Diversidade de cianobactérias em crostas biológicas e avaliação de perfil celulolítico e hemicelulolítico
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2016-10-31
Autores
Lima, Nathali Maria Machado de
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Universidade Estadual Paulista (Unesp)
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Crostas biológicas consistem de uma comunidade composta por cianobactérias, algas verdes, microfungos, bactérias, líquens e musgos. Estas ocorrem em uma variedade de solos e regiões climáticas ao redor do globo, exercendo funções importantes como a de conferir estabilidade e proteger o solo contra forças erosivas, acumular e aumentar o tempo de residência da água no solo, além de, favorecer a germinação e permitir a fixação de nitrogênio e carbono. Em crostas biológicas de regiões quentes e temperadas há o predomínio de líquens e cianobactérias, sendo que as cianobactérias são consideradas os primeiros organismos a estruturarem a crosta, sendo seguidas por outros grupos de organismos. A partir da investigação de cianobactérias em crostas biológicas, muitos novos gêneros e espécies têm sido descritos, o que também enfatiza a grande necessidade da investigação sobre tais comunidades. Devido às condições propícias do ambiente, em termos de exposição do solo e fitofisionomia, o bioma Cerrado (Savana) foi escolhido para ser o local de estudo de assembleias de cianobactérias de crostas. Além disso, devido a estudos prévios que relatam o potencial de produção de enzimas celulolíticas e hemicelulolíticas por cianobactérias heterocitadas e a presença comum deste tipo de organismos em crostas biológicas, também foram feitas análises para avaliar a produção de tais substâncias. Desse modo, os estudos objetivaram contribuir com conhecimento da biodiversidade de cianobactérias de crostas biológicas de solo e avaliar a capacidade de produção enzimática das cianobactérias encontradas em crostas biológicas de solo. Para isto, amostras de crostas foram coletadas nos Parques Nacionais da Serra da Canastra (MG) e da Serra do Cipó (MG), no Parque Estadual Furnas do Bom Jesus (SP) e na região de Caldas Novas (GO). As cianobactérias encontradas nas amostras foram estudadas morfologicamente a partir do material da natureza e também isoladas em cultivos artificiais, visando além do estudo morfológico, as análises moleculares e enzimáticas. No total, foram estudados 28 morfotipos, provenientes de 31 populações, dos quais 12 foram identificados em nível específico (destes, três ainda exigem confirmação de identidade = cf) e 16 foram identificados apenas em nível genérico. Dezenove populações foram analisadas também em nível molecular por meio do sequenciamento parcial do gene 16S rRNA, e completo da Região Espaçadora Interna Transcrita 16S-23S (ITS). Seis populações foram também investigadas quanto a produção de enzimas. Entre os gêneros encontrados, muitos são comuns em assembleias de cianobactérias de crostas biológicas em outras regiões do mundo, entretanto, algumas identificações em nível de espécie não foram possíveis por diferenças importantes com os táxons já descritos em literatura. As análises moleculares enfatizaram e contribuíram com a problemática polifilia de gêneros como Leptolyngbya, Nostoc e Calothrix, reafirmaram a existência de um novo gênero próximo a Wilmottia e Microcoleus e apresentaram um novo grupo composto até o momento por uma espécie representada por duas populações. Este grupo revelou sequências moleculares que se aproximam de Brasilonema, entretanto os indivíduos apresentam ramificações verdadeiras e, portanto, investigações mais aprofundadas são necessárias para definição da identidade das populações. Dessa forma, as análises morfológicas e moleculares demonstraram a grande diversidade não acessada em localidades e habitats pobremente investigados e enfatizaram a contribuição deste trabalho ao enfocar pela primeira vez a flora de cianobactérias de solo de Cerrado. As análises de atividade enzimática revelaram que não houve produção de tais proteínas, tendo como possíveis explicações a ausência de habilidade para produção deste tipo de enzimas por parte das cepas, ou a ineficiência do método utilizado. De qualquer forma os resultados enfatizaram a necessidade de estudo nesta área, principalmente pela dificuldade no encontro de cepas produtoras e o desconhecimento sobre quais poderiam ser os fatores a estarem influenciando certas cepas a ativarem ou selecionarem diferentemente seus metabolismos. Em linhas gerais, os resultados do presente estudo apresentam, pela primeira vez, a composição das cianobactérias de crostas biológicas do território brasileiro e indicam uma flora diversificada e, em grande parte, desconhecida. Esses resultados fornecem subsídios e abrem caminho para outras pesquisas dentro do campo do conhecimento da biodiversidade presente em crostas biológicas. Complementarmente, estes trabalhos compõem a base para estudos sobre a ecologia de ambientes mais restritivos (como a caatinga ou o próprio Cerrado), permitindo abordagens como sucessão ecológica, produtividade primária, fluxos de nutrientes e dinâmicas de solo.
Biological soil crusts (BSCs) are communities potentially composed of cyanobacteria, green algae, micro fungi, bacteria, lichens and mosses. They occur in a variety of soils and climatic regions around the world, playing important roles as giving soil stability protecting against erosive forces, accumulating and increasing residence time for water in soil, besides promoting germination and performing nitrogen and carbon fixation. Biological soil crusts from hot deserts are frequently composed of cyanobacteria and lichens, and the cyanobacteria are considered the first colonizers in structuring the crust, being followed by other groups of organisms. A lot of new genera and species have been described based on crusts investigations and this fact also emphasizes the necessity of works on those communities. Due to environmental conditions, as soil exposition and phytophysiognomy, the biome Cerrado (Savannah) was chosen to be the place for studies on cyanobacterial assemblages of biological soil crusts. Besides, due to previous studies that indicate the production capability of celulolitic and hemicelulolitic enzymes by heterocytous cyanobacteria and the common presence of this kind of cyanobacteria in biological soil crusts, analyses of such production were also conducted. In this way, the studies aimed to contribute with knowledge about cyanobacterial biodiversity in biocrusts and evaluate the bioprospection capability of cyanobacterial from these biocrusts. Therefore, BSCs were sampled at the National Parks of Serra da Canastra (Minas Gerais State) and Serra do Cipó (Minas Gerais State), at the State Park of Furnas do Bom Jesus (São Paulo State) and in the region of the municipality of Caldas Novas (Goiás State). The cyanobacteria found in the samples had their morphology analyzed from the natural and cultivated conditions. Besides, molecular and enzymatic analysis were carried out. The results summarized 28 morphotypes from 31 populations, which 12 were identified at specie level (three of them need identity confirmation = cf) and 16 were identified only at genus level. Nineteen populations also were studied with molecular methods, through partial sequencing of 16S rRNA gene and complete sequencing of the Internal Transcribed Space Region (16S-23S ITS). In addition, six populations also were explored as possible enzymatic producers. Among the found genera, many are cyanobacteria frequently found in BSCs distributed around the world, however, some identifications at specific level were not possible due to considerable divergences in comparison with described taxa. The molecular analysis reaffirmed and emphasized the polyphyletic nature of Leptolyngbya, Nostoc and Calothrix, reinforced the existence of a new genus close related with Wilmottia and Microcoleus and presented a new group composed, until now, of one species represented by two populations. This group showed molecular sequences related with Brasilonema, however, the specimens are true branched, what requires more detailed studies to confirm the identification of the populations. In this way, the morphological and molecular analyses showed the wide diversity whose has not been accessed in poorly investigated places and habitats and reinforced the contribution of this work in focusing the cyanobacterial flora of crusts of Brazilian savannah area for the first time. The enzymatic activity analysis revealed that the strains studied did not produce celulolitic or hemicelulolitic compounds, having as possible explanations the absence of the production ability or the inefficiency of the utilized method. Either way, the results emphasized the necessity of studies in this area, mainly because of the difficulty in find producer strains and the lack of knowledge about which could be the factors influencing the strains in activate or select differently their metabolism. In general, our results presented for the first time the cyanobacterial composition for BSCs from Brazil and indicated a diverse, and sometimes, unknown flora. These results provided foundation and opened doors to investigations inside the biodiversity knowledge with biological soil crusts. Complementary, these works compound the basis for investigations in the ecology of extreme environments (as Caatinga and Cerrado), permiting studies about successional ecology, primary productivity, flow of nutrients and soil dynamics.
Biological soil crusts (BSCs) are communities potentially composed of cyanobacteria, green algae, micro fungi, bacteria, lichens and mosses. They occur in a variety of soils and climatic regions around the world, playing important roles as giving soil stability protecting against erosive forces, accumulating and increasing residence time for water in soil, besides promoting germination and performing nitrogen and carbon fixation. Biological soil crusts from hot deserts are frequently composed of cyanobacteria and lichens, and the cyanobacteria are considered the first colonizers in structuring the crust, being followed by other groups of organisms. A lot of new genera and species have been described based on crusts investigations and this fact also emphasizes the necessity of works on those communities. Due to environmental conditions, as soil exposition and phytophysiognomy, the biome Cerrado (Savannah) was chosen to be the place for studies on cyanobacterial assemblages of biological soil crusts. Besides, due to previous studies that indicate the production capability of celulolitic and hemicelulolitic enzymes by heterocytous cyanobacteria and the common presence of this kind of cyanobacteria in biological soil crusts, analyses of such production were also conducted. In this way, the studies aimed to contribute with knowledge about cyanobacterial biodiversity in biocrusts and evaluate the bioprospection capability of cyanobacterial from these biocrusts. Therefore, BSCs were sampled at the National Parks of Serra da Canastra (Minas Gerais State) and Serra do Cipó (Minas Gerais State), at the State Park of Furnas do Bom Jesus (São Paulo State) and in the region of the municipality of Caldas Novas (Goiás State). The cyanobacteria found in the samples had their morphology analyzed from the natural and cultivated conditions. Besides, molecular and enzymatic analysis were carried out. The results summarized 28 morphotypes from 31 populations, which 12 were identified at specie level (three of them need identity confirmation = cf) and 16 were identified only at genus level. Nineteen populations also were studied with molecular methods, through partial sequencing of 16S rRNA gene and complete sequencing of the Internal Transcribed Space Region (16S-23S ITS). In addition, six populations also were explored as possible enzymatic producers. Among the found genera, many are cyanobacteria frequently found in BSCs distributed around the world, however, some identifications at specific level were not possible due to considerable divergences in comparison with described taxa. The molecular analysis reaffirmed and emphasized the polyphyletic nature of Leptolyngbya, Nostoc and Calothrix, reinforced the existence of a new genus close related with Wilmottia and Microcoleus and presented a new group composed, until now, of one species represented by two populations. This group showed molecular sequences related with Brasilonema, however, the specimens are true branched, what requires more detailed studies to confirm the identification of the populations. In this way, the morphological and molecular analyses showed the wide diversity whose has not been accessed in poorly investigated places and habitats and reinforced the contribution of this work in focusing the cyanobacterial flora of crusts of Brazilian savannah area for the first time. The enzymatic activity analysis revealed that the strains studied did not produce celulolitic or hemicelulolitic compounds, having as possible explanations the absence of the production ability or the inefficiency of the utilized method. Either way, the results emphasized the necessity of studies in this area, mainly because of the difficulty in find producer strains and the lack of knowledge about which could be the factors influencing the strains in activate or select differently their metabolism. In general, our results presented for the first time the cyanobacterial composition for BSCs from Brazil and indicated a diverse, and sometimes, unknown flora. These results provided foundation and opened doors to investigations inside the biodiversity knowledge with biological soil crusts. Complementary, these works compound the basis for investigations in the ecology of extreme environments (as Caatinga and Cerrado), permiting studies about successional ecology, primary productivity, flow of nutrients and soil dynamics.
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Palavras-chave
Atividade enzimática, Caracterização molecular, Caracterização morfológica, Cerrado, Brazilian savannah, Enzymatic activity, Molecular analysis, Morphological analysis