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Abstract
Aquatic plants are one of the most diverse and commercially important plants. The variety of unique morphology is one of the factors why this plant is of great
interest. Anubias species is one of the aquatic plants which have been widely used as aquarium plant for decoration. DNA barcoding is an identification
approach that may be useful in conservation strategies of the Anubias species. This method could help in identification of this species as an alternative to
morphological observation. To build a DNA barcode in selected Anubias species (Anubias nana, Anubias nana golden, Anubias nana petite, Anubias glabra,
Anubias barteri, Anubias congensis, and Anubias lanceolata), chloroplast DNA was chosen as the region for study. The PCR was performed in 10l reaction
mixture containing DNA, PCR buffer, MgCl, chloroplast DNA primer and taq polymerase. PCR product were analyzed by electrophoresis on 1 % agarose gel
and detected by EtBr. One of the region in the chloroplast genome which is matK region were selected and screened to define a universal barcoding region
across all the 7 selected species. In this research, matK region was successfully amplified from all of the 7 selected species tested. DNA sequences from matK
region from each species were analyzed to differentiate each other with haplotype. The haplotype analysis for matK region showed sufficiently high resolution to
enable differentiation between the selected Anubias species. This study has shown that the chloroplast DNA (cpDNA) region has high potential to be developed
for DNA barcoding in the Anubias species. This research is a first step towards the development of a universal DNA barcoding system for all commercial aquatic
plants in Malaysia.
PCR
AMPLIFICATION
GEL
ELECTROPHORESIS
GEL EXTRACTION
SEQUENCING
Anubias
nana
ANP
Anubias
nana
pe,te
ANG
Anubias
nana
golden
AG
Anubias
glabra
AB
Anubias
barteri
AL
Anubias
lanceolata
AC
Anubias
congensis
Figure 2: Sequences
analyzed using
Codoncode Aligner
ANALYSIS OF DATA
COD
ON
ALI CODE
GNE
R
Table 2: Haplotypes using matK region in chloroplast DNA
References:
1.
Chodon Sass,(2007). DNA Barcoding in the Cycadales: Testing the Potential of Proposed
Barcoding Markers for Species Identification of Cycads. Plos One. 11, e1154.
2.
Grierson, & Covey. Plant molecular biology. USA: Chapman & Hall.
3.
Mark W.Chase, (2005). Land Plants and DNA Barcodes: Short-term and Long-term goals. Phil.
Trans. R. Soc. B.
4.
Renaud Lahaye, (2008). DNA Barcoding the Floras of Biodiversity Hotspots. PNAS, 105(8),
2923-2928.
Acknowledgements
We would like to thank University of Malaya and Department of Fisheries for the support of the project. Special thanks to lab members Azwan,
Neena and Asmida for your support. Thanks also to Puan Siti, Kak Maz and Kak Florence for your guidance.