Seagrass leaf reddening alters the microbiome of Zostera muelleri

2020 ◽  
Vol 646 ◽  
pp. 29-44
Author(s):  
V Hurtado-McCormick ◽  
T Kahlke ◽  
D Krix ◽  
A Larkum ◽  
PJ Ralph ◽  
...  
Keyword(s):  
Significant Proportion ◽  
Amplicon Sequencing ◽  
High Irradiance ◽  
Rrna Gene ◽  
Core Microbiome ◽  
Microbiome Composition ◽  
Operational Taxonomic Units ◽  
Normal Green ◽  
White Tissue ◽  
Leaf Reddening

Seagrasses host an extremely diverse microbiome that plays fundamental roles in seagrass health and productivity but may be sensitive to shifts in host physiology. Here, we observed a leaf reddening phenomenon in Zostera muelleri and characterized bacterial assemblages associated with green and reddened leaves to determine whether this change in leaf pigmentation stimulates shifts in the seagrass microbiome. Using 16S rRNA gene amplicon sequencing, we observed that the microbiome associated with 4 different leaf pigmentation categories (i.e. green, white, purple and black) differed significantly, with substantial changes in microbiome composition when the tissue is whitened (non-pigmented). Actinobacteria, Rhodobacteraceae, Erythrobacter, Sulfitobacter and Granulosicoccus were enriched in black and/or purple tissues and discriminated these microbiomes from those associated with green leaves. Contrastingly, all ‘discriminatory’ zero-radius operational taxonomic units (zOTUs) were depleted within the communities associated with white samples. While 40% of the abundant zOTUs identified were exclusively associated with a single pigmentation category, only 3% were shared across all categories, indicating partitioning of the phyllosphere microbiome. However, a significant proportion of the ‘normal’ (green) leaf core microbiome was also retained in the core communities associated with black (70%) and purple (70%) tissues. Contrastingly, no core zOTUs were maintained in the white tissues. These results indicate that environmentally driven physiological shifts in seagrasses, such as leaf reddening expressed in response to high irradiance, can impact the seagrass leaf microbiome, resulting in significant shifts in the microbiome of reddened leaves with the most extreme expression (in white tissue of reddened leaves).

scholarly journals Handling of spurious sequences affects the outcome of high-throughput 16S rRNA gene amplicon profiling

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Sandra Reitmeier ◽  
Thomas C. A. Hitch ◽  
Nicole Treichel ◽  
Nikolaos Fikas ◽  
Bela Hausmann ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Diversity Analysis ◽  
Careful Attention ◽  
Operational Taxonomic Units ◽  
Basic Concepts ◽  
Gnotobiotic Mice ◽  
Mock Communities

Abstract16S rRNA gene amplicon sequencing is a popular approach for studying microbiomes. However, some basic concepts have still not been investigated comprehensively. We studied the occurrence of spurious sequences using defined microbial communities based on data either from the literature or generated in three sequencing facilities and analyzed via both operational taxonomic units (OTUs) and amplicon sequence variants (ASVs) approaches. OTU clustering and singleton removal, a commonly used approach, delivered approximately 50% (mock communities) to 80% (gnotobiotic mice) spurious taxa. The fraction of spurious taxa was generally lower based on ASV analysis, but varied depending on the gene region targeted and the barcoding system used. A relative abundance of 0.25% was found as an effective threshold below which the analysis of spurious taxa can be prevented to a large extent in both OTU- and ASV-based analysis approaches. Using this cutoff improved the reproducibility of analysis, i.e., variation in richness estimates was reduced by 38% compared with singleton filtering using six human fecal samples across seven sequencing runs. Beta-diversity analysis of human fecal communities was markedly affected by both the filtering strategy and the type of phylogenetic distances used for comparison, highlighting the importance of carefully analyzing data before drawing conclusions on microbiome changes. In summary, handling of artifact sequences during bioinformatic processing of 16S rRNA gene amplicon data requires careful attention to avoid the generation of misleading findings. We propose the concept of effective richness to facilitate the comparison of alpha-diversity across studies.

scholarly journals Uncovering the Core Microbiome and Distribution of Palmerolide in Synoicum adareanum Across the Anvers Island Archipelago, Antarctica

Marine Drugs ◽  
2020 ◽  
Vol 18 (6) ◽  
pp. 298
Author(s):  
Alison E. Murray ◽  
Nicole E. Avalon ◽  
Lucas Bishop ◽  
Karen W. Davenport ◽  
Erwan Delage ◽  
...  
Keyword(s):  
Marine Invertebrate ◽  
Bioactive Compound ◽  
Rrna Gene ◽  
Sequence Variants ◽  
Specific Sequence ◽  
Core Microbiome ◽  
Microbiome Composition ◽  
The Core ◽  
Synoicum Adareanum ◽  
Palmerolide A

Polar marine ecosystems hold the potential for bioactive compound biodiscovery, based on their untapped macro- and microorganism diversity. Characterization of polar benthic marine invertebrate-associated microbiomes is limited to few studies. This study was motivated by our interest in better understanding the microbiome structure and composition of the ascidian, Synoicum adareanum, in which palmerolide A (PalA), a bioactive macrolide with specificity against melanoma, was isolated. PalA bears structural resemblance to a hybrid nonribosomal peptide-polyketide that has similarities to microbially-produced macrolides. We conducted a spatial survey to assess both PalA levels and microbiome composition in S. adareanum in a region of the Antarctic Peninsula near Anvers Island (64°46′ S, 64°03′ W). PalA was ubiquitous and abundant across a collection of 21 ascidians (3 subsamples each) sampled from seven sites across the Anvers Island Archipelago. The microbiome composition (V3–V4 16S rRNA gene sequence variants) of these 63 samples revealed a core suite of 21 bacterial amplicon sequence variants (ASVs)—20 of which were distinct from regional bacterioplankton. ASV co-occurrence analysis across all 63 samples yielded subgroups of taxa that may be interacting biologically (interacting subsystems) and, although the levels of PalA detected were not found to correlate with specific sequence variants, the core members appeared to occur in a preferred optimum and tolerance range of PalA levels. These results, together with an analysis of the biosynthetic potential of related microbiome taxa, describe a conserved, high-latitude core microbiome with unique composition and substantial promise for natural product biosynthesis that likely influences the ecology of the holobiont.

scholarly journals The Wheat Microbiome Under Four Management Strategies, and Potential for Endophytes in Disease Protection

2017 ◽  
Vol 1 (3) ◽  
pp. 158-168 ◽  
Author(s):  
Kristi Gdanetz ◽  
Frances Trail
Keyword(s):  
Bacterial Communities ◽  
Management Strategy ◽  
Crop Yields ◽  
Management Strategies ◽  
Growing Season ◽  
Amplicon Sequencing ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
Operational Taxonomic Units ◽  
Plant Organ

Manipulating plant-associated microbes to reduce disease or improve crop yields requires a thorough understanding of interactions within the phytobiome. Plants were sampled from a wheat/maize/soybean crop rotation site that implements four different crop management strategies. We analyzed the fungal and bacterial communities of leaves, stems, and roots of wheat throughout the growing season using 16S and fungal internal transcribed spacer 2 rRNA gene amplicon sequencing. The most prevalent operational taxonomic units (OTUs) were shared across all samples, although levels of the low-abundance OTUs varied. Endophytes were isolated from plants, and tested for antagonistic activity toward the wheat pathogen Fusarium graminearum. Antagonistic strains were assessed for plant protective activity in seedling assays. Our results suggest that microbial communities were strongly affected by plant organ and plant age, and may be influenced by management strategy.

scholarly journals Comparison of paperpoint and curette sampling of subgingival microbiome composition as analyzed by 16S rRNA gene amplicon sequencing

2017 ◽  
Vol 9 (sup1) ◽  
pp. 1325260
Author(s):  
K. Beyer ◽  
B.W. Brandt ◽  
M.J. Buijs ◽  
J.G. Brun ◽  
W. Crielaard ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Microbiome Composition

scholarly journals A Reference Map of Microbiome Composition in Crop Production Across 8 Chinese Provinces

2021 ◽  
Author(s):  
Haiyang Hu ◽  
Yunli Liu ◽  
Zhaoyong Xu ◽  
Yiqun Huang ◽  
Hongzhi Tang
Keyword(s):  
Crop Production ◽  
Research Area ◽  
Core Microbiome ◽  
Microbiome Composition ◽  
Operational Taxonomic Units ◽  
Sampling Locations ◽  
Rdna Sequencing ◽  
Microbial Community Structures ◽  
Reference Map ◽  
New Research

Abstract Background: Although microbiome profiling of crop leaves is a relatively new research area, we know that leaf microorganism communities affect the leaf chemical composition, promote growth, and confer protection against deleterious microbes.Results: Here, we used 16S rDNA sequencing to profile the microbiomes of 78 primary dried tobacco leaf samples from plants grown at 26 locations in 8 Chinese provinces. After analyzing leaf microbial communities and identifying 4,473 operational taxonomic units (OTU) representing 1,234 species, we obtained a national core microbiome with 14 OTU representing 9 species, with functions for processes including nitrogen fixation, detoxification of diverse pollutants, and heavy-metal reduction. Although there are no obvious impacts from single environmental factors (e.g., temperature, precipitation), there are significant differences in microbial community structures among provinces and clear clustering by region. Finally, LEfSe analysis screened the significantly different species between different sampling locations.Conclusions: Our study demonstrates that both climactic and industrial factors drive microbial diversity on crop plant leaves, and many potentially useful functional bacterial resources present on tobacco leaves are still waiting for harnessing for a variety of bioprocess, agricultural, and environmental detoxification applications.

Endophytic bacterial communities and spatiotemporal variations in cotton roots in Xinjiang, China

2020 ◽  
Author(s):  
Yingwu Shi ◽  
Hongmei Yang ◽  
Min Chu ◽  
Xinxiang Niu ◽  
Xiangdong Huo ◽  
...  
Keyword(s):  
Community Structure ◽  
Endophytic Bacteria ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Bacterial Populations ◽  
Operational Taxonomic Units ◽  
Cotton Sample ◽  
Endophytic Bacterial Community ◽  
V3 Region ◽  
Illumina Amplicon Sequencing

Endophytic bacteria may be important for plant health and other ecologically relevant functions of cotton. However, the endophytic bacterial community structure and diversity in cotton is still poorly characterized. We investigated the community structure of endophytic bacteria in cotton roots growing in Xinjiang, China, using the Illumina amplicon sequencing. A total of 60.84 M effective sequences of 16S rRNA gene V3 region were obtained from cotton samples. These sequences revealed huge amount of operational taxonomic units (OTUs) in cotton, that is, 81-338 OTUs in a cotton sample, at 3% cutoff level and sequencing depth of 50000 sequences. We identified 23 classes from the resulting 2,723,384 sequences. Gammaproteobacteria were the dominant class in all cottons, followed by Alphaproteobacteria, Actinobacteria and Bacilli. A marked difference in the diversity of endophytic bacteria in cotton for different growth periods was evident. The greatest number of OTUs was detected during seedling (654 OTUs) and budding (381 OTUs). Endophytic bacteria diversity was reduced during flowering (350 OTUs) and boll-opening (351 OTUs). 217 OTUs were common to all four periods. There were more tags of Pantoea in Shihezi than other locations. While there were more tags of Erwinia in Hami than other locations. The dynamics of endophytic bacteria communities were influenced by plant growth stage. These results show the complexity of the bacterial populations present in inner tissues of cotton.

scholarly journals Specialized microbiome of the cave-dwelling sponge Plakina kanaky (Porifera, Homoscleromorpha)

2020 ◽  
Vol 96 (4) ◽  
Author(s):  
César Ruiz ◽  
Marcela Villegas-Plazas ◽  
Olivier P Thomas ◽  
Howard Junca ◽  
Thierry Pérez
Keyword(s):  
Extreme Environments ◽  
Amplicon Sequencing ◽  
Sponge Species ◽  
Rrna Gene ◽  
Microbiome Composition ◽  
16S Amplicon Sequencing ◽  
Ecological Success ◽  
Specific Community ◽  
First Time ◽  
The 16S Rrna Gene

ABSTRACT The recent description of the polychromatic sponge Plakina kanaky revealed original microsymbionts, with some morphotypes recorded for the first time in Homoscleromorpha and others never before observed in other sponge groups. Illumina 16S amplicon sequencing was used to characterize this microbial community by comparing contents of seven specimens of this Plakinidae with five other sponge species: one Homoscleromopha of the Oscarellidae family and four Demospongiae. A total of 256 458 sequences of the hypervariable V5-V6 region of the 16S rRNA gene were clustered into 2,829 OTUs at 97% similarity, with Proteobacteria, Poribacteria and Chloroflexi being the most abundant phyla. The Plakina kanaky specific community appeared to be mainly composed by five OTUs representing about 10% of the total microbiome. Among these, the filamentous bacterium Candidatus Entotheonella, which was among the dominant morphotypes previously observed in the mesohyl and the larvae of P. kanaky, was detected in all studied specimens. However, other original and dominant morphotypes could not be assigned to a known prokaryotic taxon. This cave dwelling sponge species harbors a distinctive microbiome composition of potential taxonomic and metabolic novelties that may be linked to its ecological success in such extreme environments.

scholarly journals The Oral Microbiome of Healthy Japanese People at the Age of 90

2020 ◽  
Vol 10 (18) ◽  
pp. 6450 ◽  
Author(s):  
Yoshiaki Nomura ◽  
Erika Kakuta ◽  
Noboru Kaneko ◽  
Kaname Nohno ◽  
Akihiro Yoshihara ◽  
...  
Keyword(s):  
Human Microbiome ◽  
Oral Microbiome ◽  
Rrna Gene ◽  
Core Microbiome ◽  
Operational Taxonomic Units ◽  
The Core ◽  
Hypervariable Regions ◽  
Study Population ◽  
Granulicatella Adiacens

For a healthy oral cavity, maintaining a healthy microbiome is essential. However, data on healthy microbiomes are not sufficient. To determine the nature of the core microbiome, the oral-microbiome structure was analyzed using pyrosequencing data. Saliva samples were obtained from healthy 90-year-old participants who attended the 20-year follow-up Niigata cohort study. A total of 85 people participated in the health checkups. The study population consisted of 40 male and 45 female participants. Stimulated saliva samples were obtained by chewing paraffin wax for 5 min. The V3–V4 hypervariable regions of the 16S ribosomal RNA (rRNA) gene were amplified by PCR. Pyrosequencing was performed using MiSeq. Operational taxonomic units (OTUs) were assigned on the basis of a 97% identity search in the EzTaxon-e database. Using the threshold of 100% detection on the species level, 13 species were detected: Streptococcus sinensis, Streptococcus pneumoniae, Streptococcus salivarius, KV831974_s, Streptococcus parasanguinis, Veillonella dispar, Granulicatella adiacens, Streptococcus_uc, Streptococcus peroris, KE952139_s, Veillonella parvula, Atopobium parvulum, and AFQU_vs. These species represent potential candidates for the core make-up of the human microbiome.

The effects of captivity on the microbiome of the endangered Comal Springs riffle beetle (Heterelmis comalensis)

2021 ◽  
Vol 368 (17) ◽  
Author(s):  
Zachary Mays ◽  
Amelia Hunter ◽  
Lindsay Glass Campbell ◽  
Camila Carlos-Shanley
Keyword(s):  
16S Rrna ◽  
Gut Microbiome ◽  
Amplicon Sequencing ◽  
Conservation Education ◽  
Rrna Gene ◽  
Sequence Variants ◽  
Microbiome Composition ◽  
Intrinsic Factors ◽  
Captive Populations ◽  
The 16S Rrna Gene

Abstract The gut microbiome is affected by host intrinsic factors, diet and environment, and strongly linked to host's health. Although fluctuations of microbiome composition are normal, some are due to changes in host environmental conditions. When species are moved into captive environments for conservation, education or rehabilitation, these new conditions can influence a change in gut microbiome composition. Here, we compared the microbiomes of wild and captive Comal Springs riffle beetles (Heterelmis comalensis) by using amplicon sequencing of the 16S rRNA gene. We found that the microbiome of captive beetles was more diverse than wild beetle microbiomes. We identified 24 amplicon sequence variants (ASVs) with relative abundances significantly different between the wild and captive beetles. Many of the ASVs overrepresented in captive beetle microbiomes belong to taxa linked to nitrogen-rich environments. This is one of the first studies comparing the effects of captivity on the microbiome of an endangered insect species. Our findings provide valuable information for future applications in the management of captive populations of H. comalensis.

scholarly journals Host Species Determines the Composition of the Prokaryotic Microbiota in Phlebotomus Sandflies

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 428
Author(s):  
Christos Papadopoulos ◽  
Panagiotis A. Karas ◽  
Sotirios Vasileiadis ◽  
Panagiota Ligda ◽  
Anastasios Saratsis ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Host Species ◽  
Archaeal Community ◽  
Amplicon Sequencing ◽  
Structural Effect ◽  
Rrna Gene ◽  
Host Genotype ◽  
Operational Taxonomic Units ◽  
Phlebotomine Sandflies ◽  
Leishmania Spp

Phlebotomine sandflies are vectors of the humans’ and mammals’ parasite Leishmania spp. Although the role of gut microbiome in the biological cycle of insects is acknowledged, we still know little about the factors modulating the composition of the gut microbiota of sandflies. We tested whether host species impose a strong structural effect on the gut microbiota of Phlebotomus spp. Sandflies were collected from the island of Leros, Greece, and classified to P. papatasi, P. neglectus, P. tobbi, and P. similis, all being negative to Leishmania spp. The prokaryotic gut microbiota was determined via 16S rRNA gene amplicon sequencing. Phlebotomus species supported distinct microbial communities (p < 0.001). P. papatasi microbiota was the most distinct over-dominated by three Spiroplasma, Wolbachia and Paenibacillus operational taxonomic units (OTUs), while another Wolbachia OTU prevailed in P. neglectus. Conversely, the microbiota of P. tobbi and P. similis was composed of several less dominant OTUs. Archaea showed low presence with the dominant OTUs belonging to methanogenic Euryarcheota, ammonia-oxidizing Thaumarcheota, and Nanoarchaeota. We provide first insights into the composition of the bacterial and archaeal community of Phlebotomus sandflies and showed that, in the absence of Leishmania, host genotype is the major modulator of Phlebotomus sandfly gut microbiota.

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