Isolation of methylobacterium sp. (Pink Pigmented Facultative Methylotrophs) from paddy (Oryza sativa) and oil palm (Elaeis guineensis)

Methylobacterium sp. commonly known as Pink Pigmented Facultative Methylotrops (PPFM) which able to utilize methanol and other C1 compounds as their sole carbon source. This genus of bacteria are ubiquitous in environments due to their physiology plasticity which enable them to colonize different environment. In this study, phyllosphere Methylobacterium sp. was isolated from two types of samples which are paddy leaves and oil palm leaves. Isolation was done on ammonium mineral salt supplemented with 0.5% methanol. 10 isolates labelled as EPPD1, EPPD2, EPPD3, ENPD1, ENPD2, ENPD3, EPPM1, ENPM1, ENPM2, and ENPM3 were selected for further identification and characterization based on their colony morphology. All of the isolates showed the same cellular morphology which are Gram negative and rod shape. The isolates were then undergone molecular identification by 16S rRNA sequence analysis. The genomic DNA of the isolates were extracted and proceed to polymerase chain reaction to amplify the DNA fragment by using primers 68F and 1392R. The band can be observed at range of 1000 bp to 1500 bp. Based on BLAST analysis, two Methylobacterium sp. were identified. The isolates EPPD1, EPPD4, ENPD1, ENPD3, ENPM1, ENPM2 and ENPM3 were identified as Methylobacterium radiotolerans while ENPM2 was identified as Methylobacterium aerolatum. The isolates EPPD3, EPPM1 and ENPD2 were failed to be identified due to the low quality and concentration of DNA samples.