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

Outline

• DEFINATIONS

• CACAO SOMATIC EMBRYOGENESIS (SE)

• IN VITRO PROPGATION OF PLANTAIN BANANA

• PROPAGATION OF SOME ELITE CASSAVA CULTIVARS

2016 Next Generation Cocoa Research Symposium

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

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

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

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

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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

SE CACAO

PROCEDURE OF PRODUCING CACAO PLANTLETS USING SES

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Callus

Embryos

Cacao plantletsWeaned plantlets

2016 Next Generation Cocoa Research Symposium

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

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

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

IN VITRO PROPAGATION OF BANANA

Preparation of explants

Initiation

Weaned plantlets Multiplication and Rooting

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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

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

CASSAVA

• Two methods of regeneration being evaluated

• SE and micropropagation

• High regeneration rates using micropropagation

• SE promising-preferred

2016 Next Generation Cocoa Research Symposium

MERCI POUR

VOTRE

ATTENTION

2016 Next Generation Cocoa Research Symposium