NMR Fingerprinting of Conventional and Genetically Modified Soybean Plants with AtAREB1 Transcription Factors.

Drought stress impacts soybean yields and physiological processes. However, the insertion of the activated form of the AtAREB1 gene in the soybean cultivar BR16, which is sensitive to water deficit, improved the drought response of the genetically modified plants. Thus, in this study, we used 1H NMR in solution and solid-state NMR to investigate the response of genetically modified soybean overexpressing AtAREB1 under water deficiency conditions. We achieved that drought-tolerant soybean yields high content of amino acids isoleucine, leucine, threonine, valine, proline, glutamate, aspartate, asparagine, tyrosine, and phenylalanine after 12 days of drought stress conditions, as compared to drought-sensitive soybean under the same conditions. Specific target compounds, including sugars, organic acids, and phenolic compounds, were identified as involved in controlling sensitive soybean during the vegetative stage. Solid-state NMR was used to study the impact of drought stress on starch and cellulose contents in different soybean genotypes. The findings provide insights into the metabolic adjustments of soybean overexpressing AREB transcription factors in adapting to dry climates. This study presents NMR techniques for investigating the metabolome of transgenic soybean plants in response to the water deficit. The approach allowed for the identification of physiological and morphological changes in drought-resistant and drought-tolerant soybean tissues. The findings indicate that drought stress significantly alters micro- and macromolecular metabolism in soybean plants. Differential responses were observed among roots and leaves as well as drought-tolerant and drought-sensitive cultivars, highlighting the complex interplay between overexpressed transcription factors and drought stress in soybean plants.

Influence of plant growth-promoting bacteria on leaf carbon and nitrogen metabolism of two drought-stressed neotropical tree species: a metabolomic approach.

Deforestation of Atlantic Forest has caused prolonged drought events in the last decades. The need for reforestation is growing, and the development of native seedlings that are more tolerant to drought stress is necessary. A biotechnological tool that improves plant tolerance is the use of plant growth-promoting bacteria (PGPB) as inoculants. Two species of PGPB were inoculated in drought-stressed seedlings of two neotropical tree species that have been used in environmental restoration programs: Cecropia pachystachya and Cariniana estrellensis. Biometrical, physiological, and metabolomic parameters from carbon and nitrogen pathways were evaluated. We found that the PGPB positively influenced photosynthesis and growth parameters in both trees under drought. The enzymes activities, the tricarboxylic acid cycle intermediates, the amino acids, and protein contents were also influenced by the PGPB treatments. The results allowed us to find the specific composition of secondary metabolites of each plant species. This study provides evidence that there is not a single mechanism involved in drought tolerance and that the inoculation with PGPB promotes a broad-spectrum tolerance response in Neotropical trees. The inoculation with PGPB appears as an important strategy to improve drought tolerance in Atlantic Forest native trees and enhance environmental restoration programs' success. MAIN CONCLUSION: The association with plant growth-promoting bacteria improved the tolerance to drought in Neotropical trees through biochemical, physiological, and biometrical parameters. This can enhance the success of forest restoration programs.

Machine Learning-Based Virtual Screening of Antibacterial Agents against Methicillin-Susceptible and Resistant Staphylococcus aureus.

The application of computer-aided drug discovery (CADD) approaches has enabled the discovery of new antimicrobial therapeutic agents in the past. The high prevalence of methicillin-resistantStaphylococcus aureus(MRSA) strains promoted this pathogen to a high-priority pathogen for drug development. In this sense, modern CADD techniques can be valuable tools for the search for new antimicrobial agents. We employed a combination of a series of machine learning (ML) techniques to select and evaluate potential compounds with antibacterial activity against methicillin-susceptible S. aureus (MSSA) and MRSA strains. In the present study, we describe the antibacterial activity of six compounds against MSSA and MRSA reference (American Type Culture Collection (ATCC)) strains as well as two clinical strains of MRSA. These compounds showed minimal inhibitory concentrations (MIC) in the range from 12.5 to 200 µM against the different bacterial strains evaluated. Our results constitute relevant proven ML-workflow models to distinctively screen for novel MRSA antibiotics.

Metabolic alterations in conventional and genetically modified soybean plants with GmDREB2A;2 FL and GmDREB2A;2 CA transcription factors during water deficit.

Water deficit is one of the main abiotic stress that affects plant growth and productivity. The GmDREB2A;2 (Glyma14g06080) gene is an important transcription factor involved in regulating the plants' responses under water deficit. In previous studies, soybean plants overexpressing full-length (GmDREB2A;2 FL) and constitutively active (GmDREB2A;2 CA) forms of the GmDREB2A;2 gene, presented higher tolerance to water deficit when compared with the conventional cultivar BRS 283. Therefore, identifying the changes in metabolite profile in these tolerant genotypes can contribute to the understanding of the metabolic pathways involved in the tolerance mechanism. In this work, the metabolic changes in roots and leaves of genetically modified (GM) soybean plants subjected to water deficit were elucidated by 1H-NMR spectroscopy. Three events were analyzed, one containing the gene in FL form (GmDREB2A;2 FL) and two presenting its CA form (GmDREB2A;2 CA-1 and GmDREB2A;2 CA-2) and compared with the conventional cultivar BRS 283. The results indicated different responses between leaves and roots for all genotypes. Most of these metabolic variations were related to carbohydrate and amino acid pathways. BRS 283 stood out with higher accumulation of amino acids in leaves under water deficit. The results also showed that the events GmDREB2A;2 FL and GmDREB2A;2 CA-1 presented higher concentrations of ß-glucose and fructose in leaves, whereas BRS 283 accumulated more sucrose and pinitol. In roots, the GM events accumulated higher ß-glucose, fructose, asparagine and phenylalanine, when compared with the conventional cultivar. These insights can add information on how the transcription factor (TF) DREB2A acts in soybean plants triggering and controlling a network of complex responses to drought.

Identification of primary and secondary metabolites and transcriptome profile of soybean tissues during different stages of hypoxia.

NMR and chromatography methods combined with mass spectrometry are the most important analytical techniques employed for plant metabolomics screening. Metabolomic analysis integrated to transcriptome screening add an important extra dimension to the information flow from DNA to RNA to protein. The most useful NMR experiment in metabolomics analysis is the proton spectra due the high receptivity of 1H and important structural information, through proton-proton scalar coupling. Routinely, databases have been used in identification of primary metabolites, however, there is currently no comparable data for identification of secondary metabolites, mainly, due to signal overlap in normal 1H NMR spectra and natural variation of plant. Related to spectra overlap, alternatively, better resolution can be find using 1H pure shift and 2D NMR pulse sequence in complex samples due to spreading the resonances in a second dimension. Thus, in data brief we provide a catalogue of metabolites and expression levels of genes identified in soy leaves and roots under flooding stress.

Integrating High-Resolution and Solid-State Magic Angle Spinning NMR Spectroscopy and a Transcriptomic Analysis of Soybean Tissues in Response to Water Deficiency.

INTRODUCTION: Solid-state NMR (SSNMR) spectroscopy methods provide chemical environment and ultrastructural details that are not easily accessible by other non-destructive, high-resolution spectral techniques. High-resolution magic angle spinning (HR-MAS) has been widely used to obtain the metabolic profile of a heterogeneous sample, combining the resolution enhancement provided by MAS in SSNMR with the shimming and locking procedures in liquid-state NMR. OBJECTIVE: In this work, we explored the feasibility of using the HR-MAS and SSNMR techniques to identify metabolic changes in soybean leaves subjected to water-deficient conditions. METHODOLOGY: Control and water-deficient soybean leaves were analysed using one-dimensional (1D) HR-MAS and SSNMR. Total RNA was extracted from the leaves for the transcriptomic analysis. RESULTS: The 1 H HR-MAS and CP-MAS 13 C{1 H} spectra of soybean leaves grown with and without water deficiency stress revealed striking differences in metabolites. A total of 30 metabolites were identified, and the impact of water deficiency on the metabolite profile of soybean leaves was to induce amino acid synthesis. High expression levels of genes required for amino acid biosynthesis were highly correlated with the compounds identified by 1 H HR-MAS. CONCLUSIONS: The integration of the 1 H HR-MAS and SSNMR spectra with the transcriptomic data provided a complete picture of the major changes in the metabolic profile of soybeans in response to water deficiency. Copyright © 2017 John Wiley & Sons, Ltd.

Metabolite Profiling of Sugarcane Genotypes and Identification of Flavonoid Glycosides and Phenolic Acids.

Sugarcane is an important agricultural crop in the economy of tropical regions, and Brazil has the largest cultivated acreage in the world. Sugarcane accumulates high levels of sucrose in its stalks. Other compounds produced by sugarcane are currently not of economic importance. To explore potential coproducts, we have studied the chemical diversity of sugarcane genotypes, via metabolite profiling of leaves by NMR and LC-DAD-MS. Metabolites were identified via in-house and public databases. From the analysis of 60 HPLC-fractionated extracts, LC-DAD-MS detected 144 metabolites, of which 56 were identified (MS-MS and (1)H NMR), including 19 phenolics and 25 flavones, with a predominance of isomeric flavone C-glycosides. Multivariate analysis of the profiles from genotypes utilized in Brazilian breeding programs revealed clustering according to sugar, phenolic acid, and flavone contents.

Benefícios da equitação com fins terapêuticos na reabilitação de crianças com paralisia cerebral / Benefits of hippotherapy and horseback riding in the rehabilitation of children with cerebral palsy

Este estudo surge devido à importância que a equitaçãocom fins terapêuticos tem vindo a assumir como complementona reabilitação das crianças com Paralisia Cerebral (PC). Objectivo:Verificar quais são os benefícios da equitação com fins terapêuticosnas crianças com paralisia cerebral. Material e métodos: Foram utilizadasnove bases de dados na pesquisa de artigos que abordassem estatemática. Resultados: Foram encontrados cinco artigos experimentais(um sobre equitação terapêutica e os restantes sobre hipoterapia)e um artigo de revisão sistemática. Conclusão: Foram encontradosbenefícios na função motora das crianças com PC, consequentesda utilização da equitação com fins terapêuticos...

This study emerges due to the importance ofthe horseback riding therapy as a complement of rehabilitation inchildren diagnosed with Cerebral Palsy (CP). Purpose: To verifywhich are the benefits of the horseback riding and hippotherapy inchildren diagnosed with CP. Methods: Nine databases were used tosearch articles on this theme. Results: Five experimental articles (oneabout horseback riding, and the others about hippotherapy) andone systematic literature review were found. Conclusion: We foundbenefits in the motor function of children with CP, who participatein a horseback riding therapy...

Influência da variação sazonal nos teores de flavonóides e atividade antioxidante das folhas de Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae / The Influence of seasonal variation in levels of flavonoids and antioxidant activity of the leaves of Campomanesia adamantium

Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae, é uma espécie nativa. As folhas são utilizadas na medicina popular. Os extratos hexânico e acetato de etila das folhas mostraram pouca variação na sua composição química em diferentes estações do ano associado com o desenvolvimento do vegetal, enquanto o extrato etanólico mostrou alteração significativa em relação à composição química. Todos os extratos etanólicos mostraram alta atividade antioxidante frente ao método DPPH e de moderada a alta para β-caroteno/ácido linoléico.

Campomanesia adamantium (Cambess.) O. Berg, Myrtaceae, is a native species. The extracts of ethyl acetate and hexane showed little variation in their chemical composition at different seasons of the year associated with the development of the plant, while the ethanol extract showed significant change in relationship to chemical composition. All ethanol extracts showed high antioxidant activity against DPPH method and from moderate to high antioxidant activity for β-carotene/linoleic acid.

Gas Chromatography-Mass Spectrometry (GC-MS) and evaluation of antioxidant and antimicrobial activities of essential oil of Campomanesia adamantium (Cambess.) O. Berg (Guavira) / Cromatografia Gasosa-Espectrometria de Massas (CG-EM) e avaliação da atividade antioxidante e antimicrobiana do óleo essencial de Campomanesia adamantium (Cambess.) O. Berg (guavira)

The essential oils from Campomanesia adamantium (Cambess.) O. Berg leaves, collected in the reproductive (flowering and fruit-bearing) and vegetative stages, were characterized by GC-MS (Gas Chromatography-Mass Spectrometry). A total of 95 compounds of the essential oils were identified. In the reproductive stage (flowering) the major constituents were monoterpenes (limonene, α-pinene and β-pinene) while during the vegetative stage the major constituents were the sesquiterpenes (bicyclogermacrene and globulol). The essential oil of the reproductive stage shows high antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans, and all show moderate activity against Escherichia coli. The essential oils were also evaluated for their radical-scavenging activity by DPPH. The chemogeographical variations of the oil composition from the four distinct localities studied all contained α-pinene, β-pinene, limonene, linalool, β-caryophyllene, germacrene D and bicyclogermacrene, however the samples from Jardim city contained neither limonene nor linalool.

Os óleos essenciais obtidos das folhas de Campomanesia adamantium foram caracterizados através da combinação de CG-EM e índice de retenção, sendo identificado um total de 95 compostos. Na floração as substâncias majoritárias foram monoterpenos (limoneno, α-pineno e β-pineno) e durante o estágio vegetativo as substâncias majoritárias foram sesquiterpenos (biciclogermacreno e globulol). Os óleos essenciais obtidos da floração e frutificação mostraram alta atividade contra Staphylococcus aureus, Pseudomonas aeruginosa e Candida albicans e moderada contra Escherichia coli em todos os estágios. Foi avaliada a atividade antioxidante dos óleos essenciais usando o método do DPPH. O óleo essencial das 4 cidades mostrou a presença de α-pineno, β-pineno, limoneno, linalol, β-cariofileno, germacreno D e biciclogermacreno, mas a amostra da cidade de Jardim não apresentou limoneno e linalol.