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Os microrganismos endofíticos colonizam o interior dos tecidos das plantas sem causar danos ao seu hospedeiro. A interação endófito-planta pode originar vias de síntese de metabólitos de interesse biotecnológico, contudo, para sua utilização é necessário que esses compostos sejam identificados quimicamente. Dentre as técnicas instrumentais mais utilizadas estão a cromatografia líquida de alta eficiência (CLAE), cromatografia em camada delgada (CCD), cromatografia a gás capilar (CG), cromatografia a gás capilar acoplada a espectrometria de massas (CG/MS), cromatografia de fluído supercrítico (CFS) e eletroforese capilar (EC). Desta forma, realizamos uma revisão da literatura abordando estas técnicas de identificação de metabólitos secundários de interesse biotecnológico produzidos por microrganismos endofíticos. A pesquisa revelou que estas técnicas instrumentais permitiram a identificação de coclioquinonas, isocloquinonas, dicetopiperazinas, ácido tartárico, ácido cítrico, butirilneosolaniol e outros compostos, produzidos por microrganismos endofíticos. Estas técnicas instrumentais para identificação e quantificação de metabólitos são amplamente utilizadas e aperfeiçoadas. É possível destacar que para cada característica de amostra e sua finalidade, alguns dos métodos se tornam mais eficientes e precisos. Esses métodos para análise de metabólitos devem continuar evoluindo a fim de descobrir novas moléculas e também gerar uma maior robustez para a identificação e quantificação destes compostos de origem microbiana, contribuindo para o avanço dos produtos naturais.

Bioprospecting of endophytic microorganisms has been considered a promising resource for obtaining new biomolecules with biotechnological applications. Since few studies are available on the diversity of endophytic bacteria associated with species of the Cactaceae family, current analysis isolates and characterizes the communities of endophytic bacteria associated with the species of C. peruvianus grown in vivo obtained from seed and with somaclone of the same species regenerated in vitro to prospect for their plant-growth promoting activity. Isolation of endophytic bacteria from plants obtained from seeds and from regenerated somaclone in vitro identified nine species of endophytic bacteria (Moraxela osloensis, Corynebacterium mycetoides, Micrococcus luteus, Kocuria palustris, Staphylococcus epidermidis, Staphylococcus cohnii, Pseudomonas stuzeri, Bacillus licheniformis and Microbacterium sp.) not previously described in cacti, and signalized the somaclones as the most promising source of endophytic bacteria for obtaining new biomolecules with biotechnological applications. Somaclones are indicated as the most promising source of endophytic bacteria due to their high concentration of endophytes per gram identified in fresh somaclone tissue, the identification of the highest proportion endophytic bacteria, a source of IAA, and with the highest capacity of biological nitrogen fixation, and EPS production exclusively by the endophytic bacteria isolated from the in vitro regenerated somaclone.

A família Piperaceae, é uma família de plantas que está distribuída em regiões tropicais e subtropicais, com cerca de 3.700 espécies. O gênero Piper sp. vêm destacando-se na área da química e prospecção biotecnológica por apresentarem-se capazes de produzir óleos essenciais com potencial biotecnológico para o ramo da saúde e agricultura. Considerando as problemáticas ambientais e de saúde que envolvem o uso excessivo de defensivos agrícolas e o crescente anseio de alternativas mais sustentáveis para frear os efeitos adversos no ambiente, realizamos uma revisão da literatura acerca da prospecção biotecnológica de óleos essenciais obtidos de espécies do gênero Piper L. para o setor agrícola. A pesquisa dos trabalhos científicos foi realizada por meio das plataformas online Portal CAPES, SciELO, Science Direct e Google Acadêmico. Diferentes espécies de Piper sp. foram descritas produzindo óleos essenciais com aplicação biotecnológica visando o setor agrícola, destacando-se as atividades antifúngica contra Fusarium sp. e Colletotrichum sp., ovicida contra Anticarsi agemmatalis, inseticida contra Callosobruchus maculates, acaricida contra Tetranychus urticae e nematicida sobre Meloidogyne javanica. As atividades apresentadas pelos óleos essenciais provenientes de Piper sp. estão atribuídos a diversidade de seus constituintes químicos representados em particular por terpenoides, taninos, flavonoides e polifenóis. Acreditamos que os óleos essenciais produzidos pelas espécies de Piper sp. estudadas até então, apresentam-se como uma alternativa eco-friendly para o setor agrícola possibilitando a redução do consumo e aplicação de agrotóxicos e derivados.

The endophytic fungi live inside plant tissues and do not develop any disease to their hosts. These microorganisms can produce bioactive compounds. In this work, we aimed to evaluate the leaf endophytic fungi microbiota from J. brandegeana using the culture-dependent method and multilocus sequence analysis. The endophyte's isolation was performed by the fragmentation method and the MLSA phylogeny was made using ITS, elongation factor 1-α, β-tubulin, glyceraldehyde-3-phosphate and, actin genes. For the screening selection, agar piece test was performed and, amongst the strains, JB06 isolate was chosen for metabolic extraction from the fermented culture and the evaluation of the antibacterial, antioxidant, antiproliferative activities and, phenolic compounds assay. Through the MLSA analysis, there were five genera of fungi identified, with Colletotrichum the higher isolated. For the antioxidant assay using the fungal extract obtained from Mycoleptodiscus indicus (JB06), the best results were obtained with the ABTS method. Antiproliferative effects by the crude extract of the endophyte against the tumor hepatocellular carcinoma cells (HepG2/C3A) were found. In summary, our findings revealed highly diverse endophytic fungi associated with J. brandegeana, with the bioprospection of the endophyte M. indicus with biotechnological potential for the health field.

Endophytic microorganisms live inside the plant tissues symptomless. These microorganisma may present antagonistic activity against phytopathogens or produce metabolites with antifungal activity. We aimed to evaluate in vitro antagonism and competitive interactions between endophytic fungi of Diaporthe species and Sclerotinia sclerotiorum. We also carried out in vitro screening of antifungal activity of metabolic extracts of the two endophytes against the pathogen. The antagonism was performed by the paired-culture and two promising antagonists strains were selected for metabolic extraction from the fermented culture. Metabolic extracts were obtainment using two different organic solvents (Ethyl Acetate and Hexane) and their antifungal activity was carried out using the agar diffusion test. The in vitro antagonistic index ranged from 22.1 to 59.5%, with Diaporthe sp. PL03 (59.1%), D. schini PL40 (59.5%), D. infecunda PL63 (41.8%), D. anacardii PL64 (56.8%), with inhibition by mycelial contact. The endophytes PL01 (28.6%) and PL43 (28.5%), both D. anacardii, stood out blocking mycelial growth from a distance. In the antifungal assay, D. anacardii PL01 (31.7%) and D. schini PL40 (18.2%) acetate metabolite stood out. In summary, our results indicate a few Diaporthe endophytes able to antagonize a S. sclerotiorum pathogen under in vitro conditions.

In this work, the antibacterial activity of a crude extract of the endophytic fungus Flavodon flavus (JB257), isolated from leaves of Justicia brandegeana, was evaluated against both the vegetative and sporulated forms of Alicyclobacillus acidoterrestris. The microdilution technique was performed in order to determine the antibacterial activity of the crude extract alone as well as in combination with the bacteriocin, nisin. The minimum inhibitory concentration (MIC) of the crude extract and nisin alone against A. acidoterrestris vegetative forms were 250 μg/mL and 31.5 μg/mL, respectively, while the minimum bactericidal concentrations (MBC) were 1000 μg/mL and 62.5 μg/mL,respectively. For A. acidoterrestris spores, treatment with the crude extract at a concentration of 500 μg/mL caused a 47% reduction in growth, while nisin at 62.5 μg/mL could reduce 100% of the growth. The in vitro evaluation of the crude extract combined with nisin against A. acidoterrestris by the Checkerboard method showed a synergistic interaction between the two compounds. In addition, greater selectivity towards bacterial cells over host cells, a human hepatocyte cell line, was achieved when the crude extract was combined with nisin, Using scanning electron microscopy, interferences in the cell membrane of A. acidoterrestris could be observed after treatment with the crude extract. The results presented in this study indicate that the crude extract of the endophyte F. flavus has biotechnological potential in the food industry, especially for the treatment of orange juices through the control of A. acidoterrestris.

The sugarcane borer Diatraea saccharalis is a pest that causes damage to sugarcane fields. The female lays imbricated egg masses on the leave. They cause the greatest damage as larvae, and thus, control before egg hatching is desired to prevent the development of this pest and avoid damage to the plant. Diatomaceous Earth, or diatomite, is inert dust with insecticidal properties. In this work, we aimed to test the activity of Diatomaceous Earth on viable eggs of D. saccharalis as a potential alternative control method for sugarcane borer. D. saccharalis eggs 1 day after oviposition were immersed in a Diatomaceous Earth solution (10% - w/v) for 5, 10, or 15 min and observed every 24 h for 216 h of analysis. During the first 72 h, there were no changes observed in egg morphology, which maintained an oval shape, pale yellow in color. After 96 h, chorion striations were observed in the treatments with 10 and 15 min of exposure and were not present in the chorion controls eggs. Differences between embryo's number it was observed after 96h in the 15 min treatment when compared with the untreated eggs control. Besides that, control larvae completely hatched in 192 h and the treatment with 10 min and 15 min not shown completely hatching suggesting a delay in the embryonic development of the D. saccharalis larvae. Overall, our study is the first report of the application of D. saccharalis eggs, and we believe that Diatomaceous Earth could be a potential tool for the control of Diatraea eggs, however, thorough investigations are required to elucidate the action of this powder on Diatraea and would be an alternative method to interrupt the biological cycle of the pest.

Neem (Azadirachta indica) is a plant native from India and very adaptable to hot climates. Commonly it has been found as an ornamental plant or used in popular medicine due to its herbal properties. The current work is a short review that aimed to discuss Neem' biotechnological application in agriculture, environment and pharmaceutical fields. The researches were performed using online databases and the papers that discussed the prospection of Neem in the agriculture field (as a fertilizer or playing plant protection against pests and disease), pharmaceutical industry (production of metabolites with biological activity) and environmental biotechnology (eco-friendly alternatives to mitigate chemical pesticide environ damage) have chosen. Based on Neem's properties is possible find it as a fertilizer, supplement for animal feed or playing important role in plant defenses against insects (bioinsecticide) and pathogenic microorganisms (antimicrobial activity) in the agriculture field or the pharmaceutical industry with metabolites which have antitumoral, antioxidant and anti-bactericidal activity, besides Neem has already been reported used in combination with nanoparticles. Overall, Neem has been mentioned as a great tool for biotechnological applications in different areas.

Endophytes may play important roles in agriculture. Spontaneous or induced mutant strains may increase their biotechnological properties. Seventeen Colletotrichum endophytic fungi were investigated for their plant growth-promoting characteristics (in vitro phosphate solubilization, IAA, and siderophore production). The five best strains were inoculated into bean seeds, and the most prominent isolate was selected to obtain auxotrophic mutants by Potassium Bromate Resistance System (PBRS). The plant growth-promoting ability of the mutant was also investigated. Further, 41.17% of the evaluated endophytes presented promising results for in vitro assays (C. karstii SL10, C. karstii SL28, C. karstii SL57, C. karstii SL59, C. karstii SL12, C. karstii SL40, and C. karstii SL24). The endophyte C. karstii SL57 was statistically conspicuous for plant height and root length when compared to those in control plants. Bromate-resistant mutant C. karstii SL57 increased in vitro phosphate solubilization (23%) and chlorophyll levels (Chlb 0.607 mg g-1 and Chlt 0.973 mg g-1) of bean plants when compared to the wild-type strain (Chlb 0.551 mg g-1 and Chlt 0.881 mg g-1). This is the first time an auxotrophic mutant fungus has been obtained by PBRS with a biotechnological application for the agricultural field Endophytes may play important roles in agriculture. Spontaneous or induced mutant strains may increase their biotechnological properties. Seventeen Colletotrichum endophytic fungi were investigated for their plant growth-promoting characteristics (in vitro phosphate solubilization, IAA, and siderophore production). The five best strains were inoculated into bean seeds, and the most prominent isolate was selected to obtain auxotrophic mutants by Potassium Bromate Resistance System (PBRS). The plant growth-promoting ability of the mutant was also investigated. Further, 41.17% of the evaluated endophytes presented promising results for in vitro assays (C. karstii SL10, C. karstii SL28, C. karstii SL57, C. karstii SL59, C. karstii SL12, C. karstii SL40, and C. karstii SL24). The endophyte C. karstii SL57 was statistically conspicuous for plant height and root length when compared to those in control plants. Bromate-resistant mutant C. karstii SL57 increased in vitro phosphate solubilization (23%) and chlorophyll levels (Chlb 0.607 mg g-1 and Chlt 0.973 mg g-1) of bean plants when compared to the wild-type strain (Chlb 0.551 mg g-1 and Chlt 0.881 mg g-1). This is the first time an auxotrophic mutant fungus has been obtained by PBRS with a biotechnological application for the agricultural field

Agro-industrial wastes contain high moisture content and are rich in nutrients, and can be used as useful substrates by microbes with the supplementation of nitrogen sources, thus providing an alternative tool for the industrial production of many products of economic value, such as enzymes for example. These are proteins that function as biological catalysts, responsible for carrying out various biochemical reactions, being applied in detergent, food, paper and cellulose, cosmetics, textile industries, etc. However, they are expensive raw materials, and it is worth noting that an important part of the cost of manufacturing enzymes is mainly due to the expense of means and fermentation processes. Thus, to minimize the cost of this production and in order to reduce the degradation of the environment due to agricultural waste, a variety of microorganisms and agro-industrial “by-products” can be used to facilitate the economic production of enzymes.Therefore, in this work, a review was carried out on agro-industrial residues and the main enzymes in the industrial market, as well as the use of these materials as sources to obtain enzymes produced by fungi.

Grande parte dos efluentes produzidos por indústrias e domicílios são lançados no ambiente tornando-se altamente nocivos aos seres vivos e ao ambiente. Muitos destes efluentes apresentam em sua composição metais pesados. Os metais pesados podem favorecer a inativação de algumas enzimas essenciais para o metabolismo celular causando diversos danos à saúde. Em efluentes líquidos há diversas técnicas que podem ser empregadas na remoção desses contaminantes, entretanto, alguns desses métodos apresentam custo elevado, baixa remoção do metal e alta demanda de reagentes. Uma perspectiva crescente, é o uso de materiais de origem biológica para remoção dessas substâncias nocivas. Neste trabalho, buscamos realizar uma revisão de literatura (1997-2019) acerca de métodos alternativos, especialmente os biotecnológicos como biossorção e fitorremediação, para remoção de metais pesados do meio aquoso. Tais técnicas foram escolhidas por apresentarem baixo custo associado e serem ecologicamente favoráveis. Para biossorção, encontramos diferentes subprodutos de origem vegetal capazes de remover diversos íons metálicos, como a casca de laranja, coco verde, folhas de bananeira e sementes. Fungos (Pleurotus ostreatus), bactérias (Bacillus licheniformis) e microalgas (Spirulina platensis) também foram descritos como bons removedores de metais pesados. As plantas, por meio da fitorremediação, também demonstraram-se eficazes, notadamente as espécies: Lemna aequinoctialis, Helianthus annuus, Commelina benghalensis. Zea mays, Amaranthus hybridus, Pistia stratiotes. Nesta perspectiva, podemos concluir que os processos de biossorção e fitorremediação empregando produtos/subprodutos vegetais, microbianos ou animais são viáveis pelo seu baixo custo e boa disponibilidade, além de apresentarem taxas expressivas na remoção de substâncias recalcitrantes, notadamente de metais pesados.

Endófitos são fungos e bactérias que vivem no interior das plantas sem causar nenhum dano aos seus hospedeiros. Nas últimas décadas o potencial biotecnológico desses micro-organismos tem aumentado com o advento da nanotecnologia. Estudos descobriram que fungos e bactérias são capazes de produzirem nanopartículas por meio de rotas intra ou extracelulares, processo este que passou a ser descrito como micosíntese. O interesse cada vez maior para métodos mais sustentáveis e biológicos resultou no desenvolvimento de nanopartículas não-tóxicas e comparativamente mais bioativas. Ao contrário dos métodos físicos e químicos, a micosíntese é rentável e favorável ao meio ambiente. Neste trabalho buscamos realizar uma revisão de literatura acerta dos micro-organismos endofíticos e síntese biológica de nanopartículas. Os gêneros de bactérias endofíticas mais estudados quanto a micosíntese destacam-se Bacillus e Pseudomonas, enquanto para fungos Aspergillus, Penicillium, Fusarium, Colletotrichum, Altrnaria e Trichoderma. Dentre as principais aplicações biotecnológicas destas nanopartículas oriundas da síntese microbiana temos atividades antibacterianas, citotóxicas e antioxidantes. A associação entre endófitos e nanopartículas está em fase exploratória para compreender com clareza os mecanismos responsáveis por essa interação. Entretanto, sabe-se que estas nanopartículas produzidas por meio da micosíntese já são prospectadas em algumas atividades biológicas. Espera-se que todas as espécies de plantas abriguem endófitos, os quais podem ser isolados e bioprospectados para a produção de novas ou já conhecidas nanopartículas.