V. volvacea strains were routinely grown at 32 °C on potato dextrose agar medium (PDA) (200 g.L-1 of potato tissue, 20 g.L-1 glucose,20 g.L-1 agar). Liquid cultures were prepared in potato dextrose broth by shaking at 150 rpm for 7 days.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from mycelia using pBIOZOL Plant Total RNA Extraction Reagent, according to the manufacturer’s protocol (BioFlux). The isolated RNA was treated with RNeasy plant mini kit to remove potential genomic DNA contamination, according to the manufacturer’s protocol (QIAGEN, Germany). RNA integrity and concentration were evaluated using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). RNA (6 μg) from each of the three strains was submitted to BGI-Shenzhen (Shenzhen, China) for library construction and sequencing. Sequence tag preparation was carried out using the Illumina Gene Expression Sample Prep Kit according to the manufacturer’s protocol. library strategy: 3'-tag digital gene expression (DGE)
Library strategy
RNA-Seq
Library source
transcriptomic
Library selection
cDNA
Instrument model
Illumina HiSeq 2000
Description
Sample 1
Data processing
Illumina Cluster Station and Illumina HiSeq™ 2000 instrument used for basecalling. Raw sequences have 3' adaptor fragments as well as a few low-quality sequences and several types of impurities. Raw sequences are transformed into Clean Tags after certain steps of data-processing. A virtual libraries containing all the possible CATG+17 bases length sequences of the reference gene sequences. All clean tags were mapped to the reference sequences and only 1bp mismatch is considered. Clean tags mapped to reference sequences from multiple genes were filtered. Remainder clean tags were designed as unambiguous clean tags. The number of unambiguous clean tags for each gene was calculated and then normalized to TPM (number of transcripts per million clean tags). ('t Hoen, Ariyurek, et al. 2008; Morrissy, Morin, et al. 2009). Genome_build: V.volvacea strain PYd21 genome data Supplementary_files_format_and_content: tab-delimited text files include tag sequences,tag number and TPM values for each strain.
