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First Author: LI Xi
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. RNAstoreR (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.

Contact the author: YU Xin
Page Number: 101642
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PubYear: SEP 2019
Volume: 88
Publication Name: HARMFUL ALGAE
The full text link: https://doi.org/10.1016/j.hal.2019.101642
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