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

• 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?

Food Constraints

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

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

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)

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

• 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

• 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

• 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

Marjori A. Matzke & Rebecca A. Mosher

Nature Reviews Genetics 15, 394–408 (2014)

Biological processes involving siRNAs and RdDM

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.

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

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.

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

• 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

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

Plant Microbiome

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.

- Improve Nitrogen Fixation/Assimilation ( Cook)Domestication/stress reduces efficiencies using comparative genomics to look more efficient fixersN assimilation modified pathway GDH 12% increase protein

- Modify photosynthesis - C4 ( Dehesh) -

Increased YieldsImproved efficiencies

• Increase Sucrose hydrolysis,

• Starch biosynthesis

• Increase O2 availability

Abiotic Stress:

Drought, Cold, Heat, SalinityAbiotic 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

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

malathion to control vector

Things that may disappear without biotech

.

Crops specific to AfricaCassava

• 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

Improved Nutritional ContentMany 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;

Toxins: glycoalkaloids, cyanogenic glucosides, phytohaemagglutinins

• 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?

© 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.

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