Development of a specific real-time PCR assay targeting the poly-¦Ã-glutamic acid synthesis gene, pgsB, for the quantification of Bacillus amyloliquefaciens in solid-state fermentation

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• 作者：Xiaoyu Yong^a ; ^b ; Ruifu Zhang^a ; Nan Zhang^a ; Yilu Chen^b ; Xinqi Huang^a ; Jun Zhao^a ; Qirong Shen^a ; ^{shenqirong@njau.edu.cn} ; ^{2009203043@njau.edu.cn}
• 关键词：TaqMan real-time PCR ; Poly-¦Ã-glutamic acid synthesis gene (pgsB) ; Bacillus amyloliquefaciens ; Detection ; Solid-state fermentation
• 刊名：Bioresource Technology
• 出版年：2013
• 出版时间：February, 2013
• 年：2013
• 卷：129
• 期：Complete
• 页码：477-484
• 全文大小：754 K

文摘

A TaqMan real-time PCR procedure was developed for specific detection and quantification of strains belonging to Bacillus amyloliquefaciens group. The primer pair pgsB726-f/pgsB791-r and the pgsB-probe were designed from one of the poly-¦Ã-glutamic acid synthesis gene (pgsB) of B. amyloliquefaciens. The detection limit was approximately between 10²-10³ cells/mL. A linear correlation between the log₁₀ input pMD-pgsB plasmid DNA copies and the threshold cycle values were observed with a magnitude of linearity in the range of 9.415 ¡Á 10³-10⁷ copies/mL for the standard curve, which exhibited a slope of ?3.35 and an R² value of 99.8 % . Results of validation of this method with artificially contaminated and natural solid-state fermentation samples showed that it was suitable for specific and sensitive detection and quantification for the target strains in solid-state fermentation samples. This could be more useful to understand the fermentation starting strain and the final microbiological properties of fermentation products.