Identification of Nitrogen Fixing Bacteria from Indigenous Microorganisms (IMO) Isolated From Bamboo Area at Taman Pertanian and Lepar Hilir, Pahang

Agricultural sector in Malaysia utilises high amount of chemical fertilizers as well as organic manures secreted by animals. The utilisation of the fertilizers in agricultural sector increases the level of environmental pollution. The usage of chemical fertilizers can be reduced by applying indigenous microorganisms (IMO) as beneficial microorganisms that are able to provide sufficient nutrients to various crops in agricultural sector. In this study, IMO that has been isolated from bamboo area at Taman Pertanian, Pahang and Lepar Hilir, Pahang were subjected to single colonies isolation and nitrogen fixing bacteria were screened by using several biochemical approaches including gram staining, carbohydrate fermentation and nitrate reduction tests. From the screening, a total of six bacterial isolates have been identified as potential beneficial microorganisms to fix free nitrogen to nitrate, as one of important nutrient for the growth of crops. All the six bacterial isolates were further subjected to genomic DNA isolation and followed by PCR amplification of 16S rRNA gene using 27F and 1492R primers. The PCR-amplified products were purified and further sequenced using the same primers. DNA sequence of each 16S rRNA gene of all the bacterial isolates were subjected to BLASTN analysis at the NCBI GenBank database. Interestingly, the BLASTN analysis has shown that all the bacterial isolates are homologous to strains of Bacillus spp. including B. subtilis, B. amyloliquefaciens and B. tequilensis, respectively. Phylogenic tree analysis on rRNA genes of all the bacterial isolates has shown their closely-relatedness with other strains of Bacillus spp. In conclusion, Bacillus spp. including B. subtilis, B. amyloliquefaciens and B. tequilensis isolated from the selected bamboo areas have potential to be utilised in Malaysia agricultural sector as nitrogen fixing bacteria that might reduce the usage of chemical fertilizers