Presented at the workshop "Moving Africa Towards a Knowledge based Bio-economy: How can Sweden assist?" organised by the SIANI Bio-economy Expert Group. More at: http://www.siani.se/news/siani-bioeconomy-expert-group-business

1. What Remains to be Discovered:
Unlocking the Potential of Modern
Biosciences,
Martina Newell-McGloughlin
Director, International Biotechnology
University of California, UC Davis
SEI, Stockholm, Dec 3

2. • Need to produce more on less under unprecedented
conditions Changing Climate, diminishing arable land
• Sustainable production: Less water, less fuel, less
fertilizer, less pesticides, less degradation, less
• Investing in food security is an important defense
strategy in volatile regions.
Greatest Grand
Challenge of our
time!
Population
9 billion by 2050!
70-100% More
Food Required
Why should the army be interested in Ag Biotech?

3. Food Constraints

4. Quality Traits
Improved post harvest characteristics
Shelf life, processing, taste
Improved Nutrition –Improved Functionality
Macro: protein, oils, carbs, fibre
Micro: Vitamins, minerals,
Phytochemicals – Antioxidants
Remove Antinutrients/allergens/ Toxins
Opportunities/Challenges for Biotech Crops
CO2
Agronomic Traits
Biotic Stress - pests/disease/weeds/
Abiotic Stress: Drought, heat,
salinity, submergence, marginal soils
Yield: nutrient efficiency, fossil genes
Value
Renewable Resources
Biomass conversion,
feedstocks, biofuels,
Phytoremediation
Concerns land/ water
use
Plants as Factories
Pharmaceuticals/ Industrial products
(Ventria – Rice Lactoferin Lysozyme
30% Diarrhea, recovery 3/6 days,
Concerns gene flow co-mingling

5. Agriculture is not natural!
8,000 BC
19thC
Ea 20th C
Md 20th C
1930s
1940s
1950s
1970s
1980
1990s
21st C
Cultivation
Selective Cross breeding
Cell culture
Somaclonal variation
Embryo rescue
Mutagenesis and selection
Anther culture
Recombinant DNA
Marker assisted selection
---omics - Bioinformatics
RNAi/ GEENs/
Epigenetics/network
engineering/ Novel Breeding
Synthetic biology
Systems Biology
Holistic Systems

6. Source: ISAAA
• Biotech Crops 2013: 175 million hectares, up 7 million - 3% growth
• 27 countries (19 emerging economies) 18 M farmers 90% (16.5M) resource-poor
• US 70.1Mhas (173Mac), ~90% principal biotech crops
• BT corn. HT Soybean BT Cotton (27% stacked traits world wide)

7. How to circumnavigate the “GMO” issues!
Groups of New Plant Breeding Technologies (NPBTs)
Group 1: Site specific mutagenesis Zinc Finger Nuclease (ZFN)
Meganuclease (MN) technique
Transcription Activator-Like Effector
Nuclease (TALEN) technique
Clustered Regularly Interspaced
Short Palindromic Repeats
(CRISPRs)
Oligonucleotide-Directed
Mutagenesis (ODM)
Group 2: Cisgenesis and Intragenesis Cisgenesis Intragenesis
Group 3: Breeding with transgenic inducer
line
RNA-dependent DNA methylation
(RdDM
*
)
Reverse breeding
Accelerated early flowering
Group 5: Agro-infiltration transient
expression
Agro-infiltration ‘sensu stricto’
Autonomous replicating operon

8. • Smart plants: Systems biology reductive and holistic
approaches to identify, modify, introgress and
subsequently simultaneously introduce /study
/modify/ the expression/interaction of genes and the
realtime response of plants to their environment
The Future: Smart Plants Smart Environments

10. • Potential to make major modifications to introgress desirable traits.
• Next generation sequencing, RNAseq, GWAS ( Michelmore)
• Comparative Genomics transcriptome analysis ( Beckles)
• SNAPshot High Density maps ( Dubcovsky, Van Deynze)
• “RNA family” RNA interference (RNAi) (Dubcovsky, Cook, Dehesh)
• Transcription factors (Tfs),
• Genome Editing: GEENs, MENs, Transcription activator-like
effector nucleases (TALEN) Zn Fingers, CRISPRs ( Segal)
• SNaPshot high-information-content-fingerprinting (HICF) Dvorak
• Chemical genomics ( Drakakaki)
• Novel Maternal/paternal Haploid production –
• Centromere engineering (CENH3) ( Chan)
• Mini-chromosomes Combinatorial multigene transformation
Traitup extra chromosomal transient expression operon
• Tunable Promotors
• Epigenetic modification
• Network engineering
New Tools

11. • Synthetic Nucleases
• Tools
• Zinc Fingers
• Transcription activator-like
effector nucleases (TALEN)
• CRISPR/Cas system
• Applications
• Single gene knockouts
• Subtle modification of gene functions
• Sequence-specific integration of foreign genes for gene
stacking
Genome Engineering
Puchta and Fauser (2014)
The Plant Journal 78:727-741

12. Marjori A. Matzke & Rebecca A. Mosher
Nature Reviews Genetics 15, 394–408 (2014)
Biological processes involving siRNAs and RdDM

13. Yagi et al. (2014)
The Plant Journal 78: 772–782
Custom RNA-binding proteins
• Pentatricopeptide repeat (PPR) protein
family
• Applications
• Dynamically or permanently switch
endogenous gene expression on or
off
• Track or relocate endogenous RNAs
• Track or destroy pathogenic RNAs,
notably from viruses and viroids
• Alter the sequence or coding
capacity of RNAs
• RNA relocation. By fusing a
localization tag (e.g. a nuclear
localization signal) to a PPR protein,
RNA may be transported to or
retained in different cellular locations.

14. The Role of Target of Rapamycin
Signalling Networks in Plant Growth and
Metabolism
TOR signalling in Arabidopsis and mammals. Glc-TOR-mediated transcription
networks in plants.
Xiong Y , and Sheen J Plant Physiol. 2014;164:499-512

15. Synthetic promoters for the
phytosensing of plant pathogenic bacteria.
Synthetic transcription factors for targeted gene
activation or targeted genome modification.
An example of the 'top-down' approach for the
generation of plant artificial chromosomes.
Some Additional
Novel Tools
Liu, W., J.S. Yuan and C.N. Stewart, Jr. 2013. Advanced genetic tools for
plant biotechnology. Nature Reviews Genetics 14: 781-793.

16. Expression of an Entire Bacterial
Operon (IR-PRN-GFP) in Plants
(Tomato)
Mozes-Koch R et al. Plant Physiol. 2012;158:1883-1892
Plant samples were taken 2 weeks
after administration A, GFP
fluorescence. B, masks GFP
fluorescence (chlorophyll
autofluorescence ). C, no excitation.
D, superposition of the two top
panels.
Metabolic engineering achieved by
transforming plastids with operons. IL-60 is
a platform of constructs driven from the
geminivirus Tomato yellow leaf curl virus. IL-
60 enables nontransgenic expression of an
entire bacterial operon in tomato - Delivery
to the plant is simple- rate of expressing
plants is close to 100%, eliminating the
need for selectable markers.
Trait Up Novel “transient” expression Systems

17. • In 1999, a new virulent race of stem rust was identified from wheat fields in
Uganda – known as Ug99 (Race TTKSK) after the year and country of discovery.
• Ug99 capable of infecting ~80% of all wheat varieties.
• Ug99 can kill entire week crop within a few weeks.
• UG99 evolving into more virulent forms.
• Ug99 is capable of traveling long distances via wind.
• Solution – use trait up retrofit to introduce flexible control
resitance
http://www.fao.org/agriculture/crops/rust/
stem/rust-report/stem-ug99racettksk/en/
Example Opportunity: Ug99 (Stem rust): A potential Future Driver of Famine

18. N. Johnson & J. Graham Plant Soil (2013) 363:411–419
Plant Microbiome

19. N2 + 8H+ + 8e- + 16 ATP 2NH3 + H2 + 16 ADP + 16 Pi
N2 Fixation an energy “expensive” reaction
Biological Nitrogen Fixation – bacteria harness the sun
N2 and Plant-associated bacteria in a novel glycan
Biological Nitrogen FixationPLANT MICROBE
+
Hypothesis: indigenous landraces of corn grown in
isolated regions of Mexico co-evolved diazotrophic
microbiomes that contribute to plant performance due to
nitrogen deficiency in the soil.

20. - Improve Nitrogen Fixation/Assimilation ( Cook)
Domestication/stress reduces efficiencies using
comparative genomics to look more efficient fixers
N assimilation modified pathway GDH 12% increase protein
- Modify photosynthesis - C4 ( Dehesh) -
Increased YieldsImproved efficiencies
• Increase Sucrose hydrolysis,
• Starch biosynthesis
• Increase O2 availability

22. Abiotic Stress:
Drought, Cold, Heat, Salinity
Abiotic stress limiting factor to crops
reaching genetic potential
 Drought tolerant maize ( 30% increase
in field trials under H2O stress) Fewer
crop losses -Higher yields better water
utilization
 “Resurrection” gene delay drought-
induced leaf loss and stress
 Submergence sub -1 gene produces 6X
grain - save 3 mil tons rice
 Salination: Transport protein. Grow and
fruit even in irrigation water that is >
50X saltier than normal. > 1/3 seawater.
Blumwald and Zhang)
 Nitrogen Use Efficiency (NUE) plants
equivalent yields require 30% less N
 Phosphite rather than phosphate also
control weeds
Wild type IPT gene
15 days drought, 7 days re-watered

23. Papaya
• No natural resistance so traditional breeding
will not work Removes viral reservoir thus
protects all growers
Orange Juice
• Citrus Greening – bacterial disease
• Spinach genes can save from annihilation
Bananas
• Fungal, viral & bacterial diseases threatening
clonal Cavendish banana – need biotech to
save variety
Potatoes
• Late Blight - Up to 75% can be lost Fortuna Resistant
-two genes Mexican potato - eliminate fungicide -
potential saving $4.3 B
Grapes
• Pierce's disease spread by glassy wing sharp-
shooter – Resistant gene preferable to spraying
Things that may disappear without biotech

24. .
Crops specific to Africa
Cassava
• Mosaic Virus similar protection system as
papaya working in Kenya (VIRCA project)
• Bacterial diseases cause comparable losses -
transgenes such as cecropin lytic peptide
basis for inbuilt resistance.
Banana
• $6-billion banana export just 15% of banana
production worldwide. 85% staple diet
• Many diseases – fungal, bacterial, viral,
nematodes
• Resistance to xanthmonas bacterial wilt
Cassava Brown Streak
Disease resistance

25. Improved Nutritional Content
Many common food crops not perfect for nutritional requirements.
Proteins: Maize, wheat, Sweet potato and cassava
WHO: 800 million people suffer from malnutrition, Protein-energy
malnutrition (PEM), the most lethal form, affects 1 in 4 children:
70% live in Asia, 26% Africa, 4% Latin America, Caribbean
Functional Foods: benefits beyond basic nutritional needs.
Macro:
•Protein (Better ratio, High lys/ meth, Fossil TF partitioning,
artificial)
•Carbohydrates (>complex – resistant starch )
•Fats (Higher Oleic (MUFA), Ω-3, Ω- 6 GLA, CLA, MCFA,
lower SFA, PUFA
•Fibre (low for animals, high for humans (prebiotics, FOS,
inulins, lignans)
Micro: Vitamins (Golden rice II, Golden Cassava, folate, vit C, vit E),
co-factors, minerals (Fe, Ca, Zn)
Phytochemicals: anthocyanins carotenoids, flavonoids, isoflavones,
isothiocyanates, phenolics (Sirtuins)
Anti-nutrients: Trypsin inhibitors, Phytate; caffeine
Allergens/intolerance: soy P34, peanut; gluten;

26. • Out of the world's total land area of 13 billion hectares (ha), 12%
is cultivated but the future expansion of farmland for food
production will be slower than in the past. (FAO)
• In the next 30 years, developing countries will need an additional
120 million ha for crops according to the FAO, this means, less
new land will be opened up than in the past.
• Some countries and communities will face problems related to
land scarcity. This will strengthen their dependency on food and
feed import.
• Genetically modified crop varieties are the most cost-effective
way to sustain farming in marginal areas and to restore degraded
lands to production.
• If we want to feed the world without destroying our resources,
science and technology should drive the development of modern
agriculture.
Why Biotech?

27. © UC BREP
What Sweden can help with
• Work with Regional Centers (IRRI, CORAF, IITA,
CIMMYT) And local communities, international institutes
and specialists to determine needs and best methods
• Provide resources to train world-class scientists and
develop expertise for capacity building in research and
development, Intellectual Property management,
biosafety assessment, outreach communications,
policy development
• Develop laboratory and research facilities and
capabilities pertinent to requirements of the local region
• Establish Scientist exchange and Training Programs
with leading institutes and universities
• Assist with the development of biosafety plans that
proceed concurrently with regulatory system
development
• Work with host country governments in promoting
science-based evaluation of biotechnology to increase
food security, strengthen economies, and foster trade.

28. Greatest Challenges going forward
• Technical
• Intellectual Property: PIPRA - Specialty crops – FTO
• Liability
• Biosafety: so–called – LDCs – Specialty crops
• Acceptance: - countering fear and misinformation
(ethical) - moral imperative real need v. hypothetical risk
I hope that there is nothing
genetically modified in this