use of tissue culture to alleviate the supply of …...use of tissue culture to alleviate the supply...

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


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)


SE Cont.,

Two pathways of regeneration

• Direct SE- bipolar embryos formed directly from the

explants e.g coffee

• Indirect SE-Explant- callus-embryos-plantlets


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


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


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


SE CACAO

PROCEDURE OF PRODUCING CACAO PLANTLETS USING SES

tam

ino

de

sP

eta

ls

Callus

Embryos

Cacao plantletsWeaned plantlets


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


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


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


IN VITRO PROPAGATION OF BANANA

Preparation of explants

Initiation

Weaned plantlets Multiplication and Rooting


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


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


CASSAVA

• Two methods of regeneration being evaluated

• SE and micropropagation

• High regeneration rates using micropropagation

• SE promising-preferred


MERCI POUR

VOTRE

ATTENTION


use of tissue culture to alleviate the supply of …...use of tissue culture to alleviate the supply...

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