use of tissue culture to alleviate the supply of …...use of tissue culture to alleviate the supply...
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Use of Tissue culture to alleviate the supply of quality planting
materials among cacao smallholder farmers in Cote d Ivoire
V4C Management Meeting Bassam December 2015
Jane Kahia, Siaka Kone, Georges Ngoran,
Lucien Diby.,Jean Claude Nzi and
Christophe Kouamé
2016 Next Generation Cocoa Research Symposium
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Outline
• DEFINATIONS
• CACAO SOMATIC EMBRYOGENESIS (SE)
• IN VITRO PROPGATION OF PLANTAIN BANANA
• PROPAGATION OF SOME ELITE CASSAVA CULTIVARS
2016 Next Generation Cocoa Research Symposium
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DEFINATIONS
• TISSUE CULTURE (TC)
-Production of plantlets from explants, sterile conditions and controlled environment
• SOMATIC EMBROYGENESIS
-Production of embryos from somatic cells (non sexual
cells)
2016 Next Generation Cocoa Research Symposium
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SE Cont.,
Two pathways of regeneration
• Direct SE- bipolar embryos formed directly from the
explants e.g coffee
• Indirect SE-Explant- callus-embryos-plantlets
2016 Next Generation Cocoa Research Symposium
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CACAO SE
• Conventionally propagated by seeds, cuttings and grafting.
• Propagation by cuttings is rarely used in CDI
• SE provides a feasible option for mass propagation of elite clones
• Plants derived from somatic embryos are genetically identical to their parental donor cells
2016 Next Generation Cocoa Research Symposium
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SE OF CACAO
• Unopened flowers are collected early in the morning before 9.am
• Sterilized and staminodes and petals explants extracted
• Cultured on media supplemented with cytokinins and auxins in the dark
• Callus induced after 14 days
• Embryos produced after 2 months
2016 Next Generation Cocoa Research Symposium
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SE cont.,
• Embryos germinated into plantlets in a growth room maintained 25°±2C
• Plantlets weaned in a greenhouse
• Commonly use protocol is the Penn State University protocol of 1997
2016 Next Generation Cocoa Research Symposium
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SE CACAO
PROCEDURE OF PRODUCING CACAO PLANTLETS USING SES
tam
ino
de
sP
eta
ls
Callus
Embryos
Cacao plantletsWeaned plantlets
2016 Next Generation Cocoa Research Symposium
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Table 1: Effect of 2,4-D (10 µM) and varying concentrations of Kinetin (2.5, 5 and
25 µM) on callus and embryos induction in cacao.
Genotypes 2,4-D (10 µM)
+Concentrations of
Kinetin in µM
Percent callus ± S.E Percent embryogenic
callus ± S.E
Mean No. of embryos ±
S.E
C1 Control 96.00 ± 4.00 a 14.06 ± 2.06 a 3.34 ± 0,34 a
2.597.43 ± 2 .56 a 67.52 ± 0.85 b 10.00 ± 1.15 b
5 100.00± 0.00 a 100.00 ± 0.00 c 28.00 ± 1.15 c
25 100.00 ± 0.00 a 0.00 ± 0.00 d 0.00 ± 0.00 a
C14Control
100.00 ± 0.00 a 5.33 ± 1.33 d 1.33 ± 0.33 a
2.579.09 ± 2.40 b 32.41 ± 5.29 e 7.33 ± 2.02 b
5 76.55 ± 0.36 b 6.13 ± 0.12 d 1.00 ± 0.00 a
25 96.00 ± 4.00 a 0.00 ± 0.00 d 0.00 ± 0.00 a
2016 Next Generation Cocoa Research Symposium
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Table 2: Effect of 2,4-D (20 µM) and varying concentrations of Kinetin (2.5, 5
and 25 µM) on callus and embryos induction in cacao.
Genotypes 2,4-D (20 µM)
+Concentrations of Kinetin
in µM
Percent callus ± S.E Percent embryogenic
callus ± S.E
Mean No. of embryos ±
S.E
C1 Control 96.00 ± 4.00 a 14.06 ± 2.06 a 3.33 ± 0.33 a
2.597.33 ± 2.67 a 85.95 ± 3.18 b 22.00 ± 1.73 b
5 100.00 ± 0.00 a 0.00 ± 0.00 c 0.00 ± 0.00 a
25 100.00 ± 0.00 a 0.00 ± 0.00 c 0.00 ± 0.00 a
C14Control
100.00 ± 0.00 a 5.33 ± 1.33 c 1.33± 0.33 a
2.596.29 ± 3.70 a 91.07 ± 4.49 b 22.00 ± 2.30 b
5 100.00 ± 0.00 a 16.85 ± 5.82 a 3.00 ± 1.15 a
25 92.43 ± 1.96 a 0.00 ± 0.00 c 0.00 ± 0.00 a
2016 Next Generation Cocoa Research Symposium
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IN VITRO PROPAGATION OF BANANA
• Major staple food crop for millions
• Conventionally propagated by suckers
• 9-10 suckers depending on the variety
• Banana tissue culture initiated to support CocoAction
• Using TC-1 sucker can produce upto 200 banana plantlets
2016 Next Generation Cocoa Research Symposium
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IN VITRO PROPAGATION OF BANANA
Preparation of explants
Initiation
Weaned plantlets Multiplication and Rooting
2016 Next Generation Cocoa Research Symposium
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MICROPROPAGATION OF SOME ELITE CASSAVA CULTIVARS
• Cassava (Manihot esculenta Crantz) is the fifth most important food crop in the world
• Conventionally propagated by stem cuttings
• High-quality cassava cuttings for planting are often in short supply
• In CDI, cassava production has been hampered by the use of poor-quality, disease-prone planting materials
2016 Next Generation Cocoa Research Symposium
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CASSAVA Cont.,• High-yielding, disease-tolerant improved
cassava varieties available.
• These varieties have the potential to raise the cassava productivity by 3 to 4 times
• The adoption of these elite varieties by farmers can be fast tracked by propagation using TC
Cassava plantlets
2016 Next Generation Cocoa Research Symposium
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CASSAVA
• Two methods of regeneration being evaluated
• SE and micropropagation
• High regeneration rates using micropropagation
• SE promising-preferred
2016 Next Generation Cocoa Research Symposium
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MERCI POUR
VOTRE
ATTENTION
2016 Next Generation Cocoa Research Symposium