Tissue culture production chain,Genetic basis of induced resistance: gene mining,Conventional biopesticides,Endophytes

1. Banana tissue culture:
from endophytes to market pathways
Thomas Dubois
12 October 2010
Ibadan, Nigeria

2. Benefits of tissue culture
pest- and disease free
faster and bigger yields
better marketing
faster and better distribution
sucker tissue culture
subsistence income generation

3. Tissue culture production chain
from the field multiplication rooting weaning in the field

4. Tissue culture production chain
commercial tissue culture enterprise
from the field multiplication rooting weaning in the field

5. East Africa versus Latin America
quality
size
management
markets

6. Endophytes

7. Endophyte in the tissue culture production chain
from the field multiplication rooting weaning in the field

8. Endophyte in the tissue culture production chain
from the field multiplication rooting weaning in the field
from laboratory to screenhouse

9. Endophytes pre-1997
BMZ phase I BMZ phase II
2001 - 2004 2005 - 2007
characterization of pathogenic Fusarium
isolation of endophytes molecular identification
screenhouse screening against banana weevils and nematodes
distribution, competition, in planta spread and persistence
modes of action systemic induced resistance
development of inoculation techniques
on-station performance
on-farm performance
effect on other bananas and nematodes
bio-pesticides as artifical endophytes

10. Induced resistance

11. Genetic basis of induced resistance: known genes
catalase PR-1
2.00 a
1.80
1.60
expression (ng)
1.40 a
1.20
1.00 b b
b
0.80 b b
0.60 b
0.40
0.20
0.00
Physiological and Molecular Plant Pathology (2007)
Paparu (PhD, 2007) Nematology (2010)

12. Genetic basis of induced resistance: gene mining
ABC transporter glucan synthase
18.00 a 0.80
a
16.00 0.70
14.00 0.60 a
expression (ng)
12.00
0.50
10.00
b 0.40
8.00
0.30
6.00
0.20
4.00 b b
2.00 0.10
b b
0.00 0.00
Paparu (PhD, 2007) European Journal of Plant Pathology (2011)

13. Endophytes pre-1997
BMZ phase I BMZ phase II
2001 - 2004 2005 - 2007
characterization of pathogenic Fusarium
isolation of endophytes molecular identification
screenhouse screening against banana weevils and nematodes
distribution, competition, in planta spread and persistence
modes of action systemic induced resistance
development of inoculation techniques
on-station performance
on-farm performance
effect on other bananas and nematodes
biopesticides as artifical endophytes

14. Conventional biopesticides

15. The problem with conventional biopesticides
conventional biopesticide formulation problems
costs and quantities involved for
mass production are
problematic
logistically and economically not maize bran soil
feasible for farmers
application problems
abiotic factors greatly reduce
field performance
topical
pounded rhizomes
pheromone traps

16. Conventional biopesticides as artifical endophytes?
conventional biopesticide artificial endophyte
costs and quantities involved for very low quantities needed and
mass production are the technique is easily
problematic integrated in a commercial lab
logistically and economically not farmers do not need to apply the
feasible for farmers product
abiotic factors greatly reduce the product is protected inside
field performance the plant
topical systemic

17. Beauveria bassiana as artifical endophyte
larval mortality: 23 - 89%
adult mycosis: 58 - 64%
control Beauveria bassiana
Journal of Invertebrate Pathology (2007)
MSc thesis (Akello, 2007) Crop Protection (2008)
MSc thesis (Castillo, 2007) Entomologia Experimentalis et Applicata (2008)
MSc thesis (Mwaura, 2007) Biological Control (2009)
MSc thesis (Thys, 2007) Acta Horticulturae (2009)
MSc thesis (Schnittker, 2008) Journal of Agricultural Science and Technology (2010)

18. Endophytes in other crops
100
control F. oxysporum T. asperellum
90
80
70
colonization (%)
60
50
40
30
20
10
0
MSc thesis (Mwaura, 2007)
MSc thesis (Machungo, 2007)
BSc thesis (Nedela, 2010)
MSc thesis (Waithira, 2010)

19. Endophyte inoculation: the past

20. Endophyte inoculation: semi-commercial
type of seed tray substrate spore “double-dip”
concentration
timing of and volume
drenching
duration of
drenching
dual endophyte
inoculation
MSc thesis (Hillnhütter, 2007)
MSc thesis (Hakizimana, 2011) Microbial Ecology (2007)
MSc thesis (Kato, 2011) Biocontrol Science and Technology (2010)

21. Endophyte mass production: the past

22. Endophyte mass production: semi-commercial
type of liquid quantity of water content of growth duration
medium starter culture solid medium
type of solid type of solid
medium container
pre-treatment of
solid medium
MSc thesis (Hakizimana, 2011)

23. Endophytes in the field
on station on farm
plants trials plants trials
Burundi 800 1 0 0
Kenya 500 1 12,000 3
Uganda 2,980 7 1,600 3

24. Endophytes in the field: effect on nematode populations
R. similis / 100 g (Uganda) P. goodeyi / 25 g (Kenya)
70 5,000 control
F. oxysporum Eny7.11o
60 F. oxysporum V5w2
4,000
50
40 3,000
30
2,000
20
1,000
10
0 0
0 2 4 6 8 10 12 14
months
MSc thesis (Dusabe, 2007)
MSc thesis (Machungo, 2007)
MSc thesis (Waithira, 2010)
MSc thesis (Hakizamana, 2011)
MSc thesis (Kato, 2011)

25. Endophytes in the field: effect on nematode damage
root necrosis (%) (Uganda) root necrosis (%) (Kenya)
25 70 control
F. oxysporum Eny7.11o
60 F. oxysporum V5w2
20
50
15
40
10 30
20
5
10
0 0
0 2 4 6 8 10 12 14
months
MSc thesis (Dusabe, 2007)
MSc thesis (Machungo, 2007)
MSc thesis (Waithira, 2010)
MSc thesis (Hakizamana, 2011)
MSc thesis (Kato, 2011)

26. Endophytes in the field: yield (t/ha) and revenue ($US/ha)
first plant cycle second plant cycle
3,000 3,000
2,000 2,000
farmer 1
1,000 1,000
suckers control endophyte suckers control endophyte
0 0
3,000 3,000
2,000
farmer 2
2,000
1,000 1,000
suckers control endophyte suckers control endophyte
0 0
MSc thesis (Dusabe, 2007)

27. Registration of Fusarium oxysporum V5w2 as biopesticide
Kenya
approval from Pesticide Control Product Board (PCBP)
“eco-tox” dossier: applicant, active ingredient (physical, chemical),
formulation, toxicology, residue analysis
registration field trial: stringent requirements
Burundi and Uganda
no regulatory bodies
duplication of registration field trials according to PCBP
MSc thesis (Hakizamana, 2011)
MSc thesis (Kato, 2011)

28. Transfer of endophyte-technology to private sector
Kenya: commercial bio-pesticide company
transfer of technology (strain, know-how, equipment) to RealIPM
comment: Beauveria bassiana and Trichoderma asperellum
already owned by RealIPM
RealIPM sells the bio-control product to tissue culture producers
problem: licensing and profit
Uganda: embedded in tissue culture company
transfer of technology (strain, know-how, equipment) to AGT
AGT produces its own endophyte-enhanced plants

29. BMZ (2008-2010): outputs
tissue culture laboratories nursery operators farmers
from the field multiplication rooting weaning in the field

30. BMZ (2008-2010): outputs
tissue culture laboratories nursery operators farmers
from the field multiplication rooting weaning in the field
help establish regional select intervention sites
networks and policies
strengthen nursery strengthen farmers in
operators in value chain value chain
assess impact of
tissue culture

31. Tissue culture plant flow
1,500,000 400,000
50,000 100,000
50,000
50,000
450,000
200,000
50,000

32. Tissue culture plant flow
1,500,000 400,000
50,000 100,000
50,000
50,000
450,000
200,000
Rwanda 50,000

33. Tissue culture plant flow
Sudan
1,500,000 400,000
DR Congo
50,000 100,000
50,000
50,000
450,000
200,000
Rwanda 50,000 Tanzania

34. Tissue culture producers: the ideal situation
quarantine and monitoring certification
procedures to prevent pests from procedures certifying the absence of
entering and spreading pests, esp. quarantined ones
done by governments done by governments
missing: procedures and basic missing: procedures
information
essential elements for certification
rapid and robust virus indexing protocols
independent, virus-free, virus-indexed and true-to-type mother gardens
independent virus-indexing capacity service
regional harmonization

35. Tissue culture producers: meetings and trainings
Kenya: 5 days, with commercial tissue culture producers from 3
countries
Uganda: 2 days, with plant protection and quarantine officials from
6 countries
day 1: practial day
day 2: implementation day
implemented by Lava Kumar
information on thomasdubois.110mb.com

36. Tissue culture nurseries: a bleak picture
role
distribution hub
focus centers for farmers and
farmer groups
40 nurseries
28 are functional
18 somehow NGO-independent
16 have farmer groups
MSc thesis (Burkhart, 2010)
PhD thesis (Bauer, 2011) Tropentag (2010)

37. Tissue culture nurseries: SWOT analysis
link between tissue culture producers and nurseries:
NGOs are currently the major customers of the tissue culture producers
most tissue culture producers have quality issues, plantlet delivery
issues (quantities and timeliness)
nurseries:
water access, credit and transport are the major operational issues of
all nurseries
the location of the nurseries is crucial (proximity to tissue culture
producer and market)
success of a nursery is correlated to:
good agricultural practice (seedling handling etc.)
diversification
MSc thesis (Burkhart, 2010)
PhD thesis (Bauer, 2011) Tropentag (2010)

38. Tissue culture nurseries: different models
Uganda
tissue culture laboratories nursery operators farmers
Burundi
tissue culture laboratories farmers
Kenya
tissue culture laboratories farmer groups
from the field multiplication rooting weaning in the field

39. Site location
7,000,000
gross margins (Ugsh/ha/year)
6,000,000 tissue culture
suckers
5,000,000
4,000,000
3,000,000
2,000,000
1,000,000
0
Luwero Mukono Masaka Rukungiri
further away from the market
10 km 350 km
MSc thesis (Dusabe, 2007)

40. Intervention sites in Uganda
groups participants trainings outcomes
farmer training 6 532 270 5 new farmer
groups
nursery operator training 2 57 28 5 new private
nurseries

41. Intervention sites in Burundi
groups participants trainings outcomes
farmer training 5 281 207 4 new farmer
groups
nursery operator training 3 30 71

42. Intervention sites in Kenya
groups participants trainings outcomes
farmer and nursery 6 253 75 2 new farmer
operator training groups
7 new
nurseries

43. Training: agronomy
60
tissue culture
50 suckers
yield (ton/ha/cycle)
40
30
20
10
0
low input high input
Crop Protection (2011)

44. Training: the full package
nursery operators farmers
agronomy agronomy
marketing marketing
business business
financing financing
group formation and group
dynamics

45. Training: the full package
nursery operators farmers
agronomy agronomy
marketing marketing
business business
financing financing
group formation and group
demonstration garden dynamics
dedicated market linkages

46. Training: the full package
nursery operators farmers
agronomy agronomy
marketing marketing
business business
financing financing
group formation and group
demonstration garden dynamics
dedicated market linkages
cost-benefit analysis cost-benefit analysis
4 training modules 5 training modules

47. Post-hoc impact
survey of 1,055 farmers in Kenya, Burundi and Uganda
quantifying
drivers of (non) adoption
effect on productivity, income, income distribution
Kenya:
“We find that female farmers are more likely to adopt TC and farm size does
not necessarily influence adoption. Surprisingly, we find a negative
coefficient for peer group effects on TC adoption.”
MSc thesis (Mukamba, 2010)
MSc thesis (Musuuza, 2010)
PhD thesis (Sentambu, 2010)

48. www.banana2008.com

49. Am I a good endophyte for IITA?
increased yield:
22 peer-reviewed journal articles
1 edited book, 3 book chapters
enhanced growth:
5 PhD thesis, 16 MSc theses, 2 BSc theses
Banana2008; 30 interviews/features on TV, radio, magazines and
newspapers
multiple modes of action:
from endophytes to market pathways

50. Thanks to
Students Partners IITA
PhD: Vincent Bauer, Sinnia AGT Daniel Coyne
Kapindu, Denis Ochieno, BBLF Patrick Emedong
Pamela Paparu, Nassul Bioversity Abubaker Ezale
Sentambu CIALCA Stella Kabiri
MSc: Juliet Akello, Stefan Fred Kato
ISAAA
Burkhart, Astrid Byron, John Kibalama
ISABU
Jahmna Castillo, J Dusabe, Fredrick Kimuli
JKUAT James Kisaakye
Sylvestre Hakizimana,
Christian Hillnhütter, Fred Makerere University Lava Kumar
Kato, Catherine Machungo, NARO Jim Lorenzen
Emmanuel Mukama, RealIPM Moses Lule
Patrick Musuuza, Peter University of Bonn Jane Luyiga
Mwaura, Olaf Schnittker, University of Gent Fen Beed
Jo Thys, Pieter Van University of Goettingen Valentine Nakato
Dessel, Bancy Waithira University of Hohenheim Salvator Nkurunziza
BSc: Patrick Emudong, Joshua Okonya
University of Pretoria
Amelie Nedela Emily Ouma
University of Stellenbosch
Philip Ragama
VEDCO Medat Tamale
Wageningen University Piet Van Asten
ZCA