Screening of molecular marker and rapid identification of Bacillus altitudinis 6ww6 based on comparative genomics

Abstract:

[Objective] Bacillus altitudinis 6ww6,a salt-tolerant and plant growth-promoting rhizobacterium,can be used as a preferred strain for microbial fertilizer.For protection of its intellectual property rights,it is essential to establish a method for rapid detection of this strain.[Methods] The orphan genes of strain 6ww6 were obtained through comparative genomic analysis and the elimination of homologous sequences by TBLASTn and NR library retrieval.The corresponding primers of the orphan genes were designed for PCR detection.[Results] Five specific genes (J939_13195,J9319_05960,J9319_13355,J9319_05965,and J9319_13350) were screened out for strain 6ww6.The target gene J9319_05960 of strain 6ww6 could be specifically amplified with primers 5960F and 5960R,which was confirmed to be the specific molecular marker of strain 6ww6.[Conclusion] In this study,we identified the unique identity code of strain 6ww6 and established a strain-level identification method based on comparative genomic and orphan genes analysis.This method can rapidly and accurately identify target species at the strain level,serving as a powerful tool for the protection of intellectual property rights of valuable microorganisms.