Highlights
- •Aquilaria malaccensis (Thymelaeaceae) is the main source of high-grade agarwood in Southeast Asia.
- •Aggressive collection and trade activities over the past decades have put great pressure on the natural stands.
- •This study developed DNA databases of A. malaccensis at species, population and individual levels for forensic application.
- •Two case studies are presented of how these databases were used to track A. malaccensis to original population and stump.
- •These databases are useful in providing evidence for legal proceedings and agarwood certification.
Abstract
Aquilaria malaccensis (Thymelaeaceae) is the main source of high-grade agarwood in Southeast Asia. Aggressive
collections and trade activities over the past decades have put great pressure on
the natural stands and raised concerns over the long-term survival potential of A. malaccensis. Tracking and authentication of agarwood require method with a high degree of accuracy.
Therefore, this study aimed to develop DNA databases of A. malaccensis as the tracking tools at species, population and individual levels for forensic identification
and chain of custody certification. Using two cpDNA (rbcL and matK) and an rDNA (ITS2) markers, species identification database of Aquilaria was developed to distinguish A. malaccensis from A. hirta, A. microcarpa, A. beccariana, A. crassna, A. sinensis and A. rostrata. In addition, based on 35 populations of A. malaccensis throughout Peninsular Malaysia, cpDNA haplotype and STR allele frequency databases
were developed for population and individual identification. A haplotype distribution
map based on 29 haplotypes derived from seven cpDNA showed that the A. malaccensis in Peninsular Malaysia can be associated to Kedah-Perak and Kelantan-Johor regions.
Similarly, genetic relatedness and Bayesian clustering analyses based on 10 STR markers
also divided the 35 populations into two main genetic clusters, corresponding to Kedah-Perak
and Kelantan-Johor regions. The STR allele frequency databases were established and
characterized according to these two regions. To determine the performance of the
STR allele frequency databases for population identification, independent self-assignment
tests showed that the percentage of individuals correctly assigned into the origin
population was 93.88% in Kedah-Perak and 90.29% in Kelantan-Johor. For the STR allele
frequency databases to be used for individual identification, conservativeness tests
showed that the θ should be adjusted to 0.250 and 0.200 in the Kedah-Perak and Kelantan-Johor databases,
respectively. To ensure consistency in allele calling for the dinucleotide repeat
loci across different electrophoretic platforms or laboratories, allelic ladders have
been developed for the 10 STR loci. Two case studies are presented of how these databases
were used to track A. malaccensis to the origin population and stump. These databases are ready to be used to provide
admissible forensic evidence for legal proceedings against the illegal harvesters
of agarwood and for agarwood certification to meet the consumer country regulations.
Keywords
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Article info
Publication history
Published online: December 28, 2021
Accepted:
December 23,
2021
Received in revised form:
December 20,
2021
Received:
August 10,
2021
Identification
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