Author : Diaz, Maria Genaleen Q.

Summary : The differential gene expression of the enzymes of galactomannan degradation in the developing endosperms of normal and makapuno coconut was determined by cloning, sequencing and characterization of the genes encoding alpha-galactosidase, beta-mannanase and beta-mannosidase. RNA samples were extracted from different developmental stages of the normal and makapuno endosperms using a modified method of Bugos et. al. (1995). RNA yields for the 6-7 month normal and makapuno nuts were 268 and 196 µg/5-8 g FW, respectively. The RNA yield and purity decreased as the nut matured and the makapuno relatively contained lesser amounts of RNA compared to normal endosperms in all ages sampled. The normal and makapuno RNA were reverse-transcribed. The forward and reverse primers for each of the gene were designed, synthesized and used in PCR amplification. The PCR products were cloned in TOPO TA plasmid, sequenced and analyzed. The results of the previous biochemical studies pertaining to the makapuno phenotype in comparison to the current molecular studies are further discussed.