Rapid identification for Paenibacillus mucilaginosus by PCR

Abstract:

Abstract:［Objective］ Paenibacillus mucilaginosus is widely used as microbial fertilizers，therefore，rapid identification of this species by Polymerase Chain Reaction (PCR) is necessary for detection and ecological evaluation of microbial fertilizer．［Methods］ A species-specific primer pair of P．mucilaginosus，orf06701-F (5'-ATGGAGGAAACATGGGGTGA-3')/orf06701-R (5'-TCAGGAATGAAGGCCCCCTT-3') was designed． Optimization of PCR，specificity and sensitivity determination were followed and the rapid identification was established．［Results］ A single band with 333 bp was consistently amplified from all the strains of P．mucilaginosus tested and negative results were obtained from all the reference strains，such as other strains of Paenibacillus and Bacillus． The PCR detection limitation was 400－1000 cells per assay，indicating the sensitivity of the rapid culture-PCR method．To verify the rapid identification，soil was sampled and cultured． Five of 11 soil isolates were rapidly identified as P． mucilaginosus by the method，and the similarity of 333bp sequences were 100%．［Conclusion］ The methods can be used in the rapid identification of P．mucilaginosus and the results will provide technical supports for the detection and ecological evaluation of microbial fertilizer．

Key words:Keywords: Paenibacillus mucilaginosus，PCR identification，species-specific primer pair