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. 2015 Jun 18;16(6):14039-55.
doi: 10.3390/ijms160614039.

Fungal Community Successions in Rhizosphere Sediment of Seagrasses Enhalus acoroides under PAHs Stress

Juan Ling  1   2 , Yanying Zhang  3   4 , Meilin Wu  5 , Youshao Wang  6 , Junde Dong  7   8 , Yufeng Jiang  9   10 , Qingsong Yang  11   12 , Siquan Zeng  13   14
Affiliations

Fungal Community Successions in Rhizosphere Sediment of Seagrasses Enhalus acoroides under PAHs Stress

Juan Ling et al. Int J Mol Sci. .

Abstract

Seagrass meadows represent one of the highest productive marine ecosystems and are of great ecological and economic values. Recently, they have been confronted with worldwide decline. Fungi play important roles in sustaining the ecosystem health as degraders of polycyclic aromatic hydrocarbons (PAHs), but fewer studies have been conducted in seagrass ecosystems. Hence, we investigated the dynamic variations of the fungal community succession under PAH stress in rhizosphere sediment of seagrasses Enhalus acoroides in this study. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), quantitative PCR (qPCR) and a clone library have been employed to analyze the fungal community's shifts. Sequencing results of DGGE and the clone library showed that the predominant species belong to phyla Ascomycota and Basidiomycota. The abundance of three groups decreased sharply over the incubation period, whereas they demonstrated different fungal diversity patterns. Both the exposure time and the PAH concentrations affected the microbial diversity as assessed by PCR-DGGE analysis. Redundancy analysis (RDA) indicated that significant factors driving community shifts were ammonium and pH (p < 0.05). Significant amounts of the variations (31.1%) were explained by pH and ammonium, illustrating that those two parameters were the most likely ones to influence or be influenced by the fungal communities' changes. Investigation results also indicated that fungal communities in seagrass meadow were very sensitive to PAH-induced stress and may be used as potential indicators for the PAH contamination.

Keywords: PAHs (polycyclic aromatic hydrocarbons); PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis); RDA (redundancy analysis); fungi; qPCR (quantitative PCR); seagrass.

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Figures

Figure 1
Figure 1
DGGE (denaturing gradient gel electrophoresis) profiles of sediment fungal communities exposed to different concentrations of PAH (polycyclic aromatic hydrocarbon) contamination at different incubation stages (S: Day 0; A: Day 2, B: Day 7, C: Day 14, D: Day 28; 0: control without PAH addition; 1: 100 mg/kg; 2: 1000 mg/kg) (A); neighbor-joining phylogenetic tree based on 18S rRNA gene sequences from DGGE bands. Bootstrap analysis was based on 1000 replicates. Bootstrap values from distance analysis are depicted. Bootstrap values less than 50% are not shown (B).
Figure 2
Figure 2
Shannon index for the fungal communities based on the DGGE profile (A) and the dynamics of fungal abundance 18S rDNA gene copy number during the incubation (B). (S: Day 0; A: Day 2; B: Day 7; C: Day 14; D: Day 28; 0: control without PAH addition; 1: 100 mg·kg−1; 2: 1000 mg·kg−1).
Figure 3
Figure 3
Neighbor-joining phylogenetic tree based on 18S rRNA gene sequences based on the representative clone of each Operational Taxonomic Units (OTU) (cutoff = 0.03) of Seagrass Fungi (SF) clone library Numbers in parentheses indicate clones in each OUT. Bootstrap analysis was based on 1000 replicates. Bootstrap values from distance analysis are depicted. Bootstrap values less than 50% are not shown.
Figure 4
Figure 4
DGGE band data redundancy analysis for fungal communities. Significant composting parameters are indicated by solid lines with filled arrows, while supplementary parameters are shown using gray dotted lines with unfilled arrows. Samples with the same symbol were collected on the same date.
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