Abstract
2-Methylisoborneol (MIB), along with geosmin, has been frequently associated with taste and odor (T&O) events in waters, and cyanobacterial species were considered the main producers in freshwater ecosystems. The detection and quantification of genes responsible for the synthesis of these odors on-site could be valuable for the prediction and treatment of T&O. In this study, SYBR Green and TaqMan real-time quantitative polymerase chain reaction (qPCR) assays were established on the basis of conserved regions in cyanobacterial MIB cyclase gene (mic) and applied to the quantification of potential MIB-producing cyanobacteria in waters. These two qPCR assays showed the prescribed values of amplification efficiency (0.976 and 1.023, respectively) with linearity (R 2 > 0.99) in the range of 101 to 107 mic copies. Examination on environmental samples and cyanobacterial cultures indicated that qPCRs were rarely inhibited by background biomass, and the limit of quantification was at the level of 103 mic copies L−1 in natural waters. The seasonal variations of mic abundances in Lushui Reservoir, Donghu Lake, and Qinghai Lake of China were successfully quantified using the two qPCR protocols and indicated the applicability of these assays in field monitoring. Regression analysis revealed the positive correlation between mic abundance and MIB concentration, but no significant linear correlation was found between MIB-producing cyanobacteria and total cyanobacteria in investigated waters. The qPCR assays developed in this study could rapidly and reliably detect and quantify the abundance of potential cyanobacterial MIB producers, thereby providing a valuable tool for the prediction and early detection of T&O events.
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References
Auffret M, Pilote A, Proulx E, Proulx D, Vandenberg G, Villemur R (2011) Establishment of a real-time PCR method for quantification of geosmin-producing Streptomyces spp. in recirculating aquaculture systems. Water Res 45:6753–6762
Cook D, Newcombe G, Sztajnbok P (2001) The application of powdered activated carbon for MIB and geosmin removal: predicting PAC doses in four raw waters. Water Res 35:1325–1333
Dzialowski AR, Smith VH, Huggins DG, deNoyelles F, Lim N-C, Baker DS, Beury JH (2009) Development of predictive models for geosmin-related taste and odor in Kansas, USA, drinking water reservoirs. Water Res 43:2829–2840
Giglio S, Chou WKW, Ikeda H, Cane DE, Monis PT (2011) Biosynthesis of 2-methylisoborneol in cyanobacteria. Environ Sci Technol 45:992–998
Ha JH, Hidaka T, Tsuno H (2009) Quantification of toxic Microcystis and evaluation of its dominance ratio in blooms using real-time PCR. Environ Sci Technol 43:812–818
Hurlburt B, Lloyd SW, Grimm CC (2009) Comparison of analytical techniques for detection of geosmin and 2-methylisoborneol in aqueous samples. J Chromatogr Sci 47:670–673
Ichimura T (1979) Media for blue-green algae. In: Nishizawa M, Chihara M (eds) Methods in algology. Kyoritsu, Tokyo, pp 294–304
Izaguirre G, Taylor WD (2004) A guide to geosmin- and MIB-producing cyanobacteria in the United States. Water Sci Technol 49:19–24
Jüttner F, Watson SB (2007) Biochemical and ecological control of geosmin and 2-methylisoborneol in source waters. Appl Environ Microbiol 73:4395–4406
Komatsu M, Tsuda M, Omura S, Oikawa H, Ikeda H (2008) Identification and functional analysis of genes controlling biosynthesis of 2-methylisoborneol. Proc Natl Acad Sci U S A 105:7422–7427
Kutovaya OA, Watson SB (2014) Development and application of a molecular assay to detect and monitor geosmin-producing cyanobacteria and actinomycetes in the Great Lakes. J Great Lakes Res 40:404–414
Li L, Wan N, Gan N, Xiao BD, Song LR (2007) Annual dynamics and origins of the odorous compounds in the pilot experimental area of Lake Dianchi, China. Water Sci Technol 55:43–50
Nübel U, Garcia-Pichel F, Muyzer G (1997) PCR primers to amplify 16S rRNA genes from cyanobacteria. Appl Environ Microbiol 63:3327–3332
Persson PE (1983) Off-flavours in aquatic ecosystems-an introduction. Water Sci Technol 15:1–11
Savichtcheva O, Debroas D, Kurmayer R, Villar C, Jenny JP, Arnaud F, Perga ME, Domaizon I (2011) Quantitative PCR enumeration of total/toxic Planktothrix rubescens and total cyanobacteria in preserved DNA isolated from lake sediments. Appl Environ Microbiol 77:8744–8753
Smith JL, Boyer GL, Zimba PV (2008) A review of cyanobacterial odorous and bioactive metabolites: impacts and management alternatives in aquaculture. Aquaculture 280:5–20
Srivastava A, Singh S, Ahn C-Y, Oh H-M, Asthana RK (2013) Monitoring approaches for a toxic cyanobacterial bloom. Environ Sci Technol 47:8999–9013
Su M, Gaget V, Giglio S, Burch M, An W, Yang M (2013) Establishment of quantitative PCR methods for the quantification of geosmin-producing potential and Anabaena sp. in freshwater systems. Water Res 47:3444–3454
Sugiura N, Utsumi M, Wei B, Iwami N, Okano K, Kawauchi Y, Maekawa T (2004) Assessment for the complicated occurrence of nuisance odours from phytoplankton and environmental factors in a eutrophic lake. Lakes Reserv Res Manage 9:195–201
Suurnӓkki S, Gomez-Saez GV, Rantala-Ylinen A, Jokela J, Fewer DP, Sivonen K (2015) Identification of geosmin and 2-methylisoborneol in cyanobacteria and molecular detection methods for the producers of these compounds. Water Res 68:56–66
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739
Tsao HW, Michinaka A, Yen HK, Giglio S, Hobson P, Monis P, Lin TF (2014) Monitoring of geosmin producing Anabaena circinalis using quantitative PCR. Water Res 49:416–425
Wang CM, Cane DE (2008) Biochemistry and molecular genetics of the biosynthesis of the earthy odorant methylisoborneol in Streptomyces coelicolor. J Am Chem Soc 130:8908–8909
Wang Z, Li R (2015) Effects of light and temperature on the odor production of 2-methylisoborneol-producing Pseudanabaena sp. and geosmin-producing Anabaena ucrainica (cyanobacteria). Biochem Syst Ecol. 58:219–226
Wang SH, Dzialowski AR, Lim NC, Spotts WW, Huggins DG, deNoyelles F (2005) Relationships between cyanobacterial production and the physical and chemical properties of a Midwestern reservoir, USA. Hydrobiologia 541:29–43
Wang Z, Xu Y, Shao J, Wang J, Li R (2011) Genes associated with 2-methylisoborneol biosynthesis in cyanobacteria: isolation, characterization, and expression in response to light. PLoS One 6:e18665
Wang Z, Liu Y, Xu Y, Li R (2013) The divergence of cpcBA-IGS sequences between Dolichospermum and Aphanizomenon (Cyanobacteria) and the molecular detection of Dolichospermum flos-aquae in Taihu Lake, China. Phycologia 52:447–454
Wang Z, Shao J, Xu Y, Yan B, Li R (2015) Genetic basis for geosmin production by the water bloom-forming cyanobacterium, Anabaena ucrainica. Water 7:175–187
Watanabe MM, Hiroki M (1997) NIES-Collection. List of strains, algae and protozoa. National Institute for Environmental Studies, Environment Agency, Japan, p 140
Watson SB (2004) Aquatic taste and odour: a primary signal of drinking water integrity. J Toxicol Environ Health A 67:1779–1795
Watson SB, Brownlee B, Satchwill T, Hargesheimer EE (2000) Quantitative analysis of trace levels of geosmin and MIB in source and drinking water using headspace SPME. Water Res 34:2818–2828
Watson SB, Satchwill ED, McCauley E (2001) Under-ice blooms and source-water odour in a nutrient poor reservoir: biological, ecological and applied issues. Freshw Biol 46:1553–1567
Watson SB, Charlton M, Yerubandi R, Howell T, Ridal J, Brownlee B, Marvin C, Millard S (2007) Off flavour in large waterbodies: physics, chemistry and biology in synchrony. Water Sci Technol 55:1–8
Watson SB, Ridal J, Boyer GL (2008) Taste and odor and cyanobacterial toxins: impairment, prediction, and management in the Great Lakes. Can J Fish Aquat Sci 65:1779–1796
Xu Y, Wang G, Yang W, Li R (2010) Dynamics of the water bloom-forming Microcystis and its relationship with physicochemical factors in Lake Xuanwu (China). Environ Sci Pollut Res 17:1581–1590
Zuo Y, Li L, Zhang T, Zheng L, Dai G, Liu L, Song L (2010) Contribution of Streptomyces in sediment to earthy odor in the overlying water in Xionghe Reservoir, China. Water Res 44:6085–6094
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This research was supported by the NSFC-JST Joint Fund (41261140337) and National Water Science and Technology Projects (2012ZX07105-004).
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Wang, Z., Song, G., Shao, J. et al. Establishment and field applications of real-time PCR methods for the quantification of potential MIB-producing cyanobacteria in aquatic systems. J Appl Phycol 28, 325–333 (2016). https://doi.org/10.1007/s10811-015-0529-1
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DOI: https://doi.org/10.1007/s10811-015-0529-1