Species-specific Loci of Three Indonesian Durio Inferred from ISSR Fingerprinting
Pieter Agusthinus Riupassa *
Department of Biology, Plant Biology Graduate Program, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680, Indonesia and Department of Biology, Faculty of Mathematics and Natural Sciences, Pattimura University, Ambon 97233, Indonesia.
Miftahudin .
Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680, Indonesia.
Suharsono .
Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680, Indonesia.
Tatik Chikmawati
Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680, Indonesia.
*Author to whom correspondence should be addressed.
Abstract
Aims: To assess molecular diversity and to determine species-specific loci of Indonesian durio, based on Inter-Simple Sequence Repeat (ISSR) markers.
Study Design: Genomic DNA isolation from fresh durio leaves and then DNA amplification. Electrophoresis PCR product and bands scoring. Calculating the diversity genetic parameter and identification specific loci.
Place and Duration of Study: The research was done in Plant Biology Laboratory, Biology Departments, Bogor Agricultural University, start from April 2014 until January 2016.
Methodology: Ten ISSR primers were used to reveal the genetic diversity of 58 varieties of three species (D. zibethinus, D. kutejensis, and D. tanjungpurensis) sampled from Mekarsari and Cipaku Garden, West Java, and from Sekadau Regency, West Kalimantan, Indonesia. Genetic diversity parameters were estimated based on binary data of PCR bands appearance which calculation using GenAlEx. Construction phenogram using Mega software and validating of loci specific with Chi-Square method invoked SAS® software.
Results: The diversity genetic parameter showed that the mean number of observed alleles, the mean number of effective alleles, Shannon’s Information Index, the genetic diversity, and the percentage of polymorphic loci were 1.25, 1.34, 0.30, 0.20, and 58.3%, respectively. UPGMA based cluster analysis and principal coordinate analysis classified them into three groups according to the species. We found 13 loci from 7 ISSR primers linked to species-specific loci.
Conclusion: The genetic diversity of durio can be assessed using the ISSR markers, which can also be used for species identification of Indonesian durio.
Keywords: Durio, genetic diversity, ISSR marker, fingerprinting, species-specific loci.