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The beneficial effects of bacteria on plant growth have led to the development of promising commercial formulations for sustainable agricultural intensification [Basu et al. 2021]. Azospirillum brasilense is one of the most well-studied and widely used bacteria in inoculants [Cassán et al. 2020]. However, systematic and comparative studies within the genus are remain scarce. In this work, we sought to evaluate the diversity and pathogenic potential of the genus Azospirillum. Genomes of Azospirillum spp. were selected from the evaluation of distance and identity metrics among all representatives of the Azospirillaceae family, using the MASH and fastANI tools. The identity between the genomes of the genus Azospirillum was evaluated by estimating the average nucleotide identity (ANI) with pyani. The pangenome was characterized using the Roary. The identified SNPs (single nucleotide polymorphisms) were used for phylogenetic reconstruction using IQ-TREE. The resistome and virulome of the genus were analyzed using the USEARCH tool. The evaluation with MASH and fastANI of Azospirilaceae allowed the reliable selection of 90 genomes of the genus Azospirillum and the exclusion of isolates with incorrect classification. Azospirillum genomes with ANI value >= 99.9% were clustered, consisting in a final set of 61 non-redundant genomes. We were able to determine 24 different phylogroups (ANI values >= 95%) for the genus, the largest group being represented by the species A. brasilense. The pangenome of the genus showed 72,307 genes clusters and 1,670 core genes. The virulome analysis uncovered genes related to adhesion, immune modulation, and stress resistance. The core-resistome was represented by ceoB gene, related with aminoglycosides and fluoroquinolones resistance. Collectively, our results constitute important steps to better understand Azospirillum diversity and might help prospect novel species for biotechological applications.

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#	Name
1	Isabella Oliveira-Pinheiro(isabella.op95@gmail.com)
2	Francisnei Pedrosa-Silva(francisneipedrosa@gmail.com)
3	Thiago Venâncio(tmvenancio@uenf.br)

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1	Basu, A., Prasad, P., Das, S. N., et al. (2021). Plant growth promoting rhizobacteria (PGPR) as green bioinoculants: recent developments, constraints, and prospects. Sustainability, v. 13, n. 3, p. 1140. Cassán, F., Coniglio, A., López, G., et al. (2020). Everything you must know about Azospirillum and its impact on agriculture and beyond. Biology and Fertility of Soils, v. 56, n. 4, p. 461–479.

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