Previous observations suggested that microbial communities contribute to coral health and the ecological resilience of coral reefs. However, most studies of coral microbiology focused on prokaryotes and the endosymbiotic algae . In contrast, knowledge concerning diversity of other protists is still lacking, possibly due to methodological constraints. As most eukaryotic DNA in coral samples was derived from hosts, protist diversity was missed in metagenome analyses. To tackle this issue, we designed blocking primers for Scleractinia sequences amplified with two primer sets that targeted variable loops of the 18S rRNA gene (18SV1V2 and 18SV4). These blocking primers were used on environmental colonies of from two regions with contrasting thermal regimes (Djibouti and New Caledonia). In addition to clades A/C/D, and unidentified coccidia genera were found in many samples. In particular, coccidian sequences formed a robust monophyletic clade with other protists identified in , , , , , and coral colonies. Moreover, and coccidians had different distributions between the two geographic regions. A similar pattern was observed between clades C and A/D. Although we were unable to identify factors responsible for this pattern, nor were we able to confirm that these taxa were closely associated with corals, we believe that these primer sets and the associated blocking primers offer new possibilities to describe the hidden diversity of protists within different coral species.
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http://dx.doi.org/10.3389/fmicb.2018.02043 | DOI Listing |
Environ Microbiome
January 2025
School of Natural Sciences, Bangor University, Bangor, UK.
Background: Acquiring representative bacterial 16S rRNA gene community profiles in plant microbiome studies can be challenging due to the excessive co-amplification of host chloroplast and mitochondrial rRNA gene sequences that reduce counts of plant-associated bacterial sequences. Peptide Nucleic Acid (PNA) clamps prevent this by blocking PCR primer binding or binding within the amplified region of non-target DNA to stop the function of DNA polymerase. Here, we applied a universal chloroplast (p)PNA clamp and a newly designed mitochondria (m)PNA clamp to minimise host chloroplast and mitochondria amplification in 16S rRNA gene amplicon profiles of leaf, bark and root tissue of two oak species (Quercus robur and Q.
View Article and Find Full Text PDFCommun Chem
January 2025
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo nam. 2, CZ-16000 Prague 6, Prague, Czech Republic.
Protein-RNA interactions play important biological roles and hence reactive RNA probes for cross-linking with proteins are important tools in their identification and study. To this end, we designed and synthesized 5'-O-triphosphates bearing a reactive squaramate group attached to position 5 of cytidine or position 7 of 7-deazaadenosine and used them as substrates for polymerase synthesis of modified RNA. In vitro transcription with T7 RNA polymerase or primer extension using TGK polymerase was used for synthesis of squaramate-modified RNA probes which underwent covalent bioconjugations with amine-linked fluorophore and lysine-containing peptides and proteins including several viral RNA polymerases or HIV reverse transcriptase.
View Article and Find Full Text PDFBMC Oral Health
December 2024
Faculty of Dentistry, Innovative Dental Materials and Interfaces Research Unit (URB2i), UR 4462, Paris Cité University, 1 rue Maurice Arnoux, Montrouge, 92120, France.
Objective: To evaluate the shear bond strength (SBS) and adhesive remnant index (ARI) scores of metal brackets to glazed lithium disilicate reinforced glass-ceramics and zirconia according to various surface treatment protocols.
Methods: A total of 240 lithium disilicate ceramic (LD) and 240 zirconia (Zr) blocks were randomly divided according to sandblasting, hydrofluoric acid (HF) etching, universal primer use, and the adhesive system applied. A maxillary canine metal bracket was bonded to each sample with resin cement (Transbond XT, TXT).
bioRxiv
December 2024
Division of Pulmonary and Critical Care Medicine, Zuckerberg San Francisco General Hospital and Trauma Centre, University of California, San Francisco, San Francisco, CA, USA.
Low-frequency mutations provide valuable insights in various fields, including drug resistance identification, cancer and infectious disease research. One promising strategy to enhance the sensitivity and specificity of mutation detection is the incorporation of unique molecular identifiers (UMIs) during polymerase chain reaction (PCR) amplification and before deep sequencing. However, conventional methods for UMI incorporation often necessitate multiple labor-intensive steps.
View Article and Find Full Text PDFInt J Biol Macromol
February 2025
Department of Molecular Biology, Institute of Biochemistry and Molecular Biology, Semmelweis University, Tűzoltó street 37-47., 1094 Budapest, Hungary. Electronic address:
Streptococcus mutans is a commensal oral bacterium, yet its capacity for extensive biofilm formation is a major contributor to dental caries. This study presents a novel biofilm inhibition strategy by targeting GbpC, a cornerstone protein in S. mutans biofilm architecture, with specific DNA aptamers.
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