An Analysis of Partial DNA Sequence of Meisa1 Gene on Sweet and Bitter Cassavas (Manihot esculenta Crantz.)

Abstract

Sweet and bitter taste on cassava tuber is affected by starch metabolisms. Meisa1 gene is a gene in cassava (Manihot esculenta) encoding isoamylase1 enzyme involved in starch metabolisms. This study aimed to analyze partial DNA sequences of Meisa1 gene on sweet and bitter cassavas collected by Genetics Laboratory, Department Biology, Faculty of Mathematics and Natural Sciences, Riau University, Indonesia. Methods included total DNA extraction from fresh young leaves of cassava using CTAB buffer, polymerase chain reaction (PCR), electrophoresis, and sequencing. The obtained data were analyzed using MEGA software version 5. The results showed that there were nucleotide variations in the intron region, not in the exon region. The variations were caused by the transition substitution mutation (35.39%) and transversion substitution mutation (64.61%). The genetic distance range between seven cassava genotypes was approximately 0% to 11%. Partial DNA sequence variations of Meisa1 gene located in intron region were unable to cluster seven cassava genotypes separately into two groups based on tuber taste.

Pandan (Pandanus sp), Rotan (Calamus sp), and Rengas (Gluta sp) from Kajuik Lake, Riau Province, Indonesia

“…The PCR product will be a band in electrophoresis gel. The thick and the single bands are suitable for sequencing requirement [30,31]. After that, the sequences are analyzed using BLASTn program at http://ncbi.nih.nlm.gov/.…”