Importance of messenger RNA stability of toxin synthetase genes for monitoring toxic cyanobacterial bloom |
| |
| Affiliation: | 1. Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, 361021, China;2. University of Chinese Academy of Sciences, Beijing, 100049, China;3. Fujian Provincial Investigation, Design & Research Institute of Water Conservancy & Hydropower, Fuzhou, 350001, China;4. Key Laboratory of Marine Biogenetic Resources, The Third Institute of Oceanography SOA, Xiamen, 361005, China;5. College of Petroleum Engineering, Liaoning Shihua University, Fushun, 113001, China;1. School of Environment, Harbin Institute of Technology, Harbin, 150090, China;2. Beijing Spacecrafts, Beijing, 100086, China;3. Chongqing University, Chongqing, 400044, China;4. Key Laboratory for Water Quality and Conservation of the Pearl River Delta, Ministry of Education, Institute of Environmental Research at Greater Bay, Guangzhou University, Guangzhou, 510006, China;1. Institut de Systématique, Évolution, Biodiversité, ISYEB, UMR 7205, Université des Antilles. Équipe Biologie de la mangrove. UFR SEN. 97100 Pointe-à-Pitre. France;2. Sorbonne Université, CNRS, Laboratoire d’Océanographie de Villefranche, LOV, F-06230 Villefranche-sur-Mer, France;1. Department of Aquatic Ecology, Netherlands Institute of Ecology (NIOO-KNAW), Post Office Box 50, 6700 AB Wageningen, The Netherlands;2. Microbia Environnement, Observatoire Océanologique, 66650 Banyuls/Mer, France;3. Department of Pharmacology, Faculty of Veterinary, Universidade de Santiago de Compostela, Campus Universitario, 27002 Lugo, Spain;4. Koeman & Bijkerk B.V., Post Office Box 111, 9750 AC Haren, The Netherlands;5. Marine Biological Association of the UK, The Citadel, Plymouth, PL1 2PB, UK;1. Biotechnology, Department of Biochemistry, University of Turku, Tykistökatu 6A 6th Floor, Turku 20520, Finland;2. Biochemistry, Department of Biosciences, Åbo Akademi University, Tykistökatu 6A 3rd Floor, Turku 20520, Finland;1. DISTAV, University of Genoa, C.so Europa 26, Genoa, Italy;2. CoNISMa, P.le Flaminio 9, Rome, Italy;3. CNR-IBF, via De Marini 5, Genoa, Italy;1. UNESCO Chair in Life Sciences, International Postgraduate Educational Center, Acharian 31, Avan, Yerevan 0040, Armenia;2. Ludwig-Maximilians University, Munich, Department of Biology II, Aquatic Ecology, 82152, Planegg-Martinsried, Germany;3. Auburn University, School of Fisheries, Aquaculture, and Aquatic Sciences, 203 Swingle Hall, 36849, Auburn, Alabama, United States;4. Laboratory of Catalytic - Photocatalytic Processes and Environmental Analysis, Institute of Nanoscience & Nanotechnology, National Center for Scientific Research “Demokritos”, Patriarchou Grigoriou & Neapoleos 10, 15341, Agia Paraskevi, Athens, Greece;5. Water Quality Control Department, Athens Water Supply and Sewerage Company (EYDAP SA), Oropou 156, 11146, Galatsi, Athens, Greece |
| |
| Abstract: | Toxic cyanobacterial blooms, occurring frequently worldwide, have posed serious threats to human health and aquatic ecosystem. RNA-based quantitative PCR, which could detect potential toxin-producing cyanobacteria that are actively transcribing toxin genes, is a more reliable method, compared to DNA-based qPCR. However, single-stranded mRNA is labile, and their degradation may lead to an underestimate of gene expression level, even misleading toxic risk management, and thus impeding its application. Here, the mRNA stability of microcystin synthetase genes (mcyA-J) was systematically evaluated in unicellular and colonial Microcystis with various treatments (−80 ℃, −196 ℃, 4 °C or 25 °C with RNases inhibitors). Results revealed the highly instability of toxin gene transcripts, affected by transcript structures and cell aggregation. The −196 ℃ treatment was the most effective for stabilizing these transcripts. RNAstore® (4 °C) could stabilize these transcripts effectively for a short time (less than 7 d), but their stability was strikingly reduced in colonial Microcystis. Furthermore, decay kinetics of mcyA-J transcripts in various treatments was developed, and showed that their decay rates were varied (0.0018–3.014 d−1), due to different molecular structures. The mcyH transcripts had the lowest decay rate (0.0018 d−1 at −196 ℃), attributed to the fewest AU sites and stem-loops involved in its secondary structure. Thus, mcyH was the most proper target gene for monitoring toxic cyanobacterial bloom. These findings provided new insight into mRNA stability of toxin genes, and contributed to monitoring toxic cyanobacterial blooms and water managements using RNA-based molecular techniques. |
| |
| Keywords: | mRNA stability Toxin gene Molecular structure Quantitative PCR Colonial cells Toxic cyanobacterial bloom |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
