CHARACTERS OF MITOCHONDRIAL DNA D-LOOP HYPERVARIABLE III FRAGMENTS OF INDONESIAN RHINOCEROS HORNBILL (BUCEROS RHINOCEROS) (AVES: BUCEROTIDAE)

Abstract

The rhinoceros hornbill (Buceros rhinoceros) genetic characteristics consist of nucleotide polymorphisms, haplotypes, genetic distances, and relationships which are important for their conservation effort in Indonesia. We sequenced mitochondrial DNA D-loop hypervariable III fragments from five rhinoceros hornbill individuals at Safari Park Indonesia I and Ragunan Zoo, which were isolated using Dneasy® Blood and Tissue Kit Spin-Column Protocol, Qiagen. D-loop fragment replication was done by PCR technique using DLBuce_F (5'-TGGCCTTTCTCCAAGGTCTA-3') and DLBuce_R (5'-TGAAGG AGT TCATGGGCTTAG-3') primer. Thirty SNP sites were found in 788 bp D-loop sequences of five rhinoceros hornbill individuals and each individual had a different haplotype. The average genetic distance between individuals was 3.09% and all individuals were categorized into two groups (Group I: EC6TS, EC1RG, EC2TS and Group II: EC9TS, EC10TS) with a genetic distance of 3.99%. This result indicated that the two groups were distinct subspecies. The genetic distance between Indonesian and Thai rhinoceros hornbills was 10.76%. Five Indonesian rhinoceros hornbill individuals at Safari Park Indonesia I and Ragunan Zoo probably came from different populations, ancestors, and two different islands. This study can be of use for management consideration in captive breeding effort at both zoos. The D-loop sequence obtained is a useful character to distinguish three rhinoceros hornbill subspecies in Indonesia.