DNA Barcoding Hoya lacunosa Blume dan Hoya diversifolia Blume Menggunakan Penanda Molekuler ITS2

Abstract

Hoya lacunosa and Hoya diversifolia are genera belonging to the Apocynaceae family, which grow most widely in the Southeast Asian region and also in Indonesia. To overcome the limitations of morphological identification in the two plants, this study aims to analyze the DNA barcode in H. lacunosa and H. diversifolia using the molecular marker ITS2, and perform phylogenetic construction to determine the relationship between the species of H. lacunosa and H. diversifolia with other species of hoya. The stages of this study are leaf sampling of H. lacunosa and H. diversifolia, genomic DNA isolation, amplification with PCR, visualization of PCR results, sequencing, and data analysis. This study obtained the results that a sequence length for H. lacunosa of 387 bp, and H. diversifolia of 400 bp. The sample species H. lacunosa showed the highest homology with H. lacunosa from Indonesia (OQ829456.1) with a Per.Ident of 95.91%, and the H. diversifolia sample showed the highest homology with H. diversifolia from Thailand (OQ829525.1) with Per.Ident by 90.46%. The results of the aligment in both samples showed some differences in nucleotide bases with the BLAST result sequence. The results of the phylogenetic construction obtained were that the sample H. lacunosa was in the same clade with Hoya lacunosa (OQ829456.1) with a boostrap value of 31%, while the sample Hoya diversifolia was in the same clade with Hoya diversifolia (OQ829583.1) with a boostrap value of 84%. %. The combined phylogenetic construct of the two Hoyas resulted in the two samples being separated in two different clades. Overall, this study shows that ITS2 can be a good molecular marker for DNA barcoding in H. lacunosa and H. diversifolia samples.