Genomic diversity of themoacidophilic bacteria from Los Azufres geothermal field and their implications in biotechnology and astrobiology Bolívar-Torres Hermes Hernán*, Servín-Garcidueñas Luis* *Laboratorio de Microbiómica Escuela Nacional de Estudios Superiores Unidad Morelia Universidad Nacional Autónoma de México [email protected]

Introduction Acknowledgements

The thermoacidophile microorganisms have several adaptation To Universidad Nacional Autonoma de México and Consejo mechanisms to the hostile acidic thermal environments where Nacional de Ciencia y Tecnología (CONACyT) for giving us they live. The study of this kind of microorganisms has been the resources to develop this investigation through the Re- one of the biggest challenges for understanding their evolu- search and technological innovation program (PAPIIT) key tion, and ecological function. Therefore, their metagenomic PAPIIT IA210617. study has been a way for explore new biological resources with biotechnology interest. In addition, those microorganisms usu- References ally have very primitive features, for this reason, its study could contribute to understanding the origin and evolution of life’s [1] M Sofía Urbieta, Edgardo R Donati, Kok-Gan Chan, Saleha Shahar, earth [1, 2]. Lee Li Sin, and Kian Mau Goh. Thermophiles in the genomic era: biodiversity, science, and applications. Biotechnology Advances, 33(6):633–647, 2015. [2] Guo-Qiang Chen and Xiao-Ran Jiang. Next generation industrial biotechnology based on extremophilic bacteria. Current opinion in biotechnology, 50:94–100, 2018. [3] Mario Cesar Arriaga Suarez, Mirna del Rocío Tello, and Fernando Samaniego V. Geochemical evolution of the Los Azufres, Mexico, geothermal reservoir. Figure 2:Taxonomic groups found in the MM contigs assembly. A) Domain level. B) Phylum Level. The taxonomic analysis was In Proceedings World Geothermal Congress, pages 2227–2233, 2000. developed in MG-RAST using the contigLCA Algorithm. [4] Elcia MS Brito, Norberto Villegas-Negrete, Irene A Sotelo-González, César A Caretta, Marisol Goñi-Urriza, Claire Gassie, Florence Hakil, Figure 1:Maritaro Hot springs (MM), A) Hot springs located Yannick Colin, Robert Duran, Felix Gutiérrez-Corona, et al. in the sampling zone. B & C) Microbial mats found along the Microbial diversity in los azufres geothermal field (michoacán, mexico) hot springs where the samples for this study were collected. and isolation of representative sulfate and sulfur reducers. Extremophiles, 18(2):385–398, 2014. In Mexico, the geothermal field of “Los Azufres” is located [5] Luis E Servín-Garcidueñas and Esperanza Martínez-Romero. in the TransMexican Volcanic Belt (TMVB), which contains Draft genome sequence of the sulfolobales archaeon az1, obtained several hot springs and steam vents with extreme conditions through metagenomic analysis of a mexican hot spring. such as high temperatures, low pH, and high concentrations of Genome announcements, 2(2):e00164–14, 2014. toxic compounds [3]. Microbial communities from Los Azufres [6] Anthony M Bolger, Marc Lohse, and Bjoern Usadel. have been recently analyzed [4, 5]. However, many microbial Trimmomatic: a flexible trimmer for illumina sequence data. communities and their metabolic potentials remain to be an- Bioinformatics, 30(15):2114–2120, 2014. alyzed. For this reason, the aim of this study is to show the [7] Anton Bankevich, Sergey Nurk, Dmitry Antipov, Alexey A Gurevich, advances in the metagenomic analysis of the bacterial diversity Mikhail Dvorkin, Alexander S Kulikov, Valery M Lesin, Sergey I Nikolenko, Son Pham, Andrey D Prjibelski, et al. from thermal sediments in unexplored areas on the Los Azufres Figure 3:Functional Analysis developed in MG-RAST, A) Subsystems categories. B) Categories related with Stress response subsystem. Spades: a new genome assembly algorithm and its applications to geothermal field. single-cell sequencing. C) Categories related with Virulence, Disease and Defense subsystem. Journal of computational biology, 19(5):455–477, 2012. Experimental Methodology [8] Folker Meyer, Daniel Paarmann, Mark D’Souza, Robert Olson, Elizabeth M Glass, Michael Kubal, Tobias Paczian, Alex Rodriguez, Sediment samples were collected from hot springs located in Results Conclusions Rick Stevens, Andreas Wilke, et al. the Maritaro zone (MM) in Los Azufres Geothermal field The metagenomics rast server–a public resource for the automatic phylogenetic and functional analysis of metagenomes. (Fig 1). After that, DNA was purified using the PowerMax The MM metagenome showed a dominance of Bacteria with Our results show that the Maritaro Zone contains hot springs BMC bioinformatics, 9(1):1–8, 2008. Soil Kit (QIAGEN, Carlsbad, Ca) According to the man- a lower proportion Eukarya and Archaea (Fig 2A), and high with a high bacterial taxonomic and functional diversity. The ufacturer instructions. DNA samples were sequenced with abundance of Proteobacteria, Firmicutes, Acidobacteria, microorganisms have developed widespread adaptations for dif- Illumina HiSeq 2500 technology in Macrogen Inc, for ob- Actinobacteria, Bacteroidetes, and Nitrospirae Phyla (Fig ferent stress conditions such as, High temperatures, acidic con- taining 250 pb paired-end reads. Reads were filtered us- 2B). Genes related to oxidative, osmotic, acid stress tolerance, ditions, heavy metal tolerance, and antibiotic resistance with ing Trimmomatic v. 0.39 [6]. The assembly of the reads heat|cold shock resistance, and other unknown genes were de- a great potential for obtaining microorganisms that could be was made in the SPAdes v.3.8.2 software [7] with the next tected in the MM metagenome (Fig 3B). In addition, genes used for bio-remediation, and industrial applications. Finally, k-mer combinations: (21,33,55,77,99); (33,55,77,99,111); and involved in toxic elements tolerance, especially heavy metals the detailed exploration of the genes related to stress condi- (55,77,99,111,127). The obtained metagenomes were anno- such as; Cd, As, Co, Hg, Cu, and antibiotic resistance could tions previously mentioned could help us to understand better tated using the MG-RAST v.4.0.3 pipeline [8]. be detected (Fig 3C). the evolution of microbial communities in those environments.