Using Plate-Wash PCR and High-Throughput Sequencing to Measure Cultivated Diversity for Natural Product Discovery Efforts
| Autor: | Bradley S. Stevenson, Emily N. Junkins |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
Microorganism Biology Microbiology DNA sequencing drug discovery 03 medical and health sciences chemistry.chemical_compound Plate count Microbiome 16S rRNA gene sequencing road-kill Original Research plate-wash PCR 030304 developmental biology 0303 health sciences Natural product 030306 microbiology business.industry cultivation efficiency high-throughput sequencing QR1-502 Antimicrobial drug Biotechnology cultivation chemistry 16s rrna gene sequencing Pure culture business |
| Zdroj: | Frontiers in Microbiology Frontiers in Microbiology, Vol 12 (2021) |
| ISSN: | 1664-302X |
| Popis: | Molecular techniques continue to reveal a growing disparity between the immense diversity of microbial life and the small proportion that is in pure culture. The disparity, originally dubbed “the great plate count anomaly” by Staley and Konopka, has become even more vexing given our increased understanding of the importance of microbiomes to a host and the role of microorganisms in the vital biogeochemical functions of our biosphere. Searching for novel antimicrobial drug targets often focuses on screening a broad diversity of microorganisms. If diverse microorganisms are to be screened, they need to be cultivated. Recent innovative research has used molecular techniques to assess the efficacy of cultivation efforts, providing invaluable feedback to cultivation strategies for isolating targeted and/or novel microorganisms. Here, we aimed to determine the efficiency of cultivating representative microorganisms from a non-human, mammalian microbiome, identify those microorganisms, and determine the bioactivity of isolates. Sequence-based data indicated that around 57% of the ASVs detected in the original inoculum were cultivated in our experiments, but nearly 53% of the total ASVs that were present in our cultivation experiments were not detected in the original inoculum. In light of our controls, our data suggests that when molecular tools were used to characterize our cultivation efforts, they provided a more complete and more complex, understanding of which organisms were present compared to what was eventually detected during cultivation. Lastly, about 3% of the isolates collected from our cultivation experiments showed inhibitory bioactivity against an already multidrug-resistant pathogen panel, further highlighting the importance of informing and directing future cultivation efforts with molecular tools. |
| Databáze: | OpenAIRE |
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