title: Metagenome of Fragaria x ananassa:cultivar Meahang
design: The flower and pollen samples were stored in a portable refrigerator after collection and transported to the laboratory, where each sample was weighed and adjusted to 1 g. For flower samples, 30 mL of 1 X PBS buffer (10 X PBS: 8 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, 0.24 g of KH2PO4, adjusted to a pH of 7.4 and a final volume of 1 L) was added. Pollen samples received 20 mL of 1 X PBS buffer. Then the sample tubes were sonicated at 35 kHz (20 min, 4C) to detach microbes and 3 mL of the mixture was centrifuged (12,470 x g, 10 min, 4C). The supernatant was discarded and total DNA in the pellet was purified by using a Genomic DNA isolation kit (Solgent, Korea). Gram-negative and Gram-positive bacterial DNA was purified by different protocols according to the methods of the manufacturer. To check the quality of the DNA samples and determine the composition of the communities, 16S rDNA PCR reactions were conducted. Total DNA (100 ng) was reacted with the primer pair 27 mF (5-gagtttgatcmtggctcag-3) and 518 R (5-wttaccgcggctgctgg-3), which primer pairs was amplified V1-V3 region (Yoo et al., 2016). After PCR amplification, which samples were subtracted of background and chimeric. The sample was on the PicoTiter plate and extract the raw signals. The signal was to balance with signal strengths of the different nucleotides and emulsion of background signal. After filteration signals sorting by tag sequence (barcode sequences, which was used at preparation of the sequencing libraries. The sequencing reads were checking both of the nucleotide quality scores (average Phred score >20) and read lengths (>300bp) (Supplementary Data Table 5-8). Rerefaction curve was calculated with iNEXT (version 2.0.12) in R (version 3.4.4) (Extended Data Fig 4-5). Operational taxonomic units (OTUs) were established using CD-HIT- OUT clusters rRNA tags and Mother (ver 1.33.0). And nucleotide similarity of 16sRNA sequences were flowed at smilimarity levels : phylum, >75%; class, >80%; order, >85%; family, >90%; genus, >94% ; species, >97%. Pyrosequencing was performed using a 454 GS-FLX titanium system with a Roche Genome Sequencer (GS) FLX software (v 3.0). The rarefaction curves, microbial communities, top Operational Taxonomic Units (OTUs) and Principal Coordinates Analyses (PCoA) were conducted with QIIME.