Browsing by Author "Cordeiro, Isabella Ferreira"

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    Alcaligenes faecalis associated with Mimosa calodendron rizhosphere assist plant survival in arsenic rich soils.
    (2017) Felestrino, Érica Barbosa; Assis, Renata de Almeida Barbosa; Lemes, Camila Gracyelle de Carvalho; Cordeiro, Isabella Ferreira; Fonseca, Natasha Peixoto; Villa, Morghana Marina; Vieira, Izadora Tabuso; Kamino, Luciana Hiromi Yoshino; Carmo, Flávio Fonseca do; Moreira, Leandro Marcio
    The ferruginous rupestrian grasslands (FRG) in the Iron Quadrangle (IQ) are ecosystems characterized by rocky soils with reduced availability of water and nutrients, but high levels of metals. In order to comprehend the interference of microorganisms on the adaptive process of endemic plant Mimosa calodendrum (Fabaceae), bacteria associated with its roots and rhizosphere were isolated. Fourteen isolates were obtained and subsequently grown in the presence of different concentrations of arsenic (As) species. The isolate Mc250, an Alcaligenes faecalis strain, resisted to 10 mM of As (III) and 800 mM of As (V). In the presence of this strain, atomic spectrometer detected a reduction of 55% for As (III) and 72% for As (V) respectively in 10 mM and 500 mM solution. Scanning electron microscopy of this isolate demonstrated morphological modification and EDX spectroscopy revealed the presence of both As species adsorbed on the membrane, justifying the removal observed in the in vitro assays. To validate this potential removal of As in vivo, tomato plants were used as grown model in the presence and absence of A. faecalis in soil previously contaminated with 5 mM of As (III). After 14 days, plants from contaminated soil had their growth improved when compared to untreated control plants. All these results suggest for the first time that plant-associated bacteria from FRG-IQ present potential for soil rhizoremediation and may benefit the adaptive processes of plants in extreme environments including application in recovering degraded areas.
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    Análise sobre a composição da microbiota cutânea de anfíbios anuros em ambiente contaminado por arsênio.
    (2017) Cordeiro, Isabella Ferreira; Pires, Maria Rita Silvério; Moreira, Leandro Marcio; Pires, Maria Rita Silvério; Feio, Renato Neves; Ribeiro, Sérvio Pontes
    Os microrganismos estão suscetíveis aos atributos químicos e físicos do substrato onde vivem, tais como temperatura, pH, disponibilidade de água, radiação, nutrientes, contaminantes e outros. Portanto, a microbiota presente na epiderme dos anuros expostos a estas variáveis pode ter sua composição modulada. O presente trabalho teve como objetivo analisar a composição da comunidade microbiana cultivável e sua resistência ao arsênio oriunda da pele de anfíbios da Estação Ecológica do Tripuí (EET – Ouro Preto, MG), na tentativa de compreender a importância deste fenômeno na relação e manutenção da sobrevivência do vertebrado associado. Para tanto, 214 isolados bacterianos foram obtidos a partir de 4 espécies capturadas em diferentes microambientes dentro da EET, um local naturalmente contaminado por esse elemento. Da mesma forma 68 isolados bacterianos foram obtidos a partir de 3 espécies de anuros da região do Jardim Botânico de Brasília (JBB), uma região controle, livre desta contaminação. Os resultados mostraram uma maior porcentagem de isolados resistentes a arsênio obtidos a partir de anuros da EET, além de apresentarem resistência a concentrações de 5 a 10 vezes maiores que isolados bacterianos obtidos a partir de anuros do JBB. Outro dado que chamou a atenção tem relação com a maior resistência associada a isolados obtidos a partir de anuros da EET que naturalmente estão mais expostos ao contato direto com a à água, como é o caso de Ololygon luizotavioi, frente às espécies Ischnocnema izecksohni (arbustiva) e Vitreorana uranoscopa (arborícola). Este estudo piloto estabeleceu pela primeira vez uma relação direta entre a microbiota associada a pele de anfíbios e a resistência a metais pesados do meio, pemitindo-se inferir que possam estar envolvidas com a manutenção da sobrevivência da espécie vertebrada, que classicamente é muito sensível a exposição a elementos tóxicos como o arsênio. Um provável fenômeno decorrente de coevolução entre as espécies que precisa ser melhor explorado e compreendido.
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    Arsenic resistance in cultured cutaneous microbiota is associated with anuran lifestyles in the Iron Quadrangle, Minas Gerais State, Brazil.
    (2019) Cordeiro, Isabella Ferreira; Fonseca, Natasha Peixoto; Felestrino, Érica Barbosa; Caneschi, Washington Luiz; Pires, Maria Rita Silvério; Moreira, Leandro Marcio
    The microbiota of the anuran skin contributes significantly to the maintenance and survival of these animals. In the present study, we investigated the composition of culturable microbiota on the skin of frogs from Tripui Ecological Station (TES), a locality in the Brazilian Iron Quadrangle that is naturally contaminated with arsenic. A total of 328 culturable bacterial isolates were obtained from 21 individuals belonging to seven anuran species, captured in different microenvironments within TES. The results indicate that higher numbers of culturable bacterial isolates, as well as higher resistance to arsenic and a higher capacity to produce biofilms, were associated with three frog species that had come into contact with arsenic-contaminated water during their reproductive period. These results raise the possibility that the adaptation of these anurans species to arsenicladen environments may have a direct correlation with a specialized microbiota.
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    Biotechnological potential of plant growth-promoting bacteria from the roots and rhizospheres of endemic plants in ironstone vegetation in southeastern Brazil.
    (2018) Felestrino, Érica Barbosa; Vieira, Izadora Tabuso; Caneschi, Washington Luiz; Cordeiro, Isabella Ferreira; Assis, Renata de Almeida Barbosa; Lemes, Camila Gracyelle de Carvalho; Fonseca, Natasha Peixoto; Sanchez, Angelica Bianchini; Caicedo Cepeda, Juan Carlos; Ferro, Jesus Aparecido; Garcia, Camila Carrião Machado; Carmo, Flávio Fonseca do; Kamino, Luciana Hiromi Yoshino; Moreira, Leandro Marcio
    Microorganisms associated with plants have a great biotechnological potential, but investigations of these microorganisms associated with native plants in peculiar environments has been incipient. The objective of this study was to analyze the plant growth-promoting bacteria potential of cultivable bacteria associated with rare plants from the ferruginous rocky fields of the Brazilian Iron Quadrangle. The roots and rhizospheres of nine endemic plants species and samples of a root found in a lateritiric duricrust (canga) cave were collected, the culturable bacteria isolated and prospected for distinct biotechnological and ecological potentials. Out of the 148 isolates obtained, 8 (5.4%) showed potential to promote plant growth, whereas 4 (2.7%) isolates acted as biocontrol agents against Xanthomonas citri pathotype A (Xac306), reducing the cancrotic lesions by more than 60% when co-inoculated with this phytopathogen in Citrus sinensis plants. Moreover, other 4 (2.7%) isolates were classified as potential bioremediation agents, being able to withstand high concentrations of arsenite (5 mM As3+) and arsenate (800 mM As5+), by removing up to 35% and 15% of this metalloid in solution, respectively. These same four isolates had a positive influence on the growth of both the roots and the aerial parts when inoculated with tomato seeds in the soil contaminated with arsenic. This is the first time that an investigation highlights the potentialities of bacteria associated with rare plants of ferruginous rocky fields as a reservoir of microbiota of biotechnological and ecological interest, highlighting the importance of conservation of this area that is undergoing intense anthropic activity
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    Brazilian ironstone plant communities as reservoirs of culturable bacteria with diverse biotechnological potential.
    (2018) Caneschi, Washington Luiz; Felestrino, Érica Barbosa; Fonseca, Natasha Peixoto; Villa, Morghana Marina; Lemes, Camila Gracyelle de Carvalho; Cordeiro, Isabella Ferreira; Assis, Renata de Almeida Barbosa; Sanchez, Angelica Bianchini; Vieira, Izadora Tabuso; Kamino, Luciana Hiromi Yoshino; Carmo, Flávio Fonseca do; Garcia, Camila Carrião Machado; Moreira, Leandro Marcio
    Extensive mineral extractivism in the Brazilian Iron Quadrangle (IQ) region has destroyed large areas of land, decimating plant species, and their associated microbiota. Very little is known about the microbiota of the region; hence, cultivable bacteria associated with plants of its soils were investigated for their biotechnological potential. Samples were collected from nine plant species and six soils, and 65 cultivable bacterial isolates were obtained. These represent predominantly gram-positive bacilli (70%) capable of producing amylases (55%), proteases (63%), cellulases (47%), indole acetic acid (IAA) (46%), siderophores (26%), and to solubilize phosphate (9%). In addition, 65% of these were resistant to ampicillin, 100% were sensitive to tetracycline, and 97% were tolerant to high arsenic concentrations. Three isolates were studied further: the isolate FOB3 (Rosenbergiella sp.) produced high concentrations of IAA in vitro in the absence of tryptophan – shown by the significant improvement in plant germination and growth rate where the isolate was present. For isolates C25 (Acinetobacter sp.) and FG3 (Serratia sp.), plasmids were purified and inserted into Escherichia coli cells where they modified the physiological profile of the transformed strains. The E. coli::pFG3B strain showed the highest capacity for biofilm production, as well as an increase in the replication rate, arsenic tolerance and catalase activity. Moreover, this strain increased DNA integrity in the presence of arsenic, compared to the wild-type strain. These results help to explain the importance of bacteria in maintaining plant survival in ferruginous, rocky soils, acting as plant growth promoters, and to highlight the biotechnological potential of these bacteria. IMPORTANCE The Iron Quadrangle region is responsible for ∼60% of all Brazilian iron production and, at the same time, is responsible for housing a wide diversity of landscapes, and consequently, a series of endemic plant species and dozens of rare species – all of which have been poorly studied. Studies exploring the microbiota associated with these plant species are limited and in the face of the continuous pressure of extractive action, some species along with their microbiota are being decimated. To understand the potential of this microbiota, we discovered that cultivable bacterial isolates obtained from plants in the ferruginous rocky soil of the Iron Quadrangle region have diverse biotechnological potential, revealing a genetic ancestry still unknown.
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    Complete genome sequence and analysis of Alcaligenes faecalis strain Mc250, a new potential plant bioinoculant.
    (2020) Felestrino, Érica Barbosa; Sanchez, Angelica Bianchini; Caneschi, Washington Luiz; Lemes, Camila Gracyelle de Carvalho; Assis, Renata de Almeida Barbosa; Cordeiro, Isabella Ferreira; Fonseca, Natasha Peixoto; Villa, Morghana Marina; Vieira, Izadora Tabuso; Kamino, Luciana Hiromi Yoshino; Carmo, Flávio Fonseca do; Silva, Aline Maria da; Thomas, Andrew Maltez; Patané, José Salvatore Leister; Ferreira, Fernanda Carla; Freitas, Leandro Grassi de; Varani, Alessandro de Mello; Ferro, Jesus Aparecido; Silva, Robson Soares; Almeida Junior, Nalvo Franco de; Garcia, Camila Carrião Machado; Setubal, João Carlos; Moreira, Leandro Marcio
    Here we present and analyze the complete genome of Alcaligenes faecalis strain Mc250 (Mc250), a bacterium isolated from the roots of Mimosa calodendron, an endemic plant growing in ferruginous rupestrian grasslands in Minas Gerais State, Brazil. The genome has 4,159,911 bp and 3,719 predicted protein-coding genes, in a single chromosome. Comparison of the Mc250 genome with 36 other Alcaligenes faecalis genomes revealed that there is considerable gene content variation among these strains, with the core genome representing only 39% of the protein-coding gene repertoire of Mc250. Mc250 encodes a complete denitrification pathway, a network of pathways associated with phenolic compounds degradation, and genes associated with HCN and siderophores synthesis; we also found a repertoire of genes associated with metal internalization and metabolism, sulfate/sulfonate and cysteine metabolism, oxidative stress and DNA repair. These findings reveal the genomic basis for the adaptation of this bacterium to the harsh environmental conditions from where it was isolated. Gene clusters associated with ectoine, terpene, resorcinol, and emulsan biosynthesis that can confer some competitive advantage were also found. Experimental results showed that Mc250 was able to reduce (~60%) the virulence phenotype of the plant pathogen Xanthomonas citri subsp. citri when co-inoculated in Citrus sinensis, and was able to eradicate 98% of juveniles and stabilize the hatching rate of eggs to 4% in two species of agricultural nematodes. These results reveal biotechnological potential for the Mc250 strain and warrant its further investigation as a biocontrol and plant growth-promoting bacterium.
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    Detection and identification of Xanthomonas pathotypes associated with citrus diseases using comparative genomics and multiplex PCR.
    (2019) Fonseca, Natasha Peixoto; Felestrino, Érica Barbosa; Caneschi, Washington Luiz; Sanchez, Angelica Bianchini; Cordeiro, Isabella Ferreira; Lemes, Camila Gracyelle de Carvalho; Assis, Renata de Almeida Barbosa; Carvalho, Flávia Maria de Souza; Ferro, Jesus Aparecido; Varani, Alessandro de Mello; Belasque Junior, José; Setubal, João Carlos; Telles, Guilherme Pimentel; Aguena, Deiviston da Silva; Almeida Junior, Nalvo Franco de; Moreira, Leandro Marcio
    Background. In Citrus cultures, three species of Xanthomonas are known to cause distinct diseases. X. citri subsp. citri patothype A, X. fuscans subsp. aurantifolii pathotypes B and C, and X. alfalfae subsp. citrumelonis, are the causative agents of cancrosis A, B, C, and citrus bacterial spots, respectively. Although these species exhibit different levels of virulence and aggressiveness, only limited alternatives are currently available for proper and early detection of these diseases in the fields. The present study aimed to develop a new molecular diagnostic method based on genomic sequences derived from the four species of Xanthomonas. Results. Using comparative genomics approaches, primers were synthesized for the identification of the four causative agents of citrus diseases. These primers were validated for their specificity to their target DNA by both conventional and multiplex PCR. Upon evaluation, their sensitivity was found to be 0.02 ng/µl in vitro and 1.5 × 104 CFU ml−1 in infected leaves. Additionally, none of the primers were able to generate amplicons in 19 other genomes of Xanthomonas not associated with Citrus and one species of Xylella, the causal agent of citrus variegated chlorosis (CVC). This denotes strong specificity of the primers for the different species of Xanthomonas investigated in this study. Conclusions. We demonstrated that these markers can be used as potential candidates for performing in vivo molecular diagnosis exclusively for citrus-associated Xanthomonas. The bioinformatics pipeline developed in this study to design specific genomic regions is capable of generating specific primers. It is freely available and can be utilized for any other model organism.
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    Potential bioinoculants for sustainable agriculture prospected from ferruginous caves of the Iron Quadrangle/Brazil.
    (2021) Lemes, Camila Gracyelle de Carvalho; Cordeiro, Isabella Ferreira; Fernandes, Camila Henriques de Paula; Silva, Ana K.; Carmo, Flávio Fonseca do; Kamino, Luciana Hiromi Yoshino; Carvalho, Flávia M. S.; Caicedo Cepeda, Juan Carlos; Ferro, Jesus Aparecido; Moreira, Leandro Marcio
    Biocontrol and plant growth-promoting bacteria (PGPB) are important agricultural bioinoc- ulants. This study aimed to prospect new potential bioinoculants for a more sustainable agriculture from ferruginous caves of the Brazilian Iron Quadrangle. Culturable bacteria, from seven caves and one canga soil sample, were evaluated for biocontroller activity of the phytopathogens Xanthomonas citri subsp. Citri—Xcc306 (citrus canker), Fusarium oxysporum—Fo (fusariosis), and Colletotrichum lindemuthianum—Cl89 (bean anthracnose). The ability of the superior candidates to solubilize inor- ganic phosphate, fix nitrogen, and produce hydrolytic enzymes and siderophores was then analyzed. Out of 563 isolates, 47 inhibited the growth of Xcc306 in vitro, of which 9 reduced citrus canker up to 68% when co-inoculated with the pathogen on host plants. Twenty of the 47 inhibited Fo growth directly by 51–73%, and 15 indirectly by 75–81%. These 15 inhibited Cl89 growth in vitro (up to 93% directly and 100% indirectly), fixed nitrogen, produced proteases and siderophores, showed motility ability, produced biofilm, and all but one solubilized inorganic phosphate. Therefore, 15 (2.66%) bacterial isolates, from the genera Serratia, Nissabacter, and Dickeya, act simultaneously as biocontrollers and PGPBs, and could be important candidates for future investigations in planta as an alternative to minimize the use of pesticides and chemical fertilizers through sustainable agricultural management practices.
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    Prospecção do potencial anti fúngico de isolados bacterianos obtidos a partir da epiderme de anuros do Quadrilátero ferrífero - MG.
    (2020) Fernandes, Camila Henriques de Paula; Moreira, Leandro Marcio; Sanchez, Angelica Bianchini; Cordeiro, Isabella Ferreira; Moreira, Leandro Marcio; Oliveira Júnior, Ênio Nazaré de; Cruz, Izinara Rosse da
    O controle biológico utilizando bactérias como antagonistas é uma maneira eficiente para o controle de pragas causadas por fungos. Neste cenário, o objetivo geral deste trabalho foi prospectar o potencial antifúngico de isolados bacterianos cultiváveis obtidos a partir da pele de quatro espécies de anuros coletados na EET. A Estação Ecológica do Tripuí (EET) está localizada no Quadrilátero Ferrífero, Ouro Preto, MG. Esta área possui alta biodiversidade, com destaque para a anurofauna. Assim, 192 isolados bacterianos obtidos a partir da pele das espécies Boana albopunctata, Boana faber, Rhinella crucifer e Ischnocnema izecksohni foram investigadas quanto à capacidade de atuarem como biocontroladores de Fusarium oxysporum e Colletotrichum lindemuthianum, respectivos agentes causais da fusariose e da antracnose, que acometem várias culturas agrícolas, acarretando diversos prejuízos econômicos e sociais. Para o ensaio inicial, os 192 isolados foram avaliados contra F. oxysporum, por meio de ensaio de inibição direta. Dentre os isolados analisados, 15 foram capazes de inibir em até 60% do crescimento do fungo. Estes 15 isolados foram então selecionados para o ensaio de inibição direta contra o C. lindemuthianum, seguindo o mesmo protocolo. Oito apresentaram taxa de inibição superior a 80%, dentre eles, dois com inibição de 100% do crescimento. Esses 8 isolados foram então avaliados quanto a capacidade de inibição indireta por compostos difusíveis e termoestáveis, os quais apresentaram resultados equivalentes aos testes anteriores. Porém, nenhum foi capaz de inibir por meio da produção de compostos voláteis. Além disso, foram realizados ensaios de produção de amilase, protease e celulase para estes 8 isolados, dos quais 6 apresentaram resultado positivo para todas as enzimas avaliadas. Os isolados também apresentaram resultados positivos para a produção de biofilme, auto-agregação, fixação de nitrogênio e solubilização de fosfato. Finalmente, os 8 isolados foram identificados por sequenciamento da subunidade 16S do gene ribossomal. Desta forma, conclui-se que estes isolados bacterianos apresentam potencial antagônico até então desconhecido com características fisiológicas que os apontam como potenciais agentes biocontroladores contra fungos de importância agrícola.
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    Prospecção funcional e potencial biotecnológico de bactérias associadas a anuros do Quadrilátero Ferrífero.
    (2022) Cordeiro, Isabella Ferreira; Moreira, Leandro Marcio; Moreira, Leandro Marcio; Becker, Carlos Guilherme; Blamires, Daniel; Pires, Maria Rita Silvério; Azevedo, Cristiano Schetini de
    A contaminação por metais e metaloides tóxicos, natural ou mediada por ação antrópica, tem se tornado um grave problema ambiental, acarretando danos a diversos seres vivos, em especial ao homem. Porém, alguns organismos presentes em distintos níveis tróficos são mais sensíveis a estes contaminantes, e um bom exemplo disso são os anfíbios. Por apresentarem tecido epitelial altamente permeável sofrem mais pela ação destes compostos, o que os colocam, na maioria das vezes, como potenciais bioindicadores de contaminantes. O presente trabalho teve como objetivos prospectar bactérias da pele de anfíbios anuros capazes de tolerar ao arsênio, bem como investigar possíveis relações desta tolerância como mecanismos práticos de aplicação biotecnológica. Para isso, duas investigações paralelas foram conduzidas. Num primeiro momento, isolados bacterianos advindos exclusivamente de anuros de um ambiente contaminado por metais pesados (Estação Ecológica do Tripuí - Ouro Preto, MG) foram avaliados. Para esta abordagem ficou constatado que espécies que possuiam íntimas associações com a água durante seu ciclo reprodutivo apresentam microbiota cultivável mais tolerante e ao mesmo tempo com maior potencial de produção de biofilme, contrapondo-se a microbiota advinda de animais que apresentam menor contato com a água. Considerando que os ambientes aquáticos em áreas contaminadas possuem maior disponibilidade para contato com os organismos vivos, tais resultados são indicativos que tal tolerância e adaptação fisiológica podem contribuir fortemente para a manutenção da sobrevivência da espécie anura quando em contato com estes ambientes no Quadrilátero Ferrífero (QF). Em um segundo momento, a microbiota cultivável associada a cinco espécies da região do QF foi comparada com a microbiota cultivável associada às mesmas espécies, porém advindas de um ambiente não contaminado, localizado a 320 km de distância do primeiro, no Município de João Neiva, ES. Os resultados evidenciaram que a microbiota proveniente das espécies do QF apresentam uma maior tolerância ao arsênio e foram capazes de proteger os anuros da permeabilidade deste metalóide, o que nao foi observado na microbiota advinda das espécies de ambiente não contaminado. Tais dados são indicativos da contribuição desta microbota na manutenção da sobrevivência dos anuros, reiterando os dados apresentados pela primeira abordagem. Como conclusão, os resultados auxiliaram a melhor compreensão desse modelo de interação, bem como classificam potenciais isolados com aplicações biotecnológicas que podem ser melhor exploradas quanto aos possíveis mecanismos moleculares associados a esta notória tolerância.
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    Serratia liquefaciens FG3 isolated from a metallophyte plant sheds light on the evolution and mechanisms of adaptive traits in extreme environments.
    (2019) Caneschi, Washington Luiz; Sanchez, Angelica Bianchini; Felestrino, Érica Barbosa; Lemes, Camila Gracyelle de Carvalho; Cordeiro, Isabella Ferreira; Fonseca, Natasha Peixoto; Villa, Morghana Marina; Vieira, Izadora Tabuso; Moraes, Lauro Ângelo Gonçalves de; Assis, Renata de Almeida Barbosa; Carmo, Flávio Fonseca do; Kamino, Luciana Hiromi Yoshino; Silva, Robson Soares; Ferro, Jesus Aparecido; Ferro, Maria Inês Tiraboschi; Ferreira, Rafael Marini; Santos, Vera Lúcia; Silva, Ubiana de Cássia Mourão; Almeida Junior, Nalvo Franco de; Varani, Alessandro de Mello; Garcia, Camila Carrião Machado; Setubal, João Carlos; Moreira, Leandro Marcio
    Serratia liquefaciens strain FG3 (SlFG3), isolated from the flower of Stachytarpheta glabra in the Brazilian ferruginous fields, has distinctive genomic, adaptive, and biotechnological potential. Herein, using a combination of genomics and molecular approaches, we unlocked the evolution of the adaptive traits acquired by S1FG3, which exhibits the second largest chromosome containing the largest conjugative plasmids described for Serratia. Comparative analysis revealed the presence of 18 genomic islands and 311 unique protein families involved in distinct adaptive features. S1FG3 has a diversified repertoire of genes associated with Nonribosomal peptides (NRPs/PKS), a complete and functional cluster related to cellulose synthesis, and an extensive and functional repertoire of oxidative metabolism genes. In addition, S1FG3 possesses a complete pathway related to protocatecuate and chloroaromatic degradation, and a complete repertoire of genes related to DNA repair and protection that includes mechanisms related to UV light tolerance, redox process resistance, and a laterally acquired capacity to protect DNA using phosphorothioation. These findings summarize that SlFG3 is well-adapted to different biotic and abiotic stress situations imposed by extreme conditions associated with ferruginous fields, unlocking the impact of the lateral gene transfer to adjust the genome for extreme environments, and providing insight into the evolution of prokaryotes.