Trichoderma atroviride-emitted volatiles improve growth of Arabidopsis seedlings through modulation of sucrose transport and metabolism.

Plants host a diverse microbiome and differentially react to the fungal species living as endophytes or around their roots through emission of volatiles. Here, using divided Petri plates for Arabidopsis-T. atroviride co-cultivation, we show that fungal volatiles increase endogenous sugar levels in shoots, roots and root exudates, which improve Arabidopsis root growth and branching and strengthen the symbiosis. Tissue-specific expression of three sucrose phosphate synthase-encoding genes (AtSPS1F, AtSPS2F and AtSPS3F), and AtSUC2 and SWEET transporters revealed that the gene expression signatures differ from those of the fungal pathogens Fusarium oxysporum and Alternaria alternata and that AtSUC2 is largely repressed either by increasing carbon availability or by perception of the fungal volatile 6-pentyl-2H-pyran-2-one. Our data point to Trichoderma volatiles as chemical signatures for sugar biosynthesis and exudation and unveil specific modulation of a critical, long-distance sucrose transporter in the plant.

MPK6 Kinase Regulates Plasma Membrane H+-ATPase Activity in Cold Acclimation.

Cold and freezing stresses severely affect plant growth, development, and survival rate. Some plant species have evolved a process known as cold acclimation, in which plants exposed to temperatures above 0 °C trigger biochemical and physiological changes to survive freezing. During this response, several signaling events are mediated by transducers, such as mitogen activated protein kinase (MAPK) cascades. Plasma membrane H+-ATPase is a key enzyme for the plant cell life under regular and stress conditions. Using wild type and mpk3 and mpk6 knock out mutants in Arabidopsis thaliana, we explored the transcriptional, translational, and 14-3-3 protein regulation of the plasma membrane H+-ATPase activity under the acclimation process. The kinetic analysis revealed a differential profiling of the H+-ATPase activity depending on the presence or absence of MPK3 or MPK6 under non-acclimated or acclimated conditions. Negative regulation of the plasma membrane H+-ATPase activity was found to be exerted by MPK3 in non-acclimated conditions and by MPK6 in acclimated conditions, describing a novel form of regulation of this master ATPase. The MPK6 regulation involved changes in plasma membrane fluidity. Moreover, our results indicated that MPK6 is a critical regulator in the process of cold acclimation that leads to freezing tolerance and further survival.

Role of pyrimidine salvage pathway in the maintenance of organellar and nuclear genome integrity.

Nucleotide biosynthesis proceeds through a de novo pathway and a salvage route. In the salvage route, free bases and/or nucleosides are recycled to generate the corresponding nucleotides. Thymidine kinase (TK) is the first enzyme in the salvage pathway to recycle thymidine nucleosides as it phosphorylates thymidine to yield thymidine monophosphate. The Arabidopsis genome contains two TK genes -TK1a and TK1b- that show similar expression patterns during development. In this work, we studied the respective roles of the two genes during early development and in response to genotoxic agents targeting the organellar or the nuclear genome. We found that the pyrimidine salvage pathway is crucial for chloroplast development and genome replication, as well as for the maintenance of its integrity, and is thus likely to play a crucial role during the transition from heterotrophy to autotrophy after germination. Interestingly, defects in TK activity could be partially compensated by supplementation of the medium with sugar, and this effect resulted from both the availability of a carbon source and the activation of the nucleotide de novo synthesis pathway, providing evidence for a compensation mechanism between two routes of nucleotide biosynthesis that depend on nutrient availability. Finally, we found differential roles of the TK1a and TK1b genes during the plant response to genotoxic stress, suggesting that different pools of nucleotides exist within the cells and are required to respond to different types of DNA damage. Altogether, our results highlight the importance of the pyrimidine salvage pathway, both during plant development and in response to genotoxic stress.

Effects of seed priming on germination and seedling growth of desiccation-sensitive seeds from Mexican tropical rainforest.

Seed priming increases the vigor of seeds and seedlings through metabolic and biochemical processes occurring during controlled hydration, followed by dehydration. In the field, seeds are exposed to hydration-dehydration events in and on the soil after dispersal, as in seed priming. Nevertheless, seed priming has been sparsely tested on desiccation-sensitive seeds, which are vulnerable to climate change effects. We evaluated the effect of two priming methods on seeds from two tropical rainforest species: Cupania glabra and Cymbopetalum baillonii. For hydropriming, the seeds were fully hydrated and then dehydrated to three dehydration levels. For natural priming, the seeds were buried for 12 days in either closed forest or forest gap. Primed seeds were sown in 1% agar medium and placed in an environmental chamber. The growth of the seedlings from the highest germination priming treatments was evaluated for 1 year in the field. Our results showed that for C. glabra and C. baillonii, hydroprimed seeds varied in their germination response, depending on the degree of their dehydration. However, for C. baillonii, hydropriming seems to invigorate seeds, compared to non-imbibed seeds of the same dehydration level. Natural priming increased germination speed in both species without any difference between closed forest and forest gap. Moreover, seeds with natural priming had a higher final germination percentage than seeds with hydropriming. Seedlings from seeds with natural priming showed a higher growth rate than the controls in both species, whereas hydropriming produced a similar effect in C. glabra. Both priming methods could be used for restoration practices with the studied species, natural priming being a novel method. The ecological implications of priming in desiccation sensitive seeds are discussed in this study.

Biochemical properties and subcellular localization of six members of the HXK family in maize and its metabolic contribution to embryo germination.

BACKGROUND: Seed germination is a crucial process in the plant life cycle when a dramatic variation of type and sugar content occurs just as the seed is hydrated. The production of hexose 6 phosphate is a key node in different pathways that are required for a successful germination. Hexokinase (HXK) is the only plant enzyme that phosphorylates glucose (Glc), so it is key to fueling several metabolic pathways depending on their substrate specificity, metabolite regulatory responses and subcellular localization. In maize, the HXK family is composed of nine genes, but only six of them (ZmHXK4-9) putatively encode catalytically active enzymes. Here, we cloned and functionally characterized putative catalytic enzymes to analyze their metabolic contribution during germination process. RESULTS: From the six HXKs analyzed here, only ZmHXK9 has minimal hexose phosphorylating activity even though enzymatic function of all isoforms (ZmHXK4-9) was confirmed using a yeast complementation approach. The kinetic parameters of recombinant proteins showed that ZmHXK4-7 have high catalytic efficiency for Glc, fructose (Fru) and mannose (Man), ZmHXK7 has a lower Km for ATP, and together with ZmHXK8 they have lower sensitivity to inhibition by ADP, G6P and N-acetylglucosamine than ZmHXK4-6 and ZmHXK9. Additionally, we demonstrated that ZmHXK4-6 and ZmHXK9 are located in the mitochondria and their location relies on the first 30 amino acids of the N-terminal domain. Otherwise, ZmHXK7-8 are constitutively located in the cytosol. HXK activity was detected in cytosolic and mitochondrial fractions and high Glc and Fru phosphorylating activities were found in imbibed embryos. CONCLUSIONS: Considering the biochemical characteristics, location and the expression of ZmHXK4 at onset of germination, we suggest that it is the main contributor to mitochondrial activity at early germination times, at 24 h other ZmHXKs also contribute to the total activity. While in the cytosol, ZmHXK7 could be responsible for the activity at the onset of germination, although later, ZmHXK8 also contributes to the total HXK activity. Our observations suggest that the HXKs may be redundant proteins with specific roles depending on carbon and ATP availability, metabolic needs, or sensor requirements. Further investigation is necessary to understand their specific or redundant physiological roles.

Plant Hexokinases are Multifaceted Proteins.

Sugars are the main carbon and energy source in cells, but they can also act as signaling molecules that affect the whole plant life cycle. Certain tissues can produce sugars and supply them to others, and this plant tissue heterogeneity makes sugar signaling a highly complex process that requires elements capable of perceiving changes in sugar concentrations among different tissues, cell compartments and developmental stages. In plants, the regulatory effects of glucose (Glc) have been the most studied to date. The first Glc sensor identified in plants was hexokinase (HXK), which is currently recognized as a dual-function protein. In addition to its catalytic activity, this enzyme can also repress the expression of some photosynthetic genes in response to high internal Glc concentrations. Additionally, the catalytic activity of HXKs has a profound impact on cell metabolism and other sugar signaling pathways that depend on phosphorylated hexoses and intermediate glycolytic products. HXKs are the only proteins that are able to phosphorylate Glc in plants, since no evidence has been provided to date concerning the existence of a glucokinase. Moreover, the intracellular localization of HXKs seems to be crucial to their activity and sensor functions. Recently, two new and surprising functions have been described for HXKs. In this review, we discuss the versatility of HXKs in regard to their catalytic and glucose sensor activities, intracellular location, protein-protein and hormone interactions, as well as how these HXK characteristics influence plant growth and development, in an effort to understand this enzyme's role in improving plant productivity.

Identification and Characterization of VDAC Family in Maize.

The voltage-dependent anion channel (VDAC) is the most abundant protein in the outer mitochondrial membrane (OMM) of all eukaryotes, having an important role in the communication between mitochondria and cytosol. The plant VDAC family consists of a wide variety of members that may participate in cell responses to several environmental stresses. However, there is no experimental information about the members comprising the maize VDAC (ZmVDAC) family. In this study, the ZmVDAC family was identified, and described, and its gene transcription profile was explored during the first six days of germination and under different biotic stress stimuli. Nine members were proposed as bona fide VDAC genes with a high potential to code functional VDAC proteins. Each member of the ZmVDAC family was characterized in silico, and nomenclature was proposed according to phylogenetic relationships. Transcript levels in coleoptiles showed a different pattern of expression for each ZmVDAC gene, suggesting specific roles for each one during seedling development. This expression profile changed under Fusarium verticillioides infection and salicylic acid, methyl jasmonate, and gibberellic acid treatments, suggesting no redundancy for the nine ZmVDAC genes and, thus, probably specific and diverse functions according to plant needs and environmental conditions. Nevertheless, ZmVDAC4b was significantly upregulated upon biotic stress signals, suggesting this gene's potential role during the biotic stress response.

Maize plant expresses SWEET transporters differently when interacting with Trichoderma asperellum and Fusarium verticillioides, two fungi with different lifestyles.

Most Trichoderma species are beneficial fungi that promote plant growth and resistance, while Fusarium genera cause several crop damages. During the plant-fungi interaction there is a competition for sugars in both lifestyles. Here we analyzed the plant growth promotion and biocontrol activity of T. asperellum against F. verticillioides and the effect of both fungi on the expression of the maize diffusional sugar transporters, the SWEETs. The biocontrol activity was done in two ways, the first was by observing the growth capacity of both fungus in a dual culture. The second one by analyzing the infection symptoms, the chlorophyl content and the transcript levels of defense genes determined by qPCR in plants with different developmental stages primed with T. asperellum conidia and challenged with F. verticillioides. In a dual culture, T. asperellum showed antagonist activity against F. verticillioides. In the primed plants a delay in the infection disease was observed, they sustained chlorophyll content even after the infection, and displayed upregulated defense-related genes. Additionally, the T. asperellum primed plants had longer stems than the nonprimed plants. SWEETs transcript levels were analyzed by qPCR in plants primed with either fungus. Both fungi affect the transcript levels of several maize sugar transporters differently. T. asperellum increases the expression of six SWEETs on leaves and two at the roots and causes a higher exudation of sucrose, glucose, and fructose at the roots. On the contrary, F. verticillioides reduces the expression of the SWEETs on the leaves, and more severely when a more aggressive strain is in the plant. Our results suggest that the plant is able to recognize the lifestyle of the fungi and respond accordingly by changing the expression of several genes, including the SWEETs, to establish a new sugar flux.

Fumonisin B1, a toxin produced by Fusarium verticillioides, modulates maize ß-1,3-glucanase activities involved in defense response.

Fusarium verticillioides is an important pathogen in maize that causes various diseases affecting all stages of plant development worldwide. The fungal pathogen could be seed borne or survive in soil and penetrate the germinating seed. Most F. verticillioides strains produce fumonisins, which are of concern because of their toxicity to animals and possibly humans, and because they enhance virulence against seedlings of some maize genotypes. In this work, we studied the action of fumonisin B1 (FB1) on the activity of maize ß-1,3-glucanases involved in plant defense response. In maize embryos, FB1 induced an acidic isoform while suppressing the activity of two basic isoforms. This acidic isoform was induced also with 2,6-dichloroisonicotinic acid, an analog of salicylic acid. Repression of the basic isoforms suggested a direct interaction of the enzymes with the mycotoxin as in vitro experiments showed that pure FB1 inhibited the basic ß-1,3-glucanases with an IC(50) of 53 µM. When germinating maize embryos were inoculated with F. verticillioides the same dual effect on ß-1,3-glucanase activities that we observed with the pure toxin was reproduced. Similar levels of FB1 were recovered at 24 h germination in maize tissue when they were treated with pure FB1 or inoculated with an FB1-producing strain. These results suggest that ß-1,3-glucanases are a relevant physiological target and their modulation by FB1 might contribute to F. verticillioides colonization.

Early carbon mobilization and radicle protrusion in maize germination.

Considerable amounts of information is available on the complex carbohydrates that are mobilized and utilized by the seed to support early seedling development. These events occur after radicle has protruded from the seed. However, scarce information is available on the role of the endogenous soluble carbohydrates from the embryo in the first hours of germination. The present work analysed how the soluble carbohydrate reserves in isolated maize embryos are mobilized during 6-24 h of water imbibition, an interval that exclusively embraces the first two phases of the germination process. It was found that sucrose constitutes a very significant reserve in the scutellum and that it is efficiently consumed during the time in which the adjacent embryo axis is engaged in an active metabolism. Sucrose transporter was immunolocalized in the scutellum and in vascular elements. In parallel, a cell-wall invertase activity, which hydrolyses sucrose, developed in the embryo axis, which favoured higher glucose uptake. Sucrose and hexose transporters were active in the embryo tissues, together with the plasma membrane H(+)-ATPase, which was localized in all embryo regions involved in both nutrient transport and active cell elongation to support radicle extension. It is proposed that, during the initial maize germination phases, a net flow of sucrose takes place from the scutellum towards the embryo axis and regions that undergo elongation. During radicle extension, sucrose and hexose transporters, as well as H(+)-ATPase, become the fundamental proteins that orchestrate the transport of nutrients required for successful germination.

Constructing Cell-Free Expression Systems for Low-Cost Access.

Cell-free systems for gene expression have gained attention as platforms for the facile study of genetic circuits and as highly effective tools for teaching. Despite recent progress, the technology remains inaccessible for many in low- and middle-income countries due to the expensive reagents required for its manufacturing, as well as specialized equipment required for distribution and storage. To address these challenges, we deconstructed processes required for cell-free mixture preparation and developed a set of alternative low-cost strategies for easy production and sharing of extracts. First, we explored the stability of cell-free reactions dried through a low-cost device based on silica beads, as an alternative to commercial automated freeze dryers. Second, we report the positive effect of lactose as an additive for increasing protein synthesis in maltodextrin-based cell-free reactions using either circular or linear DNA templates. The modifications were used to produce active amounts of two high-value reagents: the isothermal polymerase Bst and the restriction enzyme BsaI. Third, we demonstrated the endogenous regeneration of nucleoside triphosphates and synthesis of pyruvate in cell-free systems (CFSs) based on phosphoenol pyruvate (PEP) and maltodextrin (MDX). We exploited this novel finding to demonstrate the use of a cell-free mixture completely free of any exogenous nucleotide triphosphates (NTPs) to generate high yields of sfGFP expression. Together, these modifications can produce desiccated extracts that are 203-424-fold cheaper than commercial versions. These improvements will facilitate wider use of CFS for research and education purposes.

Moonlighting Proteins: The Case of the Hexokinases.

Moonlighting proteins are defined as proteins with two or more functions that are unrelated and independent to each other, so that inactivation of one of them should not affect the second one and vice versa. Intriguingly, all the glycolytic enzymes are described as moonlighting proteins in some organisms. Hexokinase (HXK) is a critical enzyme in the glycolytic pathway and displays a wide range of functions in different organisms such as fungi, parasites, mammals, and plants. This review discusses HXKs moonlighting functions in depth since they have a profound impact on the responses to nutritional, environmental, and disease challenges. HXKs' activities can be as diverse as performing metabolic activities, as a gene repressor complexing with other proteins, as protein kinase, as immune receptor and regulating processes like autophagy, programmed cell death or immune system responses. However, most of those functions are particular for some organisms while the most common moonlighting HXK function in several kingdoms is being a glucose sensor. In this review, we also analyze how different regulation mechanisms cause HXK to change its subcellular localization, oligomeric or conformational state, the response to substrate and product concentration, and its interactions with membrane, proteins, or RNA, all of which might impact the HXK moonlighting functions.

Fagopyrum esculentum at early stages copes with aluminum toxicity by increasing ABA levels and antioxidant system.

Aluminum toxicity (Al) is one of the main constraints for plant growth on acid soils. While most plants are sensitive to Al, some species have developed strategies to cope with this metal. Fagopyrum esculentum, Moench., var Mancan (Polygonaceae), despite being an aluminum-tolerant plant, shows root inhibition as a seedling during the first hours of exposure to Al, whereas at later times, it fully recovers. In this study, we assessed whether abscisic acid (ABA) levels and the antioxidant system might be involved in the early tolerance mechanisms of F. esculentum. The results showed that seedlings exposed to 50 µM Al for 3, 6, 12, 24, and 48 h showed decreases in the relative root growth (RRG), and there was an accumulation of Al in the root apex from 3 to 24 h. In addition, reactive oxygen species (ROS) levels increased, and were detected early after Al exposure; endogenous ABA levels increased and antioxidant enzyme activity increased, including catalase (CAT, EC1.11.1.6), glutathione reductase (GR, EC 1.6.4.2), ascorbate peroxidase (APX, EC 1.11.1.11), and superoxide dismutase (SOD, EC 1.15.1.1) activity. Seedlings treated with exogenous ABA also showed increased ROS levels and CAT and APX activity. The results suggest that after the first 12 h of Al treatment, root growth declines while ROS levels increase due to the entrance of Al into the root. However, the enzyme antioxidant system is promoted, which may impact the recovery of the root growth at later times and increasing levels of ABA might mediate this effect.

Sub-basal increases of GABA enhance the synthesis of TNF-α, TGF-ß, and IL-1ß in the immune system organs of the Nile tilapia.

The cells of the immune and neuronal systems share different receptors for cytokines or neurotransmitters, producing feedback responses between both systems. Cytokines such as IL-1ß and TNF-α can induce inflammation; however, the secretion of these molecules can be modulated by anti-inflammatory cytokines, as is the case for TGF-ß, as well as by different hormones or neurotransmitters such as the γ-aminobutyric acid (GABA). In this study, we evaluated the secretion of IL-1ß, TNF-α, and TGF-ß under basal conditions, in the head of the kidney, spleen, thymus, and serum of the Nile tilapia, as well as their release induced by different sub-basal increases of GABA. We found that at the higher dose of GABA these cytokines were synthesised at a higher concentration compared to the control group. These results may suggest that there is feedback between both systems and that GABA plays a role in the modulation of the immune response.

Phytotoxic effects of Sicyos deppei (Cucurbitaceae) in germinating tomato seeds.

The phytotoxic effect of allelochemicals is referred to as allelochemical stress and it is considered a biotic stress. Sicyos deppei G. Don (Cucurbitaceae) is an allelopathic weed that causes phytotoxicity in Lycopersicon esculentum, delaying seed germination and severely inhibiting radicle growth. This paper reports in in vitro conditions, the effects of the aqueous leachate of S. deppei-throughout tomato germination times-on (1) the dynamics of starch and sugars metabolism, (2) activity and expression of the cell wall enzymes involved in endosperm weakening that allows the protrusion of the radicle, and (3) whether abscisic acid (ABA) is involved in this altered metabolic processes. Results showed that S. deppei leachate on tomato seed germination mainly caused: (1) delay in starch degradation as well as in sucrose hydrolysis; (2) lower activity of sucrose phosphate synthase, cell wall invertase, and alpha-amylase; being sucrose phosphate synthase (SPS) gene expression down-regulated, and the last two up regulated; (3) also, lower activity of endo beta-mannanase, beta-1,3 glucanase, alpha-galactosidase, and exo-polygalacturonase with altered gene expression; and (4) higher content of ABA during all times of germination. The phytotoxic effect of S. deppei aqueous leachate is because of the sum of many metabolic processes affected during tomato seed germination that finally is evidenced by a strong inhibition of radicle growth.

SWEET Transporters for the Nourishment of Embryonic Tissues during Maize Germination.

In maize seed germination, the endosperm and the scutellum nourish the embryo axis. Here, we examined the mRNA relative amount of the SWEET protein family, which could be involved in sugar transport during germination since high [14-C]-glucose and mainly [14-C]-sucrose diffusional uptake were found in embryo tissues. We identified high levels of transcripts for SWEETs in the three phases of the germination process: ZmSWEET4c, ZmSWEET6b, ZmSWEET11, ZmSWEET13a, ZmSWEET13b, ZmSWEET14b and ZmSWEET15a, except at 0 h of imbibition where the abundance of each ZmSWEET was low. Despite the major sucrose (Suc) biosynthesis capacity of the scutellum and the high level of transcripts of the Suc symporter SUT1, Suc was not found to be accumulated; furthermore, in the embryo axis, Suc did not decrease but hexoses increased, suggesting an efficient Suc efflux from the scutellum to nourish the embryo axis. The influx of Glc into the scutellum could be mediated by SWEET4c to take up the large amount of transported sugars due to the late hydrolysis of starch. In addition, sugars regulated the mRNA amount of SWEETs at the embryo axis. These results suggest an important role for SWEETs in transporting Suc and hexoses between the scutellum and the embryo axis, and differences in SWEET transcripts between both tissues might occur because of the different sugar requirements and metabolism.

Ethnobotanical survey and antibacterial activity of plants used in the Altiplane region of Mexico for the treatment of oral cavity infections.

ETHNOPHARMACOLOGICAL RELEVANCE: The ethnobotanical survey conducted in this study showed 47 plant species used in the Altiplane region of Mexico for the treatment of dental diseases such as toothache, dental caries, periodontal disease and gingivitis. MATERIALS AND METHODS: Information was collected by performing interviews. The following data were recorded: name of the patients or herbalists with their age, sex; date and place of gathering information; pathology of persons interviewed; name of the drug (vernacular name); parts used (leaves, bark, fruits, seeds, aerial parts), mode of preparation and administration, and possible combinations. The inhibitory effects of the aqueous and ethanolic extracts of the medicinal plants detected during the survey the on the growth of Streptococcus mutans and Phrophyromonas gingivalis were determined using microdilution method. The minimum bactericidal concentrations (MBC) were determined from the wells of microplate with no visible bacterial growth. RESULTS: In total, tree places of the Altiplane region of Mexico were visited and five healers and 100 patients were questioned. 47 wild and cultivated species were recorded. The most frequent uses were to treat tooth pain, gum diseases, bad breath and cavities. Infusions were the most frequently prepared formulation. Other applied preparations mentioned with decreasing frequency were decocts, syrups, tinctures, direct application of the plant material without prior preparation and finally macerations. The ethanolic extracts of Haematoxylon brasiletto, Punica granatum, Iostephane heterophyla, Bursera simaruba, Cedrela odorata and Rhus standleyi (12.5-65.0 µg/mL) as well as water extracts of Haematoxylon brasiletto, Punica granatum, Iostephane heterophyla, Amphipterygium adstringens, Argemone mexicana, Cedrela odorata, Eysenhardtia polystachya, Persea americana, Syzygium aromaticum, Cinnamomun zeylanicum, Cnidoscolus multilobus and Rhus standleyi (10.5-78.0 µg/mL) showed the highest inhibitory effect against Streptococcus mutans and Porphyromonas gingivalis. CONCLUSIONS: Many plants are used in the Mexican traditional medicine to treat oral bacterial diseases by the healers or patients. Our study demonstrated that most of the medicinal plants showed an antibacterial effect in vitro, and justified at least in part their use in traditional medicine. These results encourage further investigations to extract and identify the active chemical compounds responsible for the antibacterial effect observed.

Isolation of the new anacardic acid 6-[16'Z-nonadecenyl]-salicylic acid and evaluation of its antimicrobial activity against Streptococcus mutans and Porphyromonas gingivalis.

A new anacardic acid, 6-[16'Z-nonadecenyl]-salicylic acid (1), along with seven known compounds, 6-[8'Z-pentadecenyl] salicylic acid (15:1 anacardic acid) (2), 6-nonadecenyl salicylic acid (anacardic acid 19:0) (3), 6-pentadecyl salicylic acid (anacardic acid 15:0) (4), masticadienonic acid (5), 3α-hydroxymasticadienonic acid (6), 3-epi-oleanolic acid (7) and ß-sitosterol, were isolated from the bark of Amphipterygium adstringens using a bioassay-guided fractionation method. The structure of the new compound (1) was elucidated by spectroscopic data interpretation. The known compounds (2-7) were identified by comparison of their spectroscopic data with reported values in the literature. Compounds 1-4 exhibited antibacterial activity against Streptococcus mutans and Porphyromonas gingivalis with minimum inhibitory concentrations ranging from 7 to 104 µg mL and from 12 to 126 µg mL, respectively.

Kinetics of the H+-ATPase from dry and 5-hours-imbibed maize embryos in its native, solubilized, and reconstituted forms.

Membranes undergo recovery upon rehydration in seed germination. Previous work has described that the plasma membrane H+-ATPase from maize embryos adopts two different forms at 0 and 5 h of imbibition. We investigated how the kinetics of these two forms could be affected by alterations in the plasma membrane (PM). In comparison to the 0-h, PMs from the 5-h imbibed embryos showed changes in glycerophospholipid composition, decrease in leakage, and increase in fluidity. Kinetics of the PM H+-ATPase from 0 and 5-h imbibed embryos showed negative cooperativity. With the removal of the membrane environment, the activity of the enzymes shifted to a more complex kinetics, displaying two enzyme components. Lipid reconstitution produced one component with positive cooperativity. In all cases, enzymes from 0 and 5-h imbibed embryos presented similar kinetics with some quantitative differences. These results indicate that the two enzyme forms have the potential ability to respond to changes in the membrane environment, but the fact that they do not show differences in the native membranes at 0 or 5 h implies that modifications in the membrane are not drastic enough to alter their kinetics, or that they are able to preserve their boundary lipids or associated proteins and thus retain the same kinetic behavior.

Fumonisin B1, a sphingoid toxin, is a potent inhibitor of the plasma membrane H+-ATPase.

Fumonisin B(1) (FB(1)) is an amphipathic toxin produced by the pathogenic fungus Fusarium verticillioides which causes stem, root and ear rot in maize (Zea mays L.). In this work, we studied the action of FB(1) on the plasma membrane H(+)-ATPase (EC 3.6.1.34) from germinating maize embryos, and on the fluidity and lipid peroxidation of these membranes. In maize embryos the toxin at 40 microM inhibited root elongation by 50% and at 30 microM decreased medium acidification by about 80%. Irrespective of the presence and absence of FB(1), the H(+)-ATPase in plasma membrane vesicles exhibited non-hyperbolic saturation kinetics by ATPH-Mg, with Hill number of 0.67. Initial velocity studies revealed that FB(1) is a total uncompetitive inhibitor of this enzyme with an inhibition constant value of 17.5+/-1 microM. Thus FB(1) decreased V(max) and increased the apparent affinity of the enzyme for ATP-Mg to the same extent. Although FB(1) increased the fluidity at the hydrophobic region of the membrane, no correlation was found with its effect on enzyme activity, since both effects showed different FB(1)-concentration dependence. Peroxidation of membrane lipids was not affected by the toxin. Our results suggest that, under in vivo conditions, the plasma membrane H(+)-ATPase is a potentially important target of the toxin, as it is inhibited not only by FB(1) but also by its structural analogs, the sphingoid intermediates, which accumulate upon the inhibition of sphinganine N-acyltransferase by this toxin.