Classification of Sweet Sorghum in Northern of Thailand Using Random Amplification of Polymorphic DNA (RAPD)

Abstract

The genetic diversity of local sweet sorghum in Northern of Thailand has been studies. Sweetsorghum is an alternative plant for fuel substitution. Genomic DNA was isolated from leaves of sweetsorghum using cetyltrimethyl ammonium bromide (CTAB) and NucleoSpin®Plant kit. The resultsshowed that genomic DNAs from both methods have given high yield and quality. Then, the genomicDNA samples from NucleoSpin® Plant kit were used as DNA templates in Random amplified polymorphicDNA. In this study, four types of RAPD primers were used which were S23 (5´AGTCAGCCAC3´), S24(5´AATCGGGCTG3´), S27 (5´GAAACGGGTG3´) and S30 (5´GTGATCGCAG3´). It was found that theoptimal annealing temperature of each primer were 38.0, 36.0, 36.4 and 36.0 ºC, respectively. In addition,the bands of PCR products were analyzed by Image Quant TL v7.01. Total PCR products of 87 bands wereobserved and 73 bands of these were polymorphic bands. Total number of polymorphic bands (TPB) wascalculated to 83%. RAPD primer S24 has given the highest polymorphic bands (24 bands, TPB = 93%)with PCR product sizes from 148 to 1850 bp, while RAPD primer S23 has showed the lowest polymorphicbands (12 bands, PPB = 66%) with size ranges from 276 to 1796 bp. These RAPD markers will be usefulfor genetic identification of sweet sorghum cultivars.

Keywords : Genomic DNA / Molecular Analysis / RAPD Technique / Genetic Diversity