Increased Crop Yield ThroughImproved Photosynthesis

4th International Conference on Agriculture and HorticultureBeijing, July 14, 2015

Andy Renz, Vice President Business Development

100% Increase in Productivity Required by 2050

Required yield increases are significantly higher than historical yield increases.

Ray et al, 2013, PLoS ONE

2nd Generation Ag Biotech – Abiotic Stress Tolerance & Yield Increase

Yield traits represent the largest opportunity in ag biotech

2025

1st generation traits

2nd generation traits

2nd Generation Ag Biotech – Abiotic Stress Tolerance & Yield Increase

• Excellent results and products from molecular breeding– AQUAmax™ corn from Pioneer– Artesian™ corn from Syngenta– Droght tolerant rice from IRRI

• Opportunity and Challenge for GM approaches:– Monsanto/BASF: largest partnership in the history of Ag Biotech

R&D: $2.5 billion(!)• HTP screens in model and crop plats• Field testing in crops (commercial germplasm)• First prducts: Droughtgard™ corn launched in 2013 (CspB)

– Benson Hill Biosystems:• Focus on yield improvement through improved photosynthesis

Far belowthreshold limits

Quite optimizedthrough breeding

Can Improvement in Photosynthesis Increase Crop Yields?

Long et al. (2006) Plant Cell Envir 29:315-30

Yield Potential: Y = 0.487 * St * ɛi * ɛp* ɛc

• 0.487: percentage of photosynthetically active radiation• St : total incident solar radiation across the growing season

• Ɛi: : light interception efficiency, i.e. ability of canopy to capture sunlight

• Ɛp : partitioning efficiency, i.e. harvest index

• Ɛc : conversion efficiency, i.e. combined gross photosynthesis of the canopy, less all plant respiratory losses

YES, by > 50% !

Recent Publications Confirming this Hypothesis (1)

Simkin et al (2015) J Exp Bot 66:4075-90

Recent Publications Confirming this Hypothesis (2)

Ambavaram et al (2014) Nature Communications 5:5302-16

Photosynthesis is the Most Promising Target to Increase Crop Productivity

I. Enhance photosynthetic efficiencyII. Increase overall energy availabilityIII. Increase photosynthetic productivity in a canopyIV. Maintain photosynthesis during abiotic stresses

Focus on rate-limiting steps of primary

metabolism

“fine tuning” to provide genetic variability that

otherwise would never occur

Benson-Calvin cycle

Carbon shuttle

Starch production

Light Harvesting

ATP + NADPH

Sugars

Energy

Requires use of multi-genic approaches and precisely controlled gene expression

Benson Hill Biosystems – Company Summary

• The Photosynthesis Company TM

• Focus: Increased Crop Yield through Improved Photosynthesis• Discovery: Integrated, systems-based approach to engineering plant primary

metabolism• One platform – multiple product opportunities:

corn, sugarcane, soybean, rice, wheat, cotton, oil palm, canola, Eucalyptus,…• GM- and non-GM product concepts• World-class plant growth and genomics facility licensed and in use• Pre-Series A company; more partnership revenues than venture capital• Partnerships with global leaders

Benson Hill Biosystems – Integrated Platform with Focus on Photosynthesis

Deep knowledge about photosynthesis

PSKbase TM – Unique omics and computational platform

State of the artphenotyping

Cropmodels

Gene regulatorynetworks

Gene-based ModificationsTransgenic &

Non-GM

Synthetic ChemistryCrop performance &

Crop protection

BiologicalsStimulants &

microbes

Field testing in crops

Aerial imaging

Network plasticityanalysis

Genomeediting

Genomic selection &Computational breeding

Partnership with Donald Danforth Plant Science Center

• IP/license access to labs of Tom Brutnell and Todd Mockler– 35+ FTEs working on photosynthesis, computational and systems biology, transcriptional

regulatory networks, novel targets and promoters, etc.

• Enablement of CapEx Lite with access to:– Tissue Culture and Transformation Facility (2,000ft2)– Potting area with soils handling room (1,900ft2)– 33 Conviron chambers and 18 Conviron rooms (3,000ft2)– 36 Greenhouses (44,000ft2)– Bioinformatics Core: 800+ processors, 3 TB memory, and a

single, high-performance 204 TB storage area network– Other Cores: Proteomics, Mass Spectrometry, Integrated Microscopy, X-Ray Crystallography– High-throughput robotics assay platforms– High-throughput plant phenotyping system

Iterative Lead Identification and Optimization

___ Lead________Transcriptomics

Association mappingGenomic selection

PhysiologyModeling

Mode of Action__Reverse geneticsOver-expression

Cell biology

Optimization__Expression profile

Protein modificationLocalization

PSKbase™

Setaria viridis as modelfor C4 crops,

rice and Brachypodiumfor C3 crops

PSKbase™ is BHB’s central vehicle for integrating and interpreting data, and a

tool for rapid curation, prioritization, and

selection of promising trait candidates

Pipeline Summary

PSKbase™: Various datasets(proprietary and publicly available)

Proprietary algorithm-based approaches

to integrate datasets

Target identification(PSKbase™ prioritization)

Lead prioritization:validation assays and in planta validation

Crop plant validation

Yield field trials

Trait Candidates Genes

>800

321 38

140 12

Licensed Genes

Discovery: PSKbaseTM and Computational Biology Platform

PSKbase™: Various datasets(proprietary and publicly available)

Proprietary algorithm-based approaches

to integrate datasets

Target identification(PSKbase™ prioritization)

Lead prioritization:validation assays and in planta validation

Crop plant validation

Yield field trials

Trait Candidates Genes

>800

321 38

140 12

Licensed Genes

Leaf development

Setaria

Section A Section B Section C

Mesophyll Bundle sheath

Example 1: C4 Bundle-Sheath and Mesophyll Cell Photosynthesis Gene Networks

Illumina RNA-seq, 3 reps for each cell population

Example 2: Wheat Leaf CO2-responsive Gene Expression Atlas

Low [CO2]

Ambient[CO2]

High [CO2]

baseline developmental and metabolic

gene expression

[CO2] – responsive functional gene candidates

Example 2: Photosynthesis Target Identification from Gene Networks

Gene Network

Photosynthetic Subnetwork

336 genesand

93 TFs

Samples from Wheat

Automated Proprietary Analytics Pipeline

Example 2: Network Plasticity Analysis Identifies Candidate Genes

[CO2]

89 wheat loci that had significant plastic gene network interactions with core photosynthesis genes

Gene X

Gene Y

Gene Z Gene A

Gene B

Our approach identifies both specific candidates and their regulatory partners that are responsive to atmospheric conditions.

Environmental Conditions and Photosynthesis are Linked

• Example project: used for cross-referencing with photosynthesis network analyses: 4,128 genes – cold, salt, drought, and heat-responsive

• Stress-associated regulons identifyco-expressed genes implicated inabiotic stress responsesand primary metabolism

Brown – DroughtBlack – SaltRed – HeatBlue – ColdPurple and Green – multiple interactions

In planta Validation of Leads

PSKbase™: Various datasets(proprietary and publicly available)

Proprietary algorithm-based approaches

to integrate datasets

Target identification(PSKbase™ prioritization)

Lead prioritization:validation assays and in planta validation

Crop plant validation

Yield field trials

Trait Candidates Genes

>800

321 38

140 12

Licensed Genes

C4-Specific Transcription Factors identified by PSKbase™

Project Objectives: Demonstrate improvements in photosynthesis, plant growth, and yield by overexpression of photosynthesis-associated transcription factors identified through developmental transcriptomics and bioinformatic analyses.

Background/Rationale:• Benson Hill Biosystem’s PSKbase™ is a

proprietary tools used for identifying and prioritizing trait candidates.

• Using PSKbase™, 8 uncharacterized maize transcription factors were identified for testing in a C3 system – Brachypodium distachyon.

• To provide for diversity of expression profiles, 4 promoters were selected and combined with each of the 8 TFs, for a total of 32 constructs.

C4-Specific Transcription Factors: High Hit Rate in Model Plants

In T1 Brachypodium plants, 5 of the 8 selected Transcription factors have phenotypes with significant increases in biomass

In each graphic: left plants are wild-type (non-transgenic) Brachypodium distachyon, and right plants are T1 Brachypodium

distachyon plants containing BHB candidate

BH15 Enhances Carbon Availability in C3 and C4 Plants

• Significantly improved photosynthetic efficiency, WUE, and yield in soybean, tobacco, and Arabidopsis

• Soybean greenhouse and field yieldincreases of >15% and >7%, respectively

• Optimizing by:– Co-expressing with genes encoding

rate-limiting steps of photosynthesis– Employing spatial- and temporal-

specific expression profile

• Advancing into maize and rice• First constructs in maize being tested

in hybrids in 2015 field season

Multi-year soybean data

BH30 Increases Water Availability and Photosynthesis

• Over-expression of BH30 results in increased hydraulic conductivity and plant growth in Arabidopsis and poplar, particularly under heat conditions

• Quantification of vessel size shows ~33% increase in vessel mean diameter

BH33 is a Well Characterized Sink Strength Lead

• Naturally-occurring, well characterized enzyme in maize grain, mutated for improved thermal stability and enzyme kinetics

• Previous mutated versions have shown promise in field trials of multiple crops• Iterative mutation has resulted in BH33, which has improved characteristics relative to

enzymes previously expressed in plants

• Maize field testing in 2014 showed up to 24% increase in ear weight relative to control (null segregants with otherwise identical genetic background)

• Inbred data in greenhouse also show positive results• Hybrid seed production completed and field trials in 2015 season ongoing

• Coupling with numerous other source-focused trait candidates• Testing initiated in multiple other crops

BH71: Increased Seed Yield and Nitrogen Utilization for C4 Crops

Low N High N0.00

0.20

0.40

0.60

0.80

1.00

1.20 Dry Weight

WTNull SegregantHomozygote

Dry

Wei

ght (

g)

Low N High N0.0

200.0

400.0

600.0

800.0

1000.0 Total Seed Weight

WTNull SegregantHomozygote

Tota

l See

d W

eigh

t (m

g)

>130%increase >400%

increase

• Significant increase of biomass and seed yield, in particular under N-limiting conditions;• Strong Lead for sugarcane, corn and sorghum;

CORNSOYBEANSCOTTONCANOLAPOTATO

SUGARCANEWHEAT

RICEALFALFA

OIL PALMTOBACCO

Computational and systems-biology

Setaria (C4),Brachypodium & rice (C3)

model plant platforms

Genetic “toolbox” for manipulating primary

metabolismPhotosynthesis

know-how and focus

PLATFORM

SORGHUMENERGY GRASSES

POPLAREUCALYPTUSOTHER TREES

TOMATOYAMS

SWEET POTATOESSUGAR BEET

CASSAVAMILLET

OTHER VEGETABLESALGAE

Product Opportunities

Improving Photosynthesis is Broadly Applicable & Enables Many Product Opportunities

Path to Commercialization – “Go to Partner”

• Seed market is consolidated,with high barriers to entry– Elite germplasm, i.e. plant genetics

• Biotech trait discovery and development is entry point for participating in the most valuable and high-growth segment

• To monetize traits, seed are used as value capture mechanism– Premium pricing for seed containing biotech trait(s)– Value sharing via royalties, which can be pre-calculated (flat rate) or based on percentage of

trait value retained by seed company

• Benson Hill Biosystems is partnering on a crop-by-crop basis– Non-exclusive deal structures in corn and soybean– Exclusive and alternative deal structures in other crops

REQUIRES PARTNERSHIPS

Thank You

• Prerequisite: most BHB lead genes are derived from crops• Validate targets identified through PSKbase™• Platform agnostic: CRISPR/Cas, TALEN, Meganucleases• Precise introduction of foreign genes• Modification of target gene expression• Replacement of genes by improved versions• Uniform expression levels (only few events required)• Modification of native genes through small insertions

– Regulatory motifs to modify gene expression– Specific base pair changes to modify activity/specificity of native enzymes

• Potential Non-GMO approach– Of particular relevance for wheat, rice, etc.

• USDA confirms BHB’s approach as non-regulated, i.e. non-GM products

Non-GM Product Concept: Genome Editing Technology