ITS-Seq of sediment: Yalujiang estuary
The  internal transcribed spacer (ITS) genes from each sample were amplified using primers ITS1-ITS4 (ITS1: TCCGTAGGTGAACCTGCGG, ITS4: TCCTCCGCTTATTGATATGC) (Berg et al., 2012; Lu et al., 2013). All PCR reactions were carried out in 15 L reactions with 7.5 L Phusion High-Fidelity PCR Master Mix (New England Biolabs), 1 L of dNTP (2.5 mM), 1 L of forward and reverse primers (10 M), and 1 L template DNA. Thermal cycling consisted of initial denaturation at 95 C for 2 min, followed by 25 cycles of denaturation at 94 C for 5 s, annealing at 55 C for 30 s, and elongation at 72 C for 30 s. The PCR products of each sample were detected by electrophoresis on 1.5% agarose gel. Then, the obtained PCR products for ITS genes were purified, quantified, and mixed in equal amounts, respectively to construct the sequencing libraries. Finally, the libraries quality were determined using Agilent Bioanalyzer 2100 system and Qubit @2.0 Fluorometer (Thermo Scientic), and sequenced on an Illumina NovaSeq platform.