Draft genome sequence of a mosquito repellent Bacillus licheniformis strain Ba1 isolated from desert soil

ABSTRACT Microbial volatile organic compounds have been shown to affect a wide insect behavior. In this paper, we report the draft genome sequence of Bacillus licheniformis strain Ba1 previously isolated from desert soil in Morocco. The assembled and annotated draft genome contains 4,726 coding genes, 6 rRNAs and 97 tRNAs.

ubiquitous distribution in the environment (1).Its versatile and expanding industrial applications are well-recognized.
Microbial volatile organic compounds (mVOCs), which are carbon-containing compounds of low molecular weight generated during microbial metabolism, have been shown to influence the behavior of numerous insects, albeit the complete extent of these interactions remains to be fully understood (2).
Remarkably, while mVOCs have been studied extensively for their impact on various insects, there is limited information regarding repellent mVOCs targeting two significant disease vectors, Anopheles gambiae and Aedes aegypti, specifically from the Bacillus genus (3).
The studied strain was initially isolated from desert soil near Merzouga, a small Saharan Village in southeast Morocco (4).The isolation process involved homogenizing a 15 g sand sample in 15 mL of sterile saline solution (0.9% NaCl w/v), followed by 10-fold dilutions and plating on tryptone soy agar (TSA) (Difco, Detroit, USA) for 96 h at 55°C.Subsequently, fresh cultures were prepared on tryptone soy broth (TSB) at 37°C and 120 rpm agitation.Strain Ba1 was present in a mixture of strains whose VOCs demonstrated repulsive effects on both Anopheles gambiae and Aedes aegypti through volatile organic compounds (VOCs) (5).
Libraries were prepared following the instructions of Ion Xpress Plus gDNA Fragment Library Preparation kit purified (AMPure XP Beads) and size-selected for 200 bp reads (Thermo-fisher Scientific, MA, USA).Whole genome sequencing was conducted using ION Proton and ION Chef technologies and following the manufacturer's instructions (Thermo-fisher Scientific, MA, USA).
As shown in Secondary metabolites biosynthesis gene cluster analysis using antiSMASH v5.2 (12) has shown that Ba1 strain is predicted to be able to produce, among others, terpenes, type III Polyketides Synthases, thiopeptides and lanthipeptides, lassopeptides and non-ribosomal peptide synthetase cluster.
The analysis of the whole genome of the mosquito repellent strain of Bacillus licheniformis is an important step in identifying the genetic factors behind such effect.

FIG 1
FIG1 The distribution of genomic annotations is visually shown by a circular graphical display in PATRIC v3.26.4.The components included within this structure, progressing from the outermost to the innermost rings, consist of contigs, coding sequences (CDS) located on the forward strand, CDS located on the reverse strand, RNA genes, CDS exhibiting homology to established antimicrobial resistance genes, CDS exhibiting homology to recognized virulence factors, as well as GC content and GC skew.The chromatic attributes of the coding sequences on both the forward and reverse strands serve as indicators of the specific subsystem to which these genes are affiliated, as per the provided list.

Table 1
, the reconstructed genome of Ba1 is 4,227,162 bp long across 171 contigs.It has a GC content of 46.37% and an N 50 of 305,417.The draft genome contains a total of 4,985 predicted genes of which 4,726 are CDS, 6 are rRNAs, and 97 are tRNAs.

TABLE 1
Assembly details a An assembled genome was submitted to the Comprehensive Genome Analysis service.This assembled genome had 171 contigs, with the total length of 4,227,162 bp and an average GC content of 46.03%. a