plant science into practice - niab innovation farm · china, india, s. africa ... puccinia graminis...
TRANSCRIPT
Yield Gains
1982 – 2007, on average 74 kg ha-1yr-1
2010 – 2050, 10 t ha-1 => 20 t ha-1
= 250 kg ha-1yr-1
Cereals of TomorrowWill they be GM?
• My talk
–Where are we now?
–“Finally Wheat”, June 1999
–Why the hold-up with wheat?
–Closing remarks
SuccessRoundup-Ready Soybean
Before RR After RR Gains and Losses
Farmer pays $25/acre for weed control
Pays $18/acre +$80m
Seed company sells 70% new crop soya
Sells 100% +$180m
Seed company makes $3 per bag of soya
Makes $5/bag +$70m
Monsanto selling no Roundup on soya
Sells 35 million litres +$300m
Monsanto – no technology fee
Makes $6.50/bag +$220m
Agchem industry – stable market in soya for selective herbicides
Market for selective herbicides devastated
-$850m
Rain Shelter Trial Corn Plot at A Monsanto Research Site
YieldGard Plus with
Roundup Ready
Corn 2
YieldGard Corn Borer with
Roundup Ready Corn 2
+ Force® insecticide
YieldGard® Corn Borer
with Roundup Ready
Corn 2
Roundup
Ready®
Corn 2
HT
Yield = 94 bu/ac
CB Protection
+HT
Yield = 113.7 bu/ac
Soil-applied RW
Protection
+CB Protection
+HT
Yield = 150 bu/ac
RW Protection
+CB Protection
+HT
Yield = 198.1 bu/ac
Gallons of Ethanol
535Pounds of Feed
3,170
Gallons of Ethanol
254Pounds of Feed
1,504
Gallons of Ethanol
307Pounds of Feed
1,819
Gallons of Ethanol
405Pounds of Feed
2,400
Positive Effects of Stress Mitigation Are
Compounded by the Stacking of Biotech Traits
Ag Biotech Driven by Lengthy Product Development
Cycle and Large Investment Process*
Year 0 1 2 4 6 8 93 5 7 10
DiscoveryGene/Trait Identification
Phase IProof of Concept
Phase IIEarly Development
Phase IVRegulatory Submission
• High
throughput
screening
• Model crop
testing
• Gene
optimization
• Crop
transformation
• Trait
development
• Pre-regulatory
data
• Large scale
transformation
• Trait
integration
• Field testing
• Regulatory
data
generation
• Regulatory
submission
• Seed bulk-up
• Pre-marketing
$2-5M(5%)
$5-10M(25%)
$10-15M(50%)
$15-30M(75%)
$20-40M(90%)
On Average:
Time to market: 8-10 years
Total expense: ~$100M
Spendin
g(P
robabilit
y o
f Success
in %
)
* Numbers (time duration, spending, and probability of success) are all estimates. The actual for individual projects could vary.
R&D TIME AND COST
Phase IIIAdvanced Development
Bt/RR2Y(Monsanto)
Low Sat(Monsanto)
Omega-3(Monsanto;
Steandonic Acid)
High
Stearate(Monsanto;
DuPont)
High Beta-
Conglycinin(Monsanto;
DuPont)
Industry Soybean Portfolio*A Steady Pipeline of New Biotech Events Nearly Every Year
Processing:
High Oil Soy(Monsanto)
High Oleic(DuPont)
Liberty Link (Bayer)
□ Agronomic
□ Quality/Food
RR2Y(Monsanto)
201X2009
Modified 7S
Protein FF (DuPont)
High Oleic, Stearate(DuPont)
Low Lin(Syngenta)
Glyphosate &
isoxazole tol. (Bayer)*
Low-
Phytate(DuPont)
Dicamba Tolerant(Monsanto)
Omega-3(EPA/DHA)
DuPont
Feed: High Protein
Soybean(Monsanto; DuPont)
Yield(Monsanto;
Pioneer)
Rust(Monsanto;
Pioneer
Antibody -
containing(against E.
coli 0157:H)
Herbicide tol.: 2,4-D (Dow) and aryloxyphenoxy
propionate herbicides
Disease(Monsanto;
Pioneer)
Soybean Cyst
NematodeMonsanto; Pioneer
GAT/Glyphosate-ALS (Pioneer)
*Estimated commercialization pipeline of soybean biotech events prepared by the American Soybean Association, November 2007.
Where we are now?
• GM crops grown on ~ 150 million hectares – 7x UK land area
– 23 countries, 95% of production in N. & S. America, China, India, S. Africa
• Dominated by – 4 crops
– 3-4 genes, < 10 transgenic events
• High development and regulatory costs
• Trait roadmap for soybean shows the way forward
Two Methods for DNA Transfer
BiolisticsI’d use this for
Functional genomicsChloroplast transformation
AgrobacteriumI’d use this for
Gene CharacterizationCommercial events
PSII images of wheat leaves three days after inoculation with either
Puccinia graminis or Puccinia sorghii
Susceptible interaction
(Hereward + P. graminis)
Race-specific resistance
Savannah + P. graminis
Non-host resistance
Savannah + P. sorghii
200 m
0 0.5 1.0
PSII
“Massive Attack”
600 NB-LRR and LRR-kinase candidates from riceFull gene amplicons (promoter, CDS, 3’UTR)Long range PCRs
Bombarded in sets of 10 + PMI selectable markerAt least 20 events/set, 4 clonal plants per event
Frequency of R-gene insertion
0
2
4
6
8
10
12
0 1 2 3 4 5 6 7 8 9 10
Number of R-genes
0
2
4
6
8
10
12
14
16
0 1 2 3 4 5 6 7 8 9 10
0
2
4
6
8
10
12
14
0 1 2 3 4 5 6 7 8 9 10
Nu
mb
er
of
ev
en
ts
Number of R-genes
Number of R-genes
Nu
mb
er
of
ev
en
tsN
um
ber
of
ev
en
ts
Set A
Set B
Set C
(b)(c)
+ - 18 21 24 54 56 57 59 61 75 92 Batch 5 Event ID 12731 +/- PCR
Result
A10
20
30
40
50
60
70
80
90
100
+ - 18 21 24 54 56 57 59 61 75 92
B
10
20
30
40
50
60
70
80
90
100
10
20
30
40
50
60
70
80
90
100
Set A
Set B
Set C
© Copyright text
Plant Science into practice
Andy Greenland • 1 September 2010
• Efficiency of wheat transformation– 16-to >30% + efficiency– Continuous method improvement
• Infrastructure– Capacity of growth facilities for donor material– Tissue culture facilities– Plant growth facilities for T0 plants
• >3000 plants to seed per year, based on 3 cycles/chamber
– Seed yield• between 90-200 seed / plant / construct • secure seed storage facility
– Pipeline Management• Analysis and Quality control• Bar-code tracking systems and Oracle database
Phytic acid biosynthesis
Glucose-6-P
Ins(3)P1
InsP5
InsP6Phytic acid
protein storage vacuole
mixed phytate salts
cytoplasm
ZmMRP4
myo-inositol3-phosphate synthase
Inositol polyphosphate kinases
myo-inositolmyo-inositol kinase
Inositol monophosphatase
Pilu et al2009
Phytate pathway mutants:Maize Rice
MIPS lpa1-241 (GM)MIK lpa3 yIPK lpa22-PGK lpa1-1ABC Transporter lpa1-241, (GM) lpa2-1
RNAi – phytic acid biosynthesis
Q-PCR:1-2 copies3-4 copies> 4 copies
Actin promoter RNAi: T1 wheat seed analysis
0
500
1000
1500
2000
2500
3000
3.1
3.2
3.3
3.4
3.5
3.6 3.7 3.8
3.9
3.1
0
3.1
1
3.1
2
3.1
3
4.1
4.2
4.3
4.4
4.5
4.6
4.7
5.1
5.2
5.3
5.4
5.5
5.6
5.7
5.8
5.9
5.1
1
5.1
2
Co
n
line
ng
P/m
g fl
ou
r
Pooled T1 seed average
0
500
1000
1500
2000
2500
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
3.1
0
3.1
1
3.1
2
3.1
3
4.1
4.2
4.3
4.4
4.5
4.6
4.7
5.1
5.2
5.3
5.4
5.5
5.6
5.7
5.8
5.9
5.1
1
5.1
2
Co
n
Line
ng
P/m
g f
lou
r
Commercial Transformation
Agrobacterium delivery
1000 events per gene
Single gene insertion
No post border integration
Acceptable selectable marker (herbicide
resistance, phosphomannose isomerase, marker segregation)
Ears are inoculated with Agrobacteriumcarrying gene(s) of interestSite of
Agrobacterium delivery
Looking Forward
• Now– Routine for many species but
imprecise
– Finding the right event is labour intensive
• 2nd generation– Predictable performance
– Very high efficiencies
– Multi-gene, large inserts
• 3rd generation (Intelligent Design)– Genome engineering
– Seamless incorporation within breeding programmes
Tissue culture allows the selection and regeneration of transformed plants
HIGH
LOW
PublicAcceptance
Delivery
SLOW
FAST
Why the hold-up with wheat?
• Uncertainties
– Technical
• Transformation efficiency – solved
• Deployment of “breakthrough genes”– Hybrids may hold the key
– Political
• Unfavourable regulatory climate/lack of commitment to change at senior levels in UK/EU government
– Despite recent Beddington/Kendall comments
• Public acceptance– Most surveys (e.g. FSA) show declining concern for GM
– See http://www.gmo-compass.org/eng/news/stories/
Wheat hybrids
• Commercial proposition–Product differentiation
• Not solely a yield target
–Platform for future delivery of novel traits• Income generation in 6-10 years
Wheat hybrids
o Production poses a number of problemso No suitable sterility mechanism
o Wheat is an inbreeder soo pollen amounts are low
o pollen is heavy and sinks
Breakthrough Genes
• Sustainability e.g.– Novel durable resistance to key pests (wheat
bulb fly, aphids) and diseases (rusts, Septoria, Fusarium head blight• Quicker, more accurate than alien introgression
– Drought, low N/P input (uptake and use efficiency)
• Dimension changing e.g.– Resistance to take-all in 2nd and 3rd wheat crops
Closing comments
• No remaining technology barriers to commercial transformation in wheat
• F1 hybrids can be a key delivery platform for novel GM traits
• Expect first products in feed wheat varieties as part of an integrated package?
GM Wheat Initiatives at NIAB
• Community Resources– 50 genes to be transformed over 5 years
– Outline proposal submitted to BBSRC BBRF
• Public Good GM Wheat programme– Showcasing beneficial trait genes
– Proposal to Technology Strategy Board
• In-house and collaborative research– Efficiency, multigene, large inserts, new genotypes
– Trait genes; e.g.disease resistance grain size, number and quality, N use, flowering time