Orschler, Laura and Agrawal, Shelesh and Lackner, Susanne (2019):
On resolving ambiguities in microbial community analysis of partial nitritation anammox reactors.
In: Scientific Reports, 9, Springer Nature, ISSN 2045-2322,
DOI: 10.1038/s41598-019-42882-8,
[Article]
Abstract
PCR-based methods have caused a surge for integration of eco-physiological approaches into research on partial nitritation anammox (PNA). However, a lack of rigorous standards for molecular analyses resulted in widespread data misinterpretation and consequently lack of consensus. Data consistency and accuracy strongly depend on the primer selection and data interpretation. An in-silico evaluation of 16S rRNA gene eubacterial primers used in PNA studies from the last ten years unraveled the difficulty of comparing ecological data from different studies due to a variation in the coverage of these primers. Our 16S amplicon sequencing approach, which includes parallel sequencing of six 16S rRNA hypervariable regions, showed that there is no perfect hypervariable region for PNA microbial communities. Using qPCR analysis, we emphasize the significance of primer choice for quantification and caution with data interpretation. We also provide a framework for PCR based analyses that will improve and assist to objectively interpret and compare such results.
| Item Type: | Article |
|---|---|
| Erschienen: | 2019 |
| Creators: | Orschler, Laura and Agrawal, Shelesh and Lackner, Susanne |
| Title: | On resolving ambiguities in microbial community analysis of partial nitritation anammox reactors |
| Language: | English |
| Abstract: | PCR-based methods have caused a surge for integration of eco-physiological approaches into research on partial nitritation anammox (PNA). However, a lack of rigorous standards for molecular analyses resulted in widespread data misinterpretation and consequently lack of consensus. Data consistency and accuracy strongly depend on the primer selection and data interpretation. An in-silico evaluation of 16S rRNA gene eubacterial primers used in PNA studies from the last ten years unraveled the difficulty of comparing ecological data from different studies due to a variation in the coverage of these primers. Our 16S amplicon sequencing approach, which includes parallel sequencing of six 16S rRNA hypervariable regions, showed that there is no perfect hypervariable region for PNA microbial communities. Using qPCR analysis, we emphasize the significance of primer choice for quantification and caution with data interpretation. We also provide a framework for PCR based analyses that will improve and assist to objectively interpret and compare such results. |
| Journal or Publication Title: | Scientific Reports |
| Journal volume: | 9 |
| Publisher: | Springer Nature |
| Divisions: | 13 Department of Civil and Environmental Engineering Sciences 13 Department of Civil and Environmental Engineering Sciences > Institute IWAR |
| Date Deposited: | 03 Nov 2019 20:56 |
| DOI: | 10.1038/s41598-019-42882-8 |
| Official URL: | https://doi.org/10.1038/s41598-019-42882-8 |
| URN: | urn:nbn:de:tuda-tuprints-92280 |
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