KERAGAMAN DURIAN BERDASARKAN FRAGMEN INTERNAL TRANSCRIBED SPACERS (ITS) DNA RIBOSOMAL MELALUI ANALISIS PCR-RFLP

Abstract

Durian adalah salah satu jenis buah-buahan tropis yang memiliki nilai ekonomi tinggi. Durian memiliki bermacam-macam kultivar dengan morfologi yang sulit untuk membedakan. jaminan identitas penting bagi informasi mendasar dalam meningkatkan efisiensi pemuliaan dan pengembangan durian. identifikasi molekuler dianggap lebih akurat dibandingkan dengan karakter morfologi. Polymerase Chain Reaction Restriction Fragment Length Polimorfism (PCR-RFLP) adalah metode untuk menganalisis hasil DNA fragmen panjang perbedaan yang mencerna menggunakan enzim restriksi dengan endonuklease. Sampel yang digunakan dalam penelitian ini adalah aksesi durian diambil secara acak di kecamatan Gunungpati, Semarang. Genomik DNA diisolasi berdasarkan protokol Kit nukleon Phytopure dengan modifikasi. ITS wilayah ribosom DNA diamplifikasi menggunakan teknik PCR-RFLP mengeksploitasi primer spesifik L ITS dan ITS 4 menghasilkan ITS panjang fragmen pada 800 bp. Amplikon yang dicerna menggunakan enam enzim restriksi AluI, Eco471, Bsp1431, BsuRI, Mph11301 dan Ade1. Hasil penelitian dari 11 aksesi durian yang diperiksa menunjukkan bahwa enzim Bsp1431 memiliki dua situs tertentu yang dipotong pada ukuran 550 bp dan 120 bp. enzim BsuR1 memiliki luka situs tertentu dalam ukuran 600 bp. Sedangkan enzim Eco471 telah mencerna situs spesifik pada ukuran 450 bp. Kesimpulan dari penelitian ini adalah durian nomor aksesi ke-12, 15, dan 17 memiliki hubungan genetik dekat dan diduga berada di salah satu spesies.Durian is one of the types of tropical fruits that has high economic value. Durian has an assortment of cultivars with morphology that are difficult to distinguish. Identity assurance is important for the fundamental information in increasing the efficiency of breeding and development of durian. Molecular identification is considered to be more accurate than the morphological characters. Polymerase Chain Reaction Restriction Fragment Length Polimorfism (PCR-RFLP) is a method to analyze the results of DNA fragment length difference that digest using restriction enzyme with endonuclease. The sample used in this study was the accession of durian taken randomly in Gunungpati district, Semarang. Genomic DNA was isolated based on Nucleon Phytopure Kit protocol with modification. ITS DNA ribosomal region was amplified using PCR-RFLP technique exploiting specific primers ITS L and ITS 4 produce ITS fragment length at 800 bp. Amplicon was digested using six restriction enzyme AluI, Eco471, Bsp1431, BsuRI, Mph11301 and Ade1. The research result of 11 accessions durian which is examined shows that the Bsp1431 enzyme has two specific sites that is cut on the size 550 bp and 120 bp. BsuR1 enzymes have a specific site cuts in the size of 600 bp. Whereas Eco471 enzyme has digesting sites specifics on size 450 bp. The conclusion of this research is durian accession number 12th, 15th, and 17th have close genetic relation and are alleged to be in one species.

Keywords

• Durian
• PCR-RFLP
• Internal Transcribed Spacers (ITS)