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Dr. Gary Peter Professor University of Florida. Southern Pines: The Bioenergy & Renewable Chemicals Star of the Southeastern US Gary Peter University.

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Presentation on theme: "Dr. Gary Peter Professor University of Florida. Southern Pines: The Bioenergy & Renewable Chemicals Star of the Southeastern US Gary Peter University."— Presentation transcript:

1 Dr. Gary Peter Professor University of Florida

2 Southern Pines: The Bioenergy & Renewable Chemicals Star of the Southeastern US Gary Peter University of Florida

3 Pine Forests of the Southeastern US Forests occupy over 200 million acres (60% of the land area), with a large fraction dominated by pines – 10 species, loblolly and slash economically important ~85% of all forestlands are privately owned About half the pine forest is planted with genetically improved seedlings – About 10 million ha / 25 million ac each Contains 12 Pg of C, 36% of the sequestered forest C in the contiguous United States Annually sequester 76 Tg C, equivalent to 13% of regional greenhouse gas emissions

4 US South: Forestry & Forest Industry Largest biomass industry in world – Produces 16% of global industrial wood supply More than any other country – Supplies 60% of US & 25% of world pulp & paper markets 43 million tons of annual capacity Generates ~2/3’s of all industrial bioenergy – Used on site Sustainability is a key focus for industry – >93% of stem is utilized Southern pulp mill location & capacity Johnson & Steppleton, 2011

5 Forest Products Supply Chain Feedstock production Feedstock logistics Biomaterials Distribution & use Scalable – Large land area – Large stable markets Sustainable – More volume growth than harvested Cost competitive for traditional products – Pulp, paper, wood

6 TimberMart-South Operating & Proposed Wood Biomass to Electric Power & Wood Pellet Facilities Approx. 30 actual or proposed plants Approx. 40 actual or proposed plants

7 Biofuel Production in the Southeast 2010 USDA biofuels report estimates that ~50% of the advanced biofuel production capacity will be located in the southeast US – Most favorable growing conditions & available land Advanced biofuel facilities that can use pine feedstock – KiOR (thermochem) – Bluefire (acid hydrolysis) – Ineos (thermochem) – Bluesugar ?

8 Stable Cost & Large Supply

9 Since 1940s, planted pine productivity has tripled, primarily due to improved genetic stock and silvicultural technology developed and disseminated by University / Government / Industry Research Cooperatives Redrawn from: Fox, T.R., E.J. Jokela and H.L. Allen The development of pine plantation silviculture in the southern United States. J. Forestry 105:

10 Quantitative Trait BLUP Variance Components P= G + E G= A + D + I -Heritability (h 2 ) -G x E -G x Age Breeding Values -Ranking genotypes -Selection Phenotype: Total Height Traditional Phenotypic Breeding with Recurrent Selection

11 CFGRP: Slash Pine Deployment Gains & Value Genetic Gains in Harvest Yields (%) Low Hazard Conservative estimate of incremental increase in stumpage value (6% interest) due to increased yields from planting genetically improved stock in FL estate Source: Greg Powell, Univ Florida

12 Breeding Testing Propagation Start Breeding Commercial Production Pine Breeding Cycle Pine Breeding is a long multi-step process Can be partitioned in three stages >30 years 1 st to 2 nd Generation Reduction of more than 10 years: -Early Selection -Smaller populations -Top-grafting White et al. 2008

13 I - QTL analysis II – Genetic association III – Genomic selection Linkage Blocks Large Small Medium Resolution Low High Medium Marker Assisted Selection B. Indirect markers based on linkage disequilibrium:

14 Validation Fit all SNPs in a prediction model Y =  SNP + e Training population Genotypes Phenotypes Define multi-loci models to predict phenotypes Genome-Wide Selection Meuwissen et al. (2001) Genetics 157:

15 Genomic Selection is operational in cattle breeding and evaluated in other animals, crops and trees Focus has been on development of methods (e.g. GBLUP, RR-BLUP, Bayes A, Bayes B, LASSO, RKHS, Machine learning, etc.) Everybody agrees that GS application depends on the accuracy of predicting phenotype with markers Theoretically accuracy depends on: – Linkage disequilibrium extent – Training population size – Heritability – Number of QTLs But also depends on the BV quality used to construct the GS model Genomic Selection “Current status in breeding”

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17 DBH & Height GWS Accuracies in CCLONES

18 BTP 10+ years 8+ years T 8+ years B 4 years P <1yr T 8+ years B 4 years P <1yr BT P 4 years 1yr BT P 4 years 1yr BT P 4 years 1yr GWS Incorporated into Pine Breeding

19 Conifer Oleoresin Canal System for Insect and Fungal Resistance The wood resin canals (vertical and horizontal) are organized into a 3D network for terpene synthesis and storage – Thin walled resin canal epithelial cells line the canal and synthesize and secrete terpenes into the lumen of the canals or duct Resin flows out of stem after wounding, typically by boring insects – Constitutive resin under positive pressure in resin canals Resin canals

20 Why Terpenes? Terpene Biosynthesis Conserved biosynthetic pathways in microbes & plants Large variety of natural terpenes with varying chemical properties – Mono, sesqui-, di- and triterpenes Terpenes as Biofuels High energy density - carbon and hydrogen rich and low in oxygen Simple & efficient chemical methods for conversion of natural terpenes to drop-in fuels suitable for blending or replacement of petroleum are available –  pinene dimers as a jet fuel replacement – Bisabolene to bisabolane a D2 diesel fuel replacement – Farensene as a diesel fuel

21 Biofuels & Co-products Extraction Sugar – Ferment to EtOH – Sugar Starch – Amylase + ferment to EtOH – Oil, animal feed Oil – Transesterification to biodiesel – Glycerin Deconstruction Lignocellulose – Sugar Platform Size reduction + degradation + fermentation Power, lignin – Gas Platform Anaerobic digestion to biogas Gasification + catalytic synthesis to liquid fuel Power – Liquid Platform Cracking / pyrolysis + upgrading 1 st GENERATION BIOFUELS2 nd GENERATION BIOFUELS

22 Conversion of Biomass to Fuel Extraction Based Compound highly concentrated in biomass that facilitates efficient recovery Starting material has high chemical uniformity High efficiency conversion with limited input costs Deconstruction Based Biomass is large & heterogeneous Starting material has relatively low chemical uniformity Requires substantial energy and/or chemical inputs to reduce Come from domesticated plants breed & selected for concentration & yield of food Non-edible parts of food plants & undomesticated grasses & trees

23 Pine Terpenes Naturally synthesize a large diversity of mono-, sesqui- and diterpenes as defense compounds against insects & fungi Terpenes accumulate in wood naturally to >20% – Constitutive synthesis – Inducible synthesis Genetic and environmental control of wood terpene content

24 Current Pine Terpene Industry Pine Wood Rosin Pulp Mill Biosynthesis Extraction Crude Products Final Products Specialty Chemicals Gum Turpentine & Rosin CTO & CST Industrial Biofuels Flavors & Fragrances Live Tree Wood Turpentine & Rosin

25 Pine Terpenes: A $3 Billion Global Industry Pine Terpene collection > 1 billion tonne/yr – Turpentine (mono- & sesquiterpene) rosin (diterpenes) – Gum terpene (60%), crude sulfated turpentine & crude tall oil (35%), wood naval stores (5%) Gum terpenes collected by tapping living trees > 850,000 tonne/yr – China, Portugal, USSR, Brazil, Indonesia, Mexico, India – China >500,000 tonne/yr [60% of global supply but little is exported] Pulp & paper industry collects terpenes as a co-product – Crude sulfated turpentine & Crude tall oil (CTO) – US south 450,000 tonne/yr of CTO

26 Phenotypes Resin canals Number of resin canals per year (averaged over triplicate samples) 543 cloned genotypes 3 sites 3 clonal replicates per site 2 years Oleoresin drymass Box-Cox transformed oleoresin drymass exuded over 24 hours 1002 cloned genotypes 3 sites 3 clonal replicates per site 3 years (one site) Wood terpene content Diterpene content in dry wood 940 cloned genotypes 2 sites 2 clonal replicates per site Total, mono- & diterpene content in wet wood 750 cloned genotypes 1 site 4 clonal replicates per site

27 Oleoresin traits are heritable H 2 resin canal number single site: 0.15 – 0.21 across sites: 0.12 H 2 oleoresin drymass single site: 0.18 – 0.34 across sites: 0.18

28 Phenotypic variation in oleoresin drymass is positively skewed Oleoresin drymass by site Xylem growth increment per year Resin canal number per year

29 Associated SNPs accurately predict additive genetic variation in oleoresin drymass

30 Estimated F1 genetic gains in oleoresin drymass under varying selection intensities siteh2h2 Fold-increase breed top 10% Fold-increase breed top 5% Fold-increase breed top 1% CUT NAS yr NAS yr PAL ALL h 2 : narrow sense heritability

31 TE-Pine Can Exceed PETRO Metrics Scalable – 13 million+ h planted pine exist – Yield gains achievable Environmentally Sustainable – High harvest index – Strong positive net energy – Strong negative CO 2 eq Economically Sustainable – Lignocellulose & terpene co- product synergy – Adds value across supply chain Adds Flexibility – No clear detrimental change in current product mix – Strengthens possibility of pine as a dedicated biofuel crop – Multiple routes to extraction Plants Engineered To Replace Oil Increase the mass of readily extractable hydrocarbons to meet technical targets at costs competitive with crude oil Technical TargetsValue Required HT- Pine 1.1-Energy density> 26.5 MJ/L (LHV) 1.2–Melting point< -40 o C <-63 o C 1.3–Boiling point> 35 o C >135 o C 1.4-Energy> 160 GJ ha -1 y Process cost< $10 GJ CO 2 useAtmospheric CO 2 Ambient 2.2- H 2 O requirement < 22 inch y -1 No irrigation 2.3- Fertilizer requirement <201 kg ha -1 y -1 N, <77 kg ha -1 y -1 P, <56 kg ha -1 y -1 K 58.5 kg ha -1 y -1 N, 7.5 kg ha -1 y -1 P

32 Genetic engineering to rapidly increase oleoresin production in pine stems Association genetics Multi-site analysis of correlated oleoresin traits in a structured clonal population Gene expression Differential expression with chemical elicitors of resinosis Tissue-specific expression in resin canals Discovery phase Validation phase Candidate genes RNAi mediated silencing Overexpression Wild-type v. mutant phenotypes

33 Project Overview To increase terpene production 5 fold Triple Resin Capacity 25% Greater Flux Activation Pathway 1.5X Faster Synthesis Enzyme Three Synergistic Strategies for Increasing Pine Terpene Synthesis & Storage Will Be Used Constitutive Resinosis Upregulate Carbon Flux to Terpenes Optimize Composition & Production of Terpenes

34 Terpenes & the Future Forest Biorefinery Issue Land Use Environmental Sustainability Conversion Efficiency Cost effective Net positive energy relative to fossil fuels Alignment Dramatic increase in GJ/ha/y Increased value to landowners sustains forest land Extracted as a co-product – lignocellulose still useful for all traditional products or energy Existing capital Flexible end product markets Strongly positive to fossil fuels

35 Acknowledgements COLLABORATORS University of Florida – John Davis, Chris Dervinis, Matias Kirst, Patricio Munoz, Marcio Resende, Alejandro Riveros-Walker, Jared Westbrook ArborGen – Will Rottmann NREL – Mark Davis, Robert Sykes University of California, Berkeley – Jim Keasling, Jim Kirby, Pamela Peralta-Yayha, Blake Simmons FUNDING DOE/ARPA-E USDA/NIFA Forest Biology Research Cooperative – Plum Creek Timber, Rayonier, Weyerhaeuser, RMS, F & W

36 Project Summary Combinatorial engineering 20% wood terpene Increased Resin canal #/volume Increased terpene synthesis Resinosis Improved enzymes Increased carbon flux Five fold increase in wood terpene Discovery Technoeconomic Modeling Forest tree growth Terpene recovery Fuel production Value Chain Analysis & Proposition Germplasm providers Landowners Harvesting/transport Wood processors Fuel synthesis Commercialization Partners Pulp & paper Biofuel Producers Wood products Bioenergy Oleochemical Refiners Flavor & Fragrances

37 Dr. Gary Peter Professor University of Florida


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