RESUMEN
Terpenes are important compounds in plant trophic interactions. A meta-analysis of GC-MS data from a diverse range of apple (Malus × domestica) genotypes revealed that apple fruit produces a range of terpene volatiles, with the predominant terpene being the acyclic branched sesquiterpene (E,E)-α-farnesene. Four quantitative trait loci (QTLs) for α-farnesene production in ripe fruit were identified in a segregating 'Royal Gala' (RG) × 'Granny Smith' (GS) population with one major QTL on linkage group 10 co-locating with the MdAFS1 (α-farnesene synthase-1) gene. Three of the four QTLs were derived from the GS parent, which was consistent with GC-MS analysis of headspace and solvent-extracted terpenes showing that cold-treated GS apples produced higher levels of (E,E)-α-farnesene than RG. Transgenic RG fruit downregulated for MdAFS1 expression produced significantly lower levels of (E,E)-α-farnesene. To evaluate the role of (E,E)-α-farnesene in fungal pathogenesis, MdAFS1 RNA interference transgenic fruit and RG controls were inoculated with three important apple post-harvest pathogens [Colletotrichum acutatum, Penicillium expansum and Neofabraea alba (synonym Phlyctema vagabunda)]. From results obtained over four seasons, we demonstrate that reduced (E,E)-α-farnesene is associated with decreased disease initiation rates of all three pathogens. In each case, the infection rate was significantly reduced 7 days post-inoculation, although the size of successful lesions was comparable with infections on control fruit. These results indicate that (E,E)-α-farnesene production is likely to be an important factor involved in fungal pathogenesis in apple fruit.
Asunto(s)
Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Malus/genética , Malus/metabolismo , Enfermedades de las Plantas/inmunología , Sesquiterpenos/metabolismo , Colletotrichum/patogenicidad , Resistencia a la Enfermedad , Regulación hacia Abajo , Hongos/patogenicidad , Cromatografía de Gases y Espectrometría de Masas , Ligamiento Genético , Genotipo , Penicillium/patogenicidad , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Sitios de Carácter Cuantitativo , Interferencia de ARN/inmunología , Terpenos/metabolismoRESUMEN
Phenylpropenes, such as eugenol and trans-anethole, are important aromatic compounds that determine flavour and aroma in many herbs and spices. Some apple varieties produce fruit with a highly desirable spicy/aromatic flavour that has been attributed to the production of estragole, a methylated phenylpropene. To elucidate the molecular basis for estragole production and its contribution to ripe apple flavour and aroma we characterised a segregating population from a Royal Gala (RG, estragole producer) × Granny Smith (GS, non-producer) apple cross. Two quantitative trait loci (QTLs; accounting for 9.2 and 24.8% of the variation) on linkage group (LG) 1 and LG2 were identified that co-located with seven candidate genes for phenylpropene O-methyltransferases (MdoOMT1-7). Of these genes, only expression of MdoOMT1 on LG1 increased strongly with ethylene and could be correlated with increasing estragole production in ripening RG fruit. Transient over-expression in tobacco showed that MdoOMT1 utilised a range of phenylpropene substrates and catalysed the conversion of chavicol to estragole. Royal Gala carried two alleles (MdoOMT1a, MdoOMT1b) whilst GS appeared to be homozygous for MdoOMT1b. MdoOMT1a showed a higher affinity and catalytic efficiency towards chavicol than MdoOMT1b, which could account for the phenotypic variation at the LG1 QTL. Multiple transgenic RG lines with reduced MdoOMT1 expression produced lower levels of methylated phenylpropenes, including estragole and methyleugenol. Differences in fruit aroma could be perceived in these fruit, compared with controls, by sensory analysis. Together these results indicate that MdoOMT1 is required for the production of methylated phenylpropenes in apple and that phenylpropenes including estragole may contribute to ripe apple fruit aroma.
Asunto(s)
Anisoles/metabolismo , Frutas/metabolismo , Malus/metabolismo , Metiltransferasas/metabolismo , Proteínas de Plantas/genética , Derivados de Alilbenceno , Etilenos/metabolismo , Eugenol/análogos & derivados , Eugenol/metabolismo , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Isoenzimas/genética , Isoenzimas/metabolismo , Malus/genética , Metiltransferasas/genética , Datos de Secuencia Molecular , Odorantes , Filogenia , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Sitios de Carácter CuantitativoRESUMEN
BACKGROUND: The chemical assignment of metabolites is crucial to understanding the relation between food composition and biological activity. OBJECTIVE: This study was designed to detect and chemically assign sulfur-containing metabolites by using LC-Fourier transform ion cyclotron resonance-mass spectrometry (FTICR-MS) in Allium plants. METHODS: Ultrahigh resolution (>250,000 full width at half-maximum) and mass accuracy (<1 mDa) by FTICR-MS allowed us to distinguish ions containing sulfur isotopes ((32)S and (34)S). RESULTS: Putative 69 S-containing monoisotopic ions (S-ions) were extracted from the metabolome data of onion (Allium cepa), green onion (Allium fistulosum), and garlic (Allium sativum) on the basis of theoretical mass differences between (32)S-ions and their (34)S-substituted counterparts and on the natural abundance of (34)S. Eight S-ions were chemically assigned by using the reference data according to the guidelines of the Metabolomics Standards Initiative. Three ions detected in garlic were assigned as derived from the isomers γ-glutamyl-S-1-propenylcysteine and γ-glutamyl-S-2-propenylcysteine and as S-2-propenylmercaptoglutathione on the basis of differences in key product ions identified in reference tandem MS spectra. CONCLUSION: The ability to discriminate between such geometric isomers will be extremely useful for the chemical assignment of unknown metabolites in MS-based metabolomics.
Asunto(s)
Cisteína/análisis , Ajo/química , Glutatión/análisis , Iones/análisis , Cebollas/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Azufre/análisis , Cromatografía Liquida/métodos , Ciclotrones , Isomerismo , Metaboloma , Metabolómica , Isótopos de Azufre/análisisRESUMEN
The 'fruity' attributes of ripe apples (Malus × domestica) arise from our perception of a combination of volatile ester compounds. Phenotypic variability in ester production was investigated using a segregating population from a 'Royal Gala' (RG; high ester production) × 'Granny Smith' (GS; low ester production) cross, as well as in transgenic RG plants in which expression of the alcohol acyl transferase 1 (AAT1) gene was reduced. In the RG × GS population, 46 quantitative trait loci (QTLs) for the production of esters and alcohols were identified on 15 linkage groups (LGs). The major QTL for 35 individual compounds was positioned on LG2 and co-located with AAT1. Multiple AAT1 gene variants were identified in RG and GS, but only two (AAT1-RGa and AAT1-GSa) were functional. AAT1-RGa and AAT1-GSa were both highly expressed in the cortex and skin of ripe fruit, but AAT1 protein was observed mainly in the skin. Transgenic RG specifically reduced in AAT1 expression showed reduced levels of most key esters in ripe fruit. Differences in the ripe fruit aroma could be perceived by sensory analysis. The transgenic lines also showed altered ratios of biosynthetic precursor alcohols and aldehydes, and expression of a number of ester biosynthetic genes increased, presumably in response to the increased substrate pool. These results indicate that the AAT1 locus is critical for the biosynthesis of esters contributing to a 'ripe apple' flavour.
Asunto(s)
Acetiltransferasas/genética , Ésteres/metabolismo , Malus/genética , Proteínas de Plantas/genética , Sitios de Carácter Cuantitativo , Acetiltransferasas/metabolismo , Acetiltransferasas/fisiología , Mapeo Cromosómico , Regulación hacia Abajo , Estudios de Asociación Genética , Ligamiento Genético , Variación Genética , Malus/metabolismo , Datos de Secuencia Molecular , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiología , Plantas Modificadas Genéticamente/metabolismoRESUMEN
Consumers of whole foods, such as fruits, demand consistent high quality and seek varieties with enhanced health properties, convenience or novel taste. We have raised the polyphenolic content of apple by genetic engineering of the anthocyanin pathway using the apple transcription factor MYB10. These apples have very high concentrations of foliar, flower and fruit anthocyanins, especially in the fruit peel. Independent lines were examined for impacts on tree growth, photosynthesis and fruit characteristics. Fruit were analysed for changes in metabolite and transcript levels. Fruit were also used in taste trials to study the consumer perception of such a novel apple. No negative taste attributes were associated with the elevated anthocyanins. Modification with this one gene provides near isogenic material and allows us to examine the effects on an established cultivar, with a view to enhancing consumer appeal independently of other fruit qualities.
Asunto(s)
Malus/crecimiento & desarrollo , Malus/metabolismo , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Antocianinas/metabolismo , Biotecnología/métodos , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Malus/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genéticaRESUMEN
The biosynthesis of anthocyanin in many plants is affected by environmental conditions. In apple (Malus × domestica Borkh.), concentrations of fruit anthocyanins are lower under hot climatic conditions. We examined the anthocyanin accumulation in the peel of maturing 'Mondial Gala' and 'Royal Gala' apples, grown in both temperate and hot climates, and using artificial heating of on-tree fruit. Heat caused a dramatic reduction of both peel anthocyanin concentration and transcripts of the genes of the anthocyanin biosynthetic pathway. Heating fruit rapidly reduced expression of the R2R3 MYB transcription factor (MYB10) responsible for coordinative regulation for red skin colour, as well as expression of other genes in the transcriptional activation complex. A single night of low temperatures is sufficient to elicit a large increase in transcription of MYB10 and consequently the biosynthetic pathway. Candidate genes that can repress anthocyanin biosynthesis did not appear to be responsible for reductions in anthocyanin content. We propose that temperature-induced regulation of anthocyanin biosynthesis is primarily caused by altered transcript levels of the activating anthocyanin regulatory complex.
Asunto(s)
Antocianinas/biosíntesis , Frutas/fisiología , Calor , Malus/fisiología , Pigmentación , Factores de Transcripción/metabolismo , Aciltransferasas/genética , Aciltransferasas/metabolismo , Clima , Clonación Molecular , Regulación hacia Abajo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Malus/genética , Filogenia , Regiones Promotoras Genéticas , Nicotiana/genética , Nicotiana/fisiología , Factores de Transcripción/genética , Activación TranscripcionalRESUMEN
The interleukin-10-deficient (IL10(-/-)) mouse develops colon inflammation in response to normal intestinal microflora and has been used as a model of Crohn's disease. Short-Column LCMS metabolite profiling of urine from IL10(-/-) and wild-type (WT) mice was used, in two independent experiments, to identify mass spectral ions differing in intensity between these two genotypes. Three differential metabolites were identified as xanthurenic acid and as the glucuronides of xanthurenic acid and of α-CEHC (2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman). The significance of several differential metabolites as potential biomarkers of colon inflammation was evaluated in an experiment which compared metabolite concentrations in IL10(-/-) and WT mice housed, either under conventional conditions and dosed with intestinal microflora, or maintained under specific pathogen-free (SPF) conditions. Concentrations of xanthurenic acid, α-CEHC glucuronide, and an unidentified metabolite m/z 495(-)/497(+) were associated with the degree of inflammation in IL10(-/-) mice and may prove useful as biomarkers of colon inflammation.
Asunto(s)
Colitis/orina , Interleucina-10/genética , Animales , Biomarcadores/orina , Cromatografía Líquida de Alta Presión , Colitis/genética , Glucurónidos/química , Glucurónidos/metabolismo , Interleucina-10/metabolismo , Iones/química , Iones/metabolismo , Masculino , Espectrometría de Masas , Metabolómica/métodos , Ratones , Ratones Transgénicos , Xanturenatos/química , Xanturenatos/metabolismoRESUMEN
Plant metabolomics within field-based food production systems is challenging owing to environmental variability and the complex architecture and metabolic growth cycles of plants. Kiwifruit cultivars of Actinidia chinensis are vigorous perennial vines grown as clones in highly structured orchard environments, intensively managed to maximize fruit yield and quality. To understand the metabolic responses of vines to orchard management practices, we needed to better understand the various sources of metabolic variability encountered in the orchard. Triplicate composite leaf, internode and fruit (mature and immature) samples were collected from each of six Actinidia chinensis var. deliciosa 'Hayward' and A. chinensis var. chinensis 'Zesy002' kiwifruit vines at three times during the growing season and measured by LC-MS. In general, there was more variation in metabolite concentrations within vines than between vines, with 'Hayward' showing a greater percentage of within-vine variability than 'Zesy002' (c. 90 vs. 70% respectively). In specific tissues, the sampler, infection by Pseudomonas syringae var. actinidiae and the rootstock also influenced metabolite variability. A similar pattern of metabolic variability was observed from quantitative analysis of specific carbohydrates and phytohormones. High within-vine metabolic variability indicates that it is more important to obtain sufficient replicate samples than to sample from multiple vines. These data provide an objective basis for optimizing metabolite sampling strategies within kiwifruit orchards.
RESUMEN
Interleukin-10 is an immunosuppressive cytokine involved in the regulation of gastrointestinal mucosal immunity toward intestinal microbiota. Interleukin-10-deficient (IL10(-/-)) mice develop Crohn's disease-like colitis unless raised in germ-free conditions. Previous gas chromatography-mass spectrometry (GC-MS) metabolomic analysis revealed urinary metabolite differences between IL10(-/-) and wildtype C57BL/6 mice. To determine which of these differences were specifically associated with intestinal inflammation arising from IL10-deficiency, urine samples from IL10(-/-) and wildtype mice, housed in either conventional or specific pathogen-free conditions, were subjected to GC-MS metabolomic analysis. Fifteen metabolite differences, including fucose, xanthurenic acid, and 5-aminovaleric acid, were associated with intestinal inflammation. Elevated urinary levels of xanthurenic acid in IL10(-/-) mice were attributed to increased production of kynurenine metabolites that may induce T-cell tolerance toward intestinal microbiota. Liquid chromatography-mass spectrometry analysis confirmed that plasma levels of kynurenine and 3-hydroxykynurenine were elevated in IL10(-/-) mice. Eleven metabolite differences, including glutaric acid, 2-hydroxyglutaric acid, and 2-hydroxyadipic acid, were unaffected by the severity of inflammation. These metabolite differences may be associated with residual genes from the embryonic stem cells of the 129P2 mouse strain that were used to create the IL10(-/-) mouse, or may indicate novel functions of IL10 unrelated to inflammation.
Asunto(s)
Enfermedad de Crohn/metabolismo , Inflamación/metabolismo , Metabolómica/métodos , Animales , Cromatografía Liquida , Análisis por Conglomerados , Enfermedad de Crohn/sangre , Enfermedad de Crohn/orina , Modelos Animales de Enfermedad , Vida Libre de Gérmenes , Interleucina-10/genética , Quinurenina/análogos & derivados , Quinurenina/sangre , Quinurenina/metabolismo , Quinurenina/orina , Masculino , Espectrometría de Masas , Metaboloma , Ratones , Ratones Transgénicos , Análisis de Componente Principal , Triptófano/sangre , Triptófano/metabolismo , Triptófano/orina , Orina/químicaRESUMEN
Runs of homozygosity (ROH) have been widely used to study population history and trait architecture in humans and livestock species, but their application in self-incompatible plants has not been reported. The distributions of ROH in 199 accessions representing Asian pears (45), European pears (109), and interspecific hybrids (45) were investigated using genotyping-by-sequencing in this study. Fruit phenotypes including fruit weight, firmness, Brix, titratable acidity, and flavor volatiles were measured for genotype-phenotype analyses. The average number of ROH and the average total genomic length of ROH were 6 and 11 Mb, respectively, in Asian accessions, and 13 and 30 Mb, respectively, in European accessions. Significant associations between genomic inbreeding coefficients (FROH) and phenotypes were observed for 23 out of 32 traits analyzed. An overlap between ROH islands and significant markers from genome-wide association analyses was observed. Previously published quantitative trait loci for fruit traits and disease resistances also overlapped with some of the ROH islands. A prominent ROH island at the bottom of linkage group 17 overlapped with a recombination-supressed genomic region harboring the self-incompatibility locus. The observed ROH patterns suggested that systematic breeding of European pears would have started earlier than of Asian pears. Our research suggest that FROH would serve as a novel tool for managing inbreeding in gene-banks of self-incompatible plant species. ROH mapping provides a complementary strategy to unravel the genetic architecture of complex traits, and to evaluate differential selection in outbred plants. This seminal work would provide foundation for the ROH research in self-incompatible plants.
RESUMEN
* High-temperature, low-light (HTLL) treatment of 35S:PAP1 Arabidopsis thaliana over-expressing the PAP1 (Production of Anthocyanin Pigment 1) gene results in reversible reduction of red colouration, suggesting the action of additional anthocyanin regulators. High-performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LCMS) and Affimetrix-based microarrays were used to measure changes in anthocyanin, flavonoids, and gene expression in response to HTLL. * HTLL treatment of control and 35S:PAP1 A. thaliana resulted in a reversible reduction in the concentrations of major anthocyanins despite ongoing over-expression of the PAP1 MYB transcription factor. Twenty-one anthocyanins including eight cis-coumaryl esters were identified by LCMS. The concentrations of nine anthocyanins were reduced and those of three were increased, consistent with a sequential process of anthocyanin degradation. Analysis of gene expression showed down-regulation of flavonol and anthocyanin biosynthesis and of transport-related genes within 24 h of HTLL treatment. No catabolic genes up-regulated by HTLL were found. * Reductions in the concentrations of anthocyanins and down-regulation of the genes of anthocyanin biosynthesis were achieved by environmental manipulation, despite ongoing over-expression of PAP1. Quantitative PCR showed reduced expression of three genes (TT8, TTG1 and EGL3) of the PAP1 transcriptional complex, and increased expression of the potential transcriptional repressors AtMYB3, AtMYB6 and AtMYBL2 coincided with HTLL-induced down-regulation of anthocyanin biosynthesis. * HTLL treatment offers a model system with which to explore anthocyanin catabolism and to discover novel genes involved in the environmental control of anthocyanins.
Asunto(s)
Arabidopsis/fisiología , Ambiente , Pigmentación , Hojas de la Planta/fisiología , Factores de Transcripción/metabolismo , Antocianinas/química , Antocianinas/metabolismo , Arabidopsis/genética , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis , Biomasa , Análisis por Conglomerados , Flavonoles/química , Flavonoles/metabolismo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Genes Reguladores , Glicósidos/química , Glicósidos/metabolismo , Luz , Proteínas Asociadas a Pancreatitis , Pigmentación/efectos de la radiación , Hojas de la Planta/efectos de la radiación , Temperatura , Factores de Transcripción/genéticaRESUMEN
Tolerance to high selenium (Se) soils in Se-hyperaccumulating plant species is correlated with the ability to biosynthesise methylselenocysteine (MeSeCys), due to the activity of selenocysteine methyltransferase (SMT). In mammals, inclusion of MeSeCys in the diet reduces the incidence of certain cancers, so increasing the range of crop plants that can produce this compound is an attractive biotechnology target. However, in the non-Se accumulator Arabidopsis, overexpression of SMT does not result in biosynthesis of MeSeCys from selenate because the rate at which selenate is reduced to selenite by ATP sulfurylase (ATPS) is low. This limitation is less problematic in other species of the Brassicaceae that can produce MeSeCys naturally. We investigated the potential for biosynthesis of MeSeCys in other plant families using Nicotiana tabacum L., a member of the Solanaceae. When plants were watered with 200 microM selenate, overexpression of a SMT transgene caused a 2- to 4-fold increase in Se accumulation (resulting in increased numbers of leaf lesions and areas of necrosis), production of MeSeCys (up to 20% of total Se) and generation of volatile dimethyl diselenide derived directly from MeSeCys. Despite the greatly increased accumulation of total Se, this did not result in increased Se toxicity effects on growth. Overexpression of ATPS did not increase Se accumulation from selenate. Accordingly, lines overexpressing both ATPS and SMT did not show a further increase in total Se accumulation or in leaf toxicity symptoms relative to overexpression of SMT alone, but directed a greater proportion of Se into MeSeCys. This work demonstrates that the production of the cancer-preventing compound MeSeCys in plants outside the Brassicaceae is possible. We conclude that while the SMT gene from Se hyperaccumulators can probably be utilised universally to increase the metabolism of Se into MeSeCys, the effects of enhancing ATPS activity will vary depending on the species involved.
Asunto(s)
Anticarcinógenos/metabolismo , Cisteína/análogos & derivados , Metiltransferasas/genética , Nicotiana/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Cisteína/biosíntesis , Compuestos de Organoselenio , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Selenocisteína/análogos & derivados , Sulfato Adenililtransferasa/metabolismo , Nicotiana/genética , Nicotiana/crecimiento & desarrollo , TransgenesRESUMEN
A recombinant alpha-farnesene synthase from apple (Malus x domestica), expressed in Escherichia coli, showed features not previously reported. Activity was enhanced 5-fold by K(+) and all four isomers of alpha-farnesene, as well as beta-farnesene, were produced from an isomeric mixture of farnesyl diphosphate (FDP). Monoterpenes, linalool, (Z)- and (E)-beta-ocimene and beta-myrcene, were synthesised from geranyl diphosphate (GDP), but at 18% of the optimised rate for alpha-farnesene synthesis from FDP. Addition of K(+) reduced monoterpene synthase activity. The enzyme also produced alpha-farnesene by a reaction involving coupling of GDP and isoprenyl diphosphate but at <1% of the rate with FDP. Mutagenesis of active site aspartate residues removed sesquiterpene, monoterpene and prenyltransferase activities suggesting catalysis through the same active site. Phylogenetic analysis clusters this enzyme with isoprene synthases rather than with other sesquiterpene synthases, suggesting that it has evolved differently from other plant sesquiterpene synthases. This is the first demonstration of a sesquiterpene synthase possessing prenyltransferase activity.
Asunto(s)
Malus/enzimología , Pirofosfatasas/metabolismo , Proteínas Recombinantes/metabolismo , Transferasas Alquil y Aril/metabolismo , Sitios de Unión , Dimetilaliltranstransferasa/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Guanosina Difosfato/metabolismo , Inosina Difosfato/metabolismo , Magnoliopsida/enzimología , Metales , Monoterpenos/análisis , Mutagénesis , Proteínas Mutantes/metabolismo , Filogenia , Fosfatos de Poliisoprenilo/análisis , Fosfatos de Poliisoprenilo/química , Pirofosfatasas/genética , Pirofosfatasas/aislamiento & purificación , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Sesquiterpenos/análisis , Sesquiterpenos/química , Factores de TiempoRESUMEN
The biosynthesis of volatile esters by Red Delicious apples was investigated by incubating fruit tissue with deuterated flavor precursors at various times after controlled atmosphere (CA) storage and measuring deuterium incorporation into branched-chain ester volatiles. 2-Methylbutyl acetate was the only volatile not significantly reduced by CA storage. Conversion of 2-methylbutanol to 2-methylbutyl acetate and of 2-methylbutanoic acid to ethyl 2-methylbutanoate and to hexyl 2-methylbutanoate was limited by the availability of 2-methylbutyl substrates but not by acetyl-CoA, ethanol, or hexanol, respectively. The enzymatic activity required for these reactions declined during CA storage. The conversion of 2-methylbutanoic acid to 2-methylbutanol was also substrate limited, but enzymic activity appeared stable in storage. Biosynthesis of both 2-methylbutanoic acid and 2-methylbutanol, from isoleucine, was severely depressed under CA storage. The reduced metabolism of isoleucine to 2-methylbutanoyl-CoA may be the primary reason for reduced branched-chain ester synthesis in CA-stored Red Delicious apples. Enantioselective gas chromatography-mass spectrometry confirmed that the chirality of (S)-2-methylbutyl acetate derives from l-isoleucine with the other enzymes in this pathway not being enantiospecific. Treatment of tissue samples with 2-methylbut-2E-enal gave only (S)-2-methylbutyl acetate, indicating that biosynthesis was not via tiglyl-CoA.
Asunto(s)
Ésteres/metabolismo , Frutas/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Malus/metabolismo , Butiratos/metabolismo , Deuterio , Conservación de Alimentos , Estereoisomerismo , VolatilizaciónRESUMEN
Selenium (Se) is an essential micronutrient for human health. Se deficiency affects hundreds of millions of people worldwide, particularly in developing countries, and there is increasing awareness that suboptimal supply of Se can also negatively affect human health. Selenium enters the diet primarily through the ingestion of plant and animal products. Although, plants are not dependent on Se they take it up from the soil through the sulphur (S) uptake and assimilation pathways. Therefore, geographic differences in the availability of soil Se and agricultural practices have a profound influence on the Se content of many foods, and there are increasing efforts to biofortify crop plants with Se. Plants from the Brassicales are of particular interest as they accumulate and synthesize Se into forms with additional health benefits, such as methylselenocysteine (MeSeCys). The Brassicaceae are also well-known to produce the glucosinolates; S-containing compounds with demonstrated human health value. Furthermore, the recent discovery of the selenoglucosinolates in the Brassicaceae raises questions regarding their potential bioefficacy. In this review we focus on Se uptake and metabolism in the Brassicaceae in the context of human health, particularly cancer prevention and immunity. We investigate the close relationship between Se and S metabolism in this plant family, with particular emphasis on the selenoglucosinolates, and consider the methodologies available for identifying and quantifying further novel Se-containing compounds in plants. Finally, we summarize the research of multiple groups investigating biofortification of the Brassicaceae and discuss which approaches might be most successful for supplying Se deficient populations in the future.
RESUMEN
Lilac alcohol epoxide (2-(5-methyl-5-(oxiran-2-yl)-tetrahydrofuran-2-yl)propan-1-ol), a previously unreported monoterpene, was identified in the solvent extract of the flowers of seven Actinidia arguta genotypes. The diastereomeric lilac alcohol epoxides co-occurred with the lilac aldehydes and alcohols. Another compound, the lilac diol (2-(5-(1-hydroxyethyl)-5-methyl-tetrahydrofuran-2-yl)propan-1-ol) was synthesised as part of our efforts to identify the lilac alcohol epoxide.
Asunto(s)
Actinidia/química , Alcoholes/química , Compuestos Epoxi/química , Óxido de Etileno/análogos & derivados , Flores/química , Furanos/química , Monoterpenos/química , Propanoles/química , Monoterpenos Acíclicos , Alcoholes/análisis , Cromatografía Líquida de Alta Presión , Compuestos Epoxi/análisis , Óxido de Etileno/química , Óxido de Etileno/aislamiento & purificación , Furanos/aislamiento & purificación , Estructura Molecular , Propanoles/aislamiento & purificaciónRESUMEN
The major anthocyanins of boysenberry fruit, a cross between Rubus loganbaccus and Rubus baileyanus Britt., were isolated by preparative high-performance liquid chromatography (HPLC). The structures of cyanidin-3-[2-(glucosyl)glucoside] (1) and cyanidin-3-[2-(glucosyl)-6-(rhamnosyl)glucoside] (2) were determined by NMR in 1% DCOOD/D(2)O. An unusually high chemical shift (delta 2.5) is reported for H-5' '' of cyanidin-3-[2-(glucosyl)glucoside].
Asunto(s)
Antocianinas/química , Frutas/química , Antocianinas/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Estructura Molecular , Resonancia Magnética Nuclear BiomolecularRESUMEN
In Brassica species, hydrolysis of (methylthio)glucosinolates produces sulfur-containing aglycons which have demonstrated anticancer benefits. Selenized Brassicaceae contain (methylseleno)glucosinolates and their selenium-containing aglycons. As a prelude to biological testing, broccoli, cauliflower, and forage rape plants were treated with sodium selenate and their tap roots, stems, leaves, and florets analyzed for selenoglucosinolates and their Se aglycons. Two new selenoglucosinolates were identified: glucoselenoraphanin in broccoli florets and glucoselenonasturtiin in forage rape roots. A new aglycon, selenoberteroin nitrile, was identified in forage rape. The major selenoglucosinolates were glucoselenoerucin in broccoli, glucoselenoiberverin in cauliflower, and glucoselenoerucin and glucoselenoberteroin in forage rape roots. In broccoli florets, the concentrations of selenglucosinolates exceeded those of their sulfur analogues. Fertilization with selenium slightly reduced (methylthio)glucosinolates and aglycons in the roots, but increased them in the florets, the leaves, and sometimes the stems. These discoveries provide a new avenue for investigating how consumption of Brassica vegetables and their organoselenides may promote human health.
Asunto(s)
Brassica/química , Glucosinolatos/análisis , Ácido Selénico/análisis , Brassica/metabolismo , Alimentos Orgánicos/análisis , Glucosinolatos/metabolismo , Humanos , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Raíces de Plantas/química , Raíces de Plantas/metabolismo , Tallos de la Planta/química , Tallos de la Planta/metabolismo , Ácido Selénico/metabolismoRESUMEN
Two syntheses of deuterated gamma-lactones for use as internal standards in stable isotope dilution assays (SIDA) were developed. [2,2,3,3-2H4]-gamma-Octa-, -gamma-deca-, and -gamma-dodecalactones with >89% deuterium incorporation were prepared in 27, 17, and 19% overall yields, respectively, by the reduction of a doubly protected hydroxypropiolic acid with deuterium gas. [3,3,4-2H3]-gamma-Octa- and -gamma-dodecalactones were prepared in 6 and 23% yields with >92% deuterium incorporation by the free radical addition of 2-iodoacetamide to [1,1,2-2H3]-1-hexene and [1,1,2-2H3]-1-decene, respectively. Reaction yields were highly dependent upon the purity of the 1-alkene starting material. The deuterated gamma-lactones were evaluated as internal standards for SIDA.
Asunto(s)
Deuterio , Técnicas de Dilución del Indicador , Lactonas/química , Marcaje IsotópicoRESUMEN
Metabolomics, comprehensive metabolite analysis, is finding increasing application as a tool to measure and enable the manipulation of the phytochemical content of foods, to identify the measures of dietary intake, and to understand human and animal responses to phytochemicals in the diet. Recent applications of metabolomics directed toward understanding the role of phytochemicals in food and nutrition are reviewed.