Evaluasi Lokus Potensial matK dan ITS2 Untuk DNA Barcoding Anggrek Bulbophyllum lobbii Lindl.
Abstract
Abstrak
Bulbophyllum lobbii Lindl. merupakan anggrek dari famili Orchidaceae yang berpotensi sebagai bahan baku obat herbal. Identikasi morfologi anggrek B. lobii memiliki keterbatasan karena kemiripan spesies dengan anggrek lain. Alternatif identifikasi secara molekuler menggunakan sekuen matK dan ITS2 sebagai barcode dalam DNA barcoding diharapkan menjadi salah satu lokus pembeda spesies anggrek B. lobbii secara akurat dan efisien. Penelitian ini bertujuan untuk mengidentifikasi sekuen matK dan ITS2 sebagai penanda molekuler yang efektif untuk anggrek B. lobbii. DNA genom B. lobbii diisolasi dengan metode Cethyl Trimethyl Ammonium Bromide (CTAB) dan amplifikasi DNA dengan PCR. Hasil penelitian menunjukkan sekuen matK dari B. lobbii memiliki tingkat homologi tinggi dengan dua spesies B. lobbii (KY966747.1 dan KY966691.1) dari China dengan nilai Per. Ident sebesar 99,20%, sedangkan sekuen ITS2 memiliki homologi tertinggi dengan nilai Per. Ident sebesar 99,76% pada spesies B. lobbii (MG253848.1) dari Polandia. Hasil analisis menunjukkan sekuen ITS2 dapat mengidentifikasi spesies dari tingkatan subspesies atau diatasnya (ordo atau genus) dan juga meningkatkan resolusi filogenetik yang baik pada hasil BLAST, sedangkan sekuen matK memberikan sedikit kontribusi dalam pengelompokkan hubungan kekerabatan antara spesies B. lobbii dengan spesies pembanding lainnya. Sekuen ITS2 dapat direkomendasikan sebagai penanda molekuler yang paling baik untuk identifikasi sampel anggrek Bulbophyllum, khususnya Bulbophyllum lobbii.
Abstract
Bulbophyllum lobbii Lindl. is an orchid from the Orchidaceae family which has potential as a raw material for herbal medicine. Morphological identification of the B. lobii orchid has limitations due to the species' similarity to other orchids. The alternative molecular identification using matK and ITS2 sequences as barcodes in DNA barcoding is expected to be one of the loci for distinguishing the B. lobbii orchid species accurately and efficiently. This study aims to identify matK and ITS2 sequences as effective molecular markers for the orchid B. lobbii. Bulbophyllum lobbii genomic DNA was isolated using the Cethyl Trimethyl Ammonium Bromide (CTAB) method and DNA amplification by PCR. The results showed that the matK sequence from B. lobbii has a high level of homology with two B. lobbii species (KY966747.1 and KY966691.1) from China with a value of Per. Ident is 99.20%, while the ITS2 sequence has the highest homology with a Per. Ident value of 99.76% with the species B. lobbii (MG253848.1) from Poland. The results of the analysis show that the ITS2 sequence can identify species from the subspecies level or above (ordo or genus) and also improves good phylogenetic resolution in BLAST results, while the matK sequence makes little contribution in grouping the relationship between the B. lobbii species and other comparison species. The ITS2 sequence can be recommended as the best molecular marker for identifying Bulbophyllum orchid samples, especially Bulbophyllum lobbii.
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