Gene Technology in Wheat

1
Gene Technology in Wheat
2
(No Transcript)
3
(No Transcript)
4
The Livestock Revolution

• Global population to increase to 7.5 billion by
  2020

• By 2020 global population projected to consume
  120 million tons of meat and 220 million tons of
  milk above 1997 consumption level.

Delgado et al. Livestock to 2020The Next Food
Revolution

• Demand for milk and meat expected
• to more than double by 2050

• Fueled by population growth, urbanisation and
  income growth in developing countries

• Additional 292 million tons of
• cereals will be used annually
• as feed by 2020

• Estimated global amount of N voided
• by animals 80 130 million tonnes
• per year, equals global annual
• N fertilizer usage

Massive Global Increase in Demand for Food of
Animal Origin
5
(No Transcript)
6
(No Transcript)
7
(No Transcript)
8
Climate Pressures

• Projected climate change implications for
  Australia suggest

2030
Summer
Annual
Summer
2070
Report to the Australian Greenhouse Office, DEH,
March 2005

• A likelihood of decreasing rainfall over most of
  Australia

Autumn

• GDP scenario losses due to reduction in
  Australian irrigation allocations estimated
  between 136 million (5 reduction) to 751
  million (20 reduction)

Winter

• Drought subtracted c. 1 from Australias GDP
  in 2002/03 equal to ca. 6.6 billion

Spring
9
Greenhouse Gas Emissions - Mitigation
Australias greenhouse gas emissions by sector
(2005)

• Agriculture accounts for 16 of net national
  emissions

• Dominant national source of
• methane and nitrous oxide

• Livestock emissions represent 70 of the
  agriculture sector emissions

• Mainly methane from enteric fermentation in
  livestock

Australian Government Department of the
Environment and Water Resources, 2005 National
Greenhouse Gas Inventory
10
(No Transcript)
11
Global Impact of GM Crops (2005)

• Cumulative US27 billion net economic benefits
  at farm level

• US13 billion for developing countries, US14
  billion for industrial countries

• 90 of beneficiary farmers in developing
  countries (6.8 million in China, 2.3 million
  India)

• Reduced pesticide spraying by 224 million kg
  active ingredient

• 15 reduced environmental footprint associated
• with pesticide use

• Significant reduction of greenhouse gas emission
  
• (equivalent to removal of 4 million cars)

• And more to come..

Brookes Barfoot, 2006
12
(No Transcript)
13
Crop Biotech Revolution
Global Planted Biotech Acres
High rate of adoption in 2006
250
canola

• North America
• Canola 79
• Soybeans 89
• Cotton 83
• Corn 61

cotton
corn
200
soybeans
150
million acres
What about GM wheat?
100
50
0
1996
1997
1998
1999
2000
2001
2002
2003
2004
2005
2006
Source National Center for Food Agriculture
Policy
14
(No Transcript)
15
Gene Technology in Wheat Methodology Development

• 1982 Establishment of embryogenic cultures
• 1990 Regeneration from long-term embryogenic
  callus, cell
• suspension and protoplast cultures
• 1991 Stable integration and expression of
  transgenes in plated cell suspension cultures
• 1992 Production of transgenic plants from
  bombardment of embryogenic cultures
• 1997 Agrobacterium-mediated transformation

16
Opportunities for GM Wheat

• Reduce crop losses caused by abiotic stress
  (drought, soil salinity, heat/cold, flooding)
• Reduce crop losses caused by pathogens
• Reduce crop losses caused by pests
• Reduce crop losses caused by weeds
• Increase crop yield
• Improve nutritional value
• Improve bread-making qualities

17
Is GM Wheat Different from other GMOs?
Wheat is a food grain, whereas corn and
soybeans are mainly used for feed
Soybean, corn canola oil largely exempt from
labelling regulations (Japan elsewhere)
Plenty of GM food now eaten in EU, Japan China
18
Background to GM Wheat in US

• Roundup Ready Wheat
• Proposed in US and Canada
• Agropolitical process
• Released deferred (indefinitely) until (at least)
  some other GM trait in wheat is commercialized
• Sought varying ways to create/conceive a system
  of segregation
• Fusarium Resistant Wheat
• Developed but not submitted for regulatory
  review
• Committed to release is 100 support at
  industry level
• Deferred
• Others drought resistance and varying forms of
  protein manipulation (all public institutions)
• Commercial plan (under development)
• Process of development and role of segregation
  In all cases
• Market segments by GM aversion
• Need for segregation
• Release not encouraged until some form of
  segregation is created

19
Status of GM Wheats

• Cost of deregulation in US and major countries
• Substantial (about 20 million) and about 2-4
  years
• Tech firms reluctant to pursue if uncertainty
  about acceptance
• Not clear how minor GM traits will be proceed
  through these steps without partners

20
Fusarium Head Blight Resistance

• World wide problem
• Development currently in HRS and CWRS in NA
• Field trials in N. America
• Value
• about 15/acre
• Several countries tightening their scab limits
  below US FDA requirements
• Penetration should be 25-50,
• must compete with mod. Resistance in conventional
  varieties
• Virtue
• Producer and consumer trait!
• Major problem for Japan costing about 20-40/mt
• Mitigate major problem in N. America, Argentina,
  Europe and China

21
Consumer Research Key Findings

• Biotech ingredients
• 70-80 of food products in US contain biotech
  ingredients
• Most important to consumers are
• sanitation, hygiene and foodborne illness are
  significant concern
• lt ½ identify food biotechnology as a food safety
  concern
• Labelling
• 76 indicate there is no information they would
  like to see added to food labels
• lt1 (and decreasing) want info on biotechnology
• Support for FDA policy to label food
  biotechnology has decreased
• Outlook on biotechnology
• 62 expect biotechnology to provide benefits over
  next 5 years
• Better health and nutrition
• Improved quality, taste variety

22
Leading Brand Bakery Snacks Already Contain GM
Ingredients

• Wheat Flour
• Partially Hydrogenated Vegetable Shortening
  (soybean, cottonseed, canola)
• Sugar
• Modified Food Starch (corn)
• Corn Syrup
• Soy Flour
• Soy Lecithin

23
Ag Biotech Product Development
(Probability of Success)
24
Summary Implications of GM Wheat

• Producers
• Reduced costs and increased convenience
• Improved disease mgmt
• Drought reduced risk of crop loss
• Grain Handlers
• Need to segregate, forward contract, and
  increased risk
• Branded product end-users
• Promoting products as GM/non-GM vs. no promotion
• Confronting non-branded competitors that may use
  GM
• Confronting risk of consumer resistance or
  boycott
• Importers/buyers with GM restrictions
• Higher cost due to higher cost technology and
  handling requirements
• End-users (domestic and importers)accepting GM
• Lower costs and increased supplies

25
(No Transcript)
26
(No Transcript)
27
Production of GM Wheat

• Biolistic transformation

• Agrobacterium transformation

28
Molecular Analysis of GM Wheat
29
Workflow in GM Wheat Production
30
Genes and Functions

• DROUGHT Arabidopsis thaliana (thale cress), Zea
  mays (maize), moss (Physcomitrella patens) and
  yeast (Saccharomyces cerevisiae). Genes encode
  proteins to enhance drought tolerance by
  regulating gene expression or modulating
  biochemical and/or signal transduction pathways

31
(No Transcript)
32
Pipeline Output
Number of
Number of Vectors
Independent Transgenic
Candidate Genes
Generated
Plants Produced
26
551
Drought Tolerance
28
964
Fungal Disease Resistance
33
850
Frost Tolerance Yield Enhancement
TOTAL
87
2365
33
(No Transcript)
34
GM Wheat for Drought Tolerance
Australias first field trial of GM wheat for
drought tolerance

• Yields of best GM wheat exceeded control under
  drought stress by up to 20

35
Pipeline Output
Number of
Number of Vectors
Independent Transgenic
Candidate Genes
Generated
Plants Produced
26
551
Drought Tolerance
28
964
Fungal Disease Resistance
33
850
Frost Tolerance Yield Enhancement
TOTAL
87
2365
36
Why Fungal Resistant Wheat?

• Stripe rust 102 million

• Crown rot 90 million

• Septoria tritici 86 million

• Septoria nodorum 83 million

• Stem rust 57 million

• Leaf rust 56 million

37
GM Wheat for Fungal Resistance
38
Pipeline Output
Number of
Number of Vectors
Independent Transgenic
Candidate Genes
Generated
Plants Produced
26
551
Drought Tolerance
28
964
Fungal Disease Resistance
33
850
Frost Tolerance Yield Enhancement
TOTAL
87
2365
39
White Clover That Lasts
Cytokinins
Senescence
40
White Clover That Lasts
Field evaluation of seed yield components in LXR
white clover
41
LXR Wheat Development
Field Evaluation
-Increased Biomass
-Increased Yield
-Drought Tolerance
Control
LXR 4
42
High Energy Grasses
Potential impact of climate change
LXRTM dallisgrass

• Pasture heat stress

Possible response

• Breeding, species change

Warm season grasses

• Approximately 40 more efficient in carbon
  accumulation

Control

• Use approximately half the water

LXRTM

• Contain less nitrogen and can be more
• N-use efficient

Report to the Australian Greenhouse Office, DEH,
March 2005
43
(No Transcript)
44
(No Transcript)
45
(No Transcript)
46
(No Transcript)
47
GM Wheat in North America Current Initiatives

• GM crops introduced in 1996
• Soon thereafter, GM wheat developed
• RRW by Monsanto
• FRW by Syngenta
• Monsanto, open and bold
• After much deliberation, chose to not release
• Indicted Will reconsider if/when another trait
  is introduced
• Concurrently Net Returns to agbiotech far
  greater in soybeans, corn and canola
• Syngenta
• Has FRW
• Will only release if/when there is complete
  consumer acceptance
• And, only if CWB concurs with release
• But, trait suffers in that
• It will require concurrent fungicide application
• There have been major advances in wheat breeding
  to reduce incidence of fusarium
• New Bayer chemicals for fungicide are twice as
  effective
• On hold (only public statement) till 2012-13 or
  beyond

48
Disease Vomitoxin

• 2005 in HRS
• Eastern districts samples averaged from 2 - 5 ppm
  DON
• Western districts had less at 0.5 ppm to 1.5 ppm
• Regulations
• FDA regulations impacting vomitoxin limits on
  products (1 ppm on products)
• Other countries Varying regulations
• Impacts on marketing and sustainability of wheat
• Highly risky for growers increased demands for
  mitigating risks (inputs)
• Premiums for low vomit increase
• Shift to competing crops where/as desirable

49
Impacts on Marketing

• Buyers specify limits on vomitoxin content to
  conform to FDA limits
• lt 2 ppm Brazil China ColumbiaDom Rep
  Ecuador, Egypt, Indonesia, Mexico, Pakistan,
  Peru, S. Africa, Taiwan
• lt1.25 EU importers (July 2006)
• lt1.2 Cyprus
• lt1.1 Japan (0 scab)
• lt1.0 China, Russia

50
Impacts of Vomitoxin are Far-Reaching

• EU Policy Increase cost about 50c/b 20/MT
• By shifting origins
• imposing risk on supplier
• Others forced to compete, thereby increasing
  their costs
• Reduced supply of low-vomit wheat available to
  domestic market, and, other import markets
• Producers
• Highly risky and costly for growers to control
• Shift to competing crops where appropriate
• Breeding seeking to develop more resistant
  varieties

51
Farmers Options to Control VomitoxinRisky
Strategies

• Crop rotation
• Reduce corn and/or reduce planting in stubble
• Varieties
• Adopt varieties that are more or moderately
  resistant to Fusarium and/or at different
  maturity dates
• Adopting MR varieties reduces fusarium
• MR varieties available, but their use declined
  after 2003
• Fungicide
• Can reduce Fus levels by 50-70 and, positive
  yield response. Common in Eastern ND about 40
  application
• Cost 9.50/ac plus application Total about
  14/ac
• Multiple applications early season and late
  season show positive yield response
• Still highly uncertain

100
80
Unknown
VS
60
S
MS
Percent Adoption
M
40
MR
R
20
0
2001
2002
2003
2004
2005
52
Biotech not a Consumer Issue in US

• Food companies insist US consumers do not care
  about biotech
• Kraft
• Consumer biotech comments down 40 from 2003
• Coca-Cola
• Of 2.5 million consumer contacts per year, fewer
  than 200 are about biotech

53
Blaine et al Public Perceptions of
Biotechnology Journal of Food Science

• International surveys
• Americans have greater support for biotechnology
  than Canadians and Europeans
• Risks
• concern for biotechnology is relatively low
  compared to other food safety issues
• Trust in regulatory systems
• Americans have a higher degree of trust in
  regulatory authorities than Europeans
• Japanese and European consumers prefer
  international regulatory agencies and have less
  trust in national regulatory agencies than
  Americans or Canadians
• Labeling
• consumers were generally not willing to pay more
  for labels (citing Angus Reid World Poll 2000).
• Surveys are generally poor predictors of actual
  consumer behavior
• consumers often say one thing and do another.

54
Yield Improvement

• Yield can be increased by improving seed
  number and/or
• weight, the latter by increasing the amount
  of starch (more than 70 of seed weight).
• Starch synthesis in cereals is regulated by
  ADP-glucose
• pyrophosphorylase (AGP), that is likely
  involved in determination of seed sink strength.
• Transgenic wheat expressing a modified maize
  Sh2 gene (Sh2r6hs) showed a 38 increase in seed
  weight/plant and a 31 increase in total biomass
  (Smidansky et al. 2002).

55
(No Transcript)
56
Global Wheat Distribution
57
A Critical Commodity ..

• Grown on about 225 m ha in more than 70 countries
  in 5 continents one-half in developing
  countries one-third in very poor countries
• With rice and maize, wheat provides 40 of food
  energy in developing countries
• Modern short-strawed wheat varieties of the Green
  Revolution saved millions from starvation,
  starting in South Asia

58
Adoption of Short-strawed Modern Varieties,1960 -
2000
59
Sources of Increased Cereal Production
Recent slow-down in wheat yield growth
60
Global Trends in Crop Areas to 1997
61
GM Events in Trials, Developing Countrieslt2004gt
Herbicides 21
Pathogens 29
Multiple 8
Abiotic 2
Quality 8
Insects 18
Other 14
62
Transforming Food Value Chains

• Shift from cereal-based diets (maize, wheat, etc)
  to energy-dense diets (meat, ..) increasing
  demand for quality and processed food
• The supermarket tsunami, setting standards for
  quality, reliability, timeliness, with global
  purchasing policies
• Growing concentration of agri-input suppliers,
  and of agricultural product markets expansion of
  contract farming
• Expanding role of regulators

63
Responses to GM Wheat
Growing concentration of the food industry and
retailing makes them key players Food
industry has sometimes adopted a conservative
approach to the commercialization of GM products,
especially in the initial stages of product
development Perceptions of labeling costs?
Risks of consumer boycotts?
64
Segregation and Labeling
Segregation (on the supply side) Widespread
in industry Operating costs depend on
tolerance/risk levels Higher start-up costs,
including establishment costs, when volumes
are low Labeling (on the demand side)
Provides choice to consumers Widespread for
many food products Frequently limited number of
ingredients
65
Global Markets

• Wheat is traded internationally more than
  other
• cereals, approx 17 of production
• Competitive markets, with small high quality
  segment
• Recent tariff escalation in flour, pasta,
  bakery
• products, and trade within trading blocks
• (e.g., EU, NAFTA)
• Global trade reforms benefits are asymmetric
• wheat prices increase approx 10-15, and big
  gainers
• would be Argentina and CIS

66
Trade Issues for GM Wheat

• Experience with other GM maize and canola crops
  shows that market access can suffer with a shift
  from conventional to GM crops (e.g., EU)
• Restricted imports of GM food crops to some
  countries and regions
• - Local food standards
• - Fears of cross-over effects on exports of other
  commodities
• Probable discounting of GM wheat in the highly
  competitive wheat markets

67
Cont

• Substantial volumes of GM maize, soybean and
  canola are traded internationally
• Estimates of market effects for GM wheat need
  to be differentiated between low and high price
  markets, different importing regions, and
  producers/consumers

68
Field Evaluations of Transgenic Wheat

• Field tests and evaluations of transgenic lines
• containing glyphosate, basta and various HMW-
• GS genes have shown clearly that transgenic
• and control lines can be regarded as
  substantially
• equivalent in terms of gene expression between
• generations and environments.
• (Vasil et al. 2001, Barro et al. 2002, Zhou et
  al. 2003, Baker et al. 2006, Baudo et al. 2006,
  Bregziter et al. 2006, Shewry et al. 2006, Blechl
  et al. 2007)

69
Wheat Improvement The Future

• Use transgenic technology only when the desired
  results cannot be obtained in a timely and useful
  fashion by traditional and/or molecular breeding.
• Establish collaborative partnerships between
  breeders, molecular biologists, biotechnologists,
  and industry.
• Use knowledge and insights gained from
  fundamental studies of plant growth and
  development, sequencing of plant genomes,
  discovery of synteny in cereal genomes, molecular
  markers, and high density chromosomal maps.

70
Pathogen Resistance The Future

• Mapping of leaf rust resistance gene Lr10
  (Feuillet et al. 2003)
• Mapping of Fhb1, a major gene controlling
  Fusarium head blight resistance (Cuthbert et al.
  2006)
• Identification and characterization of the stripe
  rust resistance gene Yr34 (Bariana et al. 2006)

71
Pest Resistance The Future

• Characterization of Hfr-3, a novel wheat
• gene encoding a putative chitin-binding
• lectin associated with resistance against
• Hessian fly (Giovanini et al. 2007).

72
Opportunity Cost of Non-Adoption in North America
73
Conclusions

• GM wheat offered potential to turn around the
  N.
• American wheat industry.
• Political Economy killed release in N.
  America, with
• producers losing.
• Release in other countries may be inevitable
  as
• research is progressing elsewhere such as
  China
• Mexico.
• Failed release of RR wheat will affect release
  of
• other GM wheats such as fusarium-resistant
  varieties.

74
Benefit-Cost Ratio Sensitivity to Segregation
Cost
7.00
0.35
Segregation Cost, /bu.
75
Sensitivity to GM Yield Increase
6
5
Benefit-Cost Ratio
4
3
2
Threshold
1
0
3
5
7
9
11
13
GM Yield Increase
76
Market Acceptance Approach Used in Analytical
Model
Market segments requiring different levels of
segregation. Segments defined in each country
as - No GM Restrictions - lt5 GM - lt.9 GM - GM
free (interpreted as nil GM) - Traceability EU
countries would be subject to yet to be
determined traceability requirements
77
Derivation Market Shares of Segments

• Regulatory approval
• - gained in each major consuming
  countries/regions
• - Sought in other importing countries that have
  formal biosafety review process
• - substantial equivalence has been established
  (Obert et al)
• Certificates of free trade Used by countries
  not having a formal biosafety review/import
  approval process
• Minor segments in each market
• - "GM Free" or nil, --2-5 of market for GM Free
• - lt.9 -facilitates trade for global product
  sku's. 8-10 of the market
• Known/anticipated regulatory thresholds
• e.g., EU .9 (with traceability), Chile 2, Korea
  3, etc.

78
U.S. Domestic Use and Exports by Segregation
9.0
8.0
5GM 5 9 GM Free
7.0
HRD Usage (Dom Exports) (MMT)
6.0
5.0
4.0
3.0
2.0
1.0
0.0
79
Canadian Exports by Segregation
80
Distribution of North American Market Segments
5GM 5 9 GM Free
81
Consumer Acceptance Summary

• Many countries regulatory processes are based on
  non- scientific processes
• Some will naturally adopt that of US, and,
  require certification (Philippines,
  Mexico)--Certificate of Free Trade
• Segments
• In nearly all mature markets, one should expect
  market segments to emerge with respect to GM
  acceptance
• Due to demand, incomes, market maturity,
  regulations, etc.
• Natural process of market maturity
• Domestic markets
• Branded product marketers vs private label
• Segregation Buyers are finding ways to make
  purchases of non-GM even though GM may be the
  predominant crop
• Numerous examples in US on corn and soybeans
• Brazil routinely serves both market segments

82
Segregation in Practice

• Segregation arises due to heterogeneity in
• Consumer/buyer demand
• Crop characteristics
• Random, or genetic
• Segregation is a process of de-commoditizatoin
• In fact, should be desired by market participants
• Segregation
• Emerges in response to buyer demands
• Segregation is used very extensively in practice
• Topics to discuss
• Modes
• Costs
• Costs and risks in GM crops

83
Major Point
Segregation Used very commonly in the US and
is well understood Buyers will choose their
procurment regime w.r.t. segregation Canada
already markets by extensive segregations -
Wharburtons etc - M. Barley - CWRS segregation
growth..substantial (60 segregations)
Australia Has same - Escalation from 2 classes
of wheat - Variety declarations - Prescribed
varieties - Golden Reward Program - Others
Mechanisms exist or near existence to faciliatate
segregation
84
Major PointHRS/CWRS
Losing agronomic competitiveness HRS/CWRS
acres is being shifted to other crops, notably
GM crops Major concern of end-users!
85
Distribution of North American Market Segments
5GM 5 9 GM Free
86
Results from Segregation Studies
87
Prospective Adoption Rates for GM Wheats in ND
Allowing RRW, GM FRW, Stacked, and
Conventional varieties Equilibrium
adoption in US HRS areas - GM FRW 34 - RRW
20 - Stacked 31 - Conventional
15
88
Value of a GM FRW Real Options

• Preliminary results
• NPV of developing FRW varies by development
  phase.
• HRS and CWRS (excl SRW)
• Tech fee 12/ac
• Adoption rates estimated
• Discounted NPV 8 years
• Observations
• Low value during discovery increases in
  subsequent phases
• Option value if gt0 ITM

Max
Min
Mean
Phase
Max
Min
Mean
Phase
Value
Value
million
million
4.3
-
3
.2
Discovery
-
Discovery
.2
4.3
3
29.6
1.6
11
Proof of
Proof of
Concept
11
29.6
1.6
Concept
79.6
35
Early
Early
Development
35
79.6
7.1
Development
141.8
27.9
75
Adv
Adv
Development
75
141.8
27.9
Development
216
71.5
130
Regulatory
Regulatory
Submission
130
216
71.5
Submission
89
Number of field test permits filed by private
agbiotech firms, 1987-2004
Seminis
Scotts
Dow
DuPont
Monsanto
Bayer
Syngenta
0
1000
2000
3000
4000
5000
6000
Number of Field Test Permits
90
Number of field test permits filed by public
institutions, 1987-2004
91
Applications for Field Trials in North Dakota, by
Crop, 1990-2004
50
Wheat
40
Sunflower
Soybean
Safflower
30
Rapeseed
Potato
Cotton
20
Corn
Beet
Barley
10
Alfalfa
0
1990
1992
1994
1996
1998
2000
2002
2004
1991
1993
1995
1997
1999
2001
2003
92
(No Transcript)
93
Commercial Trait Development

• Time for Development 8-10 years
• Cost 80-100 million (incl. 20-40 million in
  costs
• to conform to regulatory system US
  and
• ROW)
• Risks
• - Technical feasilility--proof of concept
• - Regulatory Approval--US and ROW
• - Commercial acceptance--price discounts
• - US and ROW
• - Consumers vs. buyers
• - Competitor traits and technologies
• - Patent protection--for a period
• Implication Tend to focus on crops with large
  acreage and high probability of acceptance and
  adoption

94
GM Wheat Challenge (rank orderUS)

• Continued development of GM in competing crops,
  reduces profitability of wheat
• Its a GM and ethanol world
• All major initiatives focused on ethanol,
  biofuels, GM traits for corn, soybeans and canola
• Higher returns than wheat and small grains
• Small area base relative to corn and oilseeds
  reducing incentives to develop GM
• Conforming to labeling and/or traceability
  requirements in selected import countries
• Consumer acceptance Evolving to be more
  favorable, but fragmented around the world
• Segregation/IP/Traceability Systems are
  evolving and handlers are penetrating these
  segmented markets fairly efficiently

95
Managing Transgenic Wheat Plant Development in MPB

• From Transgenes to
• Field Assessment

96
Wheat Transgenesis

• Drought Tolerance
• Fungal Disease Resistance
• A Toolbox of Promoters
• Freedom to Operate

97
Outcome Scenarios

• Enhanced Drought Tolerance
• A total of 20 million ha affected area
• Up to 60 yield impact
• 150 loss per ha
• Potential 124 - 371 million annual impact in
  Australia for wheat

• Enhanced Fungal Disease Resistance
• Estimated national trait values of diseases in
  wheat
• Stripe rust 102 million
• Crown rot 90 million
• Septoria tritici 86 million
• Septoria nodorum 83 million
• Stem rust 57 million
• Leaf rust 56 million

98
Transgenes to the Field
source material
Vector Construction
vectors
Glasshouse
info and primers
putative transgenics
Transformation and Tissue Culture
Molecular Analysis
transgenics, range of copy numbers and expression
levels
Functional Screening Glasshouse and Field
99
Vector Construction
100
Vector Construction

• Base vectors
• Run of the mill transformation vectors
  (pipeline)
• Specialty vectors, e.g.
• binary vectors
• promoter characterisation vectors
• modular vectors

101
Base Vectors

• Contain all necessary elements
• selectable marker cassette (e.g. bar)
• promoter (e.g. constitutive or inducible/tissue-sp
  ecific)
• insertion point for goi - Gateway and Gateway
  MultiSite
• terminator (e.g. 35S, nos)

102
Base Vectors
inducible
103
An Integrated Laboratory Workflow System-
Management of Wheat Transgenesis
104
Transformation Vectors
Receipt of sample
105
Transformation Vectors
Characterisation of sample
106
Transformation Vectors

• Characterisation of sample

107
Transformation Vectors

• Characterisation of sample

108
Generation of entry vector
Transformation Vectors
PCR product amplified with primers containing
attB1 and attB2 sequence

• pDONR vector
• supplied by Invitrogen
• contains in cassette
• CmR gene
• ccdB gene for negative selection (interferes with
  DNA gyrase)
• must be maintained in special E.coli strain, e.g.
  DB3.1TM

Entry vector Depository clone for all further
cloning
109
Transformation Vectors
Final vector
110
Generation of final vector
Transformation Vectors
Destination vector or Base vector can be any
vector due to availability of conversion cassette
By-product Counterselected by presence of ccdB
gene
Entry clone Obtained in Step 1
Final vector
111
Transformation Vectors

• Characterisation of final vector

112
Transformation Vectors
Final vector
113
Transformation Vectors
Maxi or Mega prep
114
Transformation Vectors

• Wide range of base vectors
• Binary vectors
• Modular vectors
• More than 80 transformation vectors
• Additional vectors for other target sequences

115
Wheat Transgenesis

• From Transgenes to the Field

116
Acknowledgments
Trent King
Allan Wenck
Katrina Meath
Daniel Isenegger
Ehab Mohamden
Heather Anderson
Ryan Prendergast
Shannon Burns
Stefanie Trabucco
Grant Hollaway
Mark Mclean
Sally Petch
Dalia Vishnudasan
Karen Fulgueras
Lalani Wijesinghe
Suzanne Georges
Nimal Wijesinghe
Helen Huxley
Ulrik John
Kerya Hang
German Spangenberg
117
(No Transcript)
118
(No Transcript)
119
Direct Economic Impacts of Scab
Barley
200
Durum
HRS
150
100
Millions
50
0
-50
1993
1995
1997
1999
2001
2003
2005