, Qiuqiang; Liou, Liang-Chun; Ren, Qun; Bao, Xiaoming; Zhang, Zhaojie

2014-03-03

The yeast cell wall plays an important role in maintaining cell morphology, cell integrity and response to environmental stresses. Here, we report that salt stress causes cell wall damage in yeast cells lacking mitochondrial DNA (ρ 0 ). Upon salt treatment, the cell wall is thickened, broken and becomes more sensitive to the cell wall-perturbing agent sodium dodecyl sulfate (SDS). Also, SCW11 mRNA levels are elevated in ρ 0 cells. Deletion of SCW11 significantly decreases the sensitivity of ρ 0 cells to SDS after salt treatment, while overexpression of SCW11 results in higher sensitivity. In addition, salt stress in ρ 0 cells induces high levels of reactive oxygen species (ROS), which further damages the cell wall, causing cells to become more sensitive towards the cell wall-perturbing agent.

Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls.

PubMed

Marcus, Susan E; Verhertbruggen, Yves; Hervé, Cécile; Ordaz-Ortiz, José J; Farkas, Vladimir; Pedersen, Henriette L; Willats, William G T; Knox, J Paul

2008-05-22

Molecular probes are required to detect cell wall polymers in-situ to aid understanding of their cell biology and several studies have shown that cell wall epitopes have restricted occurrences across sections of plant organs indicating that cell wall structure is highly developmentally regulated. Xyloglucan is the major hemicellulose or cross-linking glycan of the primary cell walls of dicotyledons although little is known of its occurrence or functions in relation to cell development and cell wall microstructure. Using a neoglycoprotein approach, in which a XXXG heptasaccharide of tamarind seed xyloglucan was coupled to BSA to produce an immunogen, we have generated a rat monoclonal antibody (designated LM15) to the XXXG structural motif of xyloglucans. The specificity of LM15 has been confirmed by the analysis of LM15 binding using glycan microarrays and oligosaccharide hapten inhibition of binding studies. The use of LM15 for the analysis of xyloglucan in the cell walls of tamarind and nasturtium seeds, in which xyloglucan occurs as a storage polysaccharide, indicated that the LM15 xyloglucan epitope occurs throughout the thickened cell walls of the tamarind seed and in the outer regions, adjacent to middle lamellae, of the thickened cell walls of the nasturtium seed. Immunofluorescence analysis of LM15 binding to sections of tobacco and pea stem internodes indicated that the xyloglucan epitope was restricted to a few cell types in these organs. Enzymatic removal of pectic homogalacturonan from equivalent sections resulted in the abundant detection of distinct patterns of the LM15 xyloglucan epitope across these organs and a diversity of occurrences in relation to the cell wall microstructure of a range of cell types. These observations support ideas that xyloglucan is associated with pectin in plant cell walls. They also indicate that documented patterns of cell wall epitopes in relation to cell development and cell differentiation may need to be re-considered in relation to the potential masking of cell wall epitopes by other cell wall components.

The Interplay between Cell Wall Mechanical Properties and the Cell Cycle in Staphylococcus aureus

PubMed Central

Bailey, Richard G.; Turner, Robert D.; Mullin, Nic; Clarke, Nigel; Foster, Simon J.; Hobbs, Jamie K.

2014-01-01

The nanoscale mechanical properties of live Staphylococcus aureus cells during different phases of growth were studied by atomic force microscopy. Indentation to different depths provided access to both local cell wall mechanical properties and whole-cell properties, including a component related to cell turgor pressure. Local cell wall properties were found to change in a characteristic manner throughout the division cycle. Splitting of the cell into two daughter cells followed a local softening of the cell wall along the division circumference, with the cell wall on either side of the division circumference becoming stiffer. Once exposed, the newly formed septum was found to be stiffer than the surrounding, older cell wall. Deeper indentations, which were affected by cell turgor pressure, did not show a change in stiffness throughout the division cycle, implying that enzymatic cell wall remodeling and local variations in wall properties are responsible for the evolution of cell shape through division. PMID:25468333

KRE5 Suppression Induces Cell Wall Stress and Alternative ER Stress Response Required for Maintaining Cell Wall Integrity in Candida glabrata

PubMed Central

Sasaki, Masato; Ito, Fumie; Aoyama, Toshio; Sato-Okamoto, Michiyo; Takahashi-Nakaguchi, Azusa; Chibana, Hiroji; Shibata, Nobuyuki

2016-01-01

The maintenance of cell wall integrity in fungi is required for normal cell growth, division, hyphae formation, and antifungal tolerance. We observed that endoplasmic reticulum stress regulated cell wall integrity in Candida glabrata, which possesses uniquely evolved mechanisms for unfolded protein response mechanisms. Tetracycline-mediated suppression of KRE5, which encodes a predicted UDP-glucose:glycoprotein glucosyltransferase localized in the endoplasmic reticulum, significantly increased cell wall chitin content and decreased cell wall β-1,6-glucan content. KRE5 repression induced endoplasmic reticulum stress-related gene expression and MAP kinase pathway activation, including Slt2p and Hog1p phosphorylation, through the cell wall integrity signaling pathway. Moreover, the calcineurin pathway negatively regulated cell wall integrity, but not the reduction of β-1,6-glucan content. These results indicate that KRE5 is required for maintaining both endoplasmic reticulum homeostasis and cell wall integrity, and that the calcineurin pathway acts as a regulator of chitin-glucan balance in the cell wall and as an alternative mediator of endoplasmic reticulum stress in C. glabrata. PMID:27548283

Imaging the Dynamics of Cell Wall Polymer Deposition in the Unicellular Model Plant, Penium margaritaceum.

PubMed

Domozych, David; Lietz, Anna; Patten, Molly; Singer, Emily; Tinaz, Berke; Raimundo, Sandra C

2017-01-01

The unicellular green alga, Penium margaritaceum, represents a novel and valuable model organism for elucidating cell wall dynamics in plants. This organism's cell wall contains several polymers that are highly similar to those found in the primary cell walls of land plants. Penium is easily grown in laboratory culture and is effectively manipulated in various experimental protocols including microplate assays and correlative microscopy. Most importantly, Penium can be live labeled with cell wall-specific antibodies or other probes and returned to culture where specific cell wall developmental events can be monitored. Additionally, live cells can be rapidly cryo-fixed and cell wall surface microarchitecture can be observed with variable pressure scanning electron microscopy. Here, we describe the methodology for maintaining Penium for experimental cell wall enzyme studies.

Cell walls of the dimorphic fungal pathogens Sporothrix schenckii and Sporothrix brasiliensis exhibit bilaminate structures and sloughing of extensive and intact layers

PubMed Central

Walker, Louise A.; Niño-Vega, Gustavo; Mora-Montes, Héctor M.; Neves, Gabriela W. P.; Villalobos-Duno, Hector; Barreto, Laura; Garcia, Karina; Franco, Bernardo; Martínez-Álvarez, José A.; Munro, Carol A.; Gow, Neil A. R.

2018-01-01

Sporotrichosis is a subcutaneous mycosis caused by pathogenic species of the Sporothrix genus. A new emerging species, Sporothrix brasiliensis, is related to cat-transmitted sporotrichosis and has severe clinical manifestations. The cell wall of pathogenic fungi is a unique structure and impacts directly on the host immune response. We reveal and compare the cell wall structures of Sporothrix schenckii and S. brasiliensis using high-pressure freezing electron microscopy to study the cell wall organization of both species. To analyze the components of the cell wall, we also used infrared and 13C and 1H NMR spectroscopy and the sugar composition was determined by quantitative high-performance anion-exchange chromatography. Our ultrastructural data revealed a bi-layered cell wall structure for both species, including an external microfibrillar layer and an inner electron-dense layer. The inner and outer layers of the S. brasiliensis cell wall were thicker than those of S. schenckii, correlating with an increase in the chitin and rhamnose contents. Moreover, the outer microfibrillar layer of the S. brasiliensis cell wall had longer microfibrils interconnecting yeast cells. Distinct from those of other dimorphic fungi, the cell wall of Sporothrix spp. lacked α-glucan component. Interestingly, glycogen α-particles were identified in the cytoplasm close to the cell wall and the plasma membrane. The cell wall structure as well as the presence of glycogen α-particles varied over time during cell culture. The structural differences observed in the cell wall of these Sporothrix species seemed to impact its uptake by monocyte-derived human macrophages. The data presented here show a unique cell wall structure of S. brasiliensis and S. schenckii during the yeast parasitic phase. A new cell wall model for Sporothrix spp. is therefore proposed that suggests that these fungi molt sheets of intact cell wall layers. This observation may have significant effects on localized and disseminated immunopathology. PMID:29522522

Effects of alternative deep bedding options on dairy cow preference, lying behavior, cleanliness, and teat end contamination.

PubMed

Wolfe, T; Vasseur, E; DeVries, T J; Bergeron, R

2018-01-01

Cows spend more time lying down when stalls are soft and dry, and bedding plays a key role in the comfort of the lying surface. The first objective of this study (experiment 1) was to compare cow preference for 2 types of alternative deep-bedding materials, switchgrass and switchgrass-lime, using wheat straw on a rubber mat as a control. Nine Holstein lactating cows were submitted in trios to a 3-choice preference test over 14 d (2 d of adaptation, 3 d of restriction to each stall, and 3 d of free access to all 3 stalls). Cows were housed individually in pens containing 3 stalls with different lying surfaces: (1) rubber mat with chopped wheat straw (WS); (2) deep-bedded switchgrass (SG); and (3) deep-bedded switchgrass, water, and lime mixture (SGL). The second objective (experiment 2) was to test, in freestall housing, the effects of these 3 types of bedding on lying behavior, cow cleanliness, and teat end bacterial contamination. Bedding treatments were compared in a 3 × 3 Latin square design using 24 cows split into groups of 8, with bedding materials being switched every 4 wk. Lying behavior was measured with data loggers in both studies. During experiment 1, cows chose to spend more time lying and had more frequent lying bouts on SG (9.4 h/d; 8.2 bouts/d) than on SGL (1.0 h/d; 0.9 bouts/d). They also spent more time standing and stood more frequently in stalls with SG (2.0 h/d; 10.1 bouts/d) than in those with SGL (0.6 h/d; 2.6 bouts/d), and stood longer in stalls with SG than with WS (0.6 h/d). In experiment 2, the total lying time, frequency of lying bouts, and mean lying bout duration were, on average, 9.7 ± 1.03 h/d, 8.2 ± 0.93 bouts/d, and 1.2 ± 0.06 h/bout, respectively, and did not differ between treatments. No treatment effects were found for cow cleanliness scores. Bedding dry matter was highest for SG (74.1%), lowest for SGL (63.5%), and intermediate for WS (68.6%) [standard error of the mean (SEM) = 1.57%]. This may explain the higher teat end count of coliforms for cows on SGL (0.92 log 10 cfu/g) compared with WS (0.13 log 10 cfu/g) (SEM = 0.144 log 10 cfu/g). In conclusion, cows preferred the deep-bedded switchgrass surface over the other 2 surfaces, and deep-bedded switchgrass appears to be a suitable bedding alternative for dairy cows. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The receptor-like kinase AtVRLK1 regulates secondary cell wall thickening.

PubMed

Huang, Cheng; Zhang, Rui; Gui, Jinshan; Zhong, Yu; Li, Laigeng

2018-04-20

During the growth and development of land plants, some specialized cells, such as tracheary elements, undergo secondary cell wall thickening. Secondary cell walls contain additional lignin, compared with primary cell walls, thus providing mechanical strength and potentially improving defenses against pathogens. However, the molecular mechanisms that initiate wall thickening are unknown. In this study, we identified an Arabidopsis thaliana leucine-rich repeat receptor-like kinase, encoded by AtVRLK1 (Vascular-Related RLK 1), that is specifically expressed in cells undergoing secondary cell wall thickening. Suppression of AtVRLK1expression resulted in a range of phenotypes that included retarded early elongation of the inflorescence stem, shorter fibers, slower root growth, and shorter flower filaments. In contrast, upregulation of AtVRLK1 led to longer fiber cells, reduced secondary cell wall thickening in fiber and vessel cells, and defects in anther dehiscence. Molecular and cellular analyses showed that downregulation of AtVRLK1 promoted secondary cell wall thickening and upregulation of AtVRLK1 enhanced cell elongation and inhibited secondary cell wall thickening. We propose that AtVRLK1 functions as a signaling component in coordinating cell elongation and cell wall thickening during growth and development. {copyright, serif} 2018 American Society of Plant Biologists. All rights reserved.

Spatio-temporal diversification of the cell wall matrix materials in the developing stomatal complexes of Zea mays.

PubMed

Giannoutsou, E; Apostolakos, P; Galatis, B

2016-11-01

The matrix cell wall materials, in developing Zea mays stomatal complexes are asymmetrically distributed, a phenomenon appearing related to the local cell wall expansion and deformation, the establishment of cell polarity, and determination of the cell division plane. In cells of developing Zea mays stomatal complexes, definite cell wall regions expand determinately and become locally deformed. This differential cell wall behavior is obvious in the guard cell mother cells (GMCs) and the subsidiary cell mother cells (SMCs) that locally protrude towards the adjacent GMCs. The latter, emitting a morphogenetic stimulus, induce polarization/asymmetrical division in SMCs. Examination of immunolabeled specimens revealed that homogalacturonans (HGAs) with a high degree of de-esterification (2F4- and JIM5-HGA epitopes) and arabinogalactan proteins are selectively distributed in the extending and deformed cell wall regions, while their margins are enriched with rhamnogalacturonans (RGAs) containing highly branched arabinans (LM6-RGA epitope). In SMCs, the local cell wall matrix differentiation constitutes the first structural event, indicating the establishment of cell polarity. Moreover, in the premitotic GMCs and SMCs, non-esterified HGAs (2F4-HGA epitope) are preferentially localized in the cell wall areas outlining the cytoplasm where the preprophase band is formed. In these areas, the forthcoming cell plate fuses with the parent cell walls. These data suggest that the described heterogeneity in matrix cell wall materials is probably involved in: (a) local cell wall expansion and deformation, (b) the transduction of the inductive GMC stimulus, and (c) the determination of the division plane in GMCs and SMCs.

Immunogold scanning electron microscopy can reveal the polysaccharide architecture of xylem cell walls

PubMed Central

Sun, Yuliang; Juzenas, Kevin

2017-01-01

Abstract Immunofluorescence microscopy (IFM) and immunogold transmission electron microscopy (TEM) are the two main techniques commonly used to detect polysaccharides in plant cell walls. Both are important in localizing cell wall polysaccharides, but both have major limitations, such as low resolution in IFM and restricted sample size for immunogold TEM. In this study, we have developed a robust technique that combines immunocytochemistry with scanning electron microscopy (SEM) to study cell wall polysaccharide architecture in xylem cells at high resolution over large areas of sample. Using multiple cell wall monoclonal antibodies (mAbs), this immunogold SEM technique reliably localized groups of hemicellulosic and pectic polysaccharides in the cell walls of five different xylem structures (vessel elements, fibers, axial and ray parenchyma cells, and tyloses). This demonstrates its important advantages over the other two methods for studying cell wall polysaccharide composition and distribution in these structures. In addition, it can show the three-dimensional distribution of a polysaccharide group in the vessel lateral wall and the polysaccharide components in the cell wall of developing tyloses. This technique, therefore, should be valuable for understanding the cell wall polysaccharide composition, architecture and functions of diverse cell types. PMID:28398585

Building a plant cell wall at a glance.

PubMed

Lampugnani, Edwin R; Khan, Ghazanfar Abbas; Somssich, Marc; Persson, Staffan

2018-01-29

Plant cells are surrounded by a strong polysaccharide-rich cell wall that aids in determining the overall form, growth and development of the plant body. Indeed, the unique shapes of the 40-odd cell types in plants are determined by their walls, as removal of the cell wall results in spherical protoplasts that are amorphic. Hence, assembly and remodeling of the wall is essential in plant development. Most plant cell walls are composed of a framework of cellulose microfibrils that are cross-linked to each other by heteropolysaccharides. The cell walls are highly dynamic and adapt to the changing requirements of the plant during growth. However, despite the importance of plant cell walls for plant growth and for applications that we use in our daily life such as food, feed and fuel, comparatively little is known about how they are synthesized and modified. In this Cell Science at a Glance article and accompanying poster, we aim to illustrate the underpinning cell biology of the synthesis of wall carbohydrates, and their incorporation into the wall, in the model plant Arabidopsis . © 2018. Published by The Company of Biologists Ltd.

Cell wall evolution and diversity

PubMed Central

Fangel, Jonatan U.; Ulvskov, Peter; Knox, J. P.; Mikkelsen, Maria D.; Harholt, Jesper; Popper, Zoë A.; Willats, William G.T.

2012-01-01

Plant cell walls display a considerable degree of diversity in their compositions and molecular architectures. In some cases the functional significance of a particular cell wall type appears to be easy to discern: secondary cells walls are often reinforced with lignin that provides durability; the thin cell walls of pollen tubes have particular compositions that enable their tip growth; lupin seed cell walls are characteristically thickened with galactan used as a storage polysaccharide. However, more frequently the evolutionary mechanisms and selection pressures that underpin cell wall diversity and evolution are unclear. For diverse green plants (chlorophytes and streptophytes) the rapidly increasing availability of transcriptome and genome data sets, the development of methods for cell wall analyses which require less material for analysis, and expansion of molecular probe sets, are providing new insights into the diversity and occurrence of cell wall polysaccharides and associated biosynthetic genes. Such research is important for refining our understanding of some of the fundamental processes that enabled plants to colonize land and to subsequently radiate so comprehensively. The study of cell wall structural diversity is also an important aspect of the industrial utilization of global polysaccharide bio-resources. PMID:22783271

Distribution of alginate and cellulose and regulatory role of calcium in the cell wall of the brown alga Ectocarpus siliculosus (Ectocarpales, Phaeophyceae).

PubMed

Terauchi, Makoto; Nagasato, Chikako; Inoue, Akira; Ito, Toshiaki; Motomura, Taizo

2016-08-01

This work investigated a correlation between the three-dimensional architecture and compound-components of the brown algal cell wall. Calcium greatly contributes to the cell wall integrity. Brown algae have a unique cell wall consisting of alginate, cellulose, and sulfated polysaccharides. However, the relationship between the architecture and the composition of the cell wall is poorly understood. Here, we investigated the architecture of the cell wall and the effect of extracellular calcium in the sporophyte and gametophyte of the model brown alga, Ectocarpus siliculosus (Dillwyn) Lyngbye, using transmission electron microscopy, histochemical, and immunohistochemical studies. The lateral cell wall of vegetative cells of the sporophyte thalli had multilayered architecture containing electron-dense and negatively stained fibrils. Electron tomographic analysis showed that the amount of the electron-dense fibrils and the junctions was different between inner and outer layers, and between the perpendicular and tangential directions of the cell wall. By immersing the gametophyte thalli in the low-calcium (one-eighth of the normal concentration) artificial seawater medium, the fibrous layers of the lateral cell wall of vegetative cells became swollen. Destruction of cell wall integrity was also induced by the addition of sorbitol. The results demonstrated that electron-dense fibrils were composed of alginate-calcium fibrous gels, and electron negatively stained fibrils were crystalline cellulose microfibrils. It was concluded that the spatial arrangement of electron-dense fibrils was different between the layers and between the directions of the cell wall, and calcium was necessary for maintaining the fibrous layers in the cell wall. This study provides insights into the design principle of the brown algal cell wall.

Cell Wall Composition and Candidate Biosynthesis Gene Expression During Rice Development.

PubMed

Lin, Fan; Manisseri, Chithra; Fagerström, Alexandra; Peck, Matthew L; Vega-Sánchez, Miguel E; Williams, Brian; Chiniquy, Dawn M; Saha, Prasenjit; Pattathil, Sivakumar; Conlin, Brian; Zhu, Lan; Hahn, Michael G; Willats, William G T; Scheller, Henrik V; Ronald, Pamela C; Bartley, Laura E

2016-10-01

Cell walls of grasses, including cereal crops and biofuel grasses, comprise the majority of plant biomass and intimately influence plant growth, development and physiology. However, the functions of many cell wall synthesis genes, and the relationships among and the functions of cell wall components remain obscure. To better understand the patterns of cell wall accumulation and identify genes that act in grass cell wall biosynthesis, we characterized 30 samples from aerial organs of rice (Oryza sativa cv. Kitaake) at 10 developmental time points, 3-100 d post-germination. Within these samples, we measured 15 cell wall chemical components, enzymatic digestibility and 18 cell wall polysaccharide epitopes/ligands. We also used quantitative reverse transcription-PCR to measure expression of 50 glycosyltransferases, 15 acyltransferases and eight phenylpropanoid genes, many of which had previously been identified as being highly expressed in rice. Most cell wall components vary significantly during development, and correlations among them support current understanding of cell walls. We identified 92 significant correlations between cell wall components and gene expression and establish nine strong hypotheses for genes that synthesize xylans, mixed linkage glucan and pectin components. This work provides an extensive analysis of cell wall composition throughout rice development, identifies genes likely to synthesize grass cell walls, and provides a framework for development of genetically improved grasses for use in lignocellulosic biofuel production and agriculture. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Screening and characterization of plant cell walls using carbohydrate microarrays.

PubMed

Sørensen, Iben; Willats, William G T

2011-01-01

Plant cells are surrounded by cell walls built largely from complex carbohydrates. The primary walls of growing plant cells consist of interdependent networks of three polysaccharide classes: cellulose, cross-linking glycans (also known as hemicelluloses), and pectins. Cellulose microfibrils are tethered together by cross-linking glycans, and this assembly forms the major load-bearing component of primary walls, which is infiltrated with pectic polymers. In the secondary walls of woody tissues, pectins are much reduced and walls are reinforced with the phenolic polymer lignin. Plant cell walls are essential for plant life and also have numerous industrial applications, ranging from wood to nutraceuticals. Enhancing our knowledge of cell wall biology and the effective use of cell wall materials is dependent to a large extent on being able to analyse their fine structures. We have developed a suite of techniques based on microarrays probed with monoclonal antibodies with specificity for cell wall components, and here we present practical protocols for this type of analysis.

Characteristic thickened cell walls of the bracts of the 'eternal flower' Helichrysum bracteatum.

PubMed

Nishikawa, Kuniko; Ito, Hiroaki; Awano, Tatsuya; Hosokawa, Munetaka; Yazawa, Susumu

2008-07-01

Helichrysum bracteatum is called an 'eternal flower' and has large, coloured, scarious bracts. These maintain their aesthetic value without wilting or discoloration for many years. There have been no research studies of cell death or cell morphology of the scarious bract, and hence the aim of this work was to elucidate these characteristics for the bract of H. bracteatum. DAPI (4'6-diamidino-2-phenylindol dihydrochloride) staining and fluorescence microscopy were used for observation of cell nuclei. Light microscopy (LM), transmission electron microscopy (TEM) and polarized light microscopy were used for observation of cells, including cell wall morphology. Cell death occurred at the bract tip during the early stage of flower development. The cell wall was the most prominent characteristic of H. bracteatum bract cells. Characteristic thickened secondary cell walls on the inside of the primary cell walls were observed in both epidermal and inner cells. In addition, the walls of all cells exhibited birefringence. Characteristic thickened secondary cell walls have orientated cellulose microfibrils as well as general secondary cell walls of the tracheary elements. For comparison, these characters were not observed in the petal and bract tissues of Chrysanthemum morifolium. Bracts at anthesis are composed of dead cells. Helichrysum bracteatum bracts have characteristic thickened secondary cell walls that have not been observed in the parenchyma of any other flowers or leaves. The cells of the H. bracteatum bract differ from other tissues with secondary cell walls, suggesting that they may be a new cell type.

Peptidoglycan turnover and recycling in Gram-positive bacteria.

PubMed

Reith, Jan; Mayer, Christoph

2011-10-01

Bacterial cells are protected by an exoskeleton, the stabilizing and shape-maintaining cell wall, consisting of the complex macromolecule peptidoglycan. In view of its function, it could be assumed that the cell wall is a static structure. In truth, however, it is steadily broken down by peptidoglycan-cleaving enzymes during cell growth. In this process, named cell wall turnover, in one generation up to half of the preexisting peptidoglycan of a bacterial cell is released from the wall. This would result in a massive loss of cell material, if turnover products were not be taken up and recovered. Indeed, in the Gram-negative model organism Escherichia coli, peptidoglycan recovery has been recognized as a complex pathway, named cell wall recycling. It involves about a dozen dedicated recycling enzymes that convey cell wall turnover products to peptidoglycan synthesis or energy pathways. Whether Gram-positive bacteria also recover their cell wall is currently questioned. Given the much larger portion of peptidoglycan in the cell wall of Gram-positive bacteria, however, recovery of the wall material would provide an even greater benefit in these organisms compared to Gram-negatives. Consistently, in many Gram-positives, orthologs of recycling enzymes were identified, indicating that the cell wall may also be recycled in these organisms. This mini-review provides a compilation of information about cell wall turnover and recycling in Gram-positive bacteria during cell growth and division, including recent findings relating to muropeptide recovery in Bacillus subtilis and Clostridium acetobutylicum from our group. Furthermore, the impact of cell wall turnover and recycling on biotechnological processes is discussed.

RADIOAUTOGRAPHIC STUDY OF CELL WALL DEPOSITION IN GROWING PLANT CELLS

PubMed Central

Ray, Peter M.

1967-01-01

Segments cut from growing oat coleoptiles and pea stems were fed glucose-3H in presence and absence of the growth hormone indoleacetic acid (IAA). By means of electron microscope radioautography it was demonstrated that new cell wall material is deposited both at the wall surface (apposition) and within the preexisting wall structure (internally). Quantitative profiles for the distribution of incorporation with position through the thickness of the wall were obtained for the thick outer wall of epidermal cells. With both oat coleoptile and pea stem epidermal outer walls, it was found that a larger proportion of the newly synthesized wall material appeared to become incorporated within the wall in the presence of IAA. Extraction experiments on coleoptile tissue showed that activity that had been incorporated into the cell wall interior represented noncellulosic constituents, mainly hemicelluloses, whereas cellulose was deposited largely or entirely by apposition. It seems possible that internal incorporation of hemicelluloses plays a role in the cell wall expansion process that is involved in cell growth. PMID:6064369

Interactions of Condensed Tannins with Saccharomyces cerevisiae Yeast Cells and Cell Walls: Tannin Location by Microscopy.

PubMed

Mekoue Nguela, Julie; Vernhet, Aude; Sieczkowski, Nathalie; Brillouet, Jean-Marc

2015-09-02

Interactions between grape tannins/red wine polyphenols and yeast cells/cell walls was previously studied within the framework of red wine aging and the use of yeast-derived products as an alternative to aging on lees. Results evidenced a quite different behavior between whole cells (biomass grown to elaborate yeast-derived products, inactivated yeast, and yeast inactivated after autolysis) and yeast cell walls (obtained from mechanical disruption of the biomass). Briefly, whole cells exhibited a high capacity to irreversibly adsorb grape and wine tannins, whereas only weak interactions were observed for cell walls. This last point was quite unexpected considering the literature and called into question the real role of cell walls in yeasts' ability to fix tannins. In the present work, tannin location after interactions between grape and wine tannins and yeast cells and cell walls was studied by means of transmission electron microscopy, light epifluorescence, and confocal microscopy. Microscopy observations evidenced that if tannins interact with cell walls, and especially cell wall mannoproteins, they also diffuse freely through the walls of dead cells to interact with their plasma membrane and cytoplasmic components.

Optimization of Southeastern Forest Biomass Crop Production: A Watershed Scale Evaluation of the Sustainability and Productivity of Dedicated Energy Crop and Woody Biomass Operations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chescheir, George M.; Nettles, Jami E,; Youssef, Mohamed

Growing switchgrass (Panicum virgatum L.) as an intercrop in managed loblolly pine (Pinus taeda L.) plantations has emerged as a potential source of bioenergy feedstock. Utilizing land resources between pine trees to produce an energy crop can potentially reduce the demand for land resources used to produce food; however, converting conventionally managed forest land to this new intercropping system constitutes changes in land use and associated management practices, which may affect the environmental and economic sustainability of the land. The overall objective of this project is to evaluate the environmental effects of large-scale forest bioenergy crop production and utilize thesemore » results to optimize cropping systems in a manner that protects the important ecosystem services provided by forests while contributing to the development of a sustainable and economically-viable biomass industry in the southeastern United States. Specific objectives are to: Quantify the hydrology of different energy crop production systems in watershed scale experiments on different landscapes in the southeast. Quantify the nutrient dynamics of energy crop production systems in watershed scale experiments to determine the impact of these systems on water quality. Evaluate the impacts of energy crop production on soil structure, fertility, and organic matter. Evaluate the response of flora and fauna populations and habitat quality to energy crop production systems. Develop watershed and regional scale models to evaluate the environmental sustainability and productivity of energy crop and woody biomass operations. Quantify the production systems in terms of bioenergy crop yield versus the energy and economic costs of production. Develop and evaluate best management practice guidelines to ensure the environmental sustainability of energy crop production systems. Watershed and plot scale studies formed the core of this research platform. Matched-watershed studies were established in North Carolina, Mississippi and Alabama. A plot scale study was also established in North Carolina to more intensive examination of the effects of biomass production on hydrology, soil properties, productivity wildlife habitat, and biodiversity on replicate 0.8 ha plots. Studies were also conducted on selected sites to define and quantify the environmental effects of biomass production on wildlife habitat, biodiversity, soil properties and productivity, and carbon storage and flux. Treatments on the sub-watersheds and plots included potential operational systems ranging from monoculture switchgrass to interplanted switchgrass to conventional managed forests as a controls. The hydrology, water quality, soil property, and productivity data collected in the watershed and plot scale experiments were used to develop process based watershed scale models. Existing models (DRAINMOD and APEX) were modified to more effectively simulate the intercropped systems. More regional scale models (DRAINMOD-INTERCROP) with GIS interface and SWAT) were used to simulate the impacts of intercropping switchgrass in pine plantations on the hydrology and water quality of larger scale watersheds. Results from the watershed and plot scale studies, and the modeling studies were used to develop Best Management Practice (BMP) guidelines to ensure environmentally sustainable bioenergy production in the forestry setting. While the results of the environmental sustainability research for this project have become publically available, many of the planning decisions and operational trial results were not public. Personnel in management, planning, operations, and logistics were interviewed to capture the important economic and operational lessons from internal operational research on approximately 30 full-scale operational tracts. This project produced a very large database documenting the impact of interplanting switchgrass with pine trees on hydrology, water quality, soil quality, and biodiversity. Some environmental impacts were observed in response to additional operations required for interplanting, but these impacts were small and short lived. Given that existing forestry BMPs provide a flexible system that can be adapted to protect water quality and biodiversity in forestry settings, interplanting switchgrass with pine trees can be considered environmentally sustainable. The project also developed models that can simulate switchgrass growth when it is in competition with pine trees as well as the hydrology and nutrient dynamics that result from this interplanted system. The models predicted switchgrass production, water use, and the quality of the water leaving the system over a range of climatological and geographic conditions. These models can be used to guide decisions toward sustainability. The project also documented the limitations of switchgrass production in the forestry setting and the challenges and increased costs arising from this practice. These challenges led to the conclusion that intercropping switchgrass with pine trees is not economically feasible in the current economic climate. Despite the barriers obstructing use of this system at this point in time, economic and technological changes may occur that will make this a feasible system for bioenergy production in the future. The data, models, BMPs and experiences documented in this report and in publications resulting from this project will be highly valuable to those implementing this system.« less

At the border: the plasma membrane-cell wall continuum.

PubMed

Liu, Zengyu; Persson, Staffan; Sánchez-Rodríguez, Clara

2015-03-01

Plant cells rely on their cell walls for directed growth and environmental adaptation. Synthesis and remodelling of the cell walls are membrane-related processes. During cell growth and exposure to external stimuli, there is a constant exchange of lipids, proteins, and other cell wall components between the cytosol and the plasma membrane/apoplast. This exchange of material and the localization of cell wall proteins at certain spots in the plasma membrane seem to rely on a particular membrane composition. In addition, sensors at the plasma membrane detect changes in the cell wall architecture, and activate cytoplasmic signalling schemes and ultimately cell wall remodelling. The apoplastic polysaccharide matrix is, on the other hand, crucial for preventing proteins diffusing uncontrollably in the membrane. Therefore, the cell wall-plasma membrane link is essential for plant development and responses to external stimuli. This review focuses on the relationship between the cell wall and plasma membrane, and its importance for plant tissue organization. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Plant and algal cell walls: diversity and functionality

PubMed Central

Popper, Zoë A.; Ralet, Marie-Christine; Domozych, David S.

2014-01-01

Background Although plants and many algae (e.g. the Phaeophyceae, brown, and Rhodophyceae, red) are only very distantly related they are united in their possession of carbohydrate-rich cell walls, which are of integral importance being involved in many physiological processes. Furthermore, wall components have applications within food, fuel, pharmaceuticals, fibres (e.g. for textiles and paper) and building materials and have long been an active topic of research. As shown in the 27 papers in this Special Issue, as the major deposit of photosynthetically fixed carbon, and therefore energy investment, cell walls are of undisputed importance to the organisms that possess them, the photosynthetic eukaryotes (plants and algae). The complexities of cell wall components along with their interactions with the biotic and abiotic environment are becoming increasingly revealed. Scope The importance of plant and algal cell walls and their individual components to the function and survival of the organism, and for a number of industrial applications, are illustrated by the breadth of topics covered in this issue, which includes papers concentrating on various plants and algae, developmental stages, organs, cell wall components, and techniques. Although we acknowledge that there are many alternative ways in which the papers could be categorized (and many would fit within several topics), we have organized them as follows: (1) cell wall biosynthesis and remodelling, (2) cell wall diversity, and (3) application of new technologies to cell walls. Finally, we will consider future directions within plant cell wall research. Expansion of the industrial uses of cell walls and potentially novel uses of cell wall components are both avenues likely to direct future research activities. Fundamentally, it is the continued progression from characterization (structure, metabolism, properties and localization) of individual cell wall components through to defining their roles in almost every aspect of plant and algal physiology that will present many of the major challenges in future cell wall research. PMID:25453142

Plant and algal cell walls: diversity and functionality.

PubMed

Popper, Zoë A; Ralet, Marie-Christine; Domozych, David S

2014-10-01

Although plants and many algae (e.g. the Phaeophyceae, brown, and Rhodophyceae, red) are only very distantly related they are united in their possession of carbohydrate-rich cell walls, which are of integral importance being involved in many physiological processes. Furthermore,wall components have applications within food, fuel, pharmaceuticals, fibres (e.g. for textiles and paper) and building materials and have long been an active topic of research. As shown in the 27 papers in this Special Issue, as the major deposit of photosynthetically fixed carbon, and therefore energy investment, cell walls are of undisputed importance to the organisms that possess them, the photosynthetic eukaryotes ( plants and algae). The complexities of cell wall components along with their interactions with the biotic and abiotic environment are becoming increasingly revealed. The importance of plant and algal cell walls and their individual components to the function and survival of the organism, and for a number of industrial applications, are illustrated by the breadth of topics covered in this issue, which includes papers concentrating on various plants and algae, developmental stages, organs, cell wall components, and techniques. Although we acknowledge that there are many alternative ways in which the papers could be categorized (and many would fit within several topics), we have organized them as follows: (1) cell wall biosynthesis and remodelling, (2) cell wall diversity, and (3) application of new technologies to cell walls. Finally, we will consider future directions within plant cell wall research. Expansion of the industrial uses of cell walls and potentially novel uses of cell wall components are both avenues likely to direct future research activities. Fundamentally, it is the continued progression from characterization (structure, metabolism, properties and localization) of individual cell wall components through to defining their roles in almost every aspect of plant and algal physiology that will present many of the major challenges in future cell wall research.

Bioenergy crop productivity and potential climate change mitigation from marginal lands in the United States: An ecosystem modeling perspective

DOE PAGES

Qin, Zhangcai; Zhuang, Qianlai; Cai, Ximing

2014-06-16

Growing biomass feedstocks from marginal lands is becoming an increasingly attractive choice for producing biofuel as an alternative energy to fossil fuels. Here, we used a biogeochemical model at ecosystem scale to estimate crop productivity and greenhouse gas (GHG) emissions from bioenergy crops grown on marginal lands in the United States. Two broadly tested cellulosic crops, switchgrass, and Miscanthus, were assumed to be grown on the abandoned land and mixed crop–vegetation land with marginal productivity. Production of biomass and biofuel as well as net carbon exchange and nitrous oxide emissions were estimated in a spatially explicit manner. We found that,more » cellulosic crops, especially Miscanthus could produce a considerable amount of biomass, and the effective ethanol yield is high on these marginal lands. For every hectare of marginal land, switchgrass and Miscanthus could produce 1.0–2.3 kl and 2.9–6.9 kl ethanol, respectively, depending on nitrogen fertilization rate and biofuel conversion efficiency. Nationally, both crop systems act as net GHG sources. Switchgrass has high global warming intensity (100–390 g CO 2eq l –1 ethanol), in terms of GHG emissions per unit ethanol produced. Miscanthus, however, emits only 21–36 g CO 2eq to produce every liter of ethanol. To reach the mandated cellulosic ethanol target in the United States, growing Miscanthus on the marginal lands could potentially save land and reduce GHG emissions in comparison to growing switchgrass. Furthermore, the ecosystem modeling is still limited by data availability and model deficiencies, further efforts should be made to classify crop–specific marginal land availability, improve model structure, and better integrate ecosystem modeling into life cycle assessment.« less

Phytoextraction of contaminated urban soils by Panicum virgatum L. enhanced with application of a plant growth regulator (BAP) and citric acid.

PubMed

Aderholt, Matthew; Vogelien, Dale L; Koether, Marina; Greipsson, Sigurdur

2017-05-01

Lead (Pb) contamination in soil represents a threat to human health. Phytoextraction has gained attention as a potential alternative to traditional remediation methods because of lower cost and minimal soil disruption. The North American native switchgrass (Panicum virgatum L.) was targeted due to its ability to produce high biomass and grow across a variety of ecozones. In this study switchgrass was chemically enhanced with applications of the soil-fungicide benomyl, chelates (EDTA and citric acid), and PGR to optimize phytoextraction of Pb and zinc (Zn) from contaminated urban soils in Atlanta, GA. Exogenous application of two plant hormones was compared in multiple concentrations to determine effects on switchgrass growth: indole-3-acetic acid (IAA), and Gibberellic Acid (GA 3 ), and one PGR benzylaminopurine (BAP), The PGR BAP (1.0 μM) was found to generate a 48% increase in biomass compared to Control plants. Chemical application of citric acid, EDTA, benomyl, and BAP were tested separately and in combination in a pot experiment in an environmentally controlled greenhouse to determine the efficacy of phtyoextraction by switchgrass. Soil acidification by citric acid application resulted in highest level of aluminum (Al) and iron (Fe) in plants foliage resulting in severe phytotoxic effects. Total Pb phytoextraction was significantly highest in plants treated with combined chemical application of B + C and B + C + H. Suppression of AMF activities by benomyl application significantly increased concentrations of Al and Fe in roots. Application of benomyl reduced AMF colonization but was also shown to dramatically increase levels of septa fungi infection as compared to Control plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

A simple and reliable multi-gene transformation method for switchgrass.

PubMed

Ogawa, Yoichi; Shirakawa, Makoto; Koumoto, Yasuko; Honda, Masaho; Asami, Yuki; Kondo, Yasuhiro; Hara-Nishimura, Ikuko

2014-07-01

A simple and reliable Agrobacterium -mediated transformation method was developed for switchgrass. Using this method, many transgenic plants carrying multiple genes-of-interest could be produced without untransformed escape. Switchgrass (Panicum virgatum L.) is a promising biomass crop for bioenergy. To obtain transgenic switchgrass plants carrying a multi-gene trait in a simple manner, an Agrobacterium-mediated transformation method was established by constructing a Gateway-based binary vector, optimizing transformation conditions and developing a novel selection method. A MultiRound Gateway-compatible destination binary vector carrying the bar selectable marker gene, pHKGB110, was constructed to introduce multiple genes of interest in a single transformation. Two reporter gene expression cassettes, GUSPlus and gfp, were constructed independently on two entry vectors and then introduced into a single T-DNA region of pHKGB110 via sequential LR reactions. Agrobacterium tumefaciens EHA101 carrying the resultant binary vector pHKGB112 and caryopsis-derived compact embryogenic calli were used for transformation experiments. Prolonged cocultivation for 7 days followed by cultivation on media containing meropenem improved transformation efficiency without overgrowth of Agrobacterium, which was, however, not inhibited by cefotaxime or Timentin. In addition, untransformed escape shoots were completely eliminated during the rooting stage by direct dipping the putatively transformed shoots into the herbicide Basta solution for a few seconds, designated as the 'herbicide dipping method'. It was also demonstrated that more than 90 % of the bar-positive transformants carried both reporters delivered from pHKGB112. This simple and reliable transformation method, which incorporates a new selection technique and the use of a MultiRound Gateway-based binary vector, would be suitable for producing a large number of transgenic lines carrying multiple genes.

The effect of biomass densification on structural sugar release and yield in biofuel feedstock and feedstock blends

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wolfrum, Edward J.; Nagle, Nicholas J.; Ness, Ryan M.

In this work, we examined the behavior of feedstock blends and the effect of a specific feedstock densification strategy (pelleting) on the release and yield of structural carbohydrates in a laboratory-scale dilute acid pretreatment (PT) and enzymatic hydrolysis (EH) assay. We report overall carbohydrate release and yield from the two-stage PT-EH assay for five single feedstocks (two corn stovers, miscanthus, switchgrass, and hybrid poplar) and three feedstock blends (corn stover-switchgrass, corn stover-switchgrass-miscanthus, and corn stover-switchgrass-hybrid poplar). We first examined the experimental results over time to establish the robustness of the PT-EH assay, which limits the precision of the experimental results.more » The use of two different control samples in the assay enabled us to identify (and correct for) a small bias in the EH portion of the combined assay for some runs. We then examined the effect of variable pretreatment reaction conditions (residence time, acid loading, and reactor temperature) on the conversion of a single feedstock (single-pass corn stover, CS-SP) in order to establish the range of pretreatment reaction conditions likely to provide optimal conversion data. Finally, we applied the assay to the 16 materials (8 feedstocks in 2 formats, loose and pelleted) over a more limited range of pretreatment experimental conditions. The four herbaceous feedstocks behaved similarly, while the hybrid poplar feedstock required higher pretreatment temperatures for optimal results. As expected, the yield data for three blended feedstocks were the average of the yield data for the individual feedstocks. As a result, the pelleting process appears to provide a slightly positive effect on overall total sugar yield.« less

The effect of biomass densification on structural sugar release and yield in biofuel feedstock and feedstock blends

DOE PAGES

Wolfrum, Edward J.; Nagle, Nicholas J.; Ness, Ryan M.; ...

2017-01-13

In this work, we examined the behavior of feedstock blends and the effect of a specific feedstock densification strategy (pelleting) on the release and yield of structural carbohydrates in a laboratory-scale dilute acid pretreatment (PT) and enzymatic hydrolysis (EH) assay. We report overall carbohydrate release and yield from the two-stage PT-EH assay for five single feedstocks (two corn stovers, miscanthus, switchgrass, and hybrid poplar) and three feedstock blends (corn stover-switchgrass, corn stover-switchgrass-miscanthus, and corn stover-switchgrass-hybrid poplar). We first examined the experimental results over time to establish the robustness of the PT-EH assay, which limits the precision of the experimental results.more » The use of two different control samples in the assay enabled us to identify (and correct for) a small bias in the EH portion of the combined assay for some runs. We then examined the effect of variable pretreatment reaction conditions (residence time, acid loading, and reactor temperature) on the conversion of a single feedstock (single-pass corn stover, CS-SP) in order to establish the range of pretreatment reaction conditions likely to provide optimal conversion data. Finally, we applied the assay to the 16 materials (8 feedstocks in 2 formats, loose and pelleted) over a more limited range of pretreatment experimental conditions. The four herbaceous feedstocks behaved similarly, while the hybrid poplar feedstock required higher pretreatment temperatures for optimal results. As expected, the yield data for three blended feedstocks were the average of the yield data for the individual feedstocks. As a result, the pelleting process appears to provide a slightly positive effect on overall total sugar yield.« less

Miscanthus and switchgrass production in central Illinois: impacts on hydrology and inorganic nitrogen leaching.

PubMed

McIsaac, Gregory F; David, Mark B; Mitchell, Corey A

2010-01-01

Biomass crops are being promoted as environmentally favorable alternatives to fossil fuels or ethanol production from maize (Zea mays L.), particularly across the Corn Belt of the United States. However, there are few if any empirical studies on inorganic N leaching losses from perennial grasses that are harvested on an annual basis, nor has there been empirical evaluation of the hydrologic consequences of perennial cropping systems. Here we report on the results of 4 yr of field measurements of soil moisture and inorganic N leaching from a conventional maize-soybean [Glycine max (L.) Merr.] system and two unfertilized perennial grasses harvested in winter for biomass: Miscanthus x giganteus and switchgrass (Panicum virgatum cv. Cave-in-Rock). All crops were grown on fertile Mollisols in east-central Illinois. Inorganic N leaching was measured with ion exchange resin lysimeters placed 50 cm below the soil surface. Maize--soybean nitrate leaching averaged 40.4 kg N ha(-1) yr(-1), whereas switchgrass and Miscanthus had values of 1.4 and 3.0 kg N ha(-1) yr(-1), respectively. Soil moisture monitoring (to a depth of 90 cm) indicated that both perennial grasses dried the soil out earlier in the growing season compared with maize-soybean. Later in the growing season, soil moisture under switchgrass tended to be greater than maize-soybean or Miscanthus, whereas the soil under Miscanthus was consistently drier than under maize--soybean. Water budget calculations indicated that evapotranspiration from Miscanthus was about 104 mm yr(-1) greater than under maize-soybean, which could reduce annual drainage water flows by 32% in central Illinois. Drainage water is a primary source of surface water flows in the region, and the impact ofextensive Miscanthus production on surface water supplies and aquatic ecosystems deserves further investigation.

Genomic diversity in switchgrass (Panicum virgatum): from the continental scale to a dune landscape

PubMed Central

Morris, Geoffrey P.; Grabowski, Paul; Borevitz, Justin O.

2011-01-01

Connecting broad-scale patterns of genetic variation and population structure to genetic diversity on a landscape is a key step towards understanding historical processes of migration and adaptation. New genomic approaches can be used to increase the resolution of phylogeographic studies while reducing locus sampling effects and circumventing ascertainment bias. Here, we use a novel approach based on high-throughput sequencing to characterize genetic diversity in complete chloroplast genomes and >10,000 nuclear loci in switchgrass, across a continental and landscape scale. Switchgrass is a North American tallgrass species, which is widely used in conservation and perennial biomass production, and shows strong ecotypic adaptation and population structure across the continental range. We sequenced 40.9 billion base pairs from 24 individuals from across the species’ range and 20 individuals from the Indiana Dunes. Analysis of plastome sequence revealed 203 variable SNP sites that define eight haplogroups, which are differentiated by 4 to 127 SNPs and confirmed by patterns of indel variation. These include three deeply divergent haplogroups, which correspond to the previously described lowland-upland ecotypic split and a novel upland haplogroup split that dates to the mid-Pleistoscene. Most of the plastome haplogroup diversity present in the northern switchgrass range, including in the Indiana Dunes, originated in the mid- or upper-Pleistocene prior to the most recent postglacial recolonization. Furthermore, a recently colonized landscape feature (~150 ya) in the Indiana Dunes contains several deeply divergent upland haplogroups. Nuclear markers also support a deep lowland-upland split, followed by limited gene flow, and show extensive gene flow in the local population of the Indiana Dunes. PMID:22060816

DBIO Best Thesis Award: Mechanics, Dynamics, and Organization of the Bacterial Cytoskeleton and Cell Wall

NASA Astrophysics Data System (ADS)

Wang, Siyuan

2012-02-01

Bacteria come in a variety of shapes. While the peptidoglycan (PG) cell wall serves as an exoskeleton that defines the static cell shape, the internal bacterial cytoskeleton mediates cell shape by recruiting PG synthesis machinery and thus defining the pattern of cell-wall synthesis. While much is known about the chemistry and biology of the cytoskeleton and cell wall, much of their biophysics, including essential aspects of the functionality, dynamics, and organization, remain unknown. This dissertation aims to elucidate the detailed biophysical mechanisms of cytoskeleton guided wall synthesis. First, I find that the bacterial cytoskeleton MreB contributes nearly as much to the rigidity of an Escherichia coli cell as the cell wall. This conclusion implies that the cytoskeletal polymer MreB applies meaningful force to the cell wall, an idea favored by theoretical modeling of wall growth, and suggests an evolutionary origin of cytoskeleton-governed cell rigidity. Second, I observe that MreB rotates around the long axis of E. coli, and the motion depends on wall synthesis. This is the first discovery of a cell-wall assembly driven molecular motor in bacteria. Third, I prove that both cell-wall synthesis and the PG network have chiral ordering, which is established by the spatial pattern of MreB. This work links the molecular structure of the cytoskeleton and of the cell wall with organismal-scale behavior. Finally, I develop a mathematical model of cytoskeleton-cell membrane interactions, which explains the preferential orientation of different cytoskeleton components in bacteria.

Wall effects in continuous microfluidic magneto-affinity cell separation.

PubMed

Wu, Liqun; Zhang, Yong; Palaniapan, Moorthi; Roy, Partha

2010-05-01

Continuous microfluidic magneto-affinity cell separator combines unique microscale flow phenomenon with advantageous nanobead properties, to isolate cells with high specificity. Owing to the comparable size of the cell-bead complexes and the microchannels, the walls of the microchannel exert a strong influence on the separation of cells by this method. We present a theoretical and experimental study that provides a quantitative description of hydrodynamic wall interactions and wall rolling velocity of cells. A transient convection model describes the transport of cells in two-phase microfluidic flow under the influence of an external magnetic field. Transport of cells along the microchannel walls is also considered via an additional equation. Results show the variation of cell flux in the fluid phases and the wall as a function of a dimensionless parameter arising in the equations. Our results suggest that conditions may be optimized to maximize cell separation while minimizing contact with the wall surfaces. Experimentally measured cell rolling velocities on the wall indicate the presence of other near-wall forces in addition to fluid shear forces. Separation of a human colon carcinoma cell line from a mixture of red blood cells, with folic acid conjugated 1 microm and 200 nm beads, is reported.

Separation of switchgrass bio-oil by water/organic solvent addition and pH adjustment

DOE PAGES

Park, Lydia Kyoung-Eun; Ren, Shoujie; Yiacoumi, Sotira; ...

2016-01-29

Applications of bio-oil are limited by its challenging properties including high moisture content, low pH, high viscosity, high oxygen content, and low heating value. Separation of switchgrass bio-oil components by adding water, organic solvents (hexadecane and octane), and sodium hydroxide may help to overcome these issues. Acetic acid and phenolic compounds were extracted in aqueous and organic phases, respectively. Polar chemicals, such as acetic acid, did not partition in the organic solvent phase. Acetic acid in the aqueous phase after extraction is beneficial for a microbial-electrolysis-cell application to produce hydrogen as an energy source for further hydrodeoxygenation of bio-oil. Organicmore » solvents extracted more chemicals from bio-oil in combined than in sequential extraction; however, organic solvents partitioned into the aqueous phase in combined extraction. When sodium hydroxide was added to adjust the pH of aqueous bio-oil, organic-phase precipitation occurred. As the pH was increased, a biphasic aqueous/organic dispersion was formed, and phase separation was optimized at approximately pH 6. The neutralized organic bio-oil had approximately 37% less oxygen and 100% increased heating value than the initial centrifuged bio-oil. In conclusion, the less oxygen content and increased heating value indicated a significant improvement of the bio-oil quality through neutralization.« less

Separation of switchgrass bio-oil by water/organic solvent addition and pH adjustment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Park, Lydia Kyoung-Eun; Ren, Shoujie; Yiacoumi, Sotira

Applications of bio-oil are limited by its challenging properties including high moisture content, low pH, high viscosity, high oxygen content, and low heating value. Separation of switchgrass bio-oil components by adding water, organic solvents (hexadecane and octane), and sodium hydroxide may help to overcome these issues. Acetic acid and phenolic compounds were extracted in aqueous and organic phases, respectively. Polar chemicals, such as acetic acid, did not partition in the organic solvent phase. Acetic acid in the aqueous phase after extraction is beneficial for a microbial-electrolysis-cell application to produce hydrogen as an energy source for further hydrodeoxygenation of bio-oil. Organicmore » solvents extracted more chemicals from bio-oil in combined than in sequential extraction; however, organic solvents partitioned into the aqueous phase in combined extraction. When sodium hydroxide was added to adjust the pH of aqueous bio-oil, organic-phase precipitation occurred. As the pH was increased, a biphasic aqueous/organic dispersion was formed, and phase separation was optimized at approximately pH 6. The neutralized organic bio-oil had approximately 37% less oxygen and 100% increased heating value than the initial centrifuged bio-oil. In conclusion, the less oxygen content and increased heating value indicated a significant improvement of the bio-oil quality through neutralization.« less

Structure, function, and biosynthesis of plant cell walls: proceedings of the seventh annual symposium in botany

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dugger, W.M.; Bartnicki-Garcia, S.

Papers in the following areas were included in these symposium proceedings: (1) cell wall chemistry and biosynthesis; (2) cell wall hydrolysis and associated physiology; (3) cellular events associated with cell wall biosynthesis; and (4) interactions of plant cell walls with pathogens and related responses. Papers have been individually abstracted for the data base. (ACR)

Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

PubMed Central

Orlean, Peter

2012-01-01

The wall gives a Saccharomyces cerevisiae cell its osmotic integrity; defines cell shape during budding growth, mating, sporulation, and pseudohypha formation; and presents adhesive glycoproteins to other yeast cells. The wall consists of β1,3- and β1,6-glucans, a small amount of chitin, and many different proteins that may bear N- and O-linked glycans and a glycolipid anchor. These components become cross-linked in various ways to form higher-order complexes. Wall composition and degree of cross-linking vary during growth and development and change in response to cell wall stress. This article reviews wall biogenesis in vegetative cells, covering the structure of wall components and how they are cross-linked; the biosynthesis of N- and O-linked glycans, glycosylphosphatidylinositol membrane anchors, β1,3- and β1,6-linked glucans, and chitin; the reactions that cross-link wall components; and the possible functions of enzymatic and nonenzymatic cell wall proteins. PMID:23135325

Retention of Proanthocyanidin in Wine-like Solution Is Conferred by a Dynamic Interaction between Soluble and Insoluble Grape Cell Wall Components.

PubMed

Bindon, Keren A; Li, Sijing; Kassara, Stella; Smith, Paul A

2016-11-09

For better understanding of the factors that impact proanthocyanidin (PA) adsorption by insoluble cell walls or interaction with soluble cell wall-derived components, application of a commercial polygalacturonase enzyme preparation was investigated to modify grape cell wall structure. Soluble and insoluble cell wall material was isolated from the skin and mesocarp components of Vitis vinifera Shiraz grapes. It was observed that significant depolymerization of the insoluble grape cell wall occurred following enzyme application to both grape cell wall fractions, with increased solubilization of rhamnogalacturonan-enriched, low molecular weight polysaccharides. However, in the case of grape mesocarp, the solubilization of protein from cell walls (in buffer) was significant and increased only slightly by the enzyme treatment. Enzyme treatment significantly reduced the adsorption of PA by insoluble cell walls, but this effect was observed only when material solubilized from grape cell walls had been removed. The loss of PA through interaction with the soluble cell wall fraction was observed to be greater for mesocarp than skin cell walls. Subsequent experiments on the soluble mesocarp cell wall fraction confirmed a role for protein in the precipitation of PA. This identified a potential mechanism by which extracted grape PA may be lost from wine during vinification, as a precipitate with solubilized grape mesocarp proteins. Although protein was a minor component in terms of total concentration, losses of PA via precipitation with proteins were in the order of 50% of available PA. PA-induced precipitation could proceed until all protein was removed from solution and may account for the very low levels of residual protein observed in red wines. The results point to a dynamic interaction of grape insoluble and soluble components in modulating PA retention in wine.

The transcription factor Rap1p is required for tolerance to cell-wall perturbing agents and for cell-wall maintenance in Saccharomyces cerevisiae.

PubMed

Azad, Gajendra Kumar; Singh, Vikash; Baranwal, Shivani; Thakare, Mayur Jankiram; Tomar, Raghuvir S

2015-01-02

Yeast repressor activator protein (Rap1p) is involved in genomic stability and transcriptional regulation. We explored the function of Rap1p in yeast physiology using Rap1p truncation mutants. Our results revealed that the N-terminal truncation of Rap1p (Rap1ΔN) leads to hypersensitivity towards elevated temperature and cell-wall perturbing agents. Cell wall analysis showed an increase in the chitin and glucan content in Rap1ΔN cells as compared with wild type cells. Accordingly, mutant cells had a twofold thicker cell wall, as observed by electron microscopy. Furthermore, Rap1ΔN cells had increased levels of phosphorylated Slt2p, a MAP kinase of the cell wall integrity pathway. Mutant cells also had elevated levels of cell wall integrity response transcripts. Taken together, our findings suggest a connection between Rap1p and cell wall homeostasis. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Bacterial cell-wall recycling

PubMed Central

Johnson, Jarrod W.; Fisher, Jed F.; Mobashery, Shahriar

2012-01-01

Many Gram-negative and Gram-positive bacteria recycle a significant proportion of the peptidoglycan components of their cell walls during their growth and septation. In many—and quite possibly all—bacteria, the peptidoglycan fragments are recovered and recycled. While cell-wall recycling is beneficial for the recovery of resources, it also serves as a mechanism to detect cell-wall–targeting antibiotics and to regulate resistance mechanisms. In several Gram-negative pathogens, anhydro-MurNAc-peptide cell-wall fragments regulate AmpC β-lactamase induction. In some Gram-positive organisms, short peptides derived from the cell wall regulate the induction of both β-lactamase and β-lactam-resistant penicillin-binding proteins. The involvement of peptidoglycan recycling with resistance regulation suggests that inhibitors of the enzymes involved in the recycling might synergize with cell-wall-targeted antibiotics. Indeed, such inhibitors improve the potency of β-lactams in vitro against inducible AmpC β-lactamase-producing bacteria. We describe the key steps of cell-wall remodeling and recycling, the regulation of resistance mechanisms by cell-wall recycling, and recent advances toward the discovery of cell-wall recycling inhibitors. PMID:23163477

Characteristic Thickened Cell Walls of the Bracts of the ‘Eternal Flower’ Helichrysum bracteatum

PubMed Central

Nishikawa, Kuniko; Ito, Hiroaki; Awano, Tatsuya; Hosokawa, Munetaka; Yazawa, Susumu

2008-01-01

Background and Aims Helichrysum bracteatum is called an ‘eternal flower’ and has large, coloured, scarious bracts. These maintain their aesthetic value without wilting or discoloration for many years. There have been no research studies of cell death or cell morphology of the scarious bract, and hence the aim of this work was to elucidate these characteristics for the bract of H. bracteatum. Methods DAPI (4'6-diamidino-2-phenylindol dihydrochloride) staining and fluorescence microscopy were used for observation of cell nuclei. Light microscopy (LM), transmission electron microscopy (TEM) and polarized light microscopy were used for observation of cells, including cell wall morphology. Key Results Cell death occurred at the bract tip during the early stage of flower development. The cell wall was the most prominent characteristic of H. bracteatum bract cells. Characteristic thickened secondary cell walls on the inside of the primary cell walls were observed in both epidermal and inner cells. In addition, the walls of all cells exhibited birefringence. Characteristic thickened secondary cell walls have orientated cellulose microfibrils as well as general secondary cell walls of the tracheary elements. For comparison, these characters were not observed in the petal and bract tissues of Chrysanthemum morifolium. Conclusions Bracts at anthesis are composed of dead cells. Helichrysum bracteatum bracts have characteristic thickened secondary cell walls that have not been observed in the parenchyma of any other flowers or leaves. The cells of the H. bracteatum bract differ from other tissues with secondary cell walls, suggesting that they may be a new cell type. PMID:18436550

Mechanical feedback coordinates cell wall expansion and assembly in yeast mating morphogenesis

PubMed Central

2018-01-01

The shaping of individual cells requires a tight coordination of cell mechanics and growth. However, it is unclear how information about the mechanical state of the wall is relayed to the molecular processes building it, thereby enabling the coordination of cell wall expansion and assembly during morphogenesis. Combining theoretical and experimental approaches, we show that a mechanical feedback coordinating cell wall assembly and expansion is essential to sustain mating projection growth in budding yeast (Saccharomyces cerevisiae). Our theoretical results indicate that the mechanical feedback provided by the Cell Wall Integrity pathway, with cell wall stress sensors Wsc1 and Mid2 increasingly activating membrane-localized cell wall synthases Fks1/2 upon faster cell wall expansion, stabilizes mating projection growth without affecting cell shape. Experimental perturbation of the osmotic pressure and cell wall mechanics, as well as compromising the mechanical feedback through genetic deletion of the stress sensors, leads to cellular phenotypes that support the theoretical predictions. Our results indicate that while the existence of mechanical feedback is essential to stabilize mating projection growth, the shape and size of the cell are insensitive to the feedback. PMID:29346368

Wall relaxation and the driving forces for cell expansive growth

NASA Technical Reports Server (NTRS)

Cosgrove, D. J.

1987-01-01

When water uptake by growing cells is prevented, the turgor pressure and the tensile stress in the cell wall are reduced by continued wall loosening. This process, termed in vivo stress relaxation, provides a new way to study the dynamics of wall loosening and to measure the wall yield threshold and the physiological wall extensibility. Stress relaxation experiments indicate that wall stress supplies the mechanical driving force for wall yielding. Cell expansion also requires water absorption. The driving force for water uptake during growth is created by wall relaxation, which lowers the water potential of the expanding cells. New techniques for measuring this driving force show that it is smaller than believed previously; in elongating stems it is only 0.3 to 0.5 bar. This means that the hydraulic resistance of the water transport pathway is small and that rate of cell expansion is controlled primarily by wall loosening and yielding.

Two endogenous proteins that induce cell wall extension in plants

NASA Technical Reports Server (NTRS)

McQueen-Mason, S.; Durachko, D. M.; Cosgrove, D. J.

1992-01-01

Plant cell enlargement is regulated by wall relaxation and yielding, which is thought to be catalyzed by elusive "wall-loosening" enzymes. By employing a reconstitution approach, we found that a crude protein extract from the cell walls of growing cucumber seedlings possessed the ability to induce the extension of isolated cell walls. This activity was restricted to the growing region of the stem and could induce the extension of isolated cell walls from various dicot stems and the leaves of amaryllidaceous monocots, but was less effective on grass coleoptile walls. Endogenous and reconstituted wall extension activities showed similar sensitivities to pH, metal ions, thiol reducing agents, proteases, and boiling in methanol or water. Sequential HPLC fractionation of the active wall extract revealed two proteins with molecular masses of 29 and 30 kD associated with the activity. Each protein, by itself, could induce wall extension without detectable hydrolytic breakdown of the wall. These proteins appear to mediate "acid growth" responses of isolated walls and may catalyze plant cell wall extension by a novel biochemical mechanism.

Pectinous cell wall thickenings formation - A common defense strategy of plants to cope with Pb.

PubMed

Krzesłowska, Magdalena; Rabęda, Irena; Basińska, Aneta; Lewandowski, Michał; Mellerowicz, Ewa J; Napieralska, Anna; Samardakiewicz, Sławomir; Woźny, Adam

2016-07-01

Lead, one of the most abundant and hazardous trace metals affecting living organisms, has been commonly detected in plant cell walls including some tolerant plants, mining ecotypes and hyperaccumulators. We have previously shown that in tip growing Funaria sp. protonemata cell wall is remodeled in response to lead by formation of thickenings rich in low-methylesterified pectins (pectin epitope JIM5 - JIM5-P) able to bind metal ions, which accumulate large amounts of Pb. Hence, it leads to the increase of cell wall capacity for Pb compartmentalization. Here we show that diverse plant species belonging to different phyla (Arabidopsis, hybrid aspen, star duckweed), form similar cell wall thickenings in response to Pb. These thickenings are formed in tip growing cells such as the root hairs, and in diffuse growing cells such as meristematic and root cap columella cells of root apices in hybrid aspen and Arabidopsis and in mesophyll cells in star duckweed fronds. Notably, all analyzed cell wall thickenings were abundant in JIM5-P and accumulated high amounts of Pb. In addition, the co-localization of JIM5-P and Pb commonly occurred in these cells. Hence, cell wall thickenings formed the extra compartment for Pb accumulation. In this way plant cells increased cell wall capacity for compartmentalization of this toxic metal, protecting protoplast from its toxicity. As cell wall thickenings occurred in diverse plant species and cell types differing in the type of growth we may conclude that pectinous cell wall thickenings formation is a widespread defense strategy of plants to cope with Pb. Moreover, detection of natural defense strategy, increasing plant cell walls capacity for metal accumulation, reveals a promising direction for enhancing plant efficiency in phytoremediation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Genetic and biochemical characterization of the GH72 family of cell wall transglycosylases in Neurospora crassa.

PubMed

Ao, Jie; Free, Stephen J

2017-04-01

The Neurospora crassa genome encodes five GH72 family transglycosylases, and four of these enzymes (GEL-1, GEL-2, GEL-3 and GEL-5) have been found to be present in the cell wall proteome. We carried out an extensive genetic analysis on the role of these four transglycosylases in cell wall biogenesis and demonstrated that the transglycosylases are required for the formation of a normal cell wall. As suggested by the proteomic analysis, we found that multiple transglycosylases were being expressed in N. crassa cells and that different combinations of the enzymes are required in different cell types. The combination of GEL-1, GEL-2 and GEL-5 is required for the growth of vegetative hyphae, while the GEL-1, GEL-2, GEL-3 combination is needed for the production of aerial hyphae and conidia. Our data demonstrates that the enzymes are redundant with partially overlapping enzymatic activities, which provides the fungus with a robust cell wall biosynthetic system. Characterization of the transglycosylase-deficient mutants demonstrated that the incorporation of cell wall proteins was severely compromised. Interestingly, we found that the transglycosylase-deficient mutant cell walls contained more β-1,3-glucan than the wild type cell wall. Our results demonstrate that the GH72 transglycosylases are not needed for the incorporation of β-1,3-glucan into the cell wall, but they are required for the incorporation of cell wall glycoprotein into the cell wall. Copyright © 2017 Elsevier Inc. All rights reserved.

A Cell Wall Proteome and Targeted Cell Wall Analyses Provide Novel Information on Hemicellulose Metabolism in Flax.

PubMed

Chabi, Malika; Goulas, Estelle; Leclercq, Celine C; de Waele, Isabelle; Rihouey, Christophe; Cenci, Ugo; Day, Arnaud; Blervacq, Anne-Sophie; Neutelings, Godfrey; Duponchel, Ludovic; Lerouge, Patrice; Hausman, Jean-François; Renaut, Jenny; Hawkins, Simon

2017-09-01

Experimentally-generated (nanoLC-MS/MS) proteomic analyses of four different flax organs/tissues (inner-stem, outer-stem, leaves and roots) enriched in proteins from 3 different sub-compartments (soluble-, membrane-, and cell wall-proteins) was combined with publically available data on flax seed and whole-stem proteins to generate a flax protein database containing 2996 nonredundant total proteins. Subsequent multiple analyses (MapMan, CAZy, WallProtDB and expert curation) of this database were then used to identify a flax cell wall proteome consisting of 456 nonredundant proteins localized in the cell wall and/or associated with cell wall biosynthesis, remodeling and other cell wall related processes. Examination of the proteins present in different flax organs/tissues provided a detailed overview of cell wall metabolism and highlighted the importance of hemicellulose and pectin remodeling in stem tissues. Phylogenetic analyses of proteins in the cell wall proteome revealed an important paralogy in the class IIIA xyloglucan endo-transglycosylase/hydrolase (XTH) family associated with xyloglucan endo-hydrolase activity.Immunolocalisation, FT-IR microspectroscopy, and enzymatic fingerprinting indicated that flax fiber primary/S1 cell walls contained xyloglucans with typical substituted side chains as well as glucuronoxylans in much lower quantities. These results suggest a likely central role of xyloglucans and endotransglucosylase/hydrolase activity in flax fiber formation and cell wall remodeling processes. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Developmental and Tissue-Specific Structural Alterations of the Cell-Wall Polysaccharides of Arabidopsis thaliana Roots.

PubMed Central

Freshour, G.; Clay, R. P.; Fuller, M. S.; Albersheim, P.; Darvill, A. G.; Hahn, M. G.

1996-01-01

The plant cell wall is a dynamic structure that plays important roles in growth and development and in the interactions of plants with their environment and other organisms. We have used monoclonal antibodies that recognize different carbohydrate epitopes present in plant cell-wall polysaccharides to locate these epitopes in roots of developing Arabidopsis thaliana seedlings. An epitope in the pectic polysaccharide rhamnogalacturonan I is observed in the walls of epidermal and cortical cells in mature parts of the root. This epitope is inserted into the walls in a developmentally regulated manner. Initially, the epitope is observed in atrichoblasts and later appears in trichoblasts and simultaneously in cortical cells. A terminal [alpha]-fucosyl-containing epitope is present in almost all of the cell walls in the root. An arabinosylated (1->6)-[beta]-galactan epitope is also found in all of the cell walls of the root with the exception of lateral root-cap cell walls. It is striking that these three polysaccharide epitopes are not uniformly distributed (or accessible) within the walls of a given cell, nor are these epitopes distributed equally across the two walls laid down by adjacent cells. Our results further suggest that the biosynthesis and differentiation of primary cell walls in plants are precisely regulated in a temporal, spatial, and developmental manner. PMID:12226270

Atomic force microscopy stiffness tomography on living Arabidopsis thaliana cells reveals the mechanical properties of surface and deep cell-wall layers during growth.

PubMed

Radotić, Ksenija; Roduit, Charles; Simonović, Jasna; Hornitschek, Patricia; Fankhauser, Christian; Mutavdžić, Dragosav; Steinbach, Gabor; Dietler, Giovanni; Kasas, Sandor

2012-08-08

Cell-wall mechanical properties play a key role in the growth and the protection of plants. However, little is known about genuine wall mechanical properties and their growth-related dynamics at subcellular resolution and in living cells. Here, we used atomic force microscopy (AFM) stiffness tomography to explore stiffness distribution in the cell wall of suspension-cultured Arabidopsis thaliana as a model of primary, growing cell wall. For the first time that we know of, this new imaging technique was performed on living single cells of a higher plant, permitting monitoring of the stiffness distribution in cell-wall layers as a function of the depth and its evolution during the different growth phases. The mechanical measurements were correlated with changes in the composition of the cell wall, which were revealed by Fourier-transform infrared (FTIR) spectroscopy. In the beginning and end of cell growth, the average stiffness of the cell wall was low and the wall was mechanically homogenous, whereas in the exponential growth phase, the average wall stiffness increased, with increasing heterogeneity. In this phase, the difference between the superficial and deep wall stiffness was highest. FTIR spectra revealed a relative increase in the polysaccharide/lignin content. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

A computational approach for inferring the cell wall properties that govern guard cell dynamics.

PubMed

Woolfenden, Hugh C; Bourdais, Gildas; Kopischke, Michaela; Miedes, Eva; Molina, Antonio; Robatzek, Silke; Morris, Richard J

2017-10-01

Guard cells dynamically adjust their shape in order to regulate photosynthetic gas exchange, respiration rates and defend against pathogen entry. Cell shape changes are determined by the interplay of cell wall material properties and turgor pressure. To investigate this relationship between turgor pressure, cell wall properties and cell shape, we focused on kidney-shaped stomata and developed a biomechanical model of a guard cell pair. Treating the cell wall as a composite of the pectin-rich cell wall matrix embedded with cellulose microfibrils, we show that strong, circumferentially oriented fibres are critical for opening. We find that the opening dynamics are dictated by the mechanical stress response of the cell wall matrix, and as the turgor rises, the pectinaceous matrix stiffens. We validate these predictions with stomatal opening experiments in selected Arabidopsis cell wall mutants. Thus, using a computational framework that combines a 3D biomechanical model with parameter optimization, we demonstrate how to exploit subtle shape changes to infer cell wall material properties. Our findings reveal that proper stomatal dynamics are built on two key properties of the cell wall, namely anisotropy in the form of hoop reinforcement and strain stiffening. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd and Society for Experimental Biology.

Virus-induced gene silencing offers a functional genomics platform for studying plant cell wall formation.

PubMed

Zhu, Xiaohong; Pattathil, Sivakumar; Mazumder, Koushik; Brehm, Amanda; Hahn, Michael G; Dinesh-Kumar, S P; Joshi, Chandrashekhar P

2010-09-01

Virus-induced gene silencing (VIGS) is a powerful genetic tool for rapid assessment of plant gene functions in the post-genomic era. Here, we successfully implemented a Tobacco Rattle Virus (TRV)-based VIGS system to study functions of genes involved in either primary or secondary cell wall formation in Nicotiana benthamiana plants. A 3-week post-VIGS time frame is sufficient to observe phenotypic alterations in the anatomical structure of stems and chemical composition of the primary and secondary cell walls. We used cell wall glycan-directed monoclonal antibodies to demonstrate that alteration of cell wall polymer synthesis during the secondary growth phase of VIGS plants has profound effects on the extractability of components from woody stem cell walls. Therefore, TRV-based VIGS together with cell wall component profiling methods provide a high-throughput gene discovery platform for studying plant cell wall formation from a bioenergy perspective.

Plant cell wall signalling and receptor-like kinases.

PubMed

Wolf, Sebastian

2017-02-15

Communication between the extracellular matrix and the cell interior is essential for all organisms as intrinsic and extrinsic cues have to be integrated to co-ordinate development, growth, and behaviour. This applies in particular to plants, the growth and shape of which is governed by deposition and remodelling of the cell wall, a rigid, yet dynamic, extracellular network. It is thus generally assumed that cell wall surveillance pathways exist to monitor the state of the wall and, if needed, elicit compensatory responses such as altered expression of cell wall remodelling and biosynthesis genes. Here, I highlight recent advances in the field of cell wall signalling in plants, with emphasis on the role of plasma membrane receptor-like kinase complexes. In addition, possible roles for cell wall-mediated signalling beyond the maintenance of cell wall integrity are discussed. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

The Specific Nature of Plant Cell Wall Polysaccharides 1

PubMed Central

Nevins, Donald J.; English, Patricia D.; Albersheim, Peter

1967-01-01

Polysaccharide compositions of cell walls were assessed by quantitative analyses of the component sugars. Cell walls were hydrolyzed in 2 n trifluoroacetic acid and the liberated sugars reduced to their respective alditols. The alditols were acetylated and the resulting alditol acetates separated by gas chromatography. Quantitative assay of the alditol acetates was accomplished by electronically integrating the detector output of the gas chromatograph. Myo-inositol, introduced into the sample prior to hydrolysis, served as an internal standard. The cell wall polysaccharide compositions of plant varieties within a given species are essentially identical. However, differences in the sugar composition were observed in cell walls prepared from different species of the same as well as of different genera. The fact that the wall compositions of different varieties of the same species are the same indicates that the biosynthesis of cell wall polysaccharides is genetically regulated. The cell walls of various morphological parts (roots, hypocotyls, first internodes and primary leaves) of bean plants were each found to have a characteristic sugar composition. It was found that the cell wall sugar composition of suspension-cultured sycamore cells could be altered by growing the cells on different carbon sources. This demonstrates that the biosynthesis of cell wall polysaccharides can be manipulated without fatal consequences. PMID:16656594

Integrative analysis and expression profiling of secondary cell wall genes in C4 biofuel model Setaria italica reveals targets for lignocellulose bioengineering

PubMed Central

Muthamilarasan, Mehanathan; Khan, Yusuf; Jaishankar, Jananee; Shweta, Shweta; Lata, Charu; Prasad, Manoj

2015-01-01

Several underutilized grasses have excellent potential for use as bioenergy feedstock due to their lignocellulosic biomass. Genomic tools have enabled identification of lignocellulose biosynthesis genes in several sequenced plants. However, the non-availability of whole genome sequence of bioenergy grasses hinders the study on bioenergy genomics and their genomics-assisted crop improvement. Foxtail millet (Setaria italica L.; Si) is a model crop for studying systems biology of bioenergy grasses. In the present study, a systematic approach has been used for identification of gene families involved in cellulose (CesA/Csl), callose (Gsl) and monolignol biosynthesis (PAL, C4H, 4CL, HCT, C3H, CCoAOMT, F5H, COMT, CCR, CAD) and construction of physical map of foxtail millet. Sequence alignment and phylogenetic analysis of identified proteins showed that monolignol biosynthesis proteins were highly diverse, whereas CesA/Csl and Gsl proteins were homologous to rice and Arabidopsis. Comparative mapping of foxtail millet lignocellulose biosynthesis genes with other C4 panicoid genomes revealed maximum homology with switchgrass, followed by sorghum and maize. Expression profiling of candidate lignocellulose genes in response to different abiotic stresses and hormone treatments showed their differential expression pattern, with significant higher expression of SiGsl12, SiPAL2, SiHCT1, SiF5H2, and SiCAD6 genes. Further, due to the evolutionary conservation of grass genomes, the insights gained from the present study could be extrapolated for identifying genes involved in lignocellulose biosynthesis in other biofuel species for further characterization. PMID:26583030

Integrative analysis and expression profiling of secondary cell wall genes in C4 biofuel model Setaria italica reveals targets for lignocellulose bioengineering.

PubMed

Muthamilarasan, Mehanathan; Khan, Yusuf; Jaishankar, Jananee; Shweta, Shweta; Lata, Charu; Prasad, Manoj

2015-01-01

Several underutilized grasses have excellent potential for use as bioenergy feedstock due to their lignocellulosic biomass. Genomic tools have enabled identification of lignocellulose biosynthesis genes in several sequenced plants. However, the non-availability of whole genome sequence of bioenergy grasses hinders the study on bioenergy genomics and their genomics-assisted crop improvement. Foxtail millet (Setaria italica L.; Si) is a model crop for studying systems biology of bioenergy grasses. In the present study, a systematic approach has been used for identification of gene families involved in cellulose (CesA/Csl), callose (Gsl) and monolignol biosynthesis (PAL, C4H, 4CL, HCT, C3H, CCoAOMT, F5H, COMT, CCR, CAD) and construction of physical map of foxtail millet. Sequence alignment and phylogenetic analysis of identified proteins showed that monolignol biosynthesis proteins were highly diverse, whereas CesA/Csl and Gsl proteins were homologous to rice and Arabidopsis. Comparative mapping of foxtail millet lignocellulose biosynthesis genes with other C4 panicoid genomes revealed maximum homology with switchgrass, followed by sorghum and maize. Expression profiling of candidate lignocellulose genes in response to different abiotic stresses and hormone treatments showed their differential expression pattern, with significant higher expression of SiGsl12, SiPAL2, SiHCT1, SiF5H2, and SiCAD6 genes. Further, due to the evolutionary conservation of grass genomes, the insights gained from the present study could be extrapolated for identifying genes involved in lignocellulose biosynthesis in other biofuel species for further characterization.

Remarkable proanthocyanidin adsorption properties of monastrell pomace cell wall material highlight its potential use as an alternative fining agent in red wine production.

PubMed

Bautista-Ortín, Ana Belén; Ruiz-García, Yolanda; Marín, Fátima; Molero, Noelia; Apolinar-Valiente, Rafael; Gómez-Plaza, Encarna

2015-01-21

The existence of interactions between the polysaccharides of vegetal cell walls and proanthocyanins makes this cell wall material an interesting option for its use as a fining agent to reduce the level of proanthocyanins in wines. Pomace wastes from the winery are widely available and a source of cell wall material, and the identification of varieties whose pomace cell walls present high proanthocyanin binding capacity and of processing methods that could enhance their adsorption properties could be of great interest. This study compared the proanthocyanin adsorption properties of pomace cell wall material from three different grape varieties (Monastrell, Cabernet Sauvignon, and Syrah), and the results were compared with those obtained using fresh grape cell walls. Also, the effect of the vinification method has been studied. Analysis of the proanthocyanidins in the solution after reaction with the cell wall material, using phloroglucinolysis and size exclusion chromatography, provided quantitative and qualitative information on the adsorbed and nonadsorbed compounds. A highlight of this study was the observation that Monastrell pomace cell wall material showed a strong affinity for proanthocyanidins, with values similar to that obtained for fresh grapes cell walls, and a preferential binding of high molecular mass proanthocyanidins, so these pomace cell walls could be used in wines to reduce astringency. The use of maceration enzymes during vinification had little effect on the retention capacity of the pomace cell walls obtained from this vinification, although an increase in the retention of low molecular mass proanthocyanidins was observed, and this might have implications for wine sensory properties.

Plant germination and growth after exposure to iron cyanide complexes.

PubMed

Kang, Dong-Hee; Hong, Lee Y; Schwab, A Paul; Banks, M Katherine

2008-05-01

Phytoremediation has been proposed for treatment of cyanide-contaminated soil. This study was conducted to identify plants with the highest potential for phytoremediation of iron cyanide contaminated soil. Multiple cultivars of two cyanogenic species, sorghum (Sorghum bicolor) and flax (Linum usitatissimum), and one non-cyanogenic species, switchgrass (Panicum virgatum L), were selected for evaluation. The cultivars were screened by quantifying germination and root elongation. Differences in germination emerged among the cultivars (P < 0.05), but these differences appeared to be unrelated to cyanide concentration. The presence of 1000 mg/kg Prussian blue tended to suppress root growth parameters of flax and switchgrass but did not affect sorghum similarly.