Molecular diversity and biogeography of benthic microeukaryotes in temperate seagrass (Zostera japonica) systems of northern China
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Abstract: The productivity and health of seagrass depend on the combined inputs of nutrients from the water and sediments in which they grow and the microbiota with which they live intimately. However, little is known about the composition and diversity pattern of single-celled benthic eukaryotes in seagrass meadows. Here, we investigated how the structure and diversity of the benthic microeukaryotic community vary with respect to season, location, and seagrass colonization, by applying 18S rRNA gene amplicon sequencing for 96 surface sediment samples that were collected from three different seagrass habitats through four seasons. We found that benthic microeukaryotic communities associated with seagrass Zostera japonica exhibited remarkable spatial and seasonal variations, as well as differences between vegetated and unvegetated sediments. Diatoms and dinoflagellates predominated in the benthic microeukaryotic communities, but they were inversely correlated and displaced each other as the dominant microbial group in different seasons or habitats. Mucoromycota was more prevalent in vegetated sediments, whereas Lobulomycetales and Chytridiales had higher proportions in unvegetated sites. Total organic carbon and total organic nitrogen were the most important environmental factors in driving the microeukaryotic assemblages and diversity. Our study expands the available knowledge on the biogeographic distribution patterns and niche preferences for benthic microeukaryotes in seagrass systems.
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Figure 1. Geographical locations of the three Zostera japonica seagrass meadows (DL: Dalian; DY: Dongying; WH: Weihai) distributing along the coast of the Yellow Sea and the Bohai Sea.
Figure 2. Venn diagrams showing the shared and unique OTU number in sediments among three sampling locations (a), four seasons (b), and seagrass vegetated (V) and unvegetated (U) samples (c). Taxonomic composition of seagrass Zostera japonica-associated microeukaryotic communities revealed by Miseq sequencing of 18S rRNA genes (d). The percentages in parentheses indicate the proportion of a given OTU number to the total. MAST: marine stramenopile.
Figure 3. Comparisons of microeukaryotic α-diversity estimators of seagrass-vegetated (V) and unvegetated (U) sediments collected from Dongying (DY), Dalian (DL), and Weihai (WH) across four seasons. The P values are given for the comparison among seasons and locations using one-way ANOVA with least significance difference post hoc, while difference between vegetated and unvegetated was examined using t-test. Different letters above the box indicate significant differences among groups.
Figure 4. Plots of non-metric multidimensional scaling (NMDS) based on the Bray-Curtis distance, showing the variations of β-diversity of the microeukaryotic community from seagrass-vegetated (solid, V) and unvegetated (hollow, U) sediments in spatial scale (a), as well as seasonal patterns within each habitat: b in Weihai (WH); c in Dongying (DY), d in Dalian (DL). The shapes of points represent different location samples: DL, diamond; DY, triangle; and WH, round. And samples are colored by their corresponding seasons: spring, green; summer, pink; autumn, blue; and winter, black.
Figure 5. Redundancy analysis (RDA) ordination diagram (a) and heatmap of Spearman’s correlation (b) showing the relationships of geochemical variables with microeukaryotic community structure and relative abundances of major taxa, respectively. Letters in sample IDs mean the vegetated (V, solid) and unvegetated (U, hollow) samples, and shapes of points represent different season samples: spring, round; summer, triangle; autumn, square; and winter, diamond. Samples are colored by habitat types: DL, Dalian (sandy coast, blue); DY, Dongying (muddy coast, red); and WH, Weihai (lagoon, black). Only vectors for environmental variables with P-values of 0.05 or smaller are shown in a and b. The values of Spearman’s correlation coefficients are indicated according to the scale bar. MAST is the abbreviation of marine stramenopile; TOC:TON, the contents ratio between total organic carbon and organic nitrogen.
Table 1. ANOSIM testing the differences of benthic microeukaryotic communities between the two sediment types and four seasons within three seagrass beds, based on Bray-Curtis distance
Groups DL DY WH R P R P R P Global test 0.553 0.001 0.445 0.001 0.381 0.001 Spring vs. Summer 0.185 0.040 0.301 0.008 0.199 0.039 Spring vs. Autumn 0.568 0.001 0.451 0.001 0.658 0.001 Spring vs. Winter 0.486 0.002 0.491 0.002 0.455 0.001 Summer vs. Autumn 0.751 0.001 0.547 0.001 0.440 0.002 Summer vs. Winter 0.316 0.011 0.693 0.001 0.458 0.002 Autumn vs. Winter 0.901 0.001 0.248 0.015 0.081 0.125 Vegetated vs. Unvegetated 0.195 0.001 0.235 0.005 0.221 0.002 Note: Significant P-values (≤0.05) are highlighted in bold. 
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Table 2. Spearman rank correlations between microeukaryotic α-diversity estimators and environment variables
Variable OTU richness Chao1 Simpson Shannon $\rho $ P $\rho $ P $\rho $ P $\rho $ P Temperature 0.35 0.001 0.23 0.029 0.33 0.001 0.37 <0.001 DO
concentration−0.22 0.034 −0.21 0.046 – – −0.21 0.043 pH −0.23 0.028 – – −0.21 0.044 −0.22 0.032 Grain size – – 0.22 0.034 – – – – ${{\rm {NH}}_4^+} $-N content −0.33 0.001 −0.34 0.001 −0.23 0.026 −0.28 0.007 ${{\rm {SO}}_4^{2-}} $ content – – – – 0.21 0.044 0.26 0.012 TOC content 0.29 0.005 0.22 0.031 0.28 0.005 0.31 0.003 TON content 0.33 0.001 0.26 0.010 0.33 0.001 0.36 <0.001 V content – – – – 0.21 0.047 – – Cr content – – – – 0.21 0.045 0.21 0.038 Note: Only the significant correlations (P≤0.05) are shown. OTU is the abbreviation of operational taxonomic unit. $\rho $ represents Spearman rank correlation coefficient; –, no data.
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