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Master's Dissertation
DOI
https://doi.org/10.11606/D.64.2020.tde-26042023-170035
Document
Author
Full name
Taiane Fernanda dos Santos Barradas
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2020
Supervisor
Committee
Fiore, Marli de Fatima (President)
Gama Junior, Watson Arantes
Sarmento, Hugo Miguel Preto de Morais
Title in Portuguese
Genômica comparativa da cianobactéria Limnospira platensis formadora de florações em lagoas do Pantanal (MS)
Keywords in Portuguese
Ambiente extremo
Filogenômica
Genes adaptativos
Genoma
Taxonomia
Abstract in Portuguese
As análises genômicas possibilitam investigar a capacidade funcional e as adaptações ambientais de numerosos microrganismos de relevância global, incluindo as cianobactérias. Nas condições extremas das lagoas salino-alcalinas do pantanal, as cianobactérias têm importância ecológica fundamental como produtores primários e engenheiras de ecossistemas. Embora o seu papel nos ciclos biogeoquímicos seja reconhecido, muito pouco se sabe sobre a composição dos genomas cianobacterianos do Pantanal. Dessa forma, o objetivo deste estudo foi realizar análises genômicas e funcionais de duas linhagens de Limnospira platensis isoladas do pantanal brasileiro visando a compreensão de sua evolução, ecologia e adaptação funcional. Para isso, as linhagens foram submetidas a um tratamento com Extran (0,1) para reduzir a abundância da comunidade de bactérias associadas e o DNA extraído das células tratadas foi sequenciado em uma plataforma HiSeq e analisado com ferramentas genômicas. O sequenciamento e a montagem dos genomas geraram 28 scaffolds para a linhagem CENA650 com conteúdo GC de 44,13% e tamanho 7.193.576 bp, enquanto que a linhagem CENA597 apresentou um (1) scaffold com conteúdo GC de 44,14% e tamanho de 7.483.160 bp. A análise filogenética baseada no gene de 16S RNAr posicionou as linhagens CENA597 e CENA650 em um clado bem suportado de Limnospira, dentro de um subclado formado somente por linhagens de Limnospira platensis. A análise filogenômica baseada no alinhamento de 120 proteínas conservadas em todas as cianobactérias posicionou as linhagens CENA597 e CENA650 no clado contendo linhagens do gênero Limnospira. Os resultados mostraram que apesar da distância geográfica (~ 21 km) entre as duas lagoas de onde as duas linhagens foram isoladas, os seus genomas são muito semelhantes, o que era esperado uma vez que as pressões seletivas são similares, reduzindo as divergências do genótipo. Entretanto, a análise genômica comparativa dessas duas linhagens com L. platensis originárias de regiões mais distantes mostrou diferenças significativas. O diagrama de Venn identificou 3649 clusters de genes que foram compartilhados por todas as linhagens analisadas, indicando a conservação destes genes. Foram encontrados genes específicos que estão envolvidos na resistência a metais pesados e em respostas celulares a estresses oxidativos, resistência a patógenos e a antibióticos. A mineração dos genomas identificou genes específicos relacionados a adaptação ao ambiente extremo estudado, o que possivelmente contribuem para a sobrevivência e a dominância da L. platensis. Os genomas sequenciados neste estudo trazem importantes contribuições para a taxonomia e conhecimento da distribuição do gênero Limnospira, além de permitir futuras explorações do potencial biotecnológico desses isolados
Title in English
Comparative genomics of bloom-forming cyanobacterium Limnospira platensis in Pantanal lakes (MS)
Keywords in English
Adaptative genes
Extreme environment
Genome
Phylogenomic
Taxonomy
Abstract in English
Genomic analyses allow to investigate the functional capacity and environmental adaptations of numerous microorganisms of global relevance, including cyanobacteria. In the harsh conditions of saline-alkaline of Pantanal lakes, cyanobacteria are of fundamental ecological importance as primary producers and ecosystem engineers. Although its role in biogeochemical cycles is recognized, little is known about the composition of cyanobacterial genomes from Pantanal. Thus, the objective of this study was to perform genomic and functional analyses of two strains of Limnospira platensis isolated from the Brazilian Pantanal in order to understanding their evolution, ecology and functional adaptation. For this, the strains were subjected to a treatment with Extran (0.1%) to reduce the abundance of associated bacteria community, and the DNA extracted from treated cells was sequenced in a HiSeq platform and analysed with genomic tools. The sequencing and assembly of the genomes generated 28 scaffolds for the strain CENA650, with 44.13% GC content and size of 7,193,576 bp, while the strain CENA597 showed one (1) scaffold, with 44.14% GC content and size of 7,483,160 bp. Phylogenetic analysis based on the 16S rRNA gene positioned the strains CENA597 and CENA650 in a well-supported Limnospira clade, within a subclade formed only by Limnospira platensis strains. Phylogenomic analysis based on the alignment of 120 conserved proteins in all cyanobacteria positioned the strains CENA597 and CENA650 in the clade containing strains of the genus Limnospira. The results showed that despite the geographic distance (~ 21 km) between the two lakes from which the two strains were isolated, their genomes are very similar, which was expected since the selective pressures are similar, reducing the divergences of the genotype. However, the comparative genomic analysis of these two strains with L. platensis originating from more distant regions showed significant differences. The Venn diagram identified 3649 clusters of genes that were shared by all strains analyzed, indicating the conservation of these genes. Specific genes involved in resistance to heavy metals and in cellular responses to oxidative stress, resistance to pathogens and antibiotics were found. The genome mining identified specific genes related to adaptation to the extreme environment studied, which possibly contribute to the survival and dominance of this cyanobacterial species. The genomes sequenced in this study bring important contributions to the taxonomy and knowledge of the distribution of the genus Limnospira, in addition to allowing future explorations of the biotechnological potential of these isolates
 
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Publishing Date
2023-05-24
 
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