دورية أكاديمية
Chloroplast sequence variation and the efficacy of peptide nucleic acids for blocking host amplification in plant microbiome studies
| العنوان: | Chloroplast sequence variation and the efficacy of peptide nucleic acids for blocking host amplification in plant microbiome studies |
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| المؤلفون: | Fitzpatrick, Connor R., Lu-Irving, Patricia, Copeland, Julia, Guttman, David S., Wang, Pauline W., Baltrus, David A., Dlugosch, Katrina M., Johnson, Marc T. J. |
| المساهمون: | Univ Arizona, Dept Ecol & Evolutionary Biol |
| المصدر: | Microbiome ; 6 ; 1 |
| بيانات النشر: | BMC |
| سنة النشر: | 2018 |
| المجموعة: | The University of Arizona: UA Campus Repository |
| مصطلحات موضوعية: | Plastid variation, PNA clamp, Host contamination, 16S amplicon sequencing, Asteraceae |
| الوصف: | Background: The ability to efficiently characterize microbial communities from host individuals can be limited by co-amplification of host organellar sequences (mitochondrial and/or plastid), which share a common ancestor and thus sequence similarity with extant bacterial lineages. One promising approach is the use of sequence-specific peptide nucleic acid (PNA) clamps, which bind to, and block amplification of, host-derived DNA. Universal PNA clamps have been proposed to block host plant-derived mitochondrial (mPNA) and plastid (pPNA) sequences at the V4 16S rRNA locus, but their efficacy across a wide range of host plant species has not been experimentally tested. Results: Using the universal PNA clamps, we amplified and sequenced root microbial communities from replicate individuals of 32 plant species with a most recent common ancestor inferred at 140 MYA. We found the average rate of host plastid contamination across plant species was 23%, however, particular lineages exhibited much higher rates (62-94%), with the highest levels of contamination occurring in the Asteraceae. We investigated chloroplast sequence variation at the V4 locus across 500 land plant species (Embryophyta) and found six lineages with mismatches between plastid and the universal pPNA sequence, including all species within the Asteraceae. Using a modified pPNA for the Asteraceae sequence, we found (1) host contamination in Asteraceae species was reduced from 65 to 23%; and (2) host contamination in non Asteraceae species was increased from 12 to 69%. These results demonstrate that even single nucleotide mismatches can lead to drastic reductions in pPNA efficacy in blocking host amplification. Importantly, we found that pPNA type (universal or modified) had no effect on the detection of individual bacterial taxa, or estimates of within and between sample bacterial diversity, suggesting that our modification did not introduce bias against particular bacterial lineages. Conclusions: When high similarity exists between host organellar DNA ... |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| تدمد: | 2049-2618 |
| العلاقة: | https://microbiomejournal.biomedcentral.com/articles/10.1186/s40168-018-0534-0Test; Fitzpatrick et al. Microbiome (2018) 6:144 https://doi.org/10.1186/s40168-018-0534-0Test; http://hdl.handle.net/10150/631155Test; MICROBIOME |
| DOI: | 10.1186/s40168-018-0534-0 |
| الإتاحة: | https://doi.org/10.1186/s40168-018-0534-0Test http://hdl.handle.net/10150/631155Test |
| حقوق: | © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License. ; https://creativecommons.org/licenses/by/4.0Test/ |
| رقم الانضمام: | edsbas.C0D97656 |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 20492618 |
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| DOI: | 10.1186/s40168-018-0534-0 |