Cantigi ungu (Vaccinium varingiifolium) merupakan tumbuhan yang teramati hanya dapat tumbuh di daerah pegunungan dengan ketinggian 1800—3340 mdpl di Indonesia. Cantigi ungu memiliki manfaat sebagai sumber makanan dan memiliki kadar antioksidan yang tinggi. Vaccinium spp. di Amerika telah didomestikasi sehingga memiliki nilai komersial. Proses domestikasi tersebut melibatkan identifikasi morfologi, tetapi identifikasi secara molekuler lebih baik digunakan agar breeding menjadi efisien dan efektif. Identifikasi molekuler dapat menggunakan DNA barcoding dan rekonstruksi filogeni. Consortium for the Barcode of Life (CBOL) telah menetapkan bahwa matK dan rbcL merupakan gen penanda (DNA barcoding) yang ideal dalam identifikasi tumbuhan terestrial. Gen matK dan rbcL merupakan gen yang berada pada kloroplas. Hasil analisis filogenetik Cantigi ungu yang ada hanya menggunakan gen penanda ITS. Tujuan dari penelitian ini adalah menganalisis dan mengonfirmasi kekerabatan sekuens Cantigi ungu yang dikoleksi dari Gunung Gede, Gunung Tangkuban Parahu dan Kawah Putih Ciwidey menggunakan gen penanda matK dan rbcL dengan rekonstruksi pohon filogeni. DNA Cantigi Ungu dari tiga tempat berbeda di Jawa Barat diisolasi menggunakan kit ekstraksi DNA tanaman kemudian dilakukan amplifikasi PCR dan optimasi suhu annealing. Suhu annealing optimal Cantigi ungu pada matK adalah 52°C dan rbcL adalah 55°C. Hasil amplifikasi PCR tersebut kemudian disekuensing, dilakukan contig sekuens, di-trimming dan diunggah pada GenBank. Hasil BLAST sekuens ketiga sampel Cantigi ungu pada kedua gen menunjukkan bahwa ketiga sampel Cantigi ungu merupakan spesies yang sama. Hasil analisis alignment menunjukkan bahwa ketiga sampel Cantigi ungu memiliki indeks similaritas 100% pada kedua gen. Hasil Rekonstruksi pohon filogeni dengan Vaccinium spp. dan out-group menunjukkan bahwa ketiga sampel berada pada cabang yang sama, mengindikasikan bahwa ketiga sampel tersebut berasal dari spesies yang sama, Vaccinium varingiifolium. ......The Purple Cantigi (Vaccinium varingiifolium) is a plant observed to only grow in mountainous areas at altitudes of 1800—3340 meters above sea level in Indonesia. The Purple Cantigi has benefits as a food source and possesses a high level of antioxidants. Vaccinium spp. in America has been domesticated, thus having commercial value. The domestication process involves morphological identification, but molecular identification is preferably used for efficient and effective breeding. Molecular identification can utilize DNA barcoding and phylogenetic reconstruction. The Consortium for the Barcode of Life (CBOL) has established that matK and rbcL are ideal marker genes (DNA barcoding) for identifying terrestrial plants. The matK and rbcL genes are located in the chloroplast. The only available results of the phylogenetic analysis of purple Cantigi use the ITS marker gene. This research aims to analyze and confirm the sequence relationships of the Purple Cantigi collected from Mount Gede, Mount Tangkuban Parahu, and Kawah Putih Ciwidey using the matK and rbcL marker genes with phylogenetic tree reconstruction. DNA of the Purple Cantigi from three different places in West Java was isolated using a plant DNA extraction kit and then subjected to PCR amplification and annealing temperature optimization. The optimal annealing temperature for the Purple Cantigi in matK was 52°C, and in rbcL was 55°C. The PCR amplification results were sequenced; contig sequences were performed, trimmed, and uploaded to GenBank. BLAST results of the sequence of the three Purple Cantigi samples on both genes showed that all three samples are the same species. Alignment analysis showed that all three Purple Cantigi samples have a 100% similarity index on both genes. Phylogenetic tree reconstruction with Vaccinium spp. and out-group showed that all three samples are on the same branch, indicating they belong to the same species, Vaccinium varingiifolium.
Kata Kunci : Cantigi ungu, Vaccinium varingifolium (Blume) Miq., Karakter Morfologi, Gen ITS, Filogenetik ......Cantigi ungu (Vaccinium varingifolium (Blume) Miq.) is an endemic plant of Indonesia that has the potential as a food and medicinal source. This plant is found abundantly in the highlands area ranging from 1800 to 3340 meters above sea level. A morphological analysis study of Cantigi ungu is necessary to determine the plant's characteristics. This research aims to identify morphological and genetic characteristics of Cantigi ungu growing in the mountainous areas of West Java province, especially in Mount Gede, Kawah Putih Ciwidey, and Mount Tangkuban Parahu. The method used in this research is direct observation, specimen photo, and morphological characteristic measurement using a ruler. The results showed that Cantigi ungu has morphologies similar to other Vaccinium species, with dark green leaves, small white flowers, and round fruits with a diameter of 8-9 mm, leaf length of 2-4 cm, leaf width of 1-1.5 cm, and tree circumference of 36-98 cm. So far, genetic data reports on this plant have yet to be widely reported. Genetic conservation is needed so that the sustainability, availability, and existence of this plant germplasm can be known, as well as its conservation status. The research was conducted on 3 Cantigi ungu plants growing in the highlands of West Java (in the Mount Gede Pangrango National Park Area, Mount Tangkuban Parahu Nature Park, and Ciwidey White Crater) and compared them based on their Internal Transcribed Spacer (ITS) gene sequence. ITS gene amplification was performed using Forward primer ITS_u1 5'-GGA AGK ARA AGT CGT AAC AAG G-3 and Reverse primer ITS_u4 5'- RGT TTC TTT TCC TCT GCT TA-'3. This study provides important information about the morphological characteristics of Cantigi ungu, which can be used for future taxonomic and plant location research purposes. Phylogenetic analysis was done using the MEGA XI method using Maximum Likelihood (ML), Minimum Evolution (ME), Neighbor-Joining (NJ), and Maximum Parsimony (MP) methods. The result shows that the Cantigi ungu in this study is the same species as the comparison species of Vaccinium varingifolium AY274564.1, Vaccinium varingifolium OR000769.1, Vaccinium varingifolium OR000770.1, Vaccinium varingifolium OR000771.1 at the Genebank database National Center for Biotechnology Information (NCBI).