Evaluation of the MIB-producing potential based on real-time qPCR in drinking water reservoirs

• Published in: Environmental research 2022-03, Vol.204 (Pt C), p.112308-112308, Article 112308
• Main Authors: Suruzzaman, Md, Cao, Tengxin, Lu, Jinping, Wang, Yongjing, Su, Ming, Yang, Min
• Format: Article
• Language: English
• Subjects: 2-Methylisoborneol (MIB); Camphanes - analysis; Cyanobacteria - genetics; Cyanobacteria - metabolism; Drinking water; Drinking Water - analysis; MIB-Synthesis (mic) gene; Odorants - analysis; Real time qPCR; Real-Time Polymerase Chain Reaction; Taste & odor
• ISSN: 0013-9351; 1096-0953
• DOI: 10.1016/j.envres.2021.112308

Cyanobacteria release 2-methylisoborneol (MIB) as a secondary metabolite. Here, we propose a reverse transcription quantitative real-time PCR (RT-qPCR) based method to evaluate the MIB-producing potential in source water by detecting the MIB-synthesis gene (mic). A MIBQSF/R primer set was designed based on 35 mic gene sequences obtained from 12 pure-cultured MIB-producing strains and 23 sequences from the NCBI database. This primer set successfully identified all known 43 MIB-producing cyanobacterial strains (12 from this study and 31 from the NCBI database), belonging to different genera, showing a wider coverage than previous primer sets. The efficiency of the method was proved by the amplification efficiency (E = 91.23%), R2 of the standard curve (0.999), the limit of detection (LOD, 5.7 fg μL−1), and the limit of quantification (LOQ, 1.86 × 104 gene copies μL−1). Further, the method was verified by the correlation between the mic gene abundance and MIB concentration 50 field samples from different reservoirs (R2 = 0.614, p