RESUMEN
Soil salinity is an increasingly global problem which hampers plant growth and crop yield. Plant productivity depends on optimal water-use efficiency and photosynthetic capacity balanced by stomatal conductance. Whether and how stomatal behavior contributes to salt sensitivity or tolerance is currently unknown. This work identifies guard cell-specific signaling networks exerted by a salt-sensitive and salt-tolerant plant under ionic and osmotic stress conditions accompanied by increasing NaCl loads. We challenged soil-grown Arabidopsis thaliana and Thellungiella salsuginea plants with short- and long-term salinity stress and monitored genome-wide gene expression and signals of guard cells that determine their function. Arabidopsis plants suffered from both salt regimes and showed reduced stomatal conductance while Thellungiella displayed no obvious stress symptoms. The salt-dependent gene expression changes of guard cells supported the ability of the halophyte to maintain high potassium to sodium ratios and to attenuate the abscisic acid (ABA) signaling pathway which the glycophyte kept activated despite fading ABA concentrations. Our study shows that salinity stress and even the different tolerances are manifested on a single cell level. Halophytic guard cells are less sensitive than glycophytic guard cells, providing opportunities to manipulate stomatal behavior and improve plant productivity.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Ácido Abscísico , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Transporte Iónico , Estomas de Plantas/metabolismo , Estrés Salino , Plantas Tolerantes a la Sal/metabolismoRESUMEN
The Venus flytrap Dionaea muscipula captures insects and consumes their flesh. Prey contacting touch-sensitive hairs trigger traveling electrical waves. These action potentials (APs) cause rapid closure of the trap and activate secretory functions of glands, which cover its inner surface. Such prey-induced haptoelectric stimulation activates the touch hormone jasmonate (JA) signaling pathway, which initiates secretion of an acidic hydrolase mixture to decompose the victim and acquire the animal nutrients. Although postulated since Darwin's pioneering studies, these secretory events have not been recorded so far. Using advanced analytical and imaging techniques, such as vibrating ion-selective electrodes, carbon fiber amperometry, and magnetic resonance imaging, we monitored stimulus-coupled glandular secretion into the flytrap. Trigger-hair bending or direct application of JA caused a quantal release of oxidizable material from gland cells monitored as distinct amperometric spikes. Spikes reminiscent of exocytotic events in secretory animal cells progressively increased in frequency, reaching steady state 1 d after stimulation. Our data indicate that trigger-hair mechanical stimulation evokes APs. Gland cells translate APs into touch-inducible JA signaling that promotes the formation of secretory vesicles. Early vesicles loaded with H+ and Cl- fuse with the plasma membrane, hyperacidifying the "green stomach"-like digestive organ, whereas subsequent ones carry hydrolases and nutrient transporters, together with a glutathione redox moiety, which is likely to act as the major detected compound in amperometry. Hence, when glands perceive the haptoelectrical stimulation, secretory vesicles are tailored to be released in a sequence that optimizes digestion of the captured animal.
Asunto(s)
Droseraceae/fisiología , Exocitosis/fisiología , Insectos , Transducción de Señal/fisiología , Animales , Droseraceae/ultraestructuraRESUMEN
Date palm Phoenix dactylifera is a desert crop well adapted to survive and produce fruits under extreme drought and heat. How are palms under such harsh environmental conditions able to limit transpirational water loss? Here, we analysed the cuticular waxes, stomata structure and function, and molecular biology of guard cells from P. dactylifera. To understand the stomatal response to the water stress phytohormone of the desert plant, we cloned the major elements necessary for guard cell fast abscisic acid (ABA) signalling and reconstituted this ABA signalosome in Xenopus oocytes. The PhoenixSLAC1-type anion channel is regulated by ABA kinase PdOST1. Energy-dispersive X-ray analysis (EDXA) demonstrated that date palm guard cells release chloride during stomatal closure. However, in Cl- medium, PdOST1 did not activate the desert plant anion channel PdSLAC1 per se. Only when nitrate was present at the extracellular face of the anion channel did the OST1-gated PdSLAC1 open, thus enabling chloride release. In the presence of nitrate, ABA enhanced and accelerated stomatal closure. Our findings indicate that, in date palm, the guard cell osmotic motor driving stomatal closure uses nitrate as the signal to open the major anion channel SLAC1. This initiates guard cell depolarization and the release of anions together with potassium.
Asunto(s)
Aniones/metabolismo , Clima Desértico , Nitratos/farmacología , Phoeniceae/fisiología , Proteínas de Plantas/metabolismo , Estomas de Plantas/fisiología , Ácido Abscísico/metabolismo , Cloruros/metabolismo , Sequías , Luz , Ósmosis , Phoeniceae/efectos de los fármacos , Phoeniceae/efectos de la radiación , Phoeniceae/ultraestructura , Estomas de Plantas/citología , Estomas de Plantas/efectos de los fármacos , Estomas de Plantas/ultraestructura , ARN de Planta/metabolismo , Fracciones Subcelulares/metabolismo , Ceras/metabolismoRESUMEN
Phenylcoumaran benzylic ether reductase (PCBER) is one of the most abundant proteins in poplar (Populus spp) xylem, but its biological role has remained obscure. In this work, metabolite profiling of transgenic poplar trees downregulated in PCBER revealed both the in vivo substrate and product of PCBER. Based on mass spectrometry and NMR data, the substrate was identified as a hexosylated 8-5-coupling product between sinapyl alcohol and guaiacylglycerol, and the product was identified as its benzyl-reduced form. This activity was confirmed in vitro using a purified recombinant PCBER expressed in Escherichia coli. Assays performed on 20 synthetic substrate analogs revealed the enzyme specificity. In addition, the xylem of PCBER-downregulated trees accumulated over 2000-fold higher levels of cysteine adducts of monolignol dimers. These compounds could be generated in vitro by simple oxidative coupling assays involving monolignols and cysteine. Altogether, our data suggest that the function of PCBER is to reduce phenylpropanoid dimers in planta to form antioxidants that protect the plant against oxidative damage. In addition to describing the catalytic activity of one of the most abundant enzymes in wood, we provide experimental evidence for the antioxidant role of a phenylpropanoid coupling product in planta.
Asunto(s)
Oxidorreductasas/metabolismo , Populus/enzimología , Xilema/enzimología , Aminoácidos/metabolismo , Pared Celular/metabolismo , Cisteína/metabolismo , Regulación hacia Abajo , Pruebas de Enzimas , Immunoblotting , Lignanos/biosíntesis , Lignanos/química , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Datos de Secuencia Molecular , Oxidación-Reducción , Estrés Oxidativo , Oxidorreductasas/química , Fenotipo , Plantas Modificadas Genéticamente , Reproducibilidad de los Resultados , Especificidad por SustratoRESUMEN
Extrafloral nectaries secrete a sweet sugar cocktail that lures predator insects for protection from foraging herbivores. Apart from sugars and amino acids, the nectar contains the anions chloride and nitrate. Recent studies with Populus have identified a type of nectary covered by apical bipolar epidermal cells, reminiscent of the secretory brush border epithelium in animals. Border epithelia operate transepithelial anion transport, which is required for membrane potential and/or osmotic adjustment of the secretory cells. In search of anion transporters expressed in extrafloral nectaries, we identified PttSLAH3 (Populus tremula × Populus tremuloides SLAC1 Homologue3), an anion channel of the SLAC/SLAH family. When expressed in Xenopus oocytes, PttSLAH3 displayed the features of a voltage-dependent anion channel, permeable to both nitrate and chloride. In contrast to the Arabidopsis SLAC/SLAH family members, the poplar isoform PttSLAH3 is independent of phosphorylation activation by protein kinases. To understand the basis for the autonomous activity of the poplar SLAH3, we generated and expressed chimera between kinase-independent PttSLAH3 and kinase-dependent Arabidopsis AtSLAH3. We identified the N-terminal tail and, to a lesser extent, the C-terminal tail as responsible for PttSLAH3 kinase-(in)dependent action. This feature of PttSLAH3 may provide the secretory cell with a channel probably controlling long-term nectar secretion.
Asunto(s)
Aniones/metabolismo , Epitelio/metabolismo , Canales Iónicos/metabolismo , Proteínas de Plantas/metabolismo , Populus/metabolismo , Proteínas Quinasas/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/metabolismo , Epitelio/efectos de los fármacos , Flores/efectos de los fármacos , Flores/metabolismo , Activación del Canal Iónico/efectos de los fármacos , Nitratos/farmacología , Néctar de las Plantas , Proteínas de Plantas/química , Populus/efectos de los fármacos , Proteínas Recombinantes de Fusión/metabolismo , Relación Estructura-ActividadRESUMEN
Leaf photosynthesis of the sensitive plant Mimosa pudica displays a transient knockout in response to electrical signals induced by heat stimulation. This study aims at clarifying the underlying mechanisms, in particular, the involvement of respiration. To this end, leaf gas exchange and light reactions of photosynthesis were assessed under atmospheric conditions largely eliminating photorespiration by either elevated atmospheric CO2 or lowered O2 concentration (i.e. 2000 µmol mol(-1) or 1%, respectively). In addition, leaf gas exchange was studied in the absence of light. Under darkness, heat stimulation caused a transient increase of respiratory CO2 release simultaneously with stomatal opening, hence reflecting direct involvement of respiratory stimulation in the drop of the net CO2 uptake rate. However, persistence of the transient decline in net CO2 uptake rate under illumination and elevated CO2 or 1% O2 makes it unlikely that photorespiration is the metabolic origin of the respiratory CO2 release. In conclusion, the transient knockout of net CO2 uptake is at least partially attributed to an increased CO2 release through mitochondrial respiration as stimulated by electrical signals. Putative CO2 limitation of Rubisco due to decreased activity of carbonic anhydrase was ruled out as the photosynthesis effect was not prevented by elevated CO2 .
Asunto(s)
Dióxido de Carbono/metabolismo , Mimosa/metabolismo , Oxígeno/metabolismo , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema II/fisiología , Transpiración de Plantas/fisiología , Anhidrasas Carbónicas/metabolismo , Respiración de la Célula/fisiología , Clorofila/metabolismo , Oscuridad , Electricidad , Calor , Luz , Mimosa/fisiología , Mimosa/efectos de la radiación , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Estomas de Plantas/fisiología , Estomas de Plantas/efectos de la radiación , Ribulosa-Bifosfato Carboxilasa/metabolismo , Agua/fisiologíaRESUMEN
Many plant species grow extrafloral nectaries and produce nectar to attract carnivore arthropods as defenders against herbivores. Two nectary types that evolved with Populus trichocarpa (Ptr) and Populus tremula × Populus tremuloides (Ptt) were studied from their ecology down to the genes and molecules. Both nectary types strongly differ in morphology, nectar composition and mode of secretion, and defense strategy. In Ptt, nectaries represent constitutive organs with continuous merocrine nectar flow, nectary appearance, nectar production, and flow. In contrast, Ptr nectaries were found to be holocrine and inducible. Neither mechanical wounding nor the application of jasmonic acid, but infestation by sucking insects, induced Ptr nectar secretion. Thus, nectaries of Ptr and Ptt seem to answer the same threat by the use of different mechanisms.
Asunto(s)
Flores/inmunología , Flores/parasitología , Herbivoria/fisiología , Néctar de las Plantas/inmunología , Néctar de las Plantas/fisiología , Populus/fisiología , Populus/parasitología , Aminoácidos/metabolismo , Animales , Análisis por Conglomerados , Exocitosis/genética , Ácidos Grasos Volátiles/metabolismo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/genética , Insectos/fisiología , Larva/fisiología , Análisis de Secuencia por Matrices de Oligonucleótidos , Especificidad de Órganos/genética , Hojas de la Planta/parasitología , Hojas de la Planta/ultraestructura , Néctar de las Plantas/metabolismo , Populus/genética , Populus/inmunología , Estrés FisiológicoRESUMEN
Understanding seasonality and longevity is a major challenge in tree biology. In woody species, growth phases and dormancy follow one another consecutively. In the oldest living individuals, the annual cycle may run for more than 1,000 years. So far, however, not much is known about the processes triggering reactivation from dormancy. In this study, we focused on wood rays, which are known to play an important role in tree development. The transition phase from dormancy to flowering in early spring was compared with the phase of active growth in summer. Rays from wood samples of poplar (Populus × canescens) were enriched by laser microdissection, and transcripts were monitored by poplar whole-genome microarrays. The resulting seasonally varying complex expression and metabolite patterns were subjected to pathway analyses. In February, the metabolic pathways related to flower induction were high, indicating that reactivation from dormancy was already taking place at this time of the year. In July, the pathways related to active growth, like lignin biosynthesis, nitrogen assimilation, and defense, were enriched. Based on "marker" genes identified in our pathway analyses, we were able to validate periodical changes in wood samples by quantitative polymerase chain reaction. These studies, and the resulting ray database, provide new insights into the steps underlying the seasonality of poplar trees.
Asunto(s)
Populus/citología , Populus/fisiología , Estaciones del Año , Árboles/fisiología , Madera/citología , Madera/fisiología , Aminoácidos/metabolismo , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/genética , Captura por Microdisección con Láser , Metaboloma/genética , Populus/genética , Populus/ultraestructura , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Tiempo , Árboles/citología , Árboles/genética , Árboles/ultraestructura , Regulación hacia Arriba/genética , Madera/genética , Madera/ultraestructuraRESUMEN
In recent years, the effect of heat-induced electrical signalling on plant photosynthetic activity has been demonstrated for many plant species. However, the underlying triggers of the resulting transient inhibition of photosynthesis still remain unknown. To further investigate on this phenomenon, we focused in our present study on soybean (Glycine max L.) on the direct effect of signal transmission in the leaf mesophyll on conductance for CO(2) diffusion in the mesophyll (g(m) ) and detected a drastic decline in g(m) following the electrical signal, whereas the photosynthetic electron transport rate (ETR) was only marginally affected. In accordance with the drop in net photosynthesis (A(N) ), energy dispersive X-ray analysis (EDXA) revealed a shift of K, Mg, O and P on leaf chloroplasts. Control experiments under elevated CO(2) conditions proved the transient reduction of A(N) , ETR, the chloroplast CO(2) concentration (C(c) ) and g(m) to be independent of the external CO(2) regime, whereas the effect of the electrical signal on stomatal conductance for CO(2) (g(s) ) turned out much less distinctive. We therefore conclude that the effect of electrical signalling on photosynthesis in soybean is triggered by its immediate effects on g(m) .
Asunto(s)
Dióxido de Carbono/metabolismo , Glycine max/metabolismo , Potenciales de la Membrana , Células del Mesófilo/metabolismo , Fotosíntesis , Cloroplastos/metabolismo , Calor , Iones/metabolismoRESUMEN
In contrast to chemical messengers, electrical signals such as action potentials and variation potentials can transmit information much faster over long distances. Electrical signals can be triggered by various abiotic stress factors and are propagated via plasmodesmata over short distances and within the phloem over long distances. Thus, in addition to assimilate transport from sources to sinks, the phloem serves as a communication highway for various types of information. Key factors for systemic signaling in the phloem are peptides, RNAs, hormones, and electrical signals. In recent years, there has been increasing evidence that rapid communication by means of electrical signals is essential for various plant physiological processes. Thus, this chapter focuses on electrical signaling and various associated physiological effects, such as regulation of leaf movements, assimilate transport, photosynthesis, and gas exchange, as well as plant water status.
Asunto(s)
Plantas , Transducción de Señal , Fotosíntesis/fisiología , Hojas de la Planta/fisiología , Floema/fisiología , Estrés FisiológicoRESUMEN
Constitutive expression of the FPF1 gene in hybrid aspen (Populus tremula L. × P. tremuloides Michx.) showed a strong effect on wood formation but no effect on flowering time. Gene expression studies showed that activity of flowering time genes PtFT1, PtCO2, and PtFUL was not increased in FPF1 transgenic plants. However, the SOC1/TM3 class gene PTM5, which has been related to wood formation and flowering time, showed a strong activity in stems of all transgenic lines studied. Wood density was lower in transgenic plants, despite significantly reduced vessel frequency which was overcompensated by thinner fibre cell walls. Chemical screening of the wood by pyrolysis GC/MS showed that FPF1 transgenics have higher fractions of cellulose and glucomannan products as well as lower lignin content. The latter observation was confirmed by UV microspectrophotometry on a cellular level. Topochemical lignin distribution revealed a slower increase of lignin incorporation in the developing xylem of the transgenics when compared with the wild-type plants. In line with the reduced wood density, micromechanical wood properties such as stiffness and ultimate stress were also significantly reduced in all transgenic lines. Thus, we provide evidence that FPF1 class genes may play a regulatory role in both wood formation and flowering in poplar.
Asunto(s)
Proteínas de Arabidopsis/química , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Populus/química , Madera/fisiología , Agrobacterium tumefaciens/química , Agrobacterium tumefaciens/genética , Proteínas de Arabidopsis/genética , Pared Celular/química , Celulosa/química , Quimera/genética , Quimera/fisiología , ADN de Plantas/genética , Flores/fisiología , Cromatografía de Gases y Espectrometría de Masas , Genes de Plantas , Lignina/química , Mananos/química , Microespectrofotometría/métodos , Células Vegetales/química , Tallos de la Planta/química , Tallos de la Planta/genética , Tallos de la Planta/fisiología , Plantas Modificadas Genéticamente/química , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/fisiología , Populus/genética , Populus/fisiología , Estrés Fisiológico , Propiedades de Superficie , Factores de Tiempo , Rayos Ultravioleta , Madera/química , Madera/genéticaRESUMEN
⢠Salinity causes osmotic stress and limits biomass production of plants. The goal of this study was to investigate mechanisms underlying hydraulic adaptation to salinity. ⢠Anatomical, ecophysiological and transcriptional responses to salinity were investigated in the xylem of a salt-sensitive (Populus × canescens) and a salt-tolerant species (Populus euphratica). ⢠Moderate salt stress, which suppressed but did not abolish photosynthesis and radial growth in P. × canescens, resulted in hydraulic adaptation by increased vessel frequencies and decreased vessel lumina. Transcript abundances of a suite of genes (FLA, COB-like, BAM, XET, etc.) previously shown to be activated during tension wood formation, were collectively suppressed in developing xylem, whereas those for stress and defense-related genes increased. A subset of cell wall-related genes was also suppressed in salt-exposed P. euphratica, although this species largely excluded sodium and showed no anatomical alterations. Salt exposure influenced cell wall composition involving increases in the lignin : carbohydrate ratio in both species. ⢠In conclusion, hydraulic stress adaptation involves cell wall modifications reciprocal to tension wood formation that result in the formation of a novel type of reaction wood in upright stems named 'pressure wood'. Our data suggest that transcriptional co-regulation of a core set of genes determines reaction wood composition.
Asunto(s)
Populus/efectos de los fármacos , Populus/fisiología , Presión , Cloruro de Sodio/farmacología , Estrés Fisiológico/efectos de los fármacos , Madera/efectos de los fármacos , Madera/fisiología , Northern Blotting , Cruzamientos Genéticos , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas/genética , Concentración de Iones de Hidrógeno/efectos de los fármacos , Mucoproteínas/genética , Mucoproteínas/metabolismo , Ósmosis/efectos de los fármacos , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Populus/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Regulón/genética , Sodio/metabolismo , Especificidad de la Especie , Estrés Fisiológico/genética , Madera/anatomía & histología , Xilema/anatomía & histología , Xilema/efectos de los fármacos , Xilema/genéticaRESUMEN
A non-targeted metabolomics-based approach using liquid chromatography high-resolution mass spectrometry was used to authenticate spruce wood (Picea abies) from two geographic source areas. The two sample sites were located in Germany and only 250â¯km apart. In order to achieve the highest possible metabolite coverage, the spruces samples were measured with four different methods using liquid chromatography high-resolution mass spectrometry. In this way, a total of approximately 4,100 features were detected, which included non-polar, polar, and intermediate-polar metabolites. Using supervised multivariate methods, a distinction between the two sample groups could be achieved on the basis of non-polar data sets. The major metabolites contributing to differentiation were identified by MS/MS experiments and were from the following classes of compounds: ceramides, fatty acids, glycerolipids, and phytosterols. Based on the soil descriptions of the two sites, it was concluded that there is probably a close relationship between nutrient availability and the differences in concentration of the marker compounds. The results show that a metabolomics-based approach is also suitable for differentiation of origin, even if the sample sites are close to each other.
Asunto(s)
Abies , Picea , Cromatografía Liquida , Metabolómica , Espectrometría de Masas en Tándem , MaderaRESUMEN
Juvenile Eucalyptus grandis were exposed to drought and elevated CO2 to evaluate the independent and interactive effects on growth, gas exchange and wood structure. Trees were grown in a greenhouse at ambient and elevated CO2 (aCO2, 410 ppm; eCO2, 950 ppm), in combination with daily irrigation and cyclic drought during one growing season. The results demonstrated that drought stress limited intercellular CO2 concentration, photosynthesis, stomatal conductance, and transpiration, which correlated with a lower increment in height, stem diameter and biomass. Drought also induced formation of frequent and narrow vessels accompanied by a reduction in vessel lumen area. Conversely, elevated CO2 increased intercellular CO2 concentration as well as photosynthesis, and partially closed stomata, leading to a more efficient water use, especially under drought. There was a clear trend towards greater biomass accumulation at eCO2, although the results did not show statistical significance for this parameter. We observed an increase in vessel diameter and vessel lumen area at eCO2, and, contrarily, the vessel frequency decreased. Thus, we conclude that eCO2 delayed the effects of drought and potentialized growth. However, results on vessel anatomy suggest that increasing vulnerability to cavitation due to formation of larger vessels may counteract the beneficial effects of eCO2 under severe drought.
RESUMEN
The stomatal complex of Zea mays is composed of two pore-forming guard cells and two adjacent subsidiary cells. For stomatal movement, potassium ions and anions are thought to shuttle between these two cell types. As potential cation transport pathways, K(+)-selective channels have already been identified and characterized in subsidiary cells and guard cells. However, so far the nature and regulation of anion channels in these cell types have remained unclear. In order to bridge this gap, we performed patch-clamp experiments with subsidiary cell and guard cell protoplasts. Voltage-independent anion channels were identified in both cell types which, surprisingly, exhibited different, cell-type specific dependencies on cytosolic Ca(2+) and pH. After impaling subsidiary cells of intact maize plants with microelectrodes and loading with BCECF [(2',7'-bis-(2-carboxyethyl)-5(and6)carboxyflurescein] as a fluorescent pH indicator, the regulation of ion channels by the cytosolic pH and the membrane voltage was further examined. Stomatal closure was found to be accompanied by an initial hyperpolarization and cytosolic acidification of subsidiary cells, while opposite responses were observed during stomatal opening. Our findings suggest that specific changes in membrane potential and cytosolic pH are likely to play a role in determining the direction and capacity of ion transport in subsidiary cells.
Asunto(s)
Canales Iónicos/metabolismo , Proteínas de Plantas/metabolismo , Estomas de Plantas/citología , Estomas de Plantas/fisiología , Zea mays/citología , Zea mays/fisiología , Ácido Abscísico/farmacología , Aniones , Calcio/farmacología , Citosol/efectos de los fármacos , Citosol/metabolismo , Citosol/efectos de la radiación , Concentración de Iones de Hidrógeno/efectos de los fármacos , Luz , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/efectos de la radiación , Especificidad de Órganos/efectos de los fármacos , Estomas de Plantas/efectos de los fármacos , Estomas de Plantas/ultraestructura , Protoplastos/efectos de los fármacos , Protoplastos/metabolismo , Protoplastos/efectos de la radiación , Zea mays/efectos de los fármacos , Zea mays/ultraestructuraRESUMEN
The determination of the geographical origin of wood can be highly relevant for several reasons: On the one hand, it can help to prevent illegal logging and timber trade, on the other hand, it is of special interest for archaeological artefacts made of wood, as well as for a variety of biological questions. For this reason, different extraction methods were first tested for the analysis of polar and non-polar metabolites using liquid chromatography coupled electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). A two-phase extraction with chloroform, methanol and water proved to be particularly successful. Subsequently, cedrela (Cedrela odorata) samples from South America were measured to distinguish geographic origin. Using multivariate data analysis, numerous origin-dependent differences could be extracted. The identification of the marker substances indicated that several metabolic pathways were affected by the geographical influences, some of them probably indicating pest infections.
Asunto(s)
Cedrela/metabolismo , Geografía , Metaboloma , Metabolómica/métodos , Espectrometría de Masa por Ionización de Electrospray/métodos , Madera/metabolismo , Acetatos/análisis , Cromatografía Líquida de Alta Presión/métodos , Ciclopentanos/análisis , Redes y Vías Metabólicas , Metanol/análisis , Oxilipinas/análisis , Análisis de Componente PrincipalRESUMEN
The SV channel encoded by the TPC1 gene represents a Ca(2+)- and voltage-dependent vacuolar cation channel. Point mutation D454N within TPC1, named fou2 for fatty acid oxygenation upregulated 2, results in increased synthesis of the stress hormone jasmonate. As wounding causes Ca2+ signals and cytosolic Ca2+ is required for SV channel function, we here studied the Ca(2+)-dependent properties of this major vacuolar cation channel with Arabidopsis thaliana mesophyll vacuoles. In patch clamp measurements, wild-type and fou2 SV channels did not exhibit differences in cytosolic Ca2+ sensitivity and Ca2+ impermeability. K+ fluxes through wild-type TPC1 were reduced or even completely faded away when vacuolar Ca2+ reached the 0.1-mm level. The fou2 protein under these conditions, however, remained active. Thus, D454N seems to be part of a luminal Ca2+ recognition site. Thereby the SV channel mutant gains tolerance towards elevated luminal Ca2+. A three-fold higher vacuolar Ca/K ratio in the fou2 mutant relative to wild-type plants seems to indicate that fou2 can accumulate higher levels of vacuolar Ca(2+) before SV channel activity vanishes and K(+) homeostasis is impaired. In response to wounding fou2 plants might thus elicit strong vacuole-derived cytosolic Ca2+ signals resulting in overproduction of jasmonate.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Canales de Calcio/metabolismo , Señalización del Calcio , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Calcio/metabolismo , Canales de Calcio/genética , Citosol/metabolismo , Regulación de la Expresión Génica de las Plantas , Potenciales de la Membrana , Técnicas de Placa-Clamp , Mutación Puntual , Potasio/metabolismo , Vacuolas/metabolismo , Vacuolas/fisiologíaRESUMEN
The slow vacuolar (SV) channel, a Ca2+-regulated vacuolar cation conductance channel, in Arabidopsis thaliana is encoded by the single-copy gene AtTPC1. Although loss-of-function tpc1 mutants were reported to exhibit a stoma phenotype, knowledge about the underlying guard cell-specific features of SV/TPC1 channels is still lacking. Here we demonstrate that TPC1 transcripts and SV current density in guard cells were much more pronounced than in mesophyll cells. Furthermore, the SV channel in motor cells exhibited a higher cytosolic Ca2+ sensitivity than in mesophyll cells. These distinct features of the guard cell SV channel therefore probably account for the published stomatal phenotype of tpc1-2.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Canales de Calcio/metabolismo , Calcio/metabolismo , Estomas de Plantas/metabolismo , Vacuolas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Canales de Calcio/genética , Fenómenos Electrofisiológicos , Células del Mesófilo/metabolismo , Mutación , Técnicas de Placa-Clamp , Hojas de la Planta/química , Potasio/análisis , Sodio/análisis , Estrés FisiológicoRESUMEN
The consequences of altered abscisic acid (ABA) sensitivity in gray poplar (Populus x canescens [Ait.] Sm.) development were examined by ectopic expression of the Arabidopsis (Arabidopsis thaliana) mutant abi1 (for abscisic acid insensitive1) gene. The expression resulted in an ABA-insensitive phenotype revealed by a strong tendency of abi1 poplars to wilt, impaired responsiveness of their stomata to ABA, and an ABA-resistant bud outgrowth. These plants therefore required cultivation under very humid conditions to prevent drought stress symptoms. Morphological alterations became evident when comparing abi1 poplars with poplars expressing Arabidopsis nonmutant ABI1 or wild-type plants. abi1 poplars showed increased stomatal size, enhanced shoot growth, and retarded leaf and root development. The increased stomatal size and its reversion to the size of wild-type plants by exogenous ABA indicate a role for ABA in regulating stomatal development. Enhanced shoot growth and retarded leaf and root development support the hypothesis that ABA acts independently from drought stress as a negative regulator of growth in shoots and as a positive regulator of growth in leaves and roots. In shoots, we observed an interaction of ABA with ethylene: abi1 poplars exhibited elevated ethylene production, and the ethylene perception inhibitor Ag(+) antagonized the enhanced shoot growth. Thus, we provide evidence that ABA acts as negative regulator of shoot growth in nonstressed poplars by restricting ethylene production. Furthermore, we show that ABA has a role in regulating shoot branching by inhibiting lateral bud outgrowth.
Asunto(s)
Ácido Abscísico/farmacología , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Mutación/genética , Fosfoproteínas Fosfatasas/genética , Estomas de Plantas/crecimiento & desarrollo , Populus/crecimiento & desarrollo , Populus/genética , Arabidopsis/efectos de los fármacos , Proteínas de Arabidopsis/metabolismo , Medios de Cultivo/farmacología , Etilenos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas/genética , Fosfoproteínas Fosfatasas/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/crecimiento & desarrollo , Estomas de Plantas/efectos de los fármacos , Plantas Modificadas Genéticamente , Populus/efectos de los fármacos , Transformación Genética/efectos de los fármacos , Agua/fisiologíaRESUMEN
Potassium and calcium are essential for tree metabolism and various physiological processes related to growth. In recent years, special interest was therefore accorded to the effect of both cations on cambial activity and xylem development. Various studies revealed a distinct correlation between potassium as well as calcium nutrition and wood formation. When poplar trees were grown under low K(+) or Ca²(+) regimes, the cambial activity as well as the seasonal rate of wood increment and the vessel size were significantly reduced. Molecular, biochemical and electrophysiological investigations indicate (i) a strong involvement of specific K(+) channels in the regulation of xylem cell expansion and (ii) a significant influence of Ca²(+) on the onset of cambial reactivation after winter dormancy as well as on wood structure and chemistry. These studies highlight the important role of potassium as well as calcium in xylogenesis. Based on that knowledge, further research will be directed towards a better understanding of the mechanisms governing K(+)- as well as Ca²(+)-dependent wood formation.