RESUMEN

The inhibitory effects on the mycelial growth of plant pathogen Botritys cinerea have been evaluated for a series of geranylphenols substituted with one, two and three methoxy groups in the aromatic ring. The results show that the antifungal activity depends on the structure of the geranylphenols, increasing from 40% to 90% by increasing the number of methoxy groups. On the other hand, the acetylation of the -OH group induces a change of activity that depends on the number of methoxy groups. The biological activity of digeranyl derivatives is lower than that exhibited by the respective monogeranyl compound. All tested geranylphenols have been synthesized by direct coupling of geraniol and the respective phenol. The effect of solvent on yields and product distribution is discussed. For monomethoxyphenols the reaction gives better yields when acetonitrile is used as a solvent and AgNO3 is used as a secondary catalyst. However, for di- and trimethoxyphenols the reaction proceeds only in dioxane.

Asunto(s)

Acetatos/química , Botrytis/efectos de los fármacos , Botrytis/crecimiento & desarrollo , Fungicidas Industriales/química , Fenoles/química , Terpenos/química , Acetatos/síntesis química , Acetatos/farmacología , Monoterpenos Acíclicos , Fungicidas Industriales/síntesis química , Fungicidas Industriales/farmacología , Tecnología Química Verde , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Fenoles/síntesis química , Fenoles/farmacología , Terpenos/síntesis química , Terpenos/farmacología

RESUMEN

KEY MESSAGE: VvMATE1 and VvMATE2 encode putative PA transporters expressed during seed development in grapevine. The subcellular localization of these MATE proteins suggests different routes for the intracellular transport of PAs. Proanthocyanidins (PAs), also called condensed tannins, protect plants against herbivores and are important quality components of many fruits. PAs biosynthesis is part of the flavonoid pathway that also produces anthocyanins and flavonols. In grape fruits, PAs are present in seeds and skin tissues. PAs are synthesized in the cytoplasm and accumulated into the vacuole and apoplast; however, little is known about the mechanisms involved in the transport of these compounds to such cellular compartments. A gene encoding a Multidrug And Toxic compound Extrusion (MATE) family protein suggested to transport anthocyanins-named VvMATE1-was used to identify a second gene of the MATE family, VvMATE2. Analysis of their deduced amino acid sequences and the phylogenetic relationship with other MATE-like proteins indicated that VvMATE1 and VvMATE2 encode putative PA transporters. Subcellular localization assays in Arabidopsis protoplasts transformed with VvMATE-GFP fusion constructs along with organelle-specific markers revealed that VvMATE1 is localized in the tonoplast whereas VvMATE2 is localized in the Golgi complex. Major expression of both genes occurs during the early stages of seed development concomitant with the accumulation of PAs. Both genes are poorly expressed in the skin of berries while VvMATE2 is also expressed in leaves. The presence of putative cis-acting elements in the promoters of VvMATE1 and VvMATE2 may explain the differential transcriptional regulation of these genes in grapevine. Altogether, these results suggest that these MATE proteins could mediate the transport and accumulation of PAs in grapevine through different routes and cellular compartments.

Asunto(s)

Frutas/crecimiento & desarrollo , Proteínas de Plantas/genética , Proantocianidinas/metabolismo , Vitis/genética , Secuencia de Aminoácidos , Arabidopsis/genética , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Aparato de Golgi/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/metabolismo , Semillas/genética , Semillas/crecimiento & desarrollo , Homología de Secuencia de Aminoácido , Vitis/crecimiento & desarrollo

RESUMEN

Natural geranyl compounds are known to exhibit important biological activities. In this work a series of geranylphenols were synthesized to evaluate their effect on the mycelial growth of Botrytis cinerea. Geranyl derivatives were synthesized by direct geranylation reactions between the corresponding phenol derivatives and geraniol, using BF3.OEt2 as catalyst and AgNO3 as secondary catalyst. Previously reported molecules [geranylhydroquinone (2), geranylhydroquinone diacetate (6) and geranylphloroglucinol (9)], and new substances [(E)-4-(3,7-dimethylocta-2,6-dienyl)benzene-1,2,3-triol (geranyl-pyrogallol, 7), (E)-4-(3,7-dimethylocta-2,6-dienyl)benzene-1,2,3-triyl triacetate (8), (E)-2-(3,7-dimethylocta-2,6-dienyl)benzene-1,3,5-triyl triacetate geranylphloroglucinol triacetate (10), 2,4-bis((E)-3,7-dimethylocta-2,6-dienyl)benzene-1,3,5-triyl triacetate (11), 2,6-bis((E)-3,7-dimethylocta-2,6-dienyl)-3,5-dihydroxyphenyl acetate (12)], were obtained. All compounds were characterized by IR, HRMS and NMR spectroscopic data. The inhibitory effect of the synthesized compounds on the mycelial growth of Botrytis cinerea was tested in vitro. Excepting compound 11, all substances constrained the mycelial growth of Botrytis cinerea. The antifungal activity depends on the chemical structure of geranylphenol derivatives. Compounds 2 and 9 were the more effective substances showing inhibition degrees higher than those obtained with the commercial fungicide Captan, even at lower concentrations. Monosubstitution on the aromatic nucleus by a geranyl chain seems to be more effective for the inhibition of mycelial growth than a double substitution. These results suggest that the new derivatives of geranylphenols have the ability to block the mycelial development of the plant pathogen B. cinerea and that this capacity depends strongly on the structural features and lipophilicity of the compounds.

Asunto(s)

Botrytis/efectos de los fármacos , Micelio/efectos de los fármacos , Fenoles/química , Plantas/microbiología , Antifúngicos/síntesis química , Antifúngicos/química , Antifúngicos/farmacología , Botrytis/crecimiento & desarrollo , Botrytis/patogenicidad , Fungicidas Industriales/síntesis química , Fungicidas Industriales/química , Fungicidas Industriales/farmacología , Micelio/crecimiento & desarrollo , Fenoles/síntesis química , Fenoles/farmacología , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/crecimiento & desarrollo , Terpenos/química , Terpenos/farmacología

RESUMEN

BACKGROUND: Zinc (Zn) deficiency is one of the most widespread mineral nutritional problems that affect normal development in plants. Because Zn cannot passively diffuse across cell membranes, it must be transported into intracellular compartments for all biological processes where Zn is required. Several members of the Zinc-regulated transporters, Iron-regulated transporter-like Protein (ZIP) gene family have been characterized in plants, and have shown to be involved in metal uptake and transport. This study describes the first putative Zn transporter in grapevine. Unravelling its function may explain an important symptom of Zn deficiency in grapevines, which is the production of clusters with fewer and usually smaller berries than normal. RESULTS: We identified and characterized a putative Zn transporter from berries of Vitis vinifera L., named VvZIP3. Compared to other members of the ZIP family identified in the Vitis vinifera L. genome, VvZIP3 is mainly expressed in reproductive tissue - specifically in developing flowers - which correlates with the high Zn accumulation in these organs. Contrary to this, the low expression of VvZIP3 in parthenocarpic berries shows a relationship with the lower Zn accumulation in this tissue than in normal seeded berries where its expression is induced by Zn. The predicted protein sequence indicates strong similarity with several members of the ZIP family from Arabidopsis thaliana and other species. Moreover, VvZIP3 complemented the growth defect of a yeast Zn-uptake mutant, ZHY3, and is localized in the plasma membrane of plant cells, suggesting that VvZIP3 has the function of a Zn uptake transporter. CONCLUSIONS: Our results suggest that VvZIP3 encodes a putative plasma membrane Zn transporter protein member of the ZIP gene family that might play a role in Zn uptake and distribution during the early reproductive development in Vitis vinifera L., indicating that the availability of this micronutrient may be relevant for reproductive development.

Asunto(s)

Proteínas Portadoras/genética , Vitis/genética , Zinc/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Transporte Biológico/genética , Proteínas Portadoras/metabolismo , ADN Complementario/genética , ADN Complementario/metabolismo , Flores/genética , Flores/crecimiento & desarrollo , Flores/metabolismo , Frutas/genética , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Prueba de Complementación Genética , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Datos de Secuencia Molecular , Mutación , Cebollas/genética , Cebollas/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , ARN de Planta/genética , Proteínas Recombinantes de Fusión , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Alineación de Secuencia , Vitis/crecimiento & desarrollo , Vitis/metabolismo , Zinc/análisis , Zinc/farmacología

RESUMEN

Vegetative propagation by stem cuttings and mini-cuttings has been used worldwide for growing Eucalyptus plants. However, clones and hybrids of this plant present a great variability in their rooting capacity, apart from a gradual decrease in the rooting potential due to the ontogenetic age of the mother plant. Several studies have demonstrated that some bacteria promote plant growth and rooting through the action of direct and indirect mechanisms that are not still completely clear. Considering this, the objective of this study was to assess the production of auxins, abscisic acid and siderophores in Bacillus subtilis and Stenotrophomona maltophilia, which in previous studies increased rooting of E. globulus cuttings. Additionally, the population of these bacteria in the rhizosphere, superficial tissues of the stem-base and callus of the mini-cuttings was identified, and quantified by real-time PCR. Only S. maltophilia produced IAA in the presence of tryptophan; none of the bacterial strains produced ABA, but both produced siderophores. A comparative analysis of the separation profiles showed that there is a diverse microbial community in the rhizosphere, and only S. maltophilia was capable of keeping its population at a density of 2.03 × 10(7) cells/mg in different tissues of the mini-cuttings. The results would indicate that the rooting stimulus in E. globulus could be related to the action of one or several mechanisms such as the production of auxins and siderophores, and it could also be associated with the ability of bacteria to stay in the rhizosphere or in plant callus tissues.

Asunto(s)

Bacillus subtilis/crecimiento & desarrollo , Eucalyptus/crecimiento & desarrollo , Eucalyptus/microbiología , Reguladores del Crecimiento de las Plantas/metabolismo , Sideróforos/metabolismo , Stenotrophomonas/crecimiento & desarrollo , Bacillus subtilis/metabolismo , Carga Bacteriana , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Tallos de la Planta/crecimiento & desarrollo , Tallos de la Planta/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Rizosfera , Microbiología del Suelo , Stenotrophomonas/metabolismo

RESUMEN

Two depsides and five depsidones, isolated from lichens, were tested to determine their in vivo protective effects on tobacco leaves challenged with tobacco mosaic virus (TMV). The results indicate that most of these compounds are able to reduce either the number and/or the size of necrotic lesions following virus infection. Pannarin, 1'-chloro-pannarin and stictic acid provided the more effective protective results, reducing by at least 45% the number and size of lesions. Real Time PCR assays were used to explore the target of action against TMV by examining the response behavior of genes involved in the plant defense mechanism. The application of the lichen substances did not lead to changes in the transcriptional levels of pathogen-related (PR1a), allene oxide synthase 2 (AOS2) or oxophytodienoate reductase (OPR3) genes. Thus, the protection observed in the tobacco leaves treated with the lichen compounds may be mediated by a mechanism which does not involved the SA- or JA-mediated defensive plant response. A possible structure-activity relationship is presented.

Asunto(s)

Depsidos/farmacología , Lactonas/farmacología , Líquenes/química , Nicotiana/microbiología , Enfermedades de las Plantas/terapia , Virus del Mosaico del Tabaco/efectos de los fármacos , Hojas de la Planta/microbiología , Relación Estructura-Actividad

RESUMEN

A callus induction and plant regeneration protocol was developed from leaf and thorn explants for the plant Ulex europaeus. Explants were incubated on 2 percent sucrose half-strength Murashige and Skoog Medium (MS) with various combinations of plant growth regulators and antioxidants. The best frequency of callus and shoot formation was obtained with 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg/l x kinetin (Kin) 0.2 mg/l (DK Medium; callus induction) and zeatin (Z) 1 mg/l (DK medium; shoot induction). Both media were supplemented with ascorbic acid 200 mg/l to prevent browning and death of the explants. The regenerated shoots transferred to rooting medium (half-strength MS Medium, 2 percent sucrose) showed rapid growth and development of roots (100 percent). Rooted plantlets were successfully transferred to soil in pots containing a 3:1 mixture of soil and vermiculite.

Asunto(s)

Regeneración , Ulex/crecimiento & desarrollo , Aclimatación , Brotes de la Planta/crecimiento & desarrollo , Fabaceae/crecimiento & desarrollo , Germinación

RESUMEN

Boron (B) is an essential micronutrient for normal development of roots, shoots and reproductive tissues in plants. Due to its role in the structure of rhamnogalacturonan II, a polysaccharide required for pollen tube growth, B deficiency has been associated with the occurrence of parthenocarpic seedless grapes in some varieties of Vitis vinifera L. Despite that, it is unclear how B is mobilized and accumulated in reproductive tissues. Here we describe the characterization of an efflux B transporter, VvBOR1, homolog to AtBOR1, which is involved in B xylem loading in Arabidopsis thaliana roots. VvBOR1-green fluorescent protein (GFP) fusion protein expressed in A. thaliana localizes in the proximal plasma membrane domain in root pericycle cells, and VvBOR1 overexpression restores the wild-type phenotype in A. thaliana bor1-3 mutant plants exposed to B deficiency. Complementation of a mutant yeast strain indicates that VvBOR1 corresponds to a B efflux transporter. Transcriptional analyses during grapevine reproductive development show that the VvBOR1 gene is preferentially expressed in flowers at anthesis and a direct correlation between the expression pattern and B content in grapes was established, suggesting the involvement of this transporter in B accumulation in grapevine berries.

Asunto(s)

Antiportadores/metabolismo , Boro/metabolismo , Proteínas de Plantas/metabolismo , Vitis/genética , Secuencia de Aminoácidos , Antiportadores/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Flores/metabolismo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Prueba de Complementación Genética , Proteínas Fluorescentes Verdes/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/genética , Raíces de Plantas/citología , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Alineación de Secuencia , Vitis/metabolismo

RESUMEN

Aqueous and ethanolic extracts obtained from nine Chilean marine macro-algae collected at different seasons were examined in vitro and in vivo for properties that reduce the growth of plant pathogens or decrease the injury severity of plant foliar tissues following pathogen infection. Particular crude aqueous or organic extracts showed effects on the growth of pathogenic bacteria whereas others displayed important effects against pathogenic fungi or viruses, either by inhibiting fungal mycelia growth or by reducing the disease symptoms in leaves caused by pathogen challenge. Organic extracts obtained from the brown-alga Lessonia trabeculata inhibited bacterial growth and reduced both the number and size of the necrotic lesion in tomato leaves following infection with Botrytis cinerea. Aqueous and ethanolic extracts from the red-alga Gracillaria chilensis prevent the growth of Phytophthora cinnamomi, showing a response which depends on doses and collecting-time. Similarly, aqueous and ethanolic extracts from the brown-alga Durvillaea antarctica were able to diminish the damage caused by tobacco mosaic virus (TMV) in tobacco leaves, and the aqueous procedure is, in addition, more effective and seasonally independent. These results suggest that macro-algae contain compounds with different chemical properties which could be considered for controlling specific plant pathogens.

Asunto(s)

Antiinfecciosos/farmacología , Fungicidas Industriales/farmacología , Phaeophyceae/química , Plantas/efectos de los fármacos , Plantas/microbiología , Rhodophyta/química , Algas Marinas/química , Antivirales/farmacología , Plantas/virología

RESUMEN

Grapevine sexual reproduction involves a seasonal separation between inflorescence primordia (flowering induction) and flower development. We hypothesized that a repression mechanism implicating epigenetic changes could play a role in the seasonal separation of these two developmental processes in grapevine. Therefore, the expression of five grapevine genes with homology to the Arabidopsis epigenetic repressor genes FERTILIZATION INDEPENDENT ENDOSPERM (FIE), EMBRYONIC FLOWER 2 (EMF2), CURLY LEAF (CLF), MULTICOPY SUPPRESSOR OF IRA 1 (MSI1) and SWINGER (SWN) was analyzed during the development of buds and vegetative and reproductive organs. During bud development, the putative grapevine epigenetic repressor genes VvCLF, VvEMF2, VvMSI1, VvSWN and VvFIE are mainly expressed in latent buds at the flowering induction period, but also detected during bud burst and inflorescence/flower development. The overlapping expression patterns of grapevine PcG-like genes in buds suggest that chromatin remodeling mechanisms could be operating during grapevine bud development for controlling processes such as seasonal flowering, dormancy and bud burst. Furthermore, the expression of grapevine PcG-like genes was also detected in fruits and vegetative organs, suggesting that epigenetic changes could be at the basis of the regulation of various proliferation-differentiation cell transitions that occur during grapevine development.

Asunto(s)

Epigénesis Genética , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas Represoras/metabolismo , Vitis/crecimiento & desarrollo , Secuencia de Aminoácidos , Clonación Molecular , Flores/genética , Flores/crecimiento & desarrollo , Frutas/genética , Frutas/crecimiento & desarrollo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Brotes de la Planta/genética , Brotes de la Planta/crecimiento & desarrollo , Proteínas Represoras/genética , Reproducción/genética , Vitis/genética

RESUMEN

Background: Several studies have shown that (-)-Jasmonic acid, (+)-7-iso-Jasmonic acid and its methyl ester, methyl jasmonate, have anti-cancer activity in vitro and in vivo, exhibiting selective cytotoxicity towards cancer cells. The degree of activity of these molecules is strongly related to their stereochemistry. The biotransformation of known compounds, natural or synthesized, related to interesting biological activities, generates new molecules displaying new improved properties compared with the original ones, increasing its value and providing new more effective products. Therefore, based on the above rationales and observations, in this work a biotransformation protocol to modify the chemical structure of the plant hormone jasmonic acid by using the fungus Gibberella fujikuroi was established. Results: The three jasmonic acid derivatives obtained, 3(S)-Hydroxy-2(R)-(2Z-pentenyl)-cyclopentane-1(R)-acetic acid (1), 3(R)-Hydroxy-2(R)-(2Z-pentenyl)-cyclopentane-1(R)-acetic acid (2), 3-Hydroxy-2(S)-(2Z-pentenyl)-cyclopentane-1(S)-acetic acid (3), were tested for cell-growth inhibition activity towards the human cancer epithelial cell line, the oral squamous carcinoma cells (KB). The results obtained show that jasmonic acid derivatives (1-3) are active on human cancer cells examined in different concentration ranges, with IC50 value less than of 25 uM. The compound 3, with the same molecular structure of compounds 1 and 2, but with different stereochemistry, was more active confirming that the activity of jasmonate compounds is related to their stereochemistry and to substituents in the cyclopentane ring. In this study, we also tested the potential proapoptotic activity of compound 3, and our data suggest that it, as other jasmonate compounds, is able to trigger apoptotic death in cancer cells. This event may be correlated at an elevation of reactive oxygen species (ROS). Administration of N-acetylcysteine (NAC) prevented compound 3 cytotoxicity...

Asunto(s)

Humanos , Apoptosis , Ciclopentanos/metabolismo , Gibberella/metabolismo , Oxilipinas/metabolismo , Antineoplásicos , Bioensayo , Biotransformación , Supervivencia Celular , Ensayo Cometa , Especies Reactivas de Oxígeno , L-Lactato Deshidrogenasa

RESUMEN

The chemical composition of any wine sample contains numerous small molecules largely derived from three different sources: the grape berry, the yeast strain used for fermentation, and the containers used for wine making and storage. The combined sum of these small molecules present in the wine, therefore, might account for all wine specific features such as cultivar, vintage, origin, and quality. Still, most wine authentication procedures rely either on subjective human measures or if they are based on measurable features, they include a limited number of compounds. In this study, which is based on an untargeted UPLC-FT-ICR-MS-based approach, we provide data, demonstrating that unbiased and objective analytical chemistry in combination with multivariate statistical methods allows to reproducible classify/distinguish wine attributes like variety, origin, vintage, and quality.

Asunto(s)

Metaboloma , Vino/análisis , Cromatografía Liquida , Espectrometría de Masas , Nanotecnología , Reproducibilidad de los Resultados

RESUMEN

A "defensin-like" antibacterial peptide from Mytilus edulis chilensis, was sub-cloned into a binary vector for expression in plant tissues. The resulting new clone was electroporated into A. tumefaciens to transform tobacco plants. The presence of the construct in transgenic tobacco lines was demonstrated through RT-PCR, Northern and Western blots. Transformed positive plants were selected and grown for challenging. Tobacco leaves were infiltrated with Pseudomonassyringae pv. syringae and visual lesions determined at different times post-exposure. Of seven plants exposed, four gave variable protection up to seven days post-infection while one of them appears to be fully protected. These results suggest that defensin-like antimicrobial peptides from molluscs are a good source to provide resistance of tobacco plants to Pseudomonassyringae pv. syringae.

RESUMEN

The development of plant transformation in the mid-1980s and of many new tools for cell biology, molecular genetics, and biochemistry has resulted in enormous progress in plant biology in the past decade. With the completion of the genome sequence of Arabidopsis thaliana just around the corner, we can expect even faster progress in the next decade. The interface between cell biology and signal transduction is emerging as a new and important field of research. In the past we thought of cell biology strictly in terms of organelles and their biogenesis and function, adn researches focused on questions such as, how do proteins enter chloroplasts? or, what is the structure of the macromolecules of the cell wall and how are the se molecules secreted? Signal transduction dealt primarily with the perception of light (photomorphogenesis) or hormones and with the effect such signals have on enhancing the activity of specific genes. Now we see that the fields of cell biology and signal transduction pathway usually involves multiple organelles of cellular structures Here are some examples to illustrate this new paradigm. How does abscisic acid (ABA) regulate stomatal closure? This pathway involves not only ABA receptors whose location is not yet known, but cation and anion channels in the plasma membrane, changes in the cytoskeleton, movement of water through water channels in the tonoplast and the plasma membrane, proteins with a farnesyl tail that can be located either in the cytosol or attached to a membrane, and probably unidentified ion channels in the tonoplast. In addition there are highly localized calcium oscillations in the cytoplasm resulting from the release of calcium stored in various compartments. The activities of all these cellular structures need to be coordinated during ABA-induced stomatal closure. For another example of the interplay between the proteins of signal transduction pathways and cytoplasmic structures, consider how plants mount defense response against pathogens. Elicitors produced by pathogens bind to receptors on the plant plasma membrane or in the cytosol and eventually activate a large number of genes. This results in the coordination of activities at the plasma membrane (production of reactive oxygen species), in the cytoskeleton, localized calcium oscillations, and the modulation of protein kinases and protein phosphatases whose locations remain to be determined. The movement of ...

Asunto(s)

Plantas/citología , Transducción de Señal

Asunto(s)

Animales , Peces/ultraestructura , Protaminas/ultraestructura