2.2 Identification of endophytic bacteria from sugarcane
The bacterial isolates were characterized by their cultural conditions, morphological and biochemical characteristics using standard methods(13). The 16S rDNA gene was PCR amplified from the genomic DNA isolated from pure bacterial colonies. The bacterial DNA extraction was performed using a MO-BIO Ultraclean Microbial DNA Isolation Kit (MoBio Laboratories, Inc., USA). The DNA was amplified with the universal primers 27F and 1522R. Sequencing of cloned DNA was undertaken by First BASE Laboratories, Malaysia. Nucleotide sequences were compared with NCBI GenBank entries and the similarities were determined by using the BLAST algorithm. Phylogenetic analysis was performed using the ClustalX programme. Distances was performed by using Bootstrap values. The results were shown by Treeview.