Two applications: Production and extraction of secondary metabolites & transient expression (Agrobacterium-mediated transformation). Their advantages and disadvantages.

1. APPLICATION OF MODERN
BIOTECHNOLOGY TO
AGRICULTURE
(COCOA)
Siti Nur Adila Binti Hamzah
2015139781

2. Theobroma
cacao
Malvaceae
Liquor, butter,
cake and powder
Well-
established and
important crop
in the world
commodities
trade
Rich in
polyphenols.
Antioxidant
compounds:
flavan-3-ols &
procyanidin
oligomers
Problems: limited
growth range for
the plant, easily
exposed to pests
and diseases.

3. Application of Modern
Biotechnology
A
Production and extraction of
secondary metabolites from
plant cell culture
B (transient expression)
Inducing somatic
embryogenesis, Agrobacterium-
mediated transformation using
the homologous BABY BOOM
transcription factor and efficient
regeneration of cocoa plants

4. A) Production and Extraction of Secondary
Metabolites from Plant Cell Culture
• Method of preparing and extracting secondary
metabolites from cocoa cell cultures grown in the
presence of monosaccharide:
1. Culturing cells sufficient to result in production of
secondary metabolites at a first rate.
2. Introducing a monosaccharide to the cells sufficient
for inducing the cells to produce higher rate of
secondary metabolites.
3. Extracting the secondary metabolites from the cells
with an extraction solution which can be ethanol-
based.

5. Secondary metabolites: Procyanidins
• Administered to a subject for therapeutic,
dietary and cosmetic purposes.
Treat/prevent atherosclerosis, cardiovascular
disease, cancer, blood pressure modulation and
hypertension.
Used in dietary compositions: orally
administered in the form of liquid that may be
water-based, milk-based, tea-based.
Improve the quality, appearance of the skin

6. Example: Glucose Addition for
Productivity Enhancement
• Increasing the procyanidin productivity of
suspension cultures of cocoa through
supplemental addition of glucose.
1. Procyanidin has been induced by changing
medium from growth production.
2. At the end of exponential growth stage,
glucose is added.
• Observation: significant improvements in
procyanidin productivity.

7. Advantages
• Independent of environmental factors, seasonal
variations, pest and microbial diseases and geographical
constraints.
• Produced under controlled conditions.
• Stable cell line maintenance.
• Induction of overproduction.
• Reliable and continuous source.

8. Disadvantages
• The use of high sugar concentration in medium
increases the cost.
• Strict aseptic conditions have to be maintained
during culture technique.

9. B) Inducing somatic embryogenesis, using the
homologous BABY BOOM transcription factor in
Agrobacterium-mediated transformation and
efficient regeneration of cocoa plants
• Relates to the manipulation of transcription
factors to enhance the formation of somatic
embryogenesis from cocoa tissue explants and
regenerating cocoa plants from somatic
embryos.
• Agrobacterium tumefaciens naturally infecting
the plant and transferring as well as integrating
the T-DNA into the plant nucleus.

10. • The gene of interest is identified and cloned
into a vector that allows the gene to be
expressed in the plant.
• Once inside the cell, the gene induces the
process of SE to improve yields or overcome
the unresponsive recalcitrant genotype

11. Advantages
• Enhance the formation of somatic embryogenesis
without compromising plant development and
avoiding GMO plant production.
• Promote the transition of somatic cacao cells from
the vegetative to embryonic state.
• Could serve as a biomarker for embryogenesis in
cocoa tissue.
• Tool for enhancement of SE efficiency, in
particular when expressed transiently

12. Disadvantage
• Ability to promote SE is dependent on the
physiological environment and the presence of
other factors in embryogenic tissue.

13. Conclusion
• Continued improvement in cocoa production
and development are needed to obtain
desirable yield and quality characteristics.
• Leads to better utilization of plant as well as its
in-vitro function. This may resulted to
production of novel compounds such as
secondary metabolites.

14. References
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The analysis of production and trade patterns in cocoa market
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2. Dianaplantsciences, Inc. (2010). Production and extraction of procyanidins
from plant cell cultures. Retrieved from
http://www.google.com/patents/EP2624704A1?cl=en
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BABY BOOM transcription factor, 30:1107–1115.

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