Complete Genome Sequence of a Pseudomonas simiae Strain with Biocontrol Potential against Aphanomyces Root Rot

Aphanomyces euteiches is a soilborne plant pathogen. It causes severe root rot in leguminous crop species. We report the complete genome sequence of a biocontrol strain, Pseudomonas simiae K-Hf-L9. The strain inhibited Aphanomyces euteiches mycelia and zoospores and suppressed root rot in field peas grown under controlled growth chamber conditions.

T he genus Pseudomonas includes species that demonstrate a great deal of metabolic diversity and are found in many habitats (1). Strains of Pseudomonas simiae are reported to exhibit biocontrol activity against fungal and oomycete pathogens via siderophore-mediated competition for iron and induction of systemic resistance in the host plant (2,3). Pseudomonas simiae strain K-Hf-L9 was isolated from the soil of a pea field crop in Saskatchewan, Canada (4).
Pseudomonas simiae strain K-Hf-L9 was inoculated from frozen stock (50% glycerol, in 280°C storage) on half-strength Trypticase soy agar plates (half-strength Difco Trypticase soy broth plus 1.5% agar) and grown for 24 h at 28°C. Genomic DNA was extracted using the FastDNA Spin kit (MP Biomedicals) according to the manufacturer's instructions. Genomic DNA library preparation and whole-genome sequencing were performed at the University of Regina (Regina, Canada). Genomic DNA was sheared using a g-TUBE (Covaris, Woburn, MA, USA) and a centrifugation protocol yielding an average DNA fragment size of approximately 8,000 bases (Eppendorf 5424 centrifuge and two 30-s centrifugations at 2,000 Â g). A library was prepared for Nanopore sequencing using the ligation sequencing kit (SQK-LSK109) together with the native barcoding expansion kit (EXP-NBD103; Oxford Nanopore Technologies). Sequencing was performed using a MinION Mk1B device and a FLO-MIN106D (R9.4.1) flow cell. The fast5 files generated by the run were base called and demultiplexed after sequencing using Guppy version 4.4.1 and the high-accuracy (HAC) model.
Data availability. This whole-genome shotgun project has been deposited in GenBank under accession number CP066169.2, BioProject number PRJNA662093, and BioSample number SAMN16076439. The raw reads are published in the SRA database under the accession number PRJNA662093.

ACKNOWLEDGMENTS
Financial support from a Saskatchewan Agriculture Development Fund (ADF) grant is gratefully acknowledged.
We thank Nikki Burnett and Kara Loos for providing technical support in DNA extraction and preparing the library for DNA sequencing.
We declare no conflicts of interest.