DNA Fingerprinting of Theobroma Cacao
Traditionally, the characterisation of Theobroma cacao is based on morphological characteristics and geographical distribution. Three major groups have been identified, namely, eriolla, Forestero and Trinitario (Wood, 1985). Crosses between these three groups have constitute the major breeding st...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English English |
| Published: |
1998
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/8398/1/FSMB_1998_21_A.pdf |
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| Summary: | Traditionally, the characterisation of Theobroma cacao is based on morphological
characteristics and geographical distribution. Three major groups have been identified,
namely, eriolla, Forestero and Trinitario (Wood, 1985). Crosses between these three
groups have constitute the major breeding strategies used during the last few years.
Currently, large numbers of cocoa clones derived from these crosses have been
introduced in cocoa plantations.
Two different techniques, Random Amplified Polymorphic DNA (RAPD) with
random primers and Restriction Fragment Length Polymorphism (RFLP) using universal
probes, has been investigated as a mean to differentiate 12 local cocoa clones with 1
imported clone as a comparison. 40 primers have been used to amplify genomic DNA
from 13 cocoa clones. Agarose and acrylamide gel electrophoresis used to separate the
RAPD products were further stained with ethidium bromide and sliver nitrate, respectively.
NTSYS-PC computer programme was used to analyse the RAPD data. Meanwhile, two types of probe were used in RFLP study, viz M13 phage DNA and a fragment from a
RAPD product.
The RAPO technique was found to be able to differentiate between local cocoa
clones. Combining the result with UPGMA analysis, 13 tested clones were determined to
fall Into 3 main clusters. A band map has been formed as a future source of reference to
determine the best combination of markers to differentiate two or more clones. M 13 DNA,
as a probe in the RFLP study, failed to give a consistence results in differentiating cocoa
clones. An RAPD product, as a probe however was able to hybridise to few digested
genomic DNA fragments. This indicates that the product is truly amplified fragment and
was confirmed to be mid-repetitive. However the probe did not produce any polymorphic
bands between the clones. |
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