Keragaman genetik pisang musa balbisiana colla di indonesia menggunakan penanda Amplified Fragment Length Polymorphism (AFLP)

Abstract

Indonesia is one of genetic diversity centers of banana, especially M. acuminata (AA genome). In addition to M. acuminata, in this area can also be found another Musa species that is Musa balbisiana (BB genome), which is known as Pisang Klutuk or Pisang Batu. M. balbisiana is needed to be studied futher because not only its resistance to some pests and diseases, but also its ability to grow well in dry condition. Molecular genetic diversity studies of M. balbisiana in Indonesia had not much been done. Therefore the aim of this research is to study genetic diversity of the banana using Amplified Fragment Length Polymorphism (AFLP). Fifteen accessions of cultivated M. balbisiana and 16 accessions of wild M. balbisiana were used for this study. Materials were collected from Banana Germ Plasm collections Cibinong Science Center Research Center for Biology-LIPI, Bogor Botanical Garden, Bogor, West Sumatera, South Sulawesi and North Sulawesi. CTAB method was used to extract total DNA of leaves. Twenty six pairs of AFLP selective primers were used for analysis and non-radioactive dye was used for visualization of AFLP bands above UV light. Densitograf ATTO CS Analyzer Ver 2.08 application was applied to determine the appearance and size of AFLP bands. NTSYS 2.02 application was utilized to calculate dissimilarity index of Nei and Li (1979) and cluster analysis. Minitab 14 application was used to perform Principal Component Analysis (PCA). The result showed that 22 pairs of 26 pairs of AFLP selective primers produced clear bands for genetic diversity analysis. A total 485 bands with 51-3206 bp in size were produced by these primers pairs. Range of bands number per primers pairs was 11-31 bands. Primer pair number 13 (E-AGC/M-CAC) created the most number of bands and primer pair number 11 (E-ACT/M-CTC) generated the least number of bands. Polymorphism level of total AFLP band was 46.18% with range of polymorphism level was 0-80% for each primer pair. The highest level of polymorphism was generated by primer pair number 7 (E-ACA/M-CTG), while the lowest was produced by primer pair number 19 (E-AGG/M-CTT). Based on AFLP bands, dissimilarity index of Nei and Li (1979) were 0.014-0.152. This result indicated that genetic diversity of the 21 accessions of M. balbisiana was 13.8%. The genetic diversity of wild M. balbisiana (12.9%) higher than genetic diversity of cultivated M. balbisiana (11.5%). Cluster analysis based on UPGMA could not seperate wild M. balbisiana and cultivated M. balbisiana in different clusters. The similarity coefficient 0.89 of dendrogram formed three clusters. The cluster pattern was similar to the grouping pattern in PCA plot, which formed three groups. Cluster I consist of two accessions of KlutukWulung-West Java. Cluster II was made up of two accessions for each of Klutuk-Jogja, Klutuk Sukun-Jogja, Cau Manggala-West Java, Klutuk Wulung-Jogja, Klutuk Hitam-West Java, Unti Batu-South Sulawesi, Pisang Pataga-North Sulawesi, M. balbisiana var liukiuensis (Matsum.) Hakkinen-Japan and an accession Klutuk-West Java. Cluster III has two accessions of Pisang Roti-West Sumatera. There were 17 AFLP bands that have the most important role in grouping of 21 accessions into three groups, i. e. three bands based on first principal component and 14 bands based on second principal component.