Abstract :
Sorghum is the fifth most important cereal crops globally after wheat, rice, maize and barley. It is potentially developed in Indonesia as a diversification of crops and raw material of biofuels. Sorghum have well adaptation in its habitat. Development of local sorghum in Indonesia is low. Genetic diversity observations of local sorgum in East Java have never been applied. In order to develop superior cultivar, identification of genetic diversity for gaining information of sorgum germplasm is necessary to be implemented as the first step of plant breeding. The present study aims to analyze the genetic diversity of eight sorghum accessions in East Java using RAPD markers.
This study was carried out at Plant Molecular Laboratory, Biotechnology Development Center/Pusat Pengembangan Bioteknologi (PUSBANG BIOTEK), University of Muhammadiyah Malang. Eight sorghum accessions was obtained from Pasuruan, Sampang, Jombang, Lamongan, Tuban, and Tulungagung. Genomic DNA was extracted from young leaf using CTAB method. The extracted DNA was qualitatively and quantitatively analyzed and was amplified by PCR using 5 RAPD primers namely OPA 02, OPA 10, OPA 13, OPA 19, and OPA 20. The efectiveness of primers was seen through the level of polymorphism. Profile of generated bands was converted as binary data for analyzing the genetic diversity using cluster analysis on NTSYSpc 2.02 program. A dendogram was constructed based on the binary data of five primers using UPGMA method with Dice similarity coefficient.
The results of present study showed that the purity levels of extracted DNA were between 1,886-2,625 and between 0,625-1,275 µg/µl for DNA concentrations. The polymorphism level reachs the number of 90%. Amplification of DNA using five RAPD primers revealed that there were high genetic diversity in eight sorgum accessions. The UPGMA dendogram devided the accessions into 2 major groups (group I and II) at 0,57 Dice similarity level and each groups contained 4 accessions.