Draft Genome Sequences of Three Rhizospheric Plant Growth- Promoting Bacteria Olubukola Oluranti Babalola, a Ayansina Segun Ayangbenro, a Oluwaseyi Samuel Olanrewaju a a Food Security and Safety Niche Area, Faculty of Natural Science and Agriculture, North-West University, Potchefstroom, South Africa ABSTRACT Here, we report the draft genome sequences of Bacillus subtilis A1, Sph- ingobacterium sp. strain A3, and Pseudomonas sp. strain A29; Sphingobacterium sp. A3 and Pseudomonas sp. A29 were identified as Bacillus velezensis strain A3 and Ba- cillus subtilis strain A29, respectively, after a quality control check of the whole- genome sequences deposited in the NCBI database. These bacteria exhibit tremen- dous production of siderophores and significant antimicrobial potential. When inoculated on maize, these isolates increase its yield. P lant growth promoting-rhizobacteria (PGPR) have become so important to agricul- tural sustainability and food security. The increasing human population and haz- ardous impact of chemical fertilizers on the environment have favored the use of PGPR as biofertilizer and biocontrol agents. Bacillus subtilis A1, Bacillus velezensis strain A3, and Bacillus subtilis strain A29, reported here, were isolated from maize rhizosphere from a maize field (25°49=S, 27°5=E) in Mafikeng, South Africa. For the isolation of rhizobacteria, 10 g of rhizospheric soil was suspended in 90 ml sterile distilled water. The serial dilution method was used for further analysis of the prepared soil suspension. Suitable dilutions (10 -2 , 10 -4 , and 10 -6 ) were plated in triplicate onto Luria-Bertani (LB) agar (Sigma-Aldrich) to isolate rhizobacteria using standard microbiological isolation techniques, and plates were incubated at 25°C for 24 h. Bacterial colonies were subcultured and purified by streaking onto fresh LB agar plates. These isolates were first identified by 16S rRNA sequencing. The genomic DNA was extracted from overnight cultures in LB medium (1) using a ZR soil microbe DNA MiniPrep extraction kit (Zymo Research, USA), following the manufacturer’s instructions. The DNA quality and quantity were determined using a NanoDrop Lite spectrophotometer (Thermo Fisher Scientific, CA, USA). The genomes of the strains were sequenced on an Illumina HiSeq sequencer at Molecular Research (MR DNA), Shallowater, TX. The libraries were prepared using Kapa HyperPlus kits (Roche), following the manufacturer’s user guide. The initial concentration of DNA was evaluated using the Qubit double-stranded DNA (dsDNA) high-sensitivity (HS) assay kit (Life Technologies). A total of 25 ng of DNA was used to prepare the libraries. The protocol starts with enzymatic fragmentation to produce dsDNA fragments, fol- lowed by end repair and A-tailing to produce end-repaired 5=-phosphorylated 3=-deoxyribosyladenine (dA)-tailed dsDNA fragments. In the adapter ligation step, dsDNA adapters are ligated to 3=-dA-tailed molecules. The final step is library amplifi- cation, which employs high-fidelity, low-bias PCR to amplify library fragments carrying appropriate adapter sequences on both ends. Following the library preparation, the final concentrations of the libraries were measured using the Qubit dsDNA HS assay kit (Life Technologies), and the average library size was determined using the Agilent 2100 Bioanalyzer (Agilent Technologies). Bacillus subtilis A1, Bacillus velezensis strain A3, and Bacillus subtilis strain A29 DNA concentrations are 114.0, 84.8, and 187.0 ng/l, respec- Citation Babalola OO, Ayangbenro AS, Olanrewaju OS. 2019. Draft genome sequences of three rhizospheric plant growth-promoting bacteria. Microbiol Resour Announc 8:e00455- 19. https://doi.org/10.1128/MRA.00455-19. Editor Steven R. Gill, University of Rochester School of Medicine and Dentistry Copyright © 2019 Babalola et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Olubukola Oluranti Babalola, Olubukola.Babalola@nwu.ac.za. Received 24 April 2019 Accepted 19 May 2019 Published 27 June 2019 GENOME SEQUENCES crossm Volume 8 Issue 26 e00455-19 mra.asm.org 1 on April 13, 2020 by guest http://mra.asm.org/ Downloaded from