Draft Genome Sequence of Fusarium sp. Strain DS 682, a Novel Fungal Isolate from the Grass Rhizosphere

The novel fungal strain, Fusarium sp. strain DS 682, was isolated from the rhizosphere of the perennial grass, Bouteloua gracilis, at the Konza Prairie Biological Station in Kansas. This fungal strain is common across North American grasslands and is resilient to environmental fluctuations. The draft genome is estimated to be 97.2% complete.

F usarium sp. strain DS 682 (DS 682) was isolated from the rhizosphere of Bouteloua gracilis, a blue grama perennial warm season (C4) grass, at the Konza Prairie Biological Station (KPBS) in Kansas. Specifically, the strain was isolated from field site 4A as part of a study to evaluate the importance of climatic, edaphic, and host plant traits as predictors of rhizobiome community structure (1). The isolation process included surface sterilization of excised root tissue of B. gracilis and plating onto malt extract agar (MEA; Difco Laboratories, MD) amended with 50 mg/liter streptomycin and tetracycline. After a 14-day incubation at room temperature, emerging colonies with different morphologies were transferred onto fresh MEA (supplemented with antibiotics) to establish pure cultures.
DS 682 was chosen for whole-genome sequencing because it is common to soil and grass rhizospheres across North America, demonstrating metabolic adaptability to environmental fluctuations. The fungal isolate was identified as a Fusarium sp. by amplicon sequencing of the internal transcribed spacer (ITS) region using the primers ITS1F (2) and ITS4 (3) and the Web resource FUSARIUM-ID (http://isolate.fusariumdb .org/blast.php). This identification was verified and refined using additional genomic markers (see below).
Genomic DNA was prepared from DS 682 mycelium grown on MEA. Cells were lysed via sonication (Branson SFX 150; Emerson Electric, MO) and extracted by using a DNeasy PowerSoil kit (Qiagen, Inc., Germany). DNA libraries were prepared by using a NEBNext Ultra DNA library prep kit (Illumina, San Diego, CA) and validated by using a DNA 1000 Chip on the Agilent TapeStation (Agilent Technologies, CA). Sequencing was performed on the Illumina HiSeq X platform (150 bp, paired end) by Genewiz, Inc. (South Plainfield, NJ).
Comparison with Fusarium oxysporum revealed 4,934 genes (;25% of the genome) in DS 682 that did not have close similarity (amino acid similarity . 60%, E value # 10 -10 ). Of these, only 664 were assigned to 355 KO families with diverse activities.
Data availability. Sequence data have been deposited at the PNNL DataHUB repository (https://data.pnnl.gov/about) and are available for download under project data DOI accession number 10.25584/KS4AIsoGFspDS682/1635527. The version described here is the first version. The data package contains raw reads, assembly, functional annotations, MIGS.eu.soil.5.0 metadata information, and a package "Read Me" file. The genome sequence information has also been deposited under BioProject number PRJNA664411 and under BioSample accession number SAMN16213216, and this whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under accession number JACYFE000000000, where the version described here is version JACYFE010000000. Gene models are available at https://www.ncbi.nlm.nih.gov/protein ?LinkName=nuccore_protein_wgs&from_uid=1910285265.