基于比较基因组学的高地芽孢杆菌6ww6分子标识筛选及其快速鉴定

【目的】高地芽孢杆菌(Bacillus altitudinis) 6ww6是一株根际耐盐促生菌，可以作为微生物肥料的优选菌种。为了加强菌种知识产权保护，建立菌株快速检测方法。【方法】本研究通过基因组比对分析、TBLASTn检索、NR库检索验证并剔除有其他同源的结果序列，最终得到菌株6ww6的孤儿基因，设计引物进行PCR鉴定。【结果】筛选出5个特异性基因J939_13195、J9319_05960、J9319_13355、J9319_05965和J9319_13350。引物5960F和5960R可以单一性扩增出菌株6ww6的目的基因J9319_05960，确定其为菌株6ww6的特异性分子标识。【结论】本研究确定了菌株6ww6的唯一性身份编码，建立了以比较基因组学和孤儿基因为基础的菌株水平上的鉴定方法。该方法具有快速、精确、能鉴定到菌株水平的优点，对于有价值微生物的知识产权保护提供了有力的抓手。

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

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.