title: Development of a specific real-time PCR assay targeting the poly-γ-glutamic acid synthesis gene, pgsB, for the quantification of Bacillus amyloliquefaciens in solid-state fermentation.
abstract: 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-γ-glutamic acid synthesis gene (pgsB) of B. amyloliquefaciens. The detection limit was approximately between 10(2)-10(3) 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 9.415×10(3)-10(7) 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.
authors.affiliation: Agricultural Ministry Key Lab of Plant Nutrition and Fertilization in Low-Middle Reaches of the Yangtze River, Jiangsu Key Lab and Engineering Center for Solid Organic Waste Utilization, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Yong
authors.forename: Ruifu
authors.initials: R
authors.lastname: Zhang
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Yilu
authors.initials: Y
authors.lastname: Chen
authors.forename: Xinqi
authors.initials: X
authors.lastname: Huang
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.forename: Qirong
authors.initials: Q
authors.lastname: Shen
mesh_terms: D001407:Bacillus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005285:Fermentation; D011099:Polyglutamic Acid / Q000096:biosynthesis* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D011099:Polyglutamic Acid