RESUMO
Aging-related processes in plant tissues are associated with changes in developmental and physiological processes relevant for stress tolerance and plant performance. While senescence-regulated processes have been extensively characterized in leaves, they remain poorly described in roots. Here, we investigated the physiological processes and molecular determinants underlying the senescence of seminal roots in hydroponically grown barley (Hordeum vulgare). Transcriptome profiling in apical and basal root tissues revealed that several NAC-, WRKY-, and APETALA2 (AP2)-type transcription factors were upregulated just before the arrest of root elongation, when root cortical cell lysis and nitrate uptake, as well as cytokinin concentrations ceased. At this time point, root abscisic acid levels peaked, suggesting that abscisic acid is involved in root aging-related processes characterized by expression changes of genes involved in oxidative stress responses. This temporal sequence of aging-related processes in roots is highly reminiscent of typical organ senescence, with the exception of evidence for the retranslocation of nutrients from roots. Supported by the identification of senescence-related transcription factors, some of which are not expressed in leaves, our study indicates that roots undergo an intrinsic genetically determined senescence program, predominantly influenced by plant age.
Assuntos
Hordeum/metabolismo , Hordeum/fisiologia , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Ácido Abscísico/metabolismo , Envelhecimento/fisiologia , Citocininas/metabolismo , Regulação da Expressão Gênica de Plantas , Hordeum/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
A lipidome is the set of lipids in a given organism, cell or cell compartment and this set reflects the organism's synthetic pathways and interactions with its environment. Recently, lipidomes of biological model organisms and cell lines were published and the number of functional studies of lipids is increasing. In this study we propose a homology metric that can quantify systematic differences in the composition of a lipidome. Algorithms were developed to 1. consistently convert lipids structure into SMILES, 2. determine structural similarity between molecular species and 3. describe a lipidome in a chemical space model. We tested lipid structure conversion and structure similarity metrics, in detail, using sets of isomeric ceramide molecules and chemically related phosphatidylinositols. Template-based SMILES showed the best properties for representing lipid-specific structural diversity. We also show that sequence analysis algorithms are best suited to calculate distances between such template-based SMILES and we adjudged the Levenshtein distance as best choice for quantifying structural changes. When all lipid molecules of the LIPIDMAPS structure database were mapped in chemical space, they automatically formed clusters corresponding to conventional chemical families. Accordingly, we mapped a pair of lipidomes into the same chemical space and determined the degree of overlap by calculating the Hausdorff distance. We named this metric the 'Lipidome jUXtaposition (LUX) score'. First, we tested this approach for estimating the lipidome similarity on four yeast strains with known genetic alteration in fatty acid synthesis. We show that the LUX score reflects the genetic relationship and growth temperature better than conventional methods although the score is based solely on lipid structures. Next, we applied this metric to high-throughput data of larval tissue lipidomes of Drosophila. This showed that the LUX score is sufficient to cluster tissues and determine the impact of nutritional changes in an unbiased manner, despite the limited information on the underlying structural diversity of each lipidome. This study is the first effort to define a lipidome homology metric based on structures that will enrich functional association of lipids in a similar manner to measures used in genetics. Finally, we discuss the significance of the LUX score to perform comparative lipidome studies across species borders.
Assuntos
Biologia Computacional/métodos , Lipídeos/química , Modelos Biológicos , Modelos Moleculares , Algoritmos , Animais , Drosophila , Isomerismo , Lipídeos/análise , Especificidade de ÓrgãosRESUMO
Antimicrobial peptides (AMPs) are important components of the innate immune system of animals, plants, fungi and bacteria and are recently under discussion as promising alternatives to conventional antibiotics. We have investigated two cecropin-like synthetic peptides, Gm1, which corresponds to the natural overall uncharged Galleria mellonella native peptide and ΔGm1, a modified overall positively charged Gm1 variant. We have analysed these peptides for their potential to inhibit the endotoxin-induced secretion of tumour necrosis factor-α (TNF-α) from human mononuclear cells. Furthermore, in a conventional microbiological assay, the ability of these peptides to inhibit the growth of the rough mutant bacteria Salmonella enterica Minnesota R60 and the polymyxin B-resistant Proteus mirabilis R45 was investigated and atomic force microscopy (AFM) measurements were performed to characterize the morphology of the bacteria treated by the two peptides. We have also studied their cytotoxic properties in a haemolysis assay to clarify potential toxic effects. Our data revealed for both peptides minor anti-inflammatory (anti-endotoxin) activity, but demonstrated antimicrobial activity with differences depending on the endotoxin composition of the respective bacteria. In accordance with the antimicrobial assay, AFM data revealed a stronger morphology change of the R45 bacteria than for the R60. Furthermore, Gm1 had a stronger effect on the bacteria than ΔGm1, leading to a different morphology regarding indentations and coalescing of bacterial structures. The findings verify the biophysical measurements with the peptides on model systems. Both peptides lack any haemolytic activity up to an amount of 100µg/ml, making them suitable as new anti-infective agents.
Assuntos
Antibacterianos , Peptídeos Catiônicos Antimicrobianos , Endotoxemia/tratamento farmacológico , Proteínas de Insetos , Leucócitos Mononucleares/metabolismo , Mariposas/química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Células Cultivadas , Endotoxemia/induzido quimicamente , Endotoxemia/metabolismo , Endotoxemia/patologia , Feminino , Humanos , Proteínas de Insetos/química , Proteínas de Insetos/farmacologia , Leucócitos Mononucleares/patologia , Lipopolissacarídeos/toxicidade , Masculino , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Little is known about the human lung lipidome, its variability in different physiological states, its alterations during carcinogenesis and the development of pulmonary emphysema. We investigated how health status might be mirrored in the lung lipidome. Tissues were sampled for both lipidomic and histological analysis. Using a screening approach, we characterised lipidomes of lung cancer tissues and corresponding tumour-free alveolar tissues. We quantified 311 lipids from 11 classes in 43 tissue samples from 26 patients. Tumour tissues exhibited elevated levels of triacylglycerols and cholesteryl esters, as well as a significantly lower abundance of phosphatidylglycerols, which are typical lung surfactant components. Adenocarcinomas and squamous cell carcinomas were distinguished with high specificity based on lipid panels. Lipidomes of tumour biopsy samples showed clear changes depending on their histology and, in particular, their proportion of active tumour cells and stroma. Partial least squares regression showed correlations between lipid profiles of tumour-free alveolar tissues and the degree of emphysema, inflammation status, and the age of patients. Unsaturated long-chain phosphatidylserines and phosphatidylinositols showed a positive correlation with a worsened emphysema status and ageing. This work provides a resource for the human lung lipidome and a systematic data analysis strategy to link clinical characteristics and histology.