RESUMEN
The plant vascular system is a key element for long-distance communication. Understanding its composition may provide valuable information on how plants grow and develop themselves. In this study, a quantitative proteome dataset of the vascular sap proteome of three commercially important Eucalyptus species was shown. Protein extraction was carried out using a pressure bomb, whereas only in silico predicted extracellular proteins were considered as part of the sap proteome. A total of 132 different proteins were identified in all three Eucalyptus species and the most abundant proteome subset within all three species was comprised of proteins involved in the carbohydrate metabolic process, proteolysis, components of membrane, and defense response. The sap proteome of the species E. grandis and E. urophylla revealed the highest similarities. Functional classification indicated that the sap proteome of E. grandis and E. urophylla are mostly comprised of proteins involved in defense response and proteolysis; whereas no prominent functional class was observed for the E. camaldulensis species. Quantitative comparison highlighted characteristic sap proteins in each of the Eucalyptus species. The results that could be found in this study can be used as a reference for the proteome sap analysis of Eucalyptus plants grown under different conditions.
Asunto(s)
Eucalyptus , Eucalyptus/metabolismo , Proteoma/metabolismoRESUMEN
Açaí palm (Euterpe oleracea Mart.) seeds are a rich source of mannans, which can be used to generate bioethanol or be converted to high-value D-mannose, in addition to being a source of polyphenols with beneficial health properties. Here, we present a quantitative proteome dataset of açaí seeds at four stages of development (S1, S2, S3, and S4 stages), in which 2465 high confidence proteins were identified and 524 of them show statistically different abundance profiles during development. Several enzymes involved in the biosynthesis of nucleotide-sugars were quantified, especially those dedicated to the formation of GDP-mannose, which showed an increase in abundance between stages S1 and S3. Our data suggest that linear mannans found abundantly in endosperm cell walls are initially deposited as galactomannans, and during development lose the galactosyl groups. Two isoforms of alpha-galactosidase enzymes showed significantly increased abundances in the S3 and S4 stages. Additionally, we quantified the enzymes participating in the central pathway of flavonoid biosynthesis responsible for the formation of catechin and epicatechin, which are subunits of procyanidins, the main class of polyphenols in the açaí seeds. These proteins showed the same pattern of deposition, in which higher abundances were seen in the S1 and S2 stages.
Asunto(s)
Euterpe , Mananos , Antioxidantes , Proteómica , Semillas/química , Polifenoles/análisis , Extractos VegetalesRESUMEN
Eucalyptus grandis and Eucalyptus globulus are important species for the Brazilian forestry industry. E. grandis plantations are mainly found in tropical regions, yet E. globulus plants are usually cultivated under moderate to low temperature conditions. As temperature seems to be a key factor for the planting of these species, we revisited our previously generated shotgun proteomics dataset to identify the main patterns of proteome regulation induced by thermal stimulus and to pinpoint specific proteins involved in the environmental response. Large-scale analysis has pointed out the different proteomic responses of E. grandis and E. globulus under temperature stimulus, with 296 proteins considered to be differentially regulated in the stems of Eucalyptus spp. grown at different temperatures. A stringent filtering approach was used to identify the most differentially regulated proteins. Through the stringent criteria, 66 proteins were found to be enriched in the plant species. Cultivation of E. globulus plants in low-temperature conditions induced the highest number of differentially regulated proteins. Additionally, metabolic proteins were mostly down-regulated, while stress-related proteins were majorly up-regulated in both species. Finally, the subset of the most differentially regulated proteins comprised new candidates of protein markers of temperature stress.
Asunto(s)
Eucalyptus/metabolismo , Tallos de la Planta/metabolismo , Proteoma , Proteómica , Temperatura , Análisis por Conglomerados , Biología Computacional , Proteómica/métodos , Estrés FisiológicoRESUMEN
The role of the cotyledonary haustorium (CH) in the mobilization of nutrient reserves in the endosperm of species of the palm family Arecaceae is a moot question. To shed light on this matter, we present here an analysis of the quantitative proteome changes associated with four developmental stages of CH and three of endosperm during germination. Together, a total of 1965 proteins were identified, being 1538 in the CH and 960 in the endosperm. Both in the CH and endosperm proteomes, we observed an increase in the diversity of hydrolases as the CH and endosperm develops. Qualitative proteomics analysis of four CH developmental stages indicated that each stage is populated by a unique set of proteins and the quantitative analysis showed an increase in the relative abundance of hydrolases, particularly mannan degrading enzymes, as development progresses. These results add weight to the hypothesis that the CH in the seeds of E. oleraceaacts both as a conduit of carbon and nitrogen sources generated by the hydrolysis of the reserves in the endosperm and as a source of hydrolases that will contribute to the mobilization of these reserves.
Asunto(s)
Euterpe/fisiología , Germinación/genética , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Semillas/crecimiento & desarrollo , Cotiledón/metabolismo , Endospermo/metabolismoRESUMEN
The phorbol esters in the seeds of Jatropha curcas are a major hindrance to the full exploitation of the potential of this oil crop as a source of raw material for the production of biodiesel. Here, various quantitative proteomic strategies are used to establish the proteomes of roots, leaves, and endosperm of two genotypes of J. curcas with contrasting levels of phorbol esters in the seeds. In total 4532, 1775, and 503 proteins are identified respectively in roots, leaves, and endosperm, comprising 5068 unique proteins; of this total, 185 are differentially abundant in roots, 72 in leaves, and 20 in the endosperm. The biosynthetic pathways for flavonoids and terpenoids are well represented in roots, including the complete set of proteins for the mevalonate and non-mevalonate/Deoxyxylulose 5-Phosphate pathways, and proteins involved in the branches which lead to the synthesis tricyclic diterpenoids and gibberellins. Also, casbene synthase which catalyzes the first committed step in the biosynthesis of tigliane-type diterpenes is identified in roots of both genotypes, but not in leaves and endosperm. This dataset will be a valuable resource to explore the biochemical basis of the low toxicity of Jatropha genotypes with low concentration of phorbol esters in the seeds.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Jatropha/metabolismo , Ésteres del Forbol/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/análisis , Proteoma/metabolismo , Semillas/metabolismo , Genotipo , Jatropha/crecimiento & desarrollo , Semillas/crecimiento & desarrolloRESUMEN
The rice hoja blanca virus (RHBV), transmitted by the planthopper insect Tagosodes orizicolus, is a disease that attacks rice and generates significant production losses in Colombia. Fedearroz 2000 and Colombia I commercial rice varieties, which have different resistance levels to the disease, were selected in this study. To identify proteins associated to the insect and virus signaling, a comparative proteomics study was performed. By comparing proteomic profiles, between virus-infected and control group plants in two-dimensional electrophoresis, proteins exhibiting significant changes in abundance were found. In another test, peptide dendrimers containing sequences conformationally restricted to α-helix from four of those rice proteins were synthesized. In the experiment, sera from mice inoculated with peptide dendrimers could recognize the corresponding native protein in ELISA assays. Reported comparative proteomic results provide new insights into the molecular mechanisms of plant response to the RHBV and comprehensive tools for the analysis of new crop varieties. Besides, results from conformational peptide dendrimer approach are promising and show that it is feasible to detect proteins as markers, and may have biological applications by decreasing the susceptibility to proteolytic degradation.
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Anticuerpos/química , Anticuerpos/inmunología , Oryza/inmunología , Péptidos/química , Proteínas de Plantas/inmunología , Conformación Proteica , Dicroismo Circular , Oryza/metabolismo , Oryza/virología , Fenotipo , Proteómica/métodos , Espectrometría de Masas en Tándem , TenuivirusRESUMEN
The natural rubber latex extracted from the bark of Hevea brasiliensis plays various important roles in modern society. Post-translational modifications (PTMs) of the latex proteins are important for the stability and functionality of the proteins. In this study, latex proteins were acquired from the C-serum, lutoids, and rubber particle layers of latex without using prior enrichment steps; they were fragmented using collision-induced dissociation (CID), higher-energy collisional dissociation (HCD), and electron-transfer dissociation (ETD) activation methods. PEAKS 7 were used to search for unspecified PTMs, followed by analysis through PTM prediction tools to crosscheck both results. There were 73 peptides in 47 proteins from H. brasiliensis protein sequences derived from UniProtKB were identified and predicted to be post-translationally modified. The peptides with PTMs identified include phosphorylation, lysine acetylation, N-terminal acetylation, hydroxylation, and ubiquitination. Most of the PTMs discovered have yet to be reported in UniProt, which would provide great assistance in the research of the functional properties of H. brasiliensis latex proteins, as well as being useful biomarkers. The data are available via the MassIVE repository with identifier MSV000082419.
Asunto(s)
Hevea/metabolismo , Látex/metabolismo , Proteínas de Plantas/metabolismo , Secuencia de Aminoácidos/fisiología , Hevea/química , Látex/química , Péptidos/metabolismo , Fosforilación , Proteínas de Plantas/genética , Procesamiento Proteico-Postraduccional/fisiología , Proteómica/métodosRESUMEN
Tymovirus is a genus of plant pathogenic viruses that infects several dicotyledonous plants worldwide, causing serious diseases in economically important crops. The known cytopathic effect on the host cell organelles involves chloroplast membrane deformation and the induction of vesicles in its periphery. These vesicles are known to be the location where tymoviral genomic RNA replication occurs. Tomato blistering mosaic virus (ToBMV) is a tymovirus recently identified in tomato plants in Brazil, which is able to infect several other plants, including tobacco. In this work, we investigated the chloroplast proteomic profile of ToBMV-infected N. benthamiana using bidimensional electrophoresis (2-DE) and mass spectrometry, aiming to study the virus-host interaction related to the virus replication and infection. A total of approximately 200 spots were resolved, out of which 36 were differentially abundant. Differential spots were identified by mass spectrometry including photosynthesis-related and defense proteins. We identified proteins that may be targets of a direct interaction with viral proteins, such as ATP synthase ß subunit, RNA polymerase beta-subunit, 50S ribosomal protein L6 and Trigger factor-like protein. The identification of these candidate proteins gives support for future protein-protein interaction studies to confirm their roles in virus replication and disease development.
Asunto(s)
Cloroplastos/metabolismo , Virus del Mosaico/fisiología , Nicotiana/metabolismo , Proteoma/metabolismo , Solanum lycopersicum , Electroforesis en Gel Bidimensional , Interacciones Huésped-Patógeno , Enfermedades de las Plantas , Proteínas de Plantas/metabolismo , Unión Proteica , Nicotiana/virología , Proteínas Virales/metabolismo , Replicación ViralRESUMEN
The natural rubber latex extracted from the bark of Hevea brasiliensis plays various important roles in today's modern society. Following ultracentrifugation, the latex can be separated into 3 layers: C-serum, lutoids, and rubber particles. Previous studies have shown that a large number of proteins are present in these 3 layers. However, a complete proteome for this important plant is still unavailable. Protein sequences have been recently translated from the completed draft genome database of H. brasiliensis, leading to the creation of annotated protein databases of the following H. brasiliensis biosynthetic pathways: photosynthesis, latex allergens, rubberwood formation, latex biosynthesis, and disease resistance. This research was conducted to identify the proteins contained within the latex by way of de novo sequencing from mass spectral data obtained from the 3 layers of the latex. Peptides from these proteins were fragmented using collision-induced dissociation, higher-energy collisional dissociation, and electron-transfer dissociation activation methods. A large percentage of proteins from the biosynthetic pathways (63% to 100%) were successfully identified. In addition, a total of 1839 unique proteins were identified from the whole translated draft genome database (AnnHBM).
Asunto(s)
Alérgenos/aislamiento & purificación , Hevea/química , Látex/química , Proteínas de Plantas/aislamiento & purificación , Proteoma/aislamiento & purificación , Alérgenos/genética , Alérgenos/inmunología , Secuencia de Aminoácidos , Fraccionamiento Químico , Resistencia a la Enfermedad/genética , Ontología de Genes , Hevea/genética , Hevea/inmunología , Anotación de Secuencia Molecular , Fotosíntesis/genética , Corteza de la Planta/química , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Proteínas de Plantas/genética , Proteínas de Plantas/inmunología , Proteómica/métodos , UltracentrifugaciónRESUMEN
Floral and extrafloral nectaries are unique organs that secrete energy rich chemical components, but their contribution for nectar production is largely unknown. Here, we present the first comparative proteome dataset of four developmental stages of the extrafloral nectaries from castor plant (Ricinus communis), an important biofuel crop. Respectively, from stage I-IV, we identified 626, 613, 449 and 356 proteins, respectively, summing up 882 nonredundant proteins. Surprisingly, we identified two isoforms of the potent toxin ricin, indicating that ricin expression is not limited to seeds, but it may serve a general defense purpose for the castor plant. To date, this is the most complete dataset of proteins either from floral or extrafloral nectaries, thus contributing to lay the foundations for investigations on their ecological and evolutionary importance.
Asunto(s)
Proteínas de Plantas/metabolismo , Ricinus/crecimiento & desarrollo , Proteínas de Plantas/análisis , Proteoma/análisis , Proteoma/metabolismo , Proteómica , Ricina/análisis , Ricina/metabolismo , Ricinus/metabolismoRESUMEN
Cowpea (Vigna unguiculata L. Walp) is an important legume species well adapted to low fertility soils and prolonged drought periods. One of the main problems that cause severe yield losses in cowpea is the root-knot nematode Meloidogyne incognita. The aim of this work was to analyze the differential expression of proteins in the contrasting cultivars of cowpea CE 31 (highly resistant) and CE 109 (slightly resistant) during early stages of M. incognita infection. Cowpea roots were collected at 3, 6, and 9 days after inoculation and used for protein extraction and 2-DE analysis. From a total of 59 differential spots, 37 proteins were identified, mostly involved in plant defense, such as spermidine synthase, patatin, proteasome component, and nitrile-specifier protein. A follow-up study was performed by quantitative RT-PCR analysis of nine selected proteins and the results revealed a very similar upregulation trend between the protein expression profiles and the corresponding transcripts. This study also identified ACT and GAPDH as a good combination of reference genes for quantitative RT-PCR analysis of the pathosystem cowpea/nematode. Additionally, an interactome analysis showed three major pathways affected by nematode infection: proteasome endopeptidase complex, oxidative phosphorylation, and flavonoid biosynthesis. Taken together, the results obtained by proteome, transcriptome, and interactome approaches suggest that oxidative stress, ubiquitination, and glucosinolate degradation may be part of cowpea CE 31 resistance mechanisms in response to nematode infection.
Asunto(s)
Fabaceae/parasitología , Interacciones Huésped-Parásitos , Raíces de Plantas/metabolismo , Raíces de Plantas/parasitología , Proteómica/métodos , Tylenchoidea/fisiología , Animales , Electroforesis en Gel Bidimensional , Fabaceae/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Estudios de Asociación Genética , Interacciones Huésped-Parásitos/genética , Espectrometría de Masas , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/parasitología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Yellow lupin (Lupinus luteus L.) is a legume crop containing a large amount of protein in its seeds. In this study, we constructed a seed-protein catalog to provide a foundation for further study of the seeds. A total of 736 proteins were identified in 341 2DE spots by nano-LC-MS/MS. Eight storage proteins were found as multiple spots in the 2DE gels. The 736 proteins correspond to 152 unique proteins as shown by UniRef50 clustering. Sixty-seven of the 152 proteins were associated with KEGG-defined pathways. Of the remaining proteins, 57 were classified according to a GO term. The functions of the remaining 28 proteins have yet to be determined. This is the first yellow lupin seed-protein catalog, and it contains considerably more data than previously reported for white lupin (L. albus L.).
Asunto(s)
Lupinus/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Proteómica/métodos , Semillas/metabolismo , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Proteínas de Plantas/genética , Semillas/química , Espectrometría de Masas en TándemRESUMEN
The main goal of this work is to evaluate some differential protein species in transgenic (T) and nontransgenic (NT) Arabidopsis thaliana plants after their cultivation in the presence or absence of sodium selenite. The transgenic line was obtained through insertion of CaMV 35S controlling nptII gene. Comparative proteomics through 2D-DIGE is carried out in four different groups (NT × T; NT × Se-NT (where Se is selenium); Se-NT × Se-T, and T × Se-T). Although no differential proteins are achieved in the T × Se-T group, for the others, 68 differential proteins (by applying a regulation factor ≥1.5) are achieved, and 27 of them accurately characterized by ESI-MS/MS. These proteins are classified into metabolism, energy, signal transduction, disease/defense categories, and some of them are involved in the glycolysis pathway-Photosystems I and II and ROS combat. Additionally, laser ablation imaging is used for evaluating the Se and sulfur distribution in leaves of different groups, corroborating some results obtained and related to proteins involved in the glycolysis pathway. From these results, it is possible to conclude that the genetic modification also confers to the plant resistance to oxidative stress.
Asunto(s)
Arabidopsis/genética , Hojas de la Planta/genética , Proteómica , Selenito de Sodio/administración & dosificación , Arabidopsis/efectos de los fármacos , Arabidopsis/crecimiento & desarrollo , Electroforesis en Gel Bidimensional/métodos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Rayos Láser , Imagen Molecular/métodos , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/crecimiento & desarrollo , Proteínas de Plantas/biosíntesis , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrolloRESUMEN
The use of cell walls to produce cellulosic ethanol from sugarcane bagasse is a new challenge. A better knowledge of proteins involved in cell wall remodelling is essential to improve the saccharification processes. Cell suspension cultures were used for this first cell wall proteomics study of sugarcane. Proteins extracted from cell walls were identified using an adapted protocol. They were extracted using 0.2 M CaCl2 and 2 M LiCl after purification of cell walls. The proteins were then identified by the innovative nanoACQUITY UPLC MS/MS technology and bioinformatics using the translated SUCEST EST cluster database of sugarcane. The experiments were reproduced three times. Since Sorghum bicolor is the closest plant with a fully sequenced genome, homologous proteins were searched for to complete the annotation of proteins, that is, prediction of subcellular localization and functional domains. Altogether, 69 different proteins predicted to be secreted were identified among 377 proteins. The reproducibility of the experiments is discussed. These proteins were distributed into eight functional classes. Oxidoreductases such as peroxidases were well represented, whereas glycoside hydrolases were scarce. This work provides information about the proteins that could be manipulated through genetic transformation, to increase second-generation ethanol production.
Asunto(s)
Pared Celular/química , Proteínas de Plantas/análisis , Saccharum/citología , Técnicas de Cultivo de Célula , Proteómica/métodos , Espectrometría de Masas en TándemRESUMEN
The root knot nematodes (RKN), Meloydogine spp., particularly Meloidogyne incognita and Meloidogyne javanica species, parasitize several plant species and are responsible for large annual yield losses all over the world. Only a few available chemical nematicides are still authorized for RKN control owing to environmental and health reasons. Thus, plant resistance is currently considered the method of choice for controlling RKN, and research performed on the molecular interactions between plants and nematodes to identify genes of interest is of paramount importance. The present work aimed to identify the differential accumulation of root proteins of a resistant cowpea genotype (CE-31) inoculated with M. incognita (Race 3) in comparison with mock-inoculated control, using 2D electrophoresis assay, mass spectrometry identification and gene expression analyses by RT-PCR. The results showed that at least 22 proteins were differentially represented in response to RKN challenge of cowpea roots mainly within 4-6 days after inoculation. Amongst the up-represented proteins were SOD, APX, PR-1, ß-1,3-glucanase, chitinases, cysteine protease, secondary metabolism enzymes, key enzymes involved in ethylene biosynthesis, proteins involved in MAPK pathway signaling and, surprisingly, leghemoglobin in non-rhizobium-bacterized cowpea. These findings show that an important rearrangement in the resistant cowpea root proteome occurred following challenge with M. incognita.
RESUMEN
Tomato, one of the most important crops cultivated worldwide, has been severely affected by begomoviruses such as the Tomato chlorotic mottle virus (ToCMoV). Virulence factor AC2 is considered crucial for a successful virus-plant interaction and is known to act as a transcriptional activator and in some begomoviruses to function as an RNA silencing suppressor factor. However, the exact functions of the AC2 protein of the begomovirus ToCMoV are not yet established. The aim of the present study was to identify differentially expressed proteins of the model plant Nicotiana benthamiana in response to the expression of the AC2 gene, isolated from ToCMoV. N. benthamiana plants were inoculated with Agrobacterium tumefaciens containing the viral vector Potato virus X (PVX) and with the PVX-AC2 construction. 2DE was performed and proteins were identified by MS. The results showed that the expression of ToCMoV AC2 alters the levels of several host proteins, which are important for normal plant development, causing an imbalance in cellular homeostasis. This study highlights the effect of AC2 in the modulation of plant defense processes by increasing the expression of several oxidative stress-related and pathogenesis-related proteins, as well as its role in modulating the proteome of the photosynthesis and energy production systems.