Research Article
Ogbulie TE and Nwaokorie FO
Abstract
This study looked at the diversity of microorganisms persistent in agricultural soil sample polluted with 100 ml of 100% Nigerian Bonny light crude oil left for four years. DNA from crude oil polluted agricultural soil sample was extraction using ZYMO soil DNA extraction Kit. DNA sequencing was performed by Next Generation Sequencing Technique [NGST] using automated PCR cycle- Genome Sequencer™ FLX System from 454 Life Sciences™ and Roche Applied. Sequence analysis and alignment was performed using Vecton NTI suite 9 (InforMax, Inc.). The resulting nucleotide sequences were compared to sequences obtained from GenBank by BLASTx analysis using CLO Bio software as well as BLASTn using NCBI. Molecular Identities of microbial community was obtained by creating different dendrograms. Gene sequencing carried out read 513 different nucleotide sequences. Seven phyla with 47 corresponding culture-dependent species and 169 culture-independent bacteria clone were obtained. The resultant tree showed cladogram of proteobacteria ( b and g - proteobacteria), bacteria/enterobacteria, firmicutes, plantomycetes, acidobacteria group/ fibrobacteres, Bacteriodetes/chlorobi Actinobacteria/high G + C and chloriflexi phyla. Furher taxonomical classification was carried out with reads of sufficient Q scores (> q30) and lengths and a total of 420 read count of top kingdom classification of 100% bacteria kingdom was obtained. Proteobacteria phyla of class betaproteobacteria, order Burkholderiales and family Comamonadaceae had the highest read count with percentage diversity of 57.14%, 53.81%, 53.81 and 53.57% respectively. The nucleotide sequences with no hit (208) was sent to Genbank for asigning of ascension number. The detection of these diverse organisms from crude oil polluted agricultural soil left for four years, depict that the organism probably, have degradative genes which aided their survival.