The Energy Biosciences Institute Realizing Cellulosic Biofuels and Benefiting the Environment.

Transcription

1 The Energy Biosciences Institute Realizing Cellulosic Biofuels and Benefiting the Environment. Steve Long, Clive Beale, Emily Heaton & Frank Dohleman, Plant Biology, Crop Sciences, Institute for Genomic Biology, National Center for Supercomputer Applications, University of Illinois. CSU 11/11/08

2 What are second generation bioenergy feedstocks?

3 In this context: sustainable crops beyond food grains - mainly perennials.

4 control ozone

5 ROADMAP Why fuels from crops? What is the ideal biomass crop? Performance of Miscanthus and switchgrass. Food vs. Fuel

6 ROADMAP Why fuels from crops? What is the ideal biomass crop? Performance of Miscanthus and switchgrass. Food vs. Fuel

7 DRIVERS FOR RENEWABLE TRANSPORTATION FUELS?

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9 Gallons per Acre Barley Wheat Corn Sugar beet Sugar cane Soy Castor Sunflower Rapeseed Jatropha Palm 2000 FUEL PER UNIT AREA OF Yield of various species varies widely LAND Worldwatch 2006 Cellulosic (Miscanthus)

10 THE ENERGY BIOSCIENCES INSTITUTE RESEARCH & DEVELOPMENT Environmentally and economically sustainable second generation feedstock and biofuel business $500M investment over 10 years by BP DIRECTOR: Chris Somerville (Berkeley) DEPUTY DIRECTOR: Steve Long (Illinois) ASSOCIATE DIRECTOR: Paul Willems (BP Group)

11 WHAT IS THE EBI? INSTITUTE R&D AREAS Thermal Treatment Syn Gas Methanogens/Others Consolidated Process Sunlight FEEDSTOCK DEVELOPMENT Biomass BIOMASS DEPOLYMERIZATION Photosynthetic Microbes Monomers BIOFUELS PRODUCTION Fuels Fossil Fuels Bio-processing & MEOR Economic, Environmental and Social Impacts Carbon & Energy Balance, Carbon Sequestration, Water use, Commercial Systems, Business Ecosystem. EDUCATION & EXTENSION Coal/Oil

12 ebi program leaders EBI PROGRAMS DISCOVERY OF ENZYMES TO DEGRADE PLANT CELL WALL LIGNOCELLULOSE Isaac Cann (Animal Sci.) ENGINEERING SUGAR FERMENTATION Huimin Zhao (Chem Eng) FEEDSTOCK PRODUCTION/ AGRONOMY PROGRAM Tom Voigt (NRES) ENGINEERING SOLUTIONS FOR BIOMASS FEEDSTOCK PRODUCTION K.C. Ting (ABE) BIOFUEL ECONOMIC AND ENVIRONMENTAL IMPACTS Madhu Khanna (ACE) ENVIRONMENTAL IMPACT AND SUSTAINABILITY OF FEEDSTOCK PRODUCTION Evan DeLucia (Plant Bio) ASSESSING IMPACT OF INSECT PESTS AND PLANT PATHOGENS ON BIOMASS PRODUCTION Michael Gray (Crop Sci) GENOMIC-DIRECTED IMPROVEMENT OF FEEDSTOCKS Stephen Moose (Crop Sci) BIOFUELS LAW AND REGULATION Jay Kesan (Law) ENGINEERING ENZYMES AND METABOLIC PATHWAYS TO DEGRADE PLANT CELL WALL LIGNOCELLULOSE John Gerlt (Biochemistry)

13 WHAT IS THE EBI? INTEGRATING PROGRAMS Under One Roof Programs Feedstock Development Feedstock Deconstruction Fuel Synthesis Environment, Economics & Policy Projects

14 WHAT IS THE EBI? ON CAMPUS ENERGY FARM

15 CELLULOSIC BIOFUELS 1.3 BILLION TON POTENTIAL FEEDSTOCK IN USA. ~ 2 t/ha Corn stover 19.9% Wheat straw 6.1% Soy 6.2% Crop residues 7.6% Grains 5.2% Perennial crops 35.2% Forest 12.8% Manure 4.1% Urban waste 2.9% From: Billion ton Study, DOE & USDA 2005

16 ROADMAP Why fuels from crops? What is the ideal biomass crop? Performance of Miscanthus and switchgrass. Basis of differences Miscanthus and potential biofuel production.

17 ebi program leaders WHICH FEEDSTOCKS?

18 POOR INFORMATION BASE

19 Interception efficiency ebi program leaders DETERMINANTS OF YIELD Total solar energy Conversion efficiency W h = S i c Harvested yield

20 THEORETICAL EFFICIENCY OF PHOTOSYNTHESIS 100% 49% 44% 37% LOSSES AT DIFFERENT STAGES IN ENERGY TRANSDUCTION 51% outside usable spectrum 5% reflected and transmitted 7% photochemical inefficiency 25.5% 5.1% 1.9% C 3 C % 6.5% 8.5% 8.5% 28.5% carbohydrate synthesis 0% photorespiration 2.5% respiration 4.6% 6.0% From Zhu, Long & Ort (2008) Current Opinion in Biotechnology 2008, 19:

21 INTERCEPTING SOLAR RADIATION ebi program leaders Solar radiation MJ d J F M A M J J A S O N D

22 ebi program leaders Why is low input a critical factor? Better energy and carbon balance. Lower or no environmental impact. Marginal lands

23 Mineral nutrients Mineral nutrients NITROGEN USE EFFICIENCY THEORY SPRING/ SUMMER FALL WINTER Translocation from rhizomes to growing shoot Translocation to rhizome as shoot senesces Lignocellulose dry shoots harvested, nutrients stay in rhizomes

24 ebi program leaders Prairie, Steppe & Savanna Life-form Prairie, Steppe and Savanna perennial grasses are sustainable if cropped or burned annually, and accumulate carbon in the soil.

25 ebi program leaders C4 PERENNIAL GRASSES USA Switchgrass (Panicum virgatum L.) The Best of Both Worlds? Europe Miscanthus (Miscanthus x giganteus Greef et Deu.)

26 THE IDEAL BIOMASS CROP? C4 photosynthesis Long canopy duration Recycles nutrients to roots Low input High water use efficiency Sterile non-invasive Can store harvest in field Easily removed No known pests/diseases Uses existing farm equipment

27 THE IDEAL BIOMASS CROP? CROP Maize C4 photosynthesis Long canopy duration Recycles nutrients to roots Low input High water use efficiency Sterile non-invasive Can store harvest in field Easily removed No known pests/diseases Uses existing farm equipment n/a

28 THE IDEAL BIOMASS CROP? CROP Corn SRC C4 photosynthesis Long canopy duration Recycles nutrients to roots Low input High water use efficiency Sterile non-invasive n/a Can store harvest in field Easily removed No known pests/diseases Uses existing farm equipment

29 THE IDEAL BIOMASS CROP? CROP Corn SRC C4PG C4 photosynthesis Long canopy duration Recycles nutrients to roots Low input High water use efficiency Sterile non-invasive n/a Can store harvest in field Easily removed No known pests/diseases Uses existing farm equipment

30 ROADMAP Why fuels from crops? What is the ideal biomass crop? Performance of Miscanthus and switchgrass. Food vs. Fuel

31 Switchgrass and Miscanthus C4 Perennial Grasses USA Switchgrass (Panicum virgatum L.) Europe Miscanthus (Miscanthus x giganteus Greef et Deu.)

32 An Inter-specific triploid hybrid hybrid Miscanthus sinensis Miscanthus sacchariflorus Miscanthus x giganteus + = Diploid 2n=2x=38 Tetraploid 2n=4x=76 Triploid 2n=3x=57 STERILE

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35 Planting in 2002

36 Randomized plot trials

37 March Pre-Emergence & Post-Harvest Miscanthus and Switchgrass

38 Early Spring (April) before maize is planted

39 Efficient Solar Radiation Capture 4 th July

40 Early August

41 Late October

42 January

43 Winter cutting (followed by baling) of demonstration plot of Miscanthus x giganteus at the South Farms, University of Illinois, Urbana, early February Harvest

44 Harvested Bales

45 ebi program leaders Miscanthus Traditional thatching material in Japan ENGINEERING ENZYMES AND METABOLIC PATHWAYS TO DEGRADE PLANT CELL WALL LIGNOCELLULOSE John Gerlt FEEDSTOCK PRODUCTION/ AGRONOMY PROGRAM Tom Voigt ENGINEERING SOLUTIONS FOR BIOMASS FEEDSTOCK PRODUCTION K.C. Ting BIOFUEL ECONOMIC AND ENVIRONMENTAL IMPACTS Madhu Khanna ENVIRONMENTAL IMPACT AND SUSTAINABILITY OF FEEDSTOCK PRODUCTION Evan DeLucia ASSESSING IMPACT OF INSECT PESTS AND PLANT PATHOGENS ON BIOMASS PRODUCTION Michael Gray GENOMIC-DIRECTED IMPROVEMENT OF FEEDSTOCKS Stephen Moose BIOFUELS LAW AND REGULATION Jay Kesan DISCOVERY OF ENZYMES TO DEGRADE PLANT CELL WALL LIGNOCELLULOSE Isaac Cann

46 ebi program leaders Miscanthus and cold-tolerance of Western Blot Analysis C4 photosynthesis. M. x giganteus Z. mays PPDK Wang, Moose, Portis & Long (2008) Plant Physiology (current issue) PEPc LS Rubisco

47 Light interception by Miscanthus and corn Champaign, IL.

48 BELOW GROUND BIOMASS MISCANTHUS AND SWITCHGRASS 5 YEARS AFTER PLANTING

49 THE BOTTOM LINE Feedstock Harvestable Dry Biomass (t/ha) Ethanol (liters/ha) Mha needed for 133 billion liters of ethanol % 2006 harvested US cropland Maize grain , Maize stover , Maize Total , Prairie mix 3.8 1, Switchgrass , Miscanthus , Source: Heaton, Dohleman & Long (2008) Global Change Biology 14,

50 ROADMAP Why fuels from crops? What is the ideal biomass crop? Performance of Miscanthus and switchgrass. Food vs. Fuel

51 Spatial Variability in Yields SPATIAL VARIABILITY IN YIELD

52 Miscanthus can be productive on marginal land MISCANTHUS IS PRODUCTIVE ON MARGINAL LAND The Miscanthus x giganteus in Cashel Co. Tipperary, Ireland photographed in September 1996 with 72HP tractor for scale.

53 WATER REQUIREMENT How much water? 10 g kpa -1 kg -1 = 0.1 t ha -1 kpa -1 mm -1 Beale, Morison & Long (1999) Agric For Met 96, mm at 1.0 kpa would allow 100 t ha -1 (assumes all water available) 500 mm at 4.0 kpa would allow 12.5 t ha -1

54 Enough land/enough water? US AVERAGE PRECIPITATION

55 Enough land/enough water? PREDICTED MISCANTHUS YIELDS t/ha Data of Dr. Fernando Miguez, EBI, Univ. Illinois.

56 US ARABLE CROPPING 1900 From: US Geological Survey -

57 US ARABLE CROPPING 1992 From: US Geological Survey -

58 WHAT IS POSSIBLE WITH THIS RETIRED LAND? Miscanthus yields: 40 dry tons/ha feasible 400 liters of ethanol / dry ton liters/ha 40 M out of 200 M ha ~640 B liters / year of ethanol US consumption (2004) = 780 B liters ethanol equivalent (excludes diesel)

59 OVER 1 BILLION ACRES OF RECENTLY ABANDONED AGRICULTURAL LAND >>A billion acres of agricultural land have been abandoned Campbell et al., Env. Sci. Technol. (2008) ASAP Article, /es800052w

60 CAN THESE CROPS BE SCALED UP? IRELAND 50 ha/day Next generation Miscanthus planter (50 acres per day).

61 MISCANTHUS FOR DRAX B

62 Looking beyond crop plants LOOKING BEYOND CURRENT CROPS

63 TRANSFORMATION SYSTEMS

64 Feedstock Genomics Program The primary aim of the EBI Feedstocks Genomics Program is to generate resources that will enable genomics-directed improvement of Saccharum and Miscanthus species as biofuels feedstocks. 1. Deep sequencing of the Miscanthus x giganteus transcriptome 2. Miscanthus and Saccharum genome sequencing 3. Assess genetic diversity within Miscanthus and Saccharum 4. Identify molecular markers for high-density genetic mapping in Miscanthus and Saccharum, and associate marker genotypes with phenotypes that contribute to biomass yield and composition in structured genetic populations. 5. Develop genome-scale expression profiling platforms for Miscanthus and Saccharum. 6. Develop an integrated bioinformatics system for functional genomics.

65 Miscanthus EST sequencing Tissue Collected Total RNA PolyA+ RNA cdna for Library Mature Leaf Yes Yes [1000+/1000ug] Yes [5+/5ug] No Post-Flowering Mature Leaf Yes No No No Emerging Shoot, Above Ground (Purple) Yes Yes [1000+/1000ug] Yes [5+/5ug] No Emerging Shoot, Above Ground (Green) Yes Yes [1000+/1000ug] Yes [5/5ug] No Emerging Shoot, Still Underground Yes Yes [1000+/1000ug] Yes [5/5ug] No Apical Meristem (Midseason) Yes No [889/1000ug] No No Apical Meristem (Pre-Flowering) Yes No No No Apical Meristem (During Flowering) Yes No No No Apical Meristem (Post-Flowering) Yes No No No Second Node Down (Midseason) Yes No [552/1000ug] No No Leaves Beginning to Open Rhizome Lateral Buds Rhizomes that had Emerging Shoots Yes Yes Yes Yes [1000+/1000ug] Yes [1000+/1000ug] No [348/1000ug] Yes [5+/5ug] Yes [5+/5ug] Post-Flowering Rhizomes Yes No No No Juvenile Roots Yes Yes [1000+/1000ug] No No [3+/5ug] Pollen Yes No No No Immature Inflorescence Yes No No No Mature Inflorescence Yes No No No No No No No Sanger sequencing approach to identify allelic variants among polyploid homeologs. Verbal commitment from JGI to sequence 150K M. x g ESTs.

66 454 Survey Sequence of M. x g genome Used flow cytometry to estimate M. x g genome at 7.5Gbp, or ~2.5 Gbp per haploid genome. Genome Shotgun Genome skim 84 Mbp of DNA in 366,448 reads average read length was 229bp 44% GC content 1.2% coverage of the Mxg nuclear subgenomes Whole chr omosome sequences 100kb aver age chunks <1kb chunks Done clone by clone Need physical map Better than nothing?

67 Agave mapisaga and A. saimiana 40 t/ha in semi-desert, Mexico DF. Nobel, Garcia- Moya & Quero (1992) Plant Cell & Environment 15,

68 Spartina alterniflora 22 t/ha, Georgia USA, in seawater Wiegert, Chalmers & Randerson (1983) Oikos 41, 1-6

69 ALTERATION Improved canopy architecture Rubisco with decreased oxygenase activity Bypassing glycine decarboxylation. Increased rate of recovery from photoprotection of photosynthesis Introduction of higher catalytic rate foreign forms of Rubisco Altered allocation of resources within the photosynthetic apparatus Long (2006) Plant Cell Env., % Increase in c relative to current value. Speculated time horizon (years) 10% (0-40%) % (5-60%) >20 20% (2-35%) % (6-40%) 5 22% (17-30%) % (0-60%) 0-5

70 % of beginning Photosynthesis Rubisco PGA Kinase GAPDH FBP aldolase FBPase Aldolase SBPase Transketolase PRK ADPGPP GCEA Kinase PGCAPase GOA Oxidase GSAT HPR reductase GGAT GDC cfbp aldolase cfbpase UDPGP SPS SPP F26BPase Zhu et al (2007) Plant Physiology 145: Evolutionary algorithm at work

71 Agronomists as seen by Plant Molecular Biologists Daily Illini 9/15/1907

72 Value of Plant Molecular Biology as viewed by Agronomists

73 WHAT IS THE EBI? INTEGRATING PROGRAMS Under One Roof Programs Feedstock Development Feedstock Deconstruction Fuel Synthesis Environment, Economics & Policy Projects

74 Meeting Global Productivity Needs or A Perfect Storm for Plant Sciences. Global Feed and Food Shortage. Replace Use of Fossil Fuels. Increased productivity solution to feed, food & fuel. Omics coming of age. Crop sequences completed & annotation advancing. Transformation technologies Cell wall, oil and carbohydrate synthetic pathways being unraveled. Plant systems biology coupled with computational biology tools emerging. Plant synthetic biology underway.

75 WHICH FEEDSTOCK Poplars Eucalypts Phylostachys Sugar cane Spartina Black Mangrove Willows Miscanthus Sorghum Haloxylon Sitka Spruce Switchgrass Echinochloa p. Casuarina Jatropha Eucalypts Poplars Crambe Arundo Willows Crambe Big Bluestem

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