| Abstrak/Abstract |
Research on genetic diversity and intra-species classification of Indonesian Dioscorea alata L. based on
morphological characters has been done, and the result shows that there are 4 sub-groups of green cultivars
group, and 5 sub-groups of purplish-red cultivars group. The objectives of this study are to determine the
phylogenetic relationship cultivars of D. alata Indonesia compared to D. bulbifera as nearest species as well as 3
cultivars from GenBank. The young leaves of 18 water yam cultivar accessions were collected from Java,
Madura, South Sumatera, South Kalimantan, Centre Celebes (Sulawesi), Ternate, West Papua, and Nusa
Tenggara islands of Indonesia. DNA Isolation was conducted using Phytopure reagent. DNA amplification was
conducted using thermocycler, predenaturation at 95 oC 5 minutes, denaturation at 95 oC 1 minutes, annealing at
60 oC 3 minutes, and elongation at 72 oC 2 minutes along with 30 cycles. The PCR products were
electrophoresed in 1.5% agarose and visualized under UV transiluminator. Fifty-five ?l of PCR products with
positive targeted band between 700-800 bp were sent to 1st Base Singapore for purification and sequencing of
18S, ITS-1, 5.8S, ITS-2, and 28S rDNA. The DNA sequences were compared and aligned by BioEdit program
(version 7.0.5.2) and MEGA programs. Comparison of entire sequences of the tested samples were aligned by
software ClustalW (version 1.83). Phylogenetic trees were based on hierarchical clustering of the alignments of
18S, ITS-1, 5.8S, ITS-2, and 28S rDNA and produced by Neigbor-Joining using MEGA 5 software of the
bootstrap values (1000 replicates). The study result shows that D. alata cultivars have high genetic variability on
ITS-1, 5.8S, and ITS-2 rDNA region. Groups of green and purplish red cultivars formed based on morphological
characters are not formed based on ITS-rDNA markers. Sub-groups were formed based on ITS-rDNA molecular
markers derived from both the green and purplish-red cultivar groups. This result revealed that two cultivar |
