PUBLIC-PRIVATE PARTNERSHIPS IN PLANT BREEDING AND BIOTECHNOLOGY

PUBLIC-PRIVATE PARTNERSHIPS IN PLANT BREEDING AND BIOTECHNOLOGY Alan B. Bennett Distinguished Professor of Plant Sciences Executive Director, Public Intellectual Property Resource

Public-Private partnerships are effective at multiple levels Genetic resource conservation Sustaining the building blocks of future crops Creating the genetic base of sustainable agricultural systems Genetic resource utilization crop improvement Biotech traits The basis for publicprivate partnerships

Plant Genetic Resource Conservation Arboreta and botanic gardens Clonal repositories Tissue culture storage Ex situ Seed banks public seed and gene banks seed storage facilities farmer / gardener collections private seed companies DNA libraries (BACs, cdnas, etc)

An ex situ conservation collection Acquisition Maintenance Evaluation Research Documentation Distribution A public-private partnership between UC Davis and the California tomato seed and processing industry.

A collection of tomato genetic stocks and wild relatives. Mutant stocks Single gene mutations Marker combinations Wild species 17 wild species of Solanum 1,200 wild populations Miscellaneous stocks Cytogenetic stocks Landrace varieties Breeding lines etc. Total 3,600 accessions

Why ex-situ genetic resource collections? Threats to Wild Tomatoes in Native Area herbivores agriculture urbanization mining

Genetic Erosion of In Situ populations Extinction of local populations or entire species; Fewer and smaller populations means less genetic variation remains; In too many countries, little is done to protect genetic resources in situ Example: of the known populations of wild tomatoes, many have already been eliminated in situ; but are preserved ex-situ

Seed Distribution Annual activity 300-500 requests from researchers in over 30 countries approx. 5,000 seed samples distributed utilization rate of ~ 100% seed provided gratis Common uses breeding genetics and genomics development stress tolerance fruit ripening, biochemistry evolutionary studies education

Utilization of genetic resources Resistances to >45 Diseases in Wild Tomatoes Root Knot Nematode Mi +

Utilization of genetic resources The jointless-2 gene from S. cheesmaniae facilitates mechanical harvest. + j-2

Utilization of genetic resources Geographic Origin and Migrations of Tomato ca. 1710 ca. 1520 mid-1900 s center of domestication center of diversity

Public-Private partnerships are effective at multiple levels Genetic resource conservation Sustaining the building blocks of future crops Creating the genetic base of sustainable agricultural systems Genetic resource utilization crop improvement Biotech traits The basis for publicprivate partnerships

100 Years of Plant Breeding in California Is the history of University of California public-private partnerships with industry There is a long documented history of plant breeding of 31 different crops at UC Davis. Each breeding program is a public-private partnership. The University hires plant breeders and the industry provides financial support to the breeding program. These programs provide the most important varieties to many of the major commodities in California (and globally). sbc.ucdavis.edu/files2/57360.pdf

Rice (Oryza sativa) Calrose variety, released in 1948 by the RES, represented 70% of CA rice area by 1975. It was the foundation variety for the growerfunded acceleration of breeding activities in 1969.

Wheat (Triticum) Wheat, barley and oat breeding began at UC in 1904. Yields then were about 1 ton/ha. Today, yields are 2.5 to 3.5 tons/ha. Funded by California wheat growers

Stopping Stripe Rust Stripe rust is a devastating disease of wheat, and the appearance of more virulent races in the year 2000 resulted in large losses that reached 25% of the California wheat crop in 2003. The availability of more precise molecular methods of selection helped provide a fast answer to the stripe rust epidemic. Today the epidemic in California is over and resistant varieties are available. Jorge Dubcovsky

Walnuts (Juglans regia) Virtually all walnut varieties in California are from UC Davis. They are planted on 113,000 ha, producing 99% of US walnuts. One UC Davis variety (Chandler), released by Harold Forde in 1979, still accounts for over 40% of production in California. The new Forde variety may eventually replace Chandler as the top walnut variety.

Alfalfa (Medicago) E.H. Stanford and W.F. Lehman released CUF 101 in 1977, which was a highly successful alfalfa variety; it is still grown on significant acreage worldwide. Highline was released in 1997 with increased pest resistance and yield and UC- Impalo-WF released in 2000 is resistant to whitefly (Bemesia). California alfalfa yields are more than double the US average, 7.7 tons/ha in 2011. California also produces one-third of the US alfalfa seed supply.

Grapes (Vitus) Harold Olmo began crossing grapes at UC Davis in 1934 and released more than 30 grape varieties, including 20 table grapes, 9 wine grapes and two rootstocks. Andy Walker continues the UC Davis breeding program, developing nematode- and disease-resistant rootstocks and varieties. The Mondavi Institute for Wine and Food Science and the Jess S. Jackson Sustainable Winery provide the most modern facilities in the world for teaching and research in grape breeding and winemaking.

Peach (Prunus persica) California produces virtually all of the processing peaches in the US (1 billion pounds). This industry depends on UC varieties - so do many other countries

Strawberry (Fragaria x ananassa) The UC has released 56 varieties. R. Bringhurst and V. Voth improved yield 4-fold between 1945-1985. More than 87% of the strawberries consumed in North America are from UC varieties Approximately 60% of global strawberries are from UC varieties. R. Bringhurst D. Shaw

North California - Australia - Portugal - United Kingdom - New Zealand Varieties developed for California are suitable for growing conditions in other countries.

Licensing Objectives A Commercialization Partnership Benefit California Industry by developing new varieties for CA Global exploitation through a licensing program in strawberry regions Revenue stream (~ US$ 12M) used, in part, to support development of new varieties Key to success are strong plant IP rights in US and globally

Licensing Strategy A Commercialization Partnership Domestic Markets: US & Canada UC licenses directly with nurseries Global Markets UC License Indirectly to a Master Licensee Non-exclusive Basis Rights to Propagate Sell to Fruit Producers Sell to Nurseries for Propagation Exclusive Rights By Territories with strong IP Systems Control bio- piracy Reduced Fees in CA Higher Fees than in CA

UC s Strawberries lead in US and globally California Strawberry Industry: $2.2 B UC varieties represent 75% of California production #1 strawberry varieties in the world In 25 years, Spain s Industry grew from 0 to $1B Transfer of Plant Varieties Know-how 70-90% UC s varieties

Public-Private partnerships are effective at multiple levels What about biotech traits? Genetic resource conservation Sustaining the building blocks of future crops Creating the genetic base of sustainable agricultural systems Genetic resource utilization crop improvement Biotech traits The basis for publicprivate partnerships

Enabling access to public innovation

Linking proprietary germplasm with a trait gene to enhance alfalfa biomass yields Trait gene Enabling technologies Proprietary Germplasm and Downstream commercialization

What about biotech traits? Developing Toolkit of Enabling Technology for Plant Transformation extensively patented in the US Traits Disease/Stress Resistance Nutritional Enhancement Stress (salt/drought) Tolerance Enabling Technologies Germplasm DNA Vectors Promoters Selectable Markers Transformation Methods Crop Varieties

Transformation technologies with FTO Scientific Criteria Validation Legal Information FTO Licensing Regulatory Information PIPRA In house IP & Regulatory Analysts, Scientists, & Pro-bono Attorneys

Towards a Patent Pool for Plant Genetic Engineering Lawyers, Ag-biotech Companies, Regulatory Experts Define technical, legal and regulatory design parameters Agrobacterium-mediated Wide range of promoters Danforth Plant Science Center, St. Louis Clear of IP blocks in US (and elsewhere) Plant product should be marker-free Desirable to have possibility of all-plant integrations (cisgenics) Crop Development Validation Scientific Criteria Licensing Information Legal Information FTO

NAVIGATING GLOBAL IP LANDSCAPE Plant constitutive promoters Plant Expression for FMV34S comparable to CaMV35S promoter in leaf and tomato fruit Constitutive Promoter FMV34S US Patent 6,051,753 Licensable from University of California Davis

NAVIGATING GLOBAL IP LANDSCAPE Plant selectable markers NPTII Technical Hurdle: Alternate plant selectable markers not as effective and broadly applicable to many crops World Australia US Germany Japan Broad Scope: Plant selection with Any DNA sequence that confers resistance to any antibiotic Expired 2008 Europe Claims narrow to promoter Used to drive selection gene Expired 2011

Flowchart of Material Transfer MTA s restrictions and access to Tangible Property may restrict developing country deployment and are typically more restrictive than patents. To Avoid MTA Restrictions, used de novo DNA Synthesis by DNA 2.0 (California) Kryder, R. D., S. P. Kowalski, et al. (2002). "The Intellectual and Technical Property Components of pro-vitamin A Rice." ISAAA Briefs 20: 1-56.

Scientific Criteria Crop Development Legal Information FTO Validation Licensing Information

Public-Private Partnership R&D Plan to Improve Alfalfa as a Plant-Based Protein Factory Alfalfa Germplasm Transformation Optimization Elite Alfalfa Germplasm Tricoli-UC Davis Transf. Facility POC Completed Marker Free Plant Transformation System PIPRA-UC Davis POC Completed Environmental Sustainability Yield Increase Cal/West Seeds Senesco PIPRA-UC Davis POC Completed Water Use Efficiency Blumwald Lab- UC Davis Discovery Pet Protein & Animal Feed Improve Nutritional Protein Content for Pets PIPRA-UC Davis MARS POC In Development Increase Rumen Stability PIPRA-UC Davis Animal Sciences POC In Development

FACTOR 5A, EIF5A3 Enhancing alfalfa biomass yields with a single trait Overexpression of PdeIF5A3 Enhanced vegetative and reproductive growth (~3.4 fold) 50-300% increase in seed yield Enhanced fitness to osmotic and nutrient stress Plant and Cell and Environment (2010) 33, 1682-1696 Factor 5A regulates senescence, programmed cell death-apoptosis, cell division, and cell survival. Factor 5A is present in all plants and has been shown to: Extend Shelf Life Increase Seed Yield Increase Biomass/Growth Rates Resistance to Environmental Stress Therapeutic to regulate cell death and induce apoptosis in cancer tumors.

Kan R ColE1 pvs1 FACTOR 5A DNA CONSTRUCTS LB NPTII FMV34S::eIF5A3::E9 RB ppipra726 Plant selection GOI cassette LB NPTII Plant selection H3::eIF5A3::E9 GOI cassette RB ppipra728 1TDNA IPT LB NPTII CodA RB ppipra720 Plant selection Segregation marker IPT LB FMV34S::eIF5A3::E9 RB ppipra717 GOI cassette IPT LB H3::eIF5A3::E9 RB ppipra723 GOI cassette Co-Transformation based system Kan R ColE1 pvs1 Figueroa-Balderas, et al.

FACTOR 5A INCREASES ALFALFA BIOMASS YIELDS BY 20-45% Wild Type R97-37-5 GE Alfalfa Factor 5A Field established 2010 Agronomic evaluation 2010, 2011, & 2012

FIELD DATA: ALFALFA BIOMASS FORAGE YIELD & PROTEIN QUALITY 2010 Field Trial established Salem, WI 2011-2012 Agronomic evaluation, forage yield & quality examined 29 alfalfa clones

Strategies to elevate Protein levels & rumen stability o Zeins are usually accumulated to high level as storage proteins. o β- and δ-zeins with high levels of methionine (10-27%) o Classified as GRAS, Generally Recognized as Safe, by the U.S. FDA. o ER localization and protein body formation help decrease rumen digestion. 41

Linking proprietary germplasm with a trait gene to enhance alfalfa biomass yields Trait gene Enabling technologies Proprietary Germplasm and Downstream commercialization

LICENSING MODEL FOR ENABLING TECHNOLOGIES Balancing Commercial and Humanitarian Access Consolidated Intellectual and Tangible Property Rights Technology Providers RESEARCH Arkansas State University Cornell University UC Berkeley UC Davis Agricultural Natural Resources, UC Davis University of Florida University of Tennessee, Knoxville USDA Virginia Polytechnic Institution and State University International Agricultural Development Initiatives Hanoi Agricultural University, Vietnam Indian Institute of Vegetable Research, India IRTA Institute of Agriculture and Food Research and Technology (Spain) The University of York, UK University of Melbourne, Australia University of Talca, Chile UPCH - Universidad Peruana Cayetano Heredia, Peru Biotechnology Companies Arcadia Biosciences Cal/West Seeds-Dow AgroSciences NovaFlora Pre-negotiated licensing terms PIPRA Patent Pool Design, test, and disseminate plant transformation technologies humanitarian use research use commercial use Royalty free Fee based Revenue flow

Public-Private-Partnerships for the Development & Delivery of Nitrogen- Use Efficient and Salt tolerant rice to small holder farmers in Africa Arcadia Biosciences, provides trait technologies PIPRA, transformation technologies under humanitarian, royalty-free terms AATF (African Agricultural Technology Foundation), facilitates collaboration USAID-Funding

Public Private Partnerships Accelerate the translation of research from Public Sector.to Private Sector for development and commercialization Many partnerships from genetic resource conservation to plant breeding and biotech trait development are possible

Jon Reich David Johnson Jay Sandman Rosa Figueroa-Balderas Xiaodong Sun Paul Jones Le Thu Hien Tamar Cohen-Davidyan Christopher De Ben David Tricoli Ed DePeters Dan Putnam Cecilia L. Chi-Ham Alan B. Bennett

Back to Alfalfa Queen of the Forages CALIFORNIA STATE Produces 9% of the nation's alfalfa hay. 7 th most valuable commodity in the State $1.7 billion/year value 2012 Provides premier forage of dairy cows. US NATIONAL Third in value among US crops, after corn and soybeans. > $8 billion/yr value. CA leading dairy state since 1993. Produce 1/5 th of country s milk. Potential as a broad-based protein source for humans or pets. not including the value of dairy or other animal products, which are the final products of alfalfa. Facts from California Alfalfa and Forage Association; http://alfalfa.ucdavis.edu/-files/pdf/alfalfafactsheet.pdf http://www.calhay.org/industry.html

Geographical Master Licensee Eurosemillas, S.A. Argentina, Chile, China, France, Korea, Japan, Poland, Morocco, Tunisia, India, Spain, UK, Mexico, Czech Republic, Slovak Republic, Romania, and E.U. for new cultivars Southern Resource Services, PTY, LTD. South Africa New Zealand Berryfruit Propagators New Zealand Toolangi Certified Strawberry Runner Growers Australia

GAMMA GLIADIN-ZEIN PROTEIN FUSION ACCUMULATES TO HIGHEST LEVEL IN TOBACCO ~1.5% of Total Soluble Protein Lane 1: γ-gliadin-zein protein fusion, 9 Lane 2: γ-gliadin-zein protein fusion, 10 Lane 3: γ-δ-zein chimeric protein fusion, 4 Lane 4: Co-expression of γ- and δ-zein, 9 Sun, Chi-Ham, and Bennett. U.S. Patent Application 62/037,730 Equal protein loaded in each lane. Relative mrna expression of three constructs about the same.

GFP mcherry Merge ER-GFP ERmCherry Three chimeric proteins localize in leaf endoplasmic reticulum and leaf protein bodies γ-δ-gfp ERmCherry γ-gliadin- Zein-GFP ER-mCherry β-zein δ-zein-gfp ERmCherry Sun, Chi-Ham, and Bennett. U.S. Patent Application 62/037,730

GAMMA GLIADIN-ZEIN WAS ALSO MORE STABLE THAN SOLUBLE PROTEINS IN IN VITRO RUMEN DIGESTION ASSAY Rubis co Gliadinzein Coomassie staining Anti-Myc blotting Sun, Chi-Ham, and Bennett. U.S. Patent Application 62/037,730