How to improve Water and Nutrient Use Efficiency,Farmer Participatory Varietal Selection,Phenotyping of Drought Resistance

1. Improving the water and nutrient use
efficiency of crops in dry savanna and
Sahel regions of Africa
S. Muranaka
International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

2. Limited Natural Resources for Small Scale Farmers
Soil moisture
- Short growing season 1200
- Rainfall 1000
Unevenly distributed
Annual rainfall (mm)
800
Unpredictable
600
Unstable
400
200
0
1960 1968 1976 1984 1992 2000 2008
Year
Soil fertility
- Low soil fertility
- Low input level
- Limited soil moisture
- Competition
Crops
Weeds
Parasitic weed

3. Introduction of Appropriate Variety
- Case study in Radi and Danja villages in Niger -
Grain yield obserbed in 4 farmers' fields of Niger Rep
Grain yield (kg / ha)
Field ID IT98K-205-8 IT99K-573-2-1 Local line
Danja 1 292 (442) 653 (988) 66 (100)
Danja 2 131 (305) 231 (539) 43 (100)
Radi 1 507 (447) 363 (319) 113 (100)
Radi 2 254 (115) 575 (261) 220 (100)
- What is “appropriate” genotype ?
- Where to get “appropriate” traits ?
- How to select “appropriate” genotypes ?

4. How Improve the Water and Nutrient Use Efficiency
Reduce water loss
Resistance to S. gesnerioides
Reduce nutrient loss
High WUE
Identification of “appropriate”
genotype in farmers’ field
Water Nutrient
High NUE
High WUE
Resistance to drought
Capturing nutrient via
root development
High N2 fixation

5. How Improve the Water and Nutrient Use Efficiency
Reduce water loss
Resistance to S. gesnerioides
Reduce nutrient loss
High WUE
Identification of “appropriate”
genotype in farmers’ field
Water Nutrient
High NUE
High WUE
Resistance to drought
Capturing nutrient via
root development
High N2 fixation

6. Farmer Participatory Varietal Selection (FPVS)
- More rapid and cost-effective way to identifying
farmer-preferred cultivars if a suitable choice of
cultivators exists
- Enhances the diffusion and adoption of varieties
developed
- Provide important feed back to breeders on their
performance under local cropping systems

7. Stability of Farmers’ Choice
Stable?
Environmental effect?
Other effect?
Strong / weak points?

8. Trial Sites in Maradi and Zinder Region of Niger Rep.
2008-2009
Sarkin housa Toumnia
Radi
Danja Tessoua
Magaria Radi
Tchadoua
Toumnia Danja
Magaria
2009
Tchadoua
Sarkin housa Minjibir
Tessoua

9. FPVS Activities
No name 3 selections x 3 replications Questionnaire

10. Materials Selected for FPVS

11. Stably Selected Genotypes
IT00K-1148
Early-medium maturing cowpea
Brown, rough, small seed with brown eye
Susceptible to S. gesnerioides
IT99K-573-2-1
Early maturing cowpea
White, rough, medium seed with black eye
Resistant to S. gesnerioides
IT90K-372-1-2
Early-medium maturing cowpea
White, rough, medium seed with brown eye
Susceptible to S. gesnerioides
IT98K-205-8
Extra early maturing cowpea
White, rough, medium seed with black eye
Resistant to S. gesnerioides

12. Specific Preference in Particular Village
Percentage of farmer selected more than 2 and 3
pairs from different replication in the FPVS
- Farmer can stably select their preferred genotype
International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

13. Selection Criteria
Focused criteria for the farmers’ selection in FPVS
- Grain yield is highly important in almost all farmers
- Relatively lower importance of seed color and size
- Higher importance of maturity in 2009?

14. Environmental Effects on Farmers’ Choice
Farmer’s preference on maturity in two years selection
Extra early maturing lines such as IT98K-205-8
gained higher importance in drought year
Farmer’s preference on seed color in two years selection
Stable criteria, such as seed color, texture, size,
plant type and so on.

15. Specific Criteria – Important, but Difficult
Farmer’s selection on the resistance to
S. gesnerioides in two years selection
- Important criteria, resistance to S. gesnerioides
may be difficult for farmers to evaluate in the field
Breeder’s decision using tools up taking
farmers’ preference such as FPVS
- Presented in 5th World Cowpea Conference
- Manuscripts will be ready shortly.

16. Further Challenges – AVEC-BF Project
Appropriate Varieties of Early maturing Cowpea for Burkina Faso
AVEC-BF sites Target country:
Sofe
Burkina Faso
Project duration:
Pathili Samboaga April 2010- March 2013
Donor:
Saria Rango
MAFF, Japan
Puni

17. AVEC-BF Framework
Focus on village level cycle for further expand to region level
SNS
Input market Input market
Grain market
Seed market
Seed producer Grain producer
Training Farmers school
- Cultivation method - Safe chemical use
- Certification system - Fertilizer application
- Variety selection - Storage method
Grouping - Basic economics
- Effective production - Various varieties
- Easier marketing Equipment support
- Splay machine
- Protection materials
Linking with markets
Social analysis Appropriate variety
FPVS
Baseline study - Selection of suitable materials
- basic understanding cowpea roll - Understanding farmers’ preference
Impact assessment Breeding for better
- Target site vs counterfactual site - Adding missing farmers’ preferable traits

18. AVEC-BF 2010
FPVS at 5 villages
- Over 400 farmers attended
Training of seed producer:
15 farmers participating (15ha)
Theoretical training at Saria in April
On-farm training with IT98K-205-8
Farmers’ school:
Trainers’ training (10 trainers)
Tow classes in each village were held
Equipment support (sprayer, protector)

19. Appropriate knowledge
Appropriate via Farmers’ school
variety
Training of
seed producer Input Higher income
(fertilizer, chemical) (150,000CFA/ha)
High yield (1500kg/ha)
High quality
Strengthened
seed production
Escape from
drought Current
Low income
(40,000CFA/ha)
Cowpea production by
small scale farmers Low yield (600kg/ha)
Drought stress
Low quality (mixed, insect damage)
Continuation of low input (no-fertilizer, no-chemical)-Low output agriculture

20. How Improve the Water and Nutrient Use Efficiency
Reduce water loss
Resistance to S. gesnerioides
Reduce nutrient loss
High WUE
Identification of “appropriate”
genotype in farmers’ field
Water Nutrient
High NUE
High WUE
Resistance to drought
Capturing nutrient via
root development
High N2 fixation

21. Marker Assisted Backcross Breeding for Striga resistance
Two SCAR markers available for Striga race SG3
developed by M. Timko’ lab
- MahSE2
- 61R
Striga susceptible, but popular in Niger
- IT00K-1148
- IT90K-372-2-1
- IT89D-574-57
- TN 246-80
F1BC1 populations developed for testing of MAB practice for Striga resitance
S/N Population P1 (female) P2 (male) BC1 (male)
1 IT09ST-1 IT98K-409-4 X IT90K-372-1-2 X IT90K-372-1-2
2 IT09ST-2 IT98K-205-8 X IT89KD-574-57 X IT89KD-574-57
3 IT09ST-3 IT98K-205-8 X IT90K-372-1-2 X IT90K-372-1-2
4 IT09ST-4 IT98K-205-8 X TN256-80 X TN256-80
5 IT09ST-5 IT00K-1148 X IT98K-409-4 X IT98K-409-4
6 IT09ST-6 IT98K-205-8 X IT00K-1148 X IT00K-1148
7 IT09ST-7 IT98K-409-4 X IT00K-1148 X IT00K-1148
8 IT09ST-8 IT89KD-574-57 X IT98K-205-8 X IT98K-205-8

22. Marker Verification
Cowpea genotypes (60) tested on
- Two Striga seeds at Nigeria
- Four Striga seeds at Niger
Current results
- Marker efficiency 76%
- All collected Striga seed are SG3
The result will be updated
- New 100 genotypes tested for SG3
- FTA method for cross check

23. No Polymorphism in Several Crosses
Popular genotypes, IT00K-1148 and IT90K-372-1-2 did not show
polymorphism with resistant parents in both MahSE2 and 61R

24. Comparison of 5 Back-Cross Pops with and without MAS
With MAS Without MAS
F2BC3
F2BC2

25. With MAS
F2BC3 populations
Results of pot evaluation for Striga resistance at F1BC3
Total plant No. of plants
S/N Population no. tested Resistant Susceptible % of Resistance
1 IT09ST-2 66 20 46 30%
2 IT09ST-3 129 20 109 16% 14% increase
F2BC2 populations
Results of pot evaluation for Striga resistance at F1BC2
Total plant No. of plants
S/N Population no. tested Resistant Susceptible % of Resistance
1 IT09ST-4 51 15 36 29%
2 IT09ST-6 83 14 69 17%
3 IT09ST-7 112 15 112 13% 12-16% increase
- Reported on GCP annual report 2009

26. Further Challenges – New markers
LG1
(middle) 9.5 SNP markers available
1_0473 3037_209 3037 9 27.9 1
ACA-CAT-8 1_1382 5720_199 5720 9 27.9 1
7.8 1_0008 6430_319 6430 9 27.9 1
AGG-CTT-2 1_0235 439_560 439 9 28 1
1.7
1.7
OA19 1_1460 13665_260 13665 9 28.4 1
OY1a 1_0752 17133_1134 17133 9 29.4 1
1.1
OE16
5.5
1_0158 1989_448 1989 9 32.3 1
AAC-CAA-5* 1_0137 3958_104 3958 9 32.3 1
1.5
Rsg 1-1 1_0276 1339_101 1339 9 46.3 1
3.7 OI2b
Rsg 2-1 1_0958 7548_1327 7548 9 46.3 1
Rsg 4-3 ACT-CAA-8*
1.1
OE9
1_1069 677_709 677 9 48.4 1
1.2 1_0948 9779_613 9779 9 48.4 1
OZ13b
1.2
ACA-CAG-4*
0.0
2.5
ATC-CTA-10*
12.2 High-throughput genotyping (SNP)
ATG-CAC-1*
6.3
5.3
AGA-CTA-9 - Phenotyping data on SG3
1.3
0.0
D1301b
- SR population for QTLs
AAC-CTT-11
0.0
1.3
AAC-CTT-13
AAC-CTT-9
- 60+100 lines for association analysis
0.5
OC16a
1.7
S05G04i
SNP markers for SG3 resistance
- Identification of probable different race
- 24 genotypes for field identification

27. Moderate Resistance to Striga gesnerioides?
“Moderate Resistance” is frequently seen
in field evaluation
- Unstable mechanisms
- High Striga seed density ?
- Environmental factor ?
High Low Break down of resistance due to high Striga
Striga hermonthica seed density hermonthica seed density had been reported
in Striga resistant Sorghum varieties
How about in cowpea?

28. Seed Density Does not Affect Striga Resistance
600mg Striga seed = 80,000 germinable seeds (at 62% germination rate)
Stable in high seed density

29. No Effect of Drought and Low Fertility Stresses
Susceptible line (IT98D-1399)
Stable mechanism under
- High seed density
- Drought
- Low soil fertility
- Enhance the effect of drought
For field evaluation
- Highly infested condition
- Importance of weed control
- Importance of purity of seed
- Wide border row
- Reported at AA annual meeting, Nov. 2009, Khartoum, Sudan
- Manuscript is under internal review for submission

30. Mechanism?
Hyper-sensitive reaction
- Any signaling via shoot such as auto-
regulation of nodule?
- Stimulation of Striga seed related?
Probably No
Left: B301 (Striga resistant)
Right: IT98K-461-4 (Striga susceptible)

31. Reciprocal Grafting Method
Shoot stock
- Striga resistant
- Striga susceptible
Root stock
- Striga resistant
- Striga susceptible
Striga attachments were observed only on the
root derived from Striga susceptible genotype

32. Double Root Grafting Method
Main plant
- Striga resistant
- Striga susceptible
Main plant
(Striga susceptible)
Grafted root
(Striga resistant)
Grafted root
- Striga resistant
- Striga susceptible
No Involvement of Shoot Mechanisms
- Reported at AA?SATREPS seminar, Sept. 2010, Awaji, Japan
- Manuscript had submitted to Journal of Plant Physiology

33. Striga Resistance - Root Localized Mechanisms

34. Further Challenges – NEDO Project Osaka University
Metabolom on Striga resistance
(Jan 2010 – Dec 2014) Low data
GC-MS
Metabolic pathway
Mini rizotron
Samples at different stages Understanding of mechanisms

35. How Improve the Water and Nutrient Use Efficiency
Reduce water loss
Resistance to S. gesnerioides
Reduce nutrient loss
High WUE
Identification of “appropriate”
genotype in farmers’ field
Water Nutrient
High NUE
High WUE
Resistance to drought
Capturing nutrient via
root development
High N2 fixation

36. Phenotyping of Drought Resistance
Phenotyping
Multi-location field trials
Field experiment at Minjibir
Pot experiment
Root cylinder experiment

37. Selection of Parents for High x High Crosses
Selected genotypes were used for breeding program

38. G x E Analysis on 50 Genotypes
Biomass yield
Wet
Dry
Ibadan condition have much
smaller environmental effect on
differentiate the genotypes

39. G x E Analysis on 16 Genotypes
Grain yield
Magaria, Maradi, Chinzana
Kano, Nampula
Ruace
Soil type?

40. Linkage Disequilibrium Mapping UCR
Plant materials
Nigeria - 339
Burkina Faso - 189
Senegal - 155
USA - 200
Phenotyping for delayed senescence at the early
vegetative stage provides sufficient precision to
map grain yield and biomass QTL
- Presented in 5th World Cowpea Conference
- Manuscript is now internal reviewing for submission

41. Mechanisms to Maintain the Growth under Drought
Grain Yield = 5.890 + (0.0508 x Stomatal conductance)
(P < 0.001)
40
R = 0.521 Maintenance of higher stomatal
30
conductance
Grain Yield (g.plant-1)
20 Deeper rooting?
10
0
0 50 100 150 200 250 300 350
Stomatal conductance (m mol m-2 s-1)

42. Root Distribution Traits
4 weeks of
stress period

43. Root Length in Various Soil Depth
A Deeper rooting is one of the
factors, but there are other factors
B B contributing to drought resistance
Root length (cm)
C
Resistant Susceptible

44. Under Root Size Limited Condition Kyushu Univ
Diurnal change of stomatal conductance
120 IT98K-205-8
Day 1
100 Day 3 Drought resistant IT98K-205-8 can
80
maintain higher stomatal conductance
% control
Day 5 under root size limited condition
Day 1
60
40
20
Day 7
0
8 10 12 14 16
Time
120 IT98K-555-1
100
Day 1 - Drastic decline in late afternoon
80
- No relation in Leaf WP - SC
% control
60
Day 3 - Cavitation in vascular bundle?
40
Day 5 - Sensitivity to ABA signaling?
20
- Water absorption mechanisms?
Day 7
0
8 10 12 14 16
Time

45. Flower Abortion - Reduction of Pod Number Kyushu Univ
IT98K-205-8 IT99K-901-5 IT98K-238-2
(34.0±2.9) (35.5±1.6) (75.2±4.0)
Reduction of peduncle water potential is linked with flower
abortion and further reduction of pod number / grain yield loss
Curently, detailed analysis with more genotypes
and different stress levels is on going
- Will be presented in 7th International AFAS Joint Symposium between Korea and Japan

46. Further Challenges
Accurate phenotyping
- Selection of more suitable parents
- Physiological trait base QTLs
- Understanding environment effect using GxE analysis
More physiological analysis
- Use of grafting method
Delta 13C of non-irrigated plants
- Root characteristics
- Utilization of stable isotopes
In collaboration with partners
UCR JIRCAS Kyushu Univ
Delta 13C of irrigated plants

47. Capacity Building
Training programs
- Introduction of Marker Assisted Selection strategy into
national cowpea breeding programs (ICCAE, 2008, Sept- Oct)
- Development of phenotyping network for drought and Striga
resistance in cowpea (ICCAE, 2009 Oct – Nov)
Trainees/students
- S. Gonne (ICCAE, Trainee, IRAD, Cameroon, 2008)
- T. Abudulaye (ICCAE, Trainee, INRAN, Niger, 2008-2009)
- F. Kusi (ICCAE, Trainee, SARI, Ghana, 2008-2009)
- M. Hayatu (Ph.D student, BUK, Nigeria, 2008-2010)
- A. Eugene (Ph.D student, Wageningen Univ., Benin, 2006-2009)
- W. Abdullahi (Gratuate student, BUK, Nigeria, 2009-2010)
- T. Sakamoto (MSc student, Kyushyu Univ, Japan, 2010)
- S. Nakamura (MSc student, Tokyo Univ Agric, Japan, 2010)
- T. Wakabayashi (MSc student, Osaka Univ, Japan, 2010)
and more.

48. For Farmers’ Happy Face
Thank You !!