Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene
In this study, a SYBR Green quantitative real-time PCR method was established and applied. Relative expression of the synthetic genes from Microcystis gas vesicles (gvpC), algal toxin genes (mcyA), and polysaccharides (espL) from water and sediments of Meiliang Bay and from the center of Lake Taihu...
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PeerJ Inc.
2019-06-01
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| Online Access: | https://peerj.com/articles/7188.pdf |
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| author | Long Yu Xiaofei Wu Yang Yu Limei Shi Min Zhang |
| author_facet | Long Yu Xiaofei Wu Yang Yu Limei Shi Min Zhang |
| author_sort | Long Yu |
| collection | DOAJ |
| description | In this study, a SYBR Green quantitative real-time PCR method was established and applied. Relative expression of the synthetic genes from Microcystis gas vesicles (gvpC), algal toxin genes (mcyA), and polysaccharides (espL) from water and sediments of Meiliang Bay and from the center of Lake Taihu were tested from January to June, 2017. Indoor Microcystis aeruginosa was used as the control group. The kit for total RNA extraction in Microcystis was optimized. Results showed that the optimized kit extracted high-concentrations and high-quality total RNA from Microcystis. The extraction purity and concentration were significantly higher than those extracted by the original kit. The transcription level of gvpC increased gradually until a peak was reached in March. However, expression of gvpC decreased continuously at the proliferating and floating stages of Cyanobacterial biomass. The maximum level of expression of gvpC in April in comparison to expression of mcyA in March occurred first. We found that the SYBR Green qRT-PCR method, which is characterized by high specificity, repeatability, is rapid, and can be used for quantitative detection of expression of gvpC, mcyA, and espL. The recruitment of cyanobacteria is the process in which cyanobacteria in the sediment began to regain their activity, started to grow and migrated to the water column. |
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| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T08:19:25Z |
| publishDate | 2019-06-01 |
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| spelling | doaj.art-477646d85a304e26a115f4adef0717c72023-12-02T21:53:54ZengPeerJ Inc.PeerJ2167-83592019-06-017e718810.7717/peerj.7188Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis geneLong Yu0Xiaofei Wu1Yang Yu2Limei Shi3Min Zhang4College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, ChinaState Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography & Limnology, Chinese Academy of Sciences, Nanjing, ChinaState Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography & Limnology, Chinese Academy of Sciences, Nanjing, ChinaState Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography & Limnology, Chinese Academy of Sciences, Nanjing, ChinaIn this study, a SYBR Green quantitative real-time PCR method was established and applied. Relative expression of the synthetic genes from Microcystis gas vesicles (gvpC), algal toxin genes (mcyA), and polysaccharides (espL) from water and sediments of Meiliang Bay and from the center of Lake Taihu were tested from January to June, 2017. Indoor Microcystis aeruginosa was used as the control group. The kit for total RNA extraction in Microcystis was optimized. Results showed that the optimized kit extracted high-concentrations and high-quality total RNA from Microcystis. The extraction purity and concentration were significantly higher than those extracted by the original kit. The transcription level of gvpC increased gradually until a peak was reached in March. However, expression of gvpC decreased continuously at the proliferating and floating stages of Cyanobacterial biomass. The maximum level of expression of gvpC in April in comparison to expression of mcyA in March occurred first. We found that the SYBR Green qRT-PCR method, which is characterized by high specificity, repeatability, is rapid, and can be used for quantitative detection of expression of gvpC, mcyA, and espL. The recruitment of cyanobacteria is the process in which cyanobacteria in the sediment began to regain their activity, started to grow and migrated to the water column.https://peerj.com/articles/7188.pdfespLmcyAgvpCqRT-PCRCyanobacterial recruitmentRelative expression |
| spellingShingle | Long Yu Xiaofei Wu Yang Yu Limei Shi Min Zhang Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene PeerJ espL mcyA gvpC qRT-PCR Cyanobacterial recruitment Relative expression |
| title | Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene |
| title_full | Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene |
| title_fullStr | Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene |
| title_full_unstemmed | Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene |
| title_short | Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene |
| title_sort | recruitment of cyanobacteria by reverse transcription quantitative real time pcr based on expression of microcystis gene |
| topic | espL mcyA gvpC qRT-PCR Cyanobacterial recruitment Relative expression |
| url | https://peerj.com/articles/7188.pdf |
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