ANALISIS KEANEKARAGAMAN KULTIVAR PISANG MENGGUNAKAN PENANDA PCR-RFLP PADA INTERNAL TRANSCRIBED SPACER (ITS) DNA RIBOSOM

Abstract

Pisang merupakan bahan makanan pokok keempat terpenting di negara berkembang yang memiliki keanekaragaman sangat tinggi. Penanda DNA mikrosatelit dapat membedakan kultivar pisang yang memiliki genom A dengan kultivar pisang bergenom B. Namun penanda mikrosatelit memiliki beberapa keterbatasan, yaitu membutuhkan primer spesifik dan membutuhkan preparasi yang lebih rumit, sehingga membutuhkan waktu dan biaya yang cukup mahal. Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) terhadap DNA internal transcribed spacer (ITS) ribosom mampu mengklasifikasikan kultivar-kultivar pisang berdasarkan pita restriksi daerah ITS yang dipotong dengan enzim RsaI. Koleksi DNA dari 15 kultivar pisang di Laboratorium Genetika dan Molekular Jurusan Biologi UNNES sudah diklasifikasikan genomnya berdasarkan mikrosatelit. DNA kultivar pisang diamplifikasi menggunakan primer ITS L dan ITS 4 menghasilkan fragmen ITS sebesar 700 pb. Pemotongan fragmen ITS DNA ribosom dengan enzim RsaI menghasilkan fragmen  530 pb yang spesifik untuk genom A, fragmen 350 pb dan 180 pb spesifik untuk genom B. Hasil perbandingan klasifikasi genomik berdasarkan mikrosatelit dan PCR-RFLP dari daerah ITS DNA ribosom menunjukkan bahwa klasifikasi genomnya serupa.

 

Banana is the fourth most important staple foods in developing countries which has very high diversity. Microsatellite markers can be able to differentiate bananas cultivars which have A and B genomes, but this marker has restrictions. It requires a specific primer which is takes time and the costs expensive enough. Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) of the ribosomal DNA internal transcribed spacer (ITS) was able to classify banana cultivars based on the restriction band ITS regions cut by RsaI enzyme. The DNA collection from 15 banana cultivars from the Laboratory of Genetics and Molecular Biology Department of Biological Science UNNES have been classified its genome based on microsatellite. Banana cultivar amplified using the primers ITS L and ITS 4 produce ITS fragment at 700 bp. The cutting of ribosomal DNA ITS fragments by RsaI enzyme produce 530 bp fragment that was unique for the A genome, the other fragment 350 bp and 180 bp genome are unique for the B genome. Comparison result of genomic classification based on microsatellite and PCR-RFLP of ribosomal DNA ITS regions showed that the genome classification was similar.