Spring Bread Wheat Improvement for Irrigated Environments

1
Spring Bread Wheat Improvement for Irrigated
Environments

• Ravi Singh, Julio Huerta, Sybil Herrera, Pawan
  Singh, Govindan Velu, Sukhwinder Singh and
  Sridhar Bhavani

2
Wheat Breeding at CIMMYT

• Mexico based
• Irrigated spring bread wheat improvement
• Rainfed spring bread wheat improvement
• Durum and triticale improvement
• Germplasm enhancement
• Regional based
• Turkey-CIMMYT-ICARDA winter and facultative wheat
  improvement for CWANA region
• CAAS-CIMMYT winter and facultative wheat
  improvement for China

3
Spring wheat mega-environments

• ME1 Irrigated (36.1 area) Temperate
• 1. High yield potential, lodging tolerance
• 2. Water and nutrient use efficiency
• 3. Resistance to three rusts
• 4. Large white grain with leavened and flat
  bread quality

4
Spring wheat mega-environments

• ME2 High rainfall gt500 mm (8.5 area) Temperate
• 1. High yield potential, lodging tolerance
• 2. Resistance to three rusts, septoria tritici
  and fusarium head blight
• 3. Large red grain with leavened bread quality

5
Spring wheat mega-environments

• ME5 Irrigated or High rainfall (7.1 area)
  Warmer
• 1. High yield potential with early maturity,
  lodging tolerance
• 2. Heat tolerance
• 3. Resistance to rusts and spot blotch for low
  rainfall areas
• Resistance to rusts and fusarium head blight
  for high rainfall areas
• 4. Large white or red grain with leavened and
  flat bread quality or noodle quality depending
  on the country

6
Irrigated Spring Bread Wheat Improvement Program-
Targeted area 45 m ha

• Irrigated Mega-environment 1 China,
  North-western India, Pakistan, Afghanistan,
  Iran, Turkey, Egypt, Mexico and Chile 30 m ha
• Irrigated (Warmer) Mega-environment 5
  North-eastern, Central and Peninsular India,
  Tarai of Nepal, Bangladesh, Southern Pakistan,
  Sudan 10 m ha
• High rainfall Mega-environment 2 West Asia and
  North Africa, Highlands of East Africa 5 m ha

7
Breeding Priorities

• High and stable yield potential
• Durable disease resistance
• Rusts- Stem (Ug99), Stripe and Leaf
• Fusarium Scab and myco-toxins
• Septoria leaf blight, Spot Blotch, Tan Spot
• Karnal bunt
• Water use efficiency/Drought tolerance
• Heat tolerance
• Appropriate end-use quality
• Enhanced Zn and Fe concentration
• Adaptation in conservation Agriculture
• Human Resource Development

8
Ciudad Obregon-Toluca/El Batan Shuttle
Breeding Backbone of CIMMYT Wheat Improvement

• Hot-spot screening- Ecuador (YR), Kenya (SR)
• International testing through yield screening
  nurseries

Cd. Obregón 39 m, High yield (irrigated) Drought
tolerance Leaf rust, stem rust
El Batán 2249 m Leaf rust, Fusarium
Mexico City
Toluca 2640 m Yellow rust Septoria
tritici Fusarium zero tillage with maize stubble
9
Why increase yield potential?

• Necessary to meet the increasing demand (2
  annual) due to population increase
• Increased production must come from the existing
  or reducing land resources
• Increased yield potential is reflected in yield
  increases in farmers field even though the
  management remains the same

10
How to protect gains in yield potential?

• Resistance to important diseases and pests
  (biotic stresses)
• Tolerance to drought, heat, salinity, etc
  (abiotic stresses)
• Resistance and tolerance to stresses in a variety
  has no cost to farmers

11
Yield stability

• Capacity of a genotype (variety) to perform well
  under a range of environments under existing
  biotic and abiotic stresses
• Environment at a location fluctuates annually
• Easiest way to determine yield stability is to
  evaluate yield performance under a range of
  environments (wide adaptation)

12
Type of Crosses

• Simple, three-way and four-way crosses an
  attempt to create new combinations of desirable
  genes (creation of a distinct genotype)
• Backcross adds a genes or few genes from a
  source into an existing genotype
• Single-backcross maintains most characteristics
  of a variety but still allows selection for
  several new genes

13
The Single-backcross Strategy

• Increases the possibility of maintaining and
  reselecting desirable genes of the recurrent
  parent
• Multiple genes or characters can be transferred
  simultaneously
• Additional genes or characters from the donor
  parents can also be selected

14
Grain yields of wheat lines developed through
traditional (Simple and 3-way crosses) and
single-backcross approach
0.8 gt Check
10.7 gt Check
Cd. Obregon 2004-2005
15
Crossing details

• Approximately 600 targeted simple crosses, 500
  single- backcross or three-way crosses per crop
  season
• Approximately 300 F2 populations from simple
  crosses and 400 from single-backcross and
  three-way crosses
• High emphasis to incorporate durable stem rust
  resistance in a range of germplasm carrying high
  yield potential, durable LR and YR resistance and
  quality characteristics

16
Selection Schemes

• Various selection schemes can be applied
• Selection schemes commonly used pedigree,
  unselected-bulk, selected-bulk, modified pedigree
  or bulk
• Our preferred strategy selected-bulk

17
Selection Method Selected Bulk(Harvest and
thresh one spike from each of the selected plants
of a population as bulk)

• Permits selection of unlimited number of plants
  that have good agronomic features and desired
  level of resistance
• Increases possibility to identify transgressive
  segregants due to larger population sizes
• Field operation is easy, fast and economic

18
Genetic gain in yield from Selected Bulk is 3.3
higher than Modified Pedigree(Source Simulation
studies- J. Wang and M. van Ginkel, Crop Science)
19
Selected bulk retained 25 more crosses in the
final selected population (Source Simulation
studies- J. Wang and M. van Ginkel)
20
Selection Strategy in Segregating Populations

• Selected bulk from F1BC1/F1Top until F4/F5
• Population sizes Space sown 400 plants in
  F1BC1/F1Top and F3-F5 1200 plants in F2 (2
  million plants/season with an average selection
  frequency of about 7-10)
• Alternate segregating generations (F2-F5) under
  zero-tillage with maize stubble in Toluca and
  normal tillage in Cd. Obregon
• Shuttling of stem rust resistance breeding F3 and
  F4 populations with Njoro, Kenya grown in
  off-season and then main season as F4 and F5. F5
  and F6 at Obregon.
• Grain selection for size (45 mg and above) and
  plumpness in each generation through sieving
• Selected plants harvested individually (or one
  spike harvested in Toluca) in F5 and F6
  generations and plants/spikes with good grain
  characteristics retained

21
Handling of Advanced Lines

• Advanced lines (F6) from individual spikes in F5
  populations harvested in Toluca planted in Cd.
  Obregon as small plots. Selected plots planted in
  Toluca and El Batan as PC. Selected lines form
  yield trials in Cd. Obregon.
• Advanced lines (F5 or F6) from individual plants
  harvested in Cd. Obregon planted as F6/F7 at El
  Batan and Toluca in small plots and selected
  lines form yield trials in Cd. Obregon.
• Yield trials-1st year (alpha-lattice design, 3
  reps) sown on raised bed system in Cd. Obregon,
  and sets of PC are grown in Cd. Obregon (leaf
  rust)
• Best lines selected based on yield and other
  traits and grain from Cd. Obregon used for
  quality analysis and for further disease and
  agronomic evaluations at Toluca, El Batan and
  Njoro (Kenya) and also multiplied in El Batan as
  Candidates for International Yield and Screening
  Nurseries (ESWYT, IBWSN, HRWYT, HRWSN)
• 2nd year of yield trials in Cd. Obregon for
  selected lines conducted under five environments
  and seed multiplied in Mexicali for International
  Nursery. Simultaneous stem rust, yellow rust and
  leaf rust testing conducted in Kenya, Ecuador and
  cd. Obregon, respectively.
• All data combined and used in selecting lines for
  International Yield Trials and Screening Nurseries

22
Yield testing of advanced lines at Cd. Obregon,
Mexico2009-2010 season

• 1st year yield trials or YT (5000 entries
  including checks) 30 entries/trial, 3 reps,
  alpha-lattice design
• raised bed 5-irrigations
• (small plots or PC planted for seed)
• 2nd year yield trials or EYT (500 entries
  including checks) 30 entries/trial on beds (20
  entries trial on Flat), 3 reps, alpha-lattice
  design
• Raised bed, zero tillage-5 irrigations (gt8 t/ha)
• Flat-5 irrigations (gt8 t/ha)
• Raised bed-2 irrigations (4-5 t/ha)
• Raised bed- drip irrigation (2.5-3 t/ha)
• Raised bed-Late (85 days delay) sown- (gt4 t/ha)
• (small plots or EPC planted for seed)

23
Characterization of EPC entries

• Diseases
• Leaf rust- seedling and field (El Batan and Cd.
  Obregon)
• Yellow rust- seedling and field (Toluca and
  Ecuador)
• Stem rust- seedling and field off- and
  main-seasons (Kenya)
• Septoria tritici- Toluca
• Fusarium- El Batan
• Karnal Bunt- Cd. Obregon
• Tan (yellow) spot- El Batan greenhouse
• Stagnospora nodorum blotch- El Batan greenhouse
• Spot blotch- Aguas Frias
• Various quality traits including grain weight
• Agronomic traits height, heading, maturity,
  lodging

24
Progress in grain-yield potential of new breeding
lines after one 5-year cycle of selection (Cd.
Obregon 2004-05 and 2009-2010)
Breeding is science, art, passion, hard work
number game
12 yield gain
2004-05 4814 entries
2009-10 4956 entries
0.6
8.9
PBW343
25
Shifting towards larger kernelsKernel weight of
1254 entries selected from 2009-2010 1st year
yield trials at Cd. Obregon, Mexico
PBW343
26
Quality profiles of newer CIMMYT wheats Changing
profiles of high and low molecular weight
glutenins in CIMMYT wheats for bread making
quality as well as reduction of 1BL.1RS
translocation
Source R.J. Peña
27
Variation for loaf volume of 486 new wheat lines
grown in Cd. Obregon during 2008-2009
28
Predicted expansion of heat-stressed wheat ME5
mega-environment in India

• Current 2050

29
Future Gains in Yield Potential and Yield
Stability under Climate Change

• Targeted improvement of high yielding, widely
  adapted wheats Identifying superior
  transgressive segregates
• Wide incorporation of white floured 7DL.7Ag alien
  segment carrying Lr19/Sr25 genes quantum jump of
  10-12 in yield potential
• Utilization of genetic resources, e.g. synthetic
  wheats
• Shifting maturity towards earliness and selecting
  under heat-stress at hot-spot sites
• Application of physiological tools in selection
• Variety mixtures must be explored as an
  alternative strategy in heat and other stressed
  environments

30
Segregating populations for selection in Toluca
in 2010
Segregating populations Plot numbers
F1 Top BWIR 1-668
F2SR BWIR 1-609
F2 FUS BWIR 610-637
F2 Harvest Plus BWIR 638-723
F3 BWIR 1-693
F3HPlus BWIR 1-43
F4Stem Rust BWIR 1-817
F5Stem Rust BWIR 1-621
31
Advanced lines for selection in Toluca El Batan
in 2010
Advanced lines Plot numbers Entries (No.)
C45IBWSN 1-1258 1258
PC BWIR (white grain) 1-5868 5868
PC BWIR (red grain) 10001-10715 715
F6 BWIR (white Grain) 1-22905 22905
F6 BWIR (red Grain) 30001-32366 2366
F6 Fus BWIR (white grain) 35001-35443 443
F6 Fus BWIR (red grain) 37001-37729 729
F5-F6 Harvest Plus 1-763 763
HP South Asia 1-251 251
HP Head Seln Lines 1-965 965
HP Head Seln Lines BWIR 1-243 243
PC Wide Cross 1-133 133
32
Future Challenges- The Population Monster
Countries with highest population in 2050 and
change relative to 2009
Rank Country Population (million) Increase (million) Change
1 India 1614 416 35
2 China 1417 71 5
3 United States of America 404 89 28
4 Pakistan 335 154 85
5 Nigeria 289 134 86
6 Indonesia 288 58 25
7 Bangladesh 222 60 37
8 Brazil 219 25 13
9 Ethiopia 174 91 110
10 Dem. Republic of Congo 148 82 124
11 Philippines 146 54 59
12 Egypt 130 47 57
13 Mexico 129 19 17
14 Russian Federation 116 -25 -18
15 Viet Nam 112 24 27
620 million more people just in South Asia by
2050 Population of USA and Brazil in 2009
33
Future challenges- Wheat Yields 2008
Average by 2020 to produce 760 mlln tons
World average 2008
UN/FAO production goal for wheat 4 tons/ha by 2020
34
Rust menace- continued fight with an old enemy
Brown (leaf) rustPuccinia triticina
Yellow (stripe) rustPuccinia striiformis
Black (stem) rustPuccinia graminis
35
Dr. Roy Johnson (1935-2002)
Durable Resistance
Resistance, which has remained effective in a
cultivar during its widespread cultivation for a
long sequence of generations or period of time in
an environment favourable to a disease or pest.
Types of Resistance

• Monogenic Race-specific Major genes
  Hypersensitive (Boom Bust)
• Polygenic Race-nonspecific Minor genes Slow
  rusting/ Partial (Durable)

36
Boom-and-Bust Race-Specific Genes for leaf
rust resistance in Northwestern Mexico
    Year Year  
Variety Resistance gene Released Breakdown Race
Bread Wheat
Yecora 70 Lr1, 13 1970 1973 ?
Tanori 71 Lr13, 17 1971 1975 ?
Jupateco 73 Lr17, 2731 1973 1977 TBD/TM
Genaro 81 Lr13, 26 1981 1984 TCB/TB
Seri 82 Lr23, 26 1982 1985 TCB/TD
Baviacora 92 Lr14b, 2731 1992 1994 MCJ/SP
Durum Wheat
Altar 84 LrAlt 1984 2001 BBG/BN
Jupare 2001 LrAlt, 2731 2001 2007 BBG/BP
37
Durable Resistance to Rust Diseases Why?

• Numerous races of rust pathogens
• Mutating and migrating nature of rust pathogens
• Annual virulence analysis and monitoring required
• Most known race-specific genes ineffective in one
  or more wheat growing regions
• Slow variety turnover in many countries
• Opportunity to break-out of Boom-and-Bust
  cycles and focus breeding for other important
  traits

38
Genes involved in durable, slow rusting
resistance to rust diseases

• Minor genes with small to intermediate effects
• Gene effects are additive
• Resistance does not involve hypersensitivity
• Genes confer slow disease progress through
• 1. Reduced infection frequency
• 2. Increased latent period
• 3. Smaller uredinia
• 4. Reduced spore production

39
Pleiotropic Slow Rusting GenesLr34 /Yr18/Pm38
and Lr46/Yr29/Pm39Lr67/Yr46/Pm?
With Lr46 Without Lr46

• Components of slow rusting are under pleiotropic
  genetic control, i.e., the same resistance
  mechanism controls all components
• Formation of cell wall appositions, instead of
  hypersensitivity

40
Leaf tip necrosis and slow rusting resistance
Leaf tip necrosis associated with Lr46

• Lr34/Yr18/Pm38, Lr46/Yr29/Pm39 and Lr67/Yr46/Pm?
  linked to some level of leaf tip necrosis
  expression
• Slow rusting resistance without leaf tip necrosis
  also known

LalbahadurLr46
Lalbahadur
41
Identification and characterization ofslow
rusting resistance

• High or susceptible infection type in the
  seedling growth stage
• Lower disease severity or rate of disease
  progress in the field compared to susceptible
  check
• Brown rust High (compatible) infection type in
  the field
• Yellow rust Infection type not a reliable
  criteria due to systemic growth habit
• Stem rust Variable size of pustules- bigger near
  nodes

42
Genetic basis of durable resistance to rust
diseases of wheat
Rust
Susceptible
100
80
1 to 2 minor genes
60
40
2 to 3 minor genes
20
4 to 5 minor genes
0
20
0
10
30
50
40
Days data recorded

• Relatively few additive genes, each having small
  to intermediate effects, required for
  satisfactory disease control
• Near-immunity (trace to 5 severity) can be
  achieved even under high disease pressure by
  combining 4-5 additive genes

43
Advances in Molecular Mapping of Slow Rusting
Resistance Genes

• Several Genomic locations (QTLs) known
• Developing and characterizing mapping populations
  that segregate for single resistance genes
• Single gene based populations for 2 or 3
  undesignated genes now available at CIMMYT
• Very difficult to characterize populations
  segregating for minor genes that have relatively
  small effects
• Gene-based markers for relatively larger effect
  slow rusting genes becoming reality
• Gene Lr34/Yr18/Pm38 cloned and gene-based marker
  available
• Significant progress made towards cloning of
  Lr46/Yr29/Pm39

44
Durable pleiotropic resistance gene Lr34/Yr18/Pm38
Perfect marker for Lr34 -veLr34sp
veLr34spA (multiplex)
ABC (ATP Binding Cassette) transporter of PDR
(Pleiotropic Drug Resistance) subfamily
1 2 3 4 5 6

• Cloning of Lr34/Yr18/Pm38
• Single gene based fine mapping populations
• Gamma-ray induced deletion stocks
• Azide-induced mutations
• Precision phenotyping
• Partnership (CIMMYT, CSIRO and Univ. of Zurich)

1. Lalbahadur
2. LalbahadurLr34
3. Thatcher
4. RL6058 (ThatcherLr34)
5. Chinese Spring (Lr34)
6. Lr34 deletion mutant

Krattinger et al. Science 2009
45
Advances in breeding for slow rusting resistance
to brown and yellow rusts at CIMMYT

• 1970s Wheat lines with intermediate levels of
  slow rusting resistance selected.
• 1990s Wheat lines with near-immune level of
  resistance developed through intercrossing
  diverse sources of resistance followed by
  selection of transgressive segregants.
• 2000s Targeted introgression of resistance into
  adapted cultivars and genotypes resulting in
  high-yielding wheats with high levels of
  resistance.

46
Controlled field epidemics remain the best
tool for selecting slow rusting resistance
47
Adult plant leaf rust responses of 144
race-specific gene carrying and 360 seedling
susceptible new elite entries in El Batan, Mexico
2009
Susceptible checks 100 severity
0-15 severity
48
Variation in resistance to yellow rust in 504 new
elite entries tested during 2009
Severity of susceptible checks 100S (N)
49
Ug99 migration and evolution current status
Iran
2007
Pakistan

• 1988 Uganda
• 2002 Kenya
• 2003 Ethiopia
• 2006 Yemen and Sudan
• 2006 Sr24 virulent mutant-Kenya
• 2007 Iran
• 2007 Sr36 virulent mutant-Kenya
• 2007 Sr24 virulent mutant-caused epidemic in
  Kenya
• 2008 2009 Similar races found in South Africa

2006
Yemen
Sudan
2006
2003
1998
2002
50
Why Ug99 is a threat to wheat producing countries?

• Historical importance of stem rust
• Span of susceptible wheat varieties on gt80 area
• Favorable environment (dew/rain and temperatures)
• Mountains and other areas for off-season survival
• Continued evolution
• Early epidemics can cause gt70 losses
• If measures not taken, estimated 10 losses in
  production in South Asian countries alone can be
  worth approx. US1.5 billion and will provoke
  sharp increases in wheat prices

51
Borlaug Global Rust Initiative
A multi-institutional partnership for
systematically reducing vulnerability of global
wheat crop to wheat rusts

• Durable Rust Resistance in Wheat Project-
  Objectives

1. Planning for the Threat of Emerging Wheat Rust
   Variants
2. Advocating and Coordinating Global Cooperation
3. Tracking Wheat Rust Pathogens
4. Supporting Critical Rust Screening Facilities in
   East Africa
5. Breeding to Produce Rust Resistant Varieties
6. Developing and Optimizing Markers for Rust
   Resistance
7. Reducing Linkage Drag
8. Discovering New Sources of Rust Resistance
9. Exploring Rice Immunity to Rust

52
Durable Rust Resistance in Wheat
53
Methodology used for identifying adult plant
resistance to Ug99 in current wheat materials

• Field evaluation of advanced breeding lines in
  Kenya/Ethiopia
• Greenhouse seedling tests for susceptibility to
  Ug99 at USDA-ARS Lab. in St. Paul, Minnesota, US
• Characterization of pseudo-black chaff phenotype
  and application of Sr2 molecular marker
• Identified APR Sources Kingbird, Kiritati,
  Juchi, Pavon, Parula, Picaflor, Danphe, Chonte

Kingbird-the best source of APR
54
Pseudo black-chaff
Durable adult-plant resistance (APR) to stem rust

• Sr2-Complex
• (Sr2 and other minor genes)
• Sr2 transferred to wheat from Yaroslav emmer in
  1920s by McFadden
• Sr2 is linked to pseudo-black chaff
• Sr2 confers only moderate levels of resistance
  (about 30 reduction in disease severity)
• Adequate resistance achieved when Sr2 combined
  with other unknown genes
• Essential to reduce/curtail the evolution of Ug99
  in East Africa and other high risk areas

Sr2 present
Sr2 absent
55
Breeding for durable, adult-plant resistance at
CIMMYTMexico (Cd. Obregon-Toluca/El Batan)-
Kenya International Shuttle Breeding a
five-year breeding cycle)
Cd. Obregón 39 masl High yield (irrigated),
Water-use efficiency, Heat tolerance, Leaf rust,
stem rust (not Ug99)
Njoro, Kenya 2185 masl Stem rust (Ug99
group) Yellow rust
El Batán 2249 masl Leaf rust, Fusarium
Toluca 2640 masl Yellow rust Septoria
tritici Fusarium Zero tillage

• Shuttle breeding between Mexico and Kenya
  initiated in 2006
• gt1000 F3/F4 populations undergo Mexico-Kenya
  shuttle
• High yielding, resistant lines from 1st cycle of
  Mexico-Kenya shuttle under seed multiplication
  for international distribution in 2010

56
Grain yield performance comparison Mexico
Shuttle vs. Mexico-Kenya Shuttle Breeding, Cd.
Obregon 2009-2010
Mexico shuttle n3903
Mexico-Kenya shuttle N1053
8-9 entries
PBW343
No effect of selection in Kenya on grain yield
performance
57
Alternative approaches

• Use effective race-specific resistance genes in
  combinations aided by molecular markers
  (short-term) few useful genes with markers
• Develop cassettes of durable or unutilized
  race-specific resistance genes- GMO solution
  (long-term) need a strong collaborative cloning
  effort

58
Diversity and utilization race-specific
resistance genes effective to Ug99 group of races
Seeding infection types

• About 20 resistance genes have potential (Sr13,
  14, 22, 25, 26, 27, 28, 29, 33, 35, 39, 40, 43,
  44, 45, Tmp, 1A.1R, Sha7 and a few more)
• Virulence known in other races for seven genes
  (Sr13, 14, 27, 25, 28, Tmp, 1A.1R)
• Immediate value Sr22, 26, 35, Huw234, Sha7 and
  Sr13, 14, 25, 1A.1R and Tmp for use in
  combinations
• Translocations being shortened to reduce the
  negative effects and new genes being searched
• Molecular markers essential for selecting gene
  combinations

Susceptible
-------Resistant--------
59
Ug99 Stem Rust Resistance in 728 new CIMMYT wheat
lines developed after one cycle of breeding
(2006-2010)
  Stem rust Entries Entries
Resistance category severity and reaction Number Percent
Adult-plant resistance
Near-immune resistant 1 MS-S 120 16.5
Resistant 5-10 MS-S 178 24.5
Resistant-moderately resistant 15-20 MS-S 199 27.3
Moderately resistant 30 MS-S 63 8.7
Mod. resistant-mod. susceptible 40 MS-S 34 4.7
Moderately susceptible 50-60 MS-S 27 3.7
Mod. susceptible-susceptible 70-80 MS-S 5 0.7
Susceptible 100 S 2 0.3
CACUKE (Susceptible check) 100 S (N)
Race-specific genes
Sr25 1-10 R-MR 17 2.3
Sr26 5-10 R-MR 9 1.2
SrTmp 1-40 MR-MS 49 6.7
SrHuw234 30 MR-MS 1 0.1
SrSha7 1-10 R-MR 19 2.6
Other unknown genes 1-5 R-MR 5 0.7
60
Grain-yield performance of 298 entries with APR
(NIR and R categories) to Ug99 stem rust compared
with all 728 lines retained after one
5-year-breeding-cycle (Cd. Obregon 2009-2010)
39.3
31.2
11.1
6.7
About 90 lines also highly resistant to leaf
rust and yellow rust and resistance of about 60
lines based on APR
61
Acknowledging agencies supporting bread wheat
improvement rust research
Bill and Melinda Gates Foundation through
DRRW Project CSISA Project Harvest Plus
Project Syngenta Foundation
Governments ICAR, India USAID, USA USDA-ARS,
USA SDC, Switzerland ACIAR, Australia
Farmers organizations Agrovegetal,
Spain Cofupro, Mexico GRDC, Australia Patronato-So
nora, Mexico
Thank you