RESUMEN
Inorganic phosphorus (Pi) fertilizers are expected to become scarce in the near future; so, breeding for improved Pi acquisition-related root traits would decrease the need for fertilizer application. This work aimed to decipher the physiological and molecular mechanisms underlying the differences between two commercial wheat cultivars (Crac and Tukan) with contrasting Pi acquisition efficiencies (PAE). For that, four independent experiments with different growth conditions were conducted. When grown under non-limiting Pi conditions, both cultivars performed similarly. Crac was less affected by Pi starvation than Tukan, presenting higher biomass production, and an enhanced root development, root:shoot ratio, and root efficiency for Pi uptake under this condition. Higher PAE in Crac correlated with enhanced expression of the Pi transporter genes TaPht1;2 and TaPht1;10. Crac also presented a faster and higher modulation of the IPS1-miR399-PHO2 pathway upon Pi starvation. Interestingly, Crac showed increased levels of strigolactones, suggesting a direct relationship between this phytohormone and plant P responses. Based on these findings, we propose that higher PAE of the cultivar Crac is associated with an improved P signalling through a fine-tuning modulation of PHO2 activity, which seems to be regulated by strigolactones. This knowledge will help to develop new strategies for improved plant performance under P stress conditions.
Asunto(s)
Fosfatos/metabolismo , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Triticum/metabolismo , Regulación de la Expresión Génica de las Plantas , Compuestos Heterocíclicos con 3 Anillos/metabolismo , Lactonas/metabolismo , Proteínas de Plantas/metabolismoRESUMEN
Background: Hyalodendriella sp. Ponipodef12, an endophytic fungus from a poplar hybrid, was a high producer of botrallin and TMC-264 with various bioactivities. In this study, the influences of eight metal ions (i.e.,Mn2+,Na+, Mg2+,Zn2+,Cu2+,Fe2+,Fe3+ and Al3+) on botrallin and TMC-264 production in liquid culture of the endophytic fungus Hyalodendriella sp. Ponipodef12 were investigated. Results: Three most effective metal ions (Zn2+,Cu2+ and Mg2+) along with their optimum concentrations were screened. The optimum addition time and concentrations of Zn2+,Cu2+ and Mg2+ were further obtained respectively for improving botrallin and TMC-264 production. The combination effects of Zn2+,Cu2+ and Mg2+ on the production of botrallin and TMC-264 by employing statistical method based on the central composite design (CCD) and response surface methodology (RSM) were evaluated, and two quadratic predictive models were developed for botrallin and TMC-264 production. The yields of botrallin and TMC-264, which were predicted as 144.12 mg/L and 36.04 mg/L respectively, were validated to be 146.51 mg/L and 36.63 mg/L accordingly with the optimum concentrations of Zn2+ at 0.81 mmol/L, Cu2+ at 0.20 mmol/L, and Mg2+ at 0.13 mmol/L in medium. Conclusion: The results indicated that the enhancement of botrallin and TMC-264 accumulation in liquid culture of the endophytic fungus Hyalodendriella sp. Ponipodef12 by the metal ions and their combination should be an effective strategy.
Asunto(s)
Ascomicetos/metabolismo , Compuestos Heterocíclicos con 3 Anillos/metabolismo , Pironas/metabolismo , Ascomicetos/efectos de los fármacos , Compuestos Heterocíclicos con 3 Anillos/química , Metales/farmacología , Pironas/químicaRESUMEN
Trypanosoma cruzi is the causative agent of Chagas' disease, which is a major endemic disease in Latin America and is recognized by the WHO as one of the 17 neglected tropical diseases in the world. Psilostachyin and psilostachyin C, two sesquiterpene lactones isolated from Ambrosia spp., have been demonstrated to have trypanocidal activity. Considering both the potential therapeutic targets present in the parasite, and the several mechanisms of action proposed for sesquiterpene lactones, the aim of this work was to characterize the mode of action of psilostachyin and psilostachyin C on Trypanosoma cruzi and to identify the possible targets for these molecules. Psilostachyin and psilostachyin C were isolated from Ambrosia tenuifolia and Ambrosia scabra, respectively. Interaction of sesquiterpene lactones with hemin, the induction of oxidative stress, the inhibition of cruzipain and trypanothione reductase and their ability to inhibit sterol biosynthesis were evaluated. The induction of cell death by apoptosis was also evaluated by analyzing phosphatidylserine exposure detected using annexin-V/propidium iodide, decreased mitochondrial membrane potential, assessed with Rhodamine 123 and nuclear DNA fragmentation evaluated by the TUNEL assay. Both STLs were capable of interacting with hemin. Psilostachyin increased about 5 times the generation of reactive oxygen species in Trypanosoma cruzi after a 4h treatment, unlike psilostachyin C which induced an increase in reactive oxygen species levels of only 1.5 times. Only psilostachyin C was able to inhibit the biosynthesis of ergosterol, causing an accumulation of squalene. Both sesquiterpene lactones induced parasite death by apoptosis. Upon evaluating the combination of both compounds, and additive trypanocidal effect was observed. Despite their structural similarity, both sesquiterpene lactones exerted their anti-T. cruzi activity through interaction with different targets. Psilostachyin accomplished its antiparasitic effect by interacting with hemin, while psilostachyin C interfered with sterol synthesis.
Asunto(s)
Enfermedad de Chagas/tratamiento farmacológico , Compuestos Heterocíclicos con 3 Anillos/farmacología , Lactonas/farmacología , Pironas/farmacología , Sesquiterpenos/farmacología , Trypanosoma cruzi/efectos de los fármacos , Ambrosia/química , Apoptosis/efectos de los fármacos , Enfermedad de Chagas/parasitología , Hemina/metabolismo , Compuestos Heterocíclicos con 3 Anillos/química , Compuestos Heterocíclicos con 3 Anillos/metabolismo , Humanos , Lactonas/química , Lactonas/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , Pironas/química , Pironas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Sesquiterpenos/química , Sesquiterpenos/metabolismo , Trypanosoma cruzi/patogenicidadRESUMEN
Ca²âº release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP3Rs) plays a relevant role in numerous physiological processes. IP3R-mediated Ca²âº signals involve Ca²âº-induced Ca²âº-release (CICR) whereby Ca²âº release through one open IP3R induces the opening of other channels. IP3Rs are apparently organized in clusters. The signals can remain localized (i.e., Ca²âº puffs) if CICR is limited to one cluster or become waves that propagate between clusters. Ca²âº puffs are the building blocks of Ca²âº waves. Thus, there is great interest in determining puff properties, especially in view of the current controversy on the spatial distribution of activatable IP3Rs. Ca²âº puffs have been observed in intact cells with optical techniques proving that they are intrinsically Ca²âº dyes, slow exogenous buffers (e.g., EGTA) to disrupt inter-cluster CICR and UV-photolyzable caged IP3. Single-wavelength dyes increase their fluorescence upon calcium binding producing images that are strongly dependent on their kinetic, transport and photophysical properties. Determining the artifacts that the imaging setting introduces is particularly relevant when trying to analyze the smallest Ca²âº signals. In this paper we introduce a method to estimate the expected signal-to-noise ratio of Ca²âº imaging experiments that use single-wavelength dyes. The method is based on the Number and rightness technique. It involves the performance of a series of experiments and their subsequent analysis in terms of a fluorescence fluctuation model with which the model parameters are quantified. Using the model, the expected signal-to-noise ratio is then computed. Equivalence classes between different experimental conditions that produce images with similar signal-to-noise ratios can then be established. The method may also be used to estimate the smallest signals that can reliably be observed with each setting.
Asunto(s)
Calcio/metabolismo , Imagen Óptica/métodos , Animales , Colorantes Fluorescentes/metabolismo , Compuestos Heterocíclicos con 3 Anillos/metabolismo , Procesamiento de Imagen Asistido por Computador , Relación Señal-Ruido , Xenopus laevis/metabolismoRESUMEN
High-K(+)-cardioplegia (CPG) and pyruvate (Pyr) are used as cardioprotective agents. Considering that mitochondria play a critical role in cardiac dysfunction, we investigated the effect of CPG on mitochondrial Ca(2+) uptake and sarcorreticular (SR) calcium handling. Cytosolic and mitochondrial Ca(2+), as well as mitochondrial membrane potential (ΔΨm) were assessed in rat cardiomyocytes by confocal microscopy. Mechano-calorimetrical correlation was studied in perfused hearts. CPG did not modify JC-1 (ΔΨm), but transiently increased, by up to 1.8 times, the Fura-2 (intracellular Ca concentration, [Ca(2+)]i) and Rhod-2 (mitochondrial free Ca concentration [Ca(2+)]m) fluorescence of resting cells, with exponential decays. The addition of 5 µmol·L(-1) thapsigargin (Tpg) increased the Rhod-2 fluorescence in a group of cells without any effect on the Fura-2 signal. In rat hearts perfused with CPG, 1 µmol·L(-1) Tpg decreased resting heat rate (ΔH(r): -0.44 ± 0.07 mW·g(-1)), while the addition of 5 µmol·L(-1) KB-R7943 increased resting pressure (ΔrLVP by +5.26 ± 1.10 mm Hg; 1 mm Hg = 133.322 Pa). The addition of 10 mmol·L(-1) Pyr to CPG increased H(r) (+3.30 ± 0.24 mW·g(-1)) and ΔrLVP (+2.2 ± 0.4 mm Hg), which are effects potentiated by KB-R7943. The results suggest that under CPG, (i) there was an increase in [Ca(2+)]i and [Ca(2+)]m (without changing ΔΨm) that decayed by exothermic removal mechanisms; (ii) mitochondrial Ca(2+) uptake contributed to the removal of cytosolic Ca(2+), in a process that was potentiated by inhibition of sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA), and reduced by KB-R7943; (iii) under these conditions, SERCA represents the main energetic consumer; (iv) Pyr increased the energetic performance of hearts,mainly by inducing mitochondrial metabolism.