Africa Biofortified Sorghum (ABS)

Project Update—Sept 2012 Dr. Florence Wambugu, Africa Harvest—ABS Project Co-PI

Dr. Marc C. Albertsen, DuPont Pioneer—ABS Project PI

Dr. Silas Obukosia, Africa Harvest ABS Project Manager

Dr. Zho-yu Zhao, DuPont Pioneer ABS Project Lead Scientist

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Biofortified Sorghum Project Sorghum

– A staple food for 300 million people in

Africa

• Fifth most important grain for food

use

– Center of origin in Africa

– Primary cereal in arid and semi-arid

geographies

• Importance will grow as climate

change unfolds

Sorghum Grain and Food Quality

– Very low pro-vitamin A content

– Low iron and zinc bio-availability because of high phytate

– Poor protein digestibility – even worse after cooking

SIGNIFICANT IMPACT ON HUMAN HEALTH, PARTICULARLY IN INFANTS AND YOUNG CHILDREN

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Original ABS Consortium Concept and Orientation

• ABS Consortium was established with 13 organizations, most of which were African-based

• The Consortium was coordinated by Africa Harvest, an African NGO

• Management was by adherence to Milestones and specific Activities

• In addition to technology development, a critical component was development of biosafety

protocols and regulations that were African-centric

• Enabled eventual achievement of world’s first confined field trial of transgenic sorghum in

Africa

Technology transfer

and capacity building

were additional critical

Project aspects

Initial phase funded by

The Bill and Melinda

Gates Foundation

(2005-2010) as part of

their Global Challenge

for nutritional

improvement of critical

staple crops

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Overall Key Biofortified Sorghum Project Objectives

The targeted traits • Beta carotene increased AND stabilized • Iron and zinc bioavailability increased • Protein digestibility improved after cooking

Technology access and application to sorghum • Improved sorghum transformation technology • Nutritional improvement • Trait efficacy • Product development feasibility

• Technology transfer from USA to Africa

• Capacity building in Africa • Research, seed systems, regulatory,

communication, institutional

• Biosafety policy/regulations developed/executed • Conduct Confined Field Trials (CFTs) in Africa

• Provide pathway for biotechnology traits

• Biotech acceptance advanced in key countries – farmers, consumers

• Improved product development and delivery systems for Africa

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ABS Project Progress: Confined Field Trials

Successfully Conducted in Africa

Confined Field Trial of Transgenic

Sorghum In Kenya Managed by

KARI and AH

Confined Field Trial of Transgenic

Sorghum In Nigeria Managed by

IAR and AH

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ABS Project Progress: Gene Flow Studies

• Collaboration underway amongst Kenya Ag Research Institute (KARI) and USDA (Univ. of Neb) researchers • Evaluating weediness and fitness of

accessions

• No surprises thus far, none expected

• First round of work will be complete 4Q12

• Next round will be conducted with final transgenic event

• Publication anticipated. Will be led by KARI and Africa Harvest.

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Overview of ABS Project Phase I and II

Phase – I Phase - II

Time Frame July 1, 2005 - June 30, 2010 April 1, 2011 – March 30, 2015

Funding Bill and Melinda Gates Foundation Howard Buffet Research Foundation

Fund Recipient Coordinator

Africa Harvest Biotech International Foundation Danforth Center

Major Collaborators ABS Consortium (13 members), Biosafety and Regulatory Network (BRN) and Harvest Plus (HP)

•Phase I no-cost extension with Gates Foundation (BRN & HP) •Phase II with the Danforth Center

•PI/Co PI •Lead Scientist

•P. Anderson (M. Albertsen)/Florence Wambugu •Zuo-Yu Zhao

•M. Albertsen/Florence Wambugu •Zuo-Yu Zhao

Trait Targets •Protein quality and digestibility •Fe & Zn bioavailability •Pro-vitamin A bioavailability

•Increased levels of pro-vitamin A •Enhanced Fe & Zn bioavailability •Protein digestibility after cooking

Other Collaborations

Bioassays-UCD, Purdue, Connell, USDA via HP; Pollen Flow-USDA; Regulatory Core Package- EnviroLogix, Inc. via BRN

•African activities coordinated through Africa Harvest •Collaborations continued through the Phase I no-cost extension (ended 30 June 2012)

Target Countries •Tier-1: BF, Nigeria & Kenya •Tier-2: South Africa & Egypt

•Continuation of established emphasis •CFT’s conducted through Africa Harvest in Nigeria & Kenya; planning for CGT in Burkina Faso

Training Duty Trained 13 African scientists and breeders NA

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Phase II Biofortified Sorghum Project Countries

• Strategically positioned anchor

countries for development and

deployment

– Primary targets: Kenya, Nigeria

• Burkina Faso?

• Project activities supported by

in-country enthusiasm from Ministers

of Agriculture and NARS

– Biosafety acts enacted or being

enacted

• Countries will become focal points

from which seed will be distributed

and disseminated

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Phase II Deliverables—Stage-1 and Stage-2:

• Phase II, Stage-1: Deliver a transgenic sorghum donor line that can provide

30-50% of the RDA (300 – 900 µg RAE*/day) for vitamin A in children during

4-6 months of ambient air storage

– Increase accumulation of β-carotene and Carotenoids

– Increase stability of β-carotene and Carotenoids through co-expression of vitamin E

• Phase II, Stage-2: Identify event(s) for Fe/Zn bioavailability increases of 50%

and 35% that can be stacked with genes for beta carotene

improvement/stability and improved protein digestibility

– Validate experimental results showing enhanced Fe/Zn bioavailability in phytate-

reduced transgenic seeds

– Develop construct(s) to achieve phytate reduction without negative impact on seed

germination rate or seed weight

– Identify genes and constructs to meet beta carotene and micronutrient

bioavailability goals and stack with previously developed construct for improved

protein digestibility improvement

*RAE = Rentinol Activity Equivalents

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Stage-1 Progress: Identified a leading transgenic line with

potential for first generation product development

• Over 250 events have been produced and are being analyzed for

enhanced carotenoid levels, beta-carotene stability, and field

performance

– Have achieved 50% improvement in beta-carotene half-life stability from

3 weeks to 7.5 weeks

• Additional constructs with combinations of new genes to further

improve beta-carotene levels and stability are either in

transformation or in early generation analysis

WT ABS203 WT ABS203 Color differences demonstrate enhancement of beta-carotene in transgenics

Vs wild type control

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ABS Project Phase II Progress:

Communications

• Communications Strategy to Date

– Align overall communications strategy with Africa Harvest

– Ensure communications does not get ahead of R&D progress

– Managed information surrounding CFTs

– Scenario planning and issues management

– Internal stakeholders up-to-date

– Establish regular communications between Pioneer and Africa Harvest

• Key Messages and Positioning

– African-led initiative and collaboration

– High research standards and adherence to country regulatory mandates

– Significant milestones

– There is a significant need for ABS initiative to be successful

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ABS Project: Next Steps

• Determine final β-carotene gene composition and optimize

final construct

• Establish pro-vitamin A conversion rate of transgenic

sorghum in animal model systems

• Determine final Fe & Zn bioavailability gene composition

and optimize final construct

• Establish Fe & Zn bioavailability of transgenic sorghum in

animal model systems

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ABS Project: Next Steps

• Continue stewardship and compliance training

• Continue CFTs in African target countries (Kenya, Nigeria)

– Initiate greenhouse study in Burkina Faso

• Build capacity for Phase II trait integration into local

sorghum varieties

• Continue to define Communication needs for ABS product

development in Africa

• Continue dialogue, education, and awareness with

regulatory authorities

– Demonstrate local interest and investment

– Continually refine and update core regulatory package needs

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Phase II Summary

1. Meaningful targets with ex-ante data have been established as guidelines for

trait improvement

2. The quantity and stability of Pro-vitamin A (beta carotene) has been improved

• The levels of carotenoids and β-carotene are increased ~20X compared with wild type.

• Preliminary bioavailability data from Caco-2 cell analysis are encouraging.

• Beta carotene stability upon storage has been doubled.

• Collaboration underway with ISU scientist to estimate conversion rate of beta carotene to

retinol.

3. Essential micronutrient bioavailability has been increased

• Phytate-reduced sorghum showed increased bioavailability of iron and zinc.

• Alternative approaches for phytate reduction & mineral bioavailability are underway.

4. Significant progress in regulatory framework development

• First transgenic sorghum field trials grown in targeted countries in Africa (Kenya and Nigeria)

• Gene flow studies being conducted in Kenya and Nebraska have not uncovered any issues

5. Strong African partner relationship established

• Sustained funding for African partners is critical to maintain continued regulatory involvement

and development.

• Successful CFT execution and GMO acceptance in Africa is crucial for project success.

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ICRISAT/ABS: Potential for Alignment

• African varieties for eventual introgression of ABS improved material

• Share information on Fe and Zn efforts • ICRISAT native traits program

• ABS transgenic trait program

• Potential for collaboration

• Plans for sorghum hybrid development • Desire to ramp up efforts?

• What might this look like?

• Seed production capacity in Africa • How can we work together to make it happen

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ABS Project: Urgent Requirement for

Continued, New Funding

• Require a new source of funding to continue CFTs in African target

countries to maintain momentum in developing robust biosafety

procedures and processes

• Continued funding will enable capacity building for trait integration into

local sorghum varieties

• New funding will help support communication needs for initial product

development and release in Africa

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2012 2013 2014 2015 2016 2017 2018

• Lead Event Development

• US Field Trials

• African Field Trials &

Capacity Building

• Evaluate Protein

Digestibility and Pro-

vitamin A Bioavailability

• Nutrition Profile Analysis

Howard Buffet Research Foundation

Pioneer

Foundation

Funding

• Trait Integration &

Efficacy in Africa

• Efficacy Testing

• Produce

Homozygous Seeds

Obtain

Funding for

Phase 3

• Permits

• Seed to Target Countries

• FTO management

• CFTs in US

Phase 3 Funding

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ACKNOWLEGDEMENTS • Gates Foundation for Phase-I funding and no cost extension • Howard Buffet Foundation for Phase-II research funding • Africa Harvest for project management and CFT development in Africa

•Florence Wambugu •Silas Obukosia •Daniel Kamanga

• Danforth Plant Science Center for coordination of Phase-II research • Harvest Plus for coordination of Bioavailability analysis • UCD, Purdue and Cornell/USDA for collaboration on Bioavailability analysis • BRN for coordination of Core Package • Original ABS Consortium members • Pioneer ABS Team

• Marc Albertsen (PI) • Zuo-Yu Zhao (Science Lead) • Jim Gaffney (Regulatory Lead) • Lonnetta Ragland (Communication Lead) • Ping Che (Scientist) • Kimberly Glassman (Scientist)