Implementing high-throughput insect barcoding in microbiome studies: impact of non-destructive DNA extraction on microbiome reconstruction DOI
Veronika Andriienko, Mateusz Buczek, Rudolf Meier

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 30, 2024

Abstract Background Symbiotic relationships with diverse microorganisms are crucial for many aspects of insect biology. However, while our understanding taxonomic diversity and the distribution species in natural communities is limited, we know much less about their microbiota. In era rapid biodiversity declines, as researchers increasingly turn towards DNA-based monitoring, developing broadly implementing approaches high-throughput cost-effective characterization both insect-associated microbial essential. We need to verify whether such barcoding, a powerful tool identifying wild insects, would permit subsequent microbiota reconstruction these specimens. Methods High-throughput barcoding (“megabarcoding”) methods often rely on non-destructive obtaining template DNA PCR amplification by leaching out specimens using alkaline buffers HotSHOT. This study investigated impact HotSHOT abundance estimates reconstructed bacterial community profiles. addressed this question comparing quantitative 16S rRNA amplicon sequencing data HotSHOT-treated or untreated 16 representing six orders selected based expectation limited variation among individuals. Results find that 13 species, treatment significantly reduced estimates, corresponding an estimated 15-fold decrease amplifiable average. On other hand, pre-treatment had effect composition. The presence abundant bacteria known significant effects was not affected. observed changes low-abundance microbes, those close reliable detection threshold. Alpha beta analyses showed compositional differences only few species. Conclusion Our results indicate pre-treated remain suitable composition reconstruction, even if may be hard estimate. These can cost-effectively combine across communities. Thus, voucher obtained megabarcoding studies targeted at characterizing used microbiome characterizations. substantially aid speeding up accumulation knowledge microbiomes hyperdiverse

Language: Английский

Implementing high-throughput insect barcoding in microbiome studies: impact of non-destructive DNA extraction on microbiome reconstruction DOI
Veronika Andriienko, Mateusz Buczek, Rudolf Meier

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 30, 2024

Abstract Background Symbiotic relationships with diverse microorganisms are crucial for many aspects of insect biology. However, while our understanding taxonomic diversity and the distribution species in natural communities is limited, we know much less about their microbiota. In era rapid biodiversity declines, as researchers increasingly turn towards DNA-based monitoring, developing broadly implementing approaches high-throughput cost-effective characterization both insect-associated microbial essential. We need to verify whether such barcoding, a powerful tool identifying wild insects, would permit subsequent microbiota reconstruction these specimens. Methods High-throughput barcoding (“megabarcoding”) methods often rely on non-destructive obtaining template DNA PCR amplification by leaching out specimens using alkaline buffers HotSHOT. This study investigated impact HotSHOT abundance estimates reconstructed bacterial community profiles. addressed this question comparing quantitative 16S rRNA amplicon sequencing data HotSHOT-treated or untreated 16 representing six orders selected based expectation limited variation among individuals. Results find that 13 species, treatment significantly reduced estimates, corresponding an estimated 15-fold decrease amplifiable average. On other hand, pre-treatment had effect composition. The presence abundant bacteria known significant effects was not affected. observed changes low-abundance microbes, those close reliable detection threshold. Alpha beta analyses showed compositional differences only few species. Conclusion Our results indicate pre-treated remain suitable composition reconstruction, even if may be hard estimate. These can cost-effectively combine across communities. Thus, voucher obtained megabarcoding studies targeted at characterizing used microbiome characterizations. substantially aid speeding up accumulation knowledge microbiomes hyperdiverse

Language: Английский

Citations

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