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Sample records for arabidopsis sucrose transporters

  1. Functional Analysis of Arabidopsis Sucrose Transporters

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    John M. Ward

    2009-03-31

    Sucrose is the main photosynthetic product that is transported in the vasculature of plants. The long-distance transport of carbohydrates is required to support the growth and development of net-importing (sink) tissues such as fruit, seeds and roots. This project is focused on understanding the transport mechanism sucrose transporters (SUTs). These are proton-coupled sucrose uptake transporters (membrane proteins) that are required for transport of sucrose in the vasculature and uptake into sink tissues. The accomplishments of this project included: 1) the first analysis of substrate specificity for any SUT. This was accomplished using electrophysiology to analyze AtSUC2, a sucrose transporter from companion cells in Arabidopsis. 2) the first analysis of the transport activity for a monocot SUT. The transport kinetics and substrate specificity of HvSUT1 from barley were studied. 3) the first analysis of a sucrose transporter from sugarcane. and 4) the first analysis of transport activity of a sugar alcohol transporter homolog from plants, AtPLT5. During this period four primary research papers, funded directly by the project, were published in refereed journals. The characterization of several sucrose transporters was essential for the current effort in the analysis of structure/function for this gene family. In particular, the demonstration of strong differences in substrate specificity between type I and II SUTs was important to identify targets for site-directed mutagenesis.

  2. Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system

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    Lalonde Sylvie

    2003-03-01

    Full Text Available Abstract Background The Arabidopsis genome contains nine sucrose transporter paralogs falling into three clades: SUT1-like, SUT2 and SUT4. The carriers differ in their kinetic properties. Many transport proteins are known to exist as oligomers. The yeast-based split ubiquitin system can be used to analyze the ability of membrane proteins to interact. Results Promoter-GUS fusions were used to analyze the cellular expression of the three transporter genes in transgenic Arabidopsis plants. All three fusion genes are co-expressed in companion cells. Protein-protein interactions between Arabidopsis sucrose transporters were tested using the split ubiquitin system. Three paralogous sucrose transporters are capable of interacting as either homo- or heteromers. The interactions are specific, since a potassium channel and a glucose transporter did not show interaction with sucrose transporters. Also the biosynthetic and metabolizing enzymes, sucrose phosphate phosphatase and sucrose synthase, which were found to be at least in part bound to the plasma membrane, did not specifically interact with sucrose transporters. Conclusions The split-ubiquitin system provides a powerful tool to detect potential interactions between plant membrane proteins by heterologous expression in yeast, and can be used to screen for interactions with membrane proteins as baits. Like other membrane proteins, the Arabidopsis sucrose transporters are able to form oligomers. The biochemical approaches are required to confirm the in planta interaction.

  3. Arabidopsis Sucrose Transporter SUT4 Interacts with Cytochrome b5-2 to Regulate Seed Germination in Response to Sucrose and Glucose

    Institute of Scientific and Technical Information of China (English)

    Yan Li; Ling-Li Li; Ren-Chun Fan; Chang-Cao Peng; Hai-Li Sun; Sai-Yong Zhu; Xiao-Fang Wang; Ling-Yun Zhang; Da-Peng Zhang

    2012-01-01

    It remains unknown whether a sucrose transporter mediates sugar signaling.Here,we report that the Arabidopsis (Arabidopsis thaliana) sucrose transporter SUT4 interacts with five members of the Arabidopsis cytochrome b5(Cyb5) family,and sucrose represses the interaction between SUT4 and a Cyb5 member Cyb5-2/A.We observed that downregulation of SUT4 and three cytochrome b5 members (Cyb5-2,Cyb5-4,and Cyb5-6) confers the sucrose-and glucoseinsensitive phenotypes in the sucrose/glucose-induced inhibition of seed germination.The sut4 cyb5-2 double mutant displays slightly stronger sucrose/glucose-insensitive phenotypes than either the sut4 or cyb5-2 single mutant.We showed that the SUT4/Cyb5-2-mediated signaling in the sucrose/glucose-induced inhibition of seed germination does not require ABA or the currently known ABI2/ABI4/ABI5-mediated signaling pathway(s).These data provide evidence that the sucrose transporter SUT4 interacts with Cyb5 to positively mediate sucrose and glucose signaling in the sucrose/glucose-induced inhibition of seed germination.

  4. Water Deficit Enhances C Export to the Roots in Arabidopsis thaliana Plants with Contribution of Sucrose Transporters in Both Shoot and Roots.

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    Durand, Mickaël; Porcheron, Benoît; Hennion, Nils; Maurousset, Laurence; Lemoine, Rémi; Pourtau, Nathalie

    2016-03-01

    Root high plasticity is an adaptation to its changing environment. Water deficit impairs growth, leading to sugar accumulation in leaves, part of which could be available to roots via sucrose (Suc) phloem transport. Phloem loading is widely described in Arabidopsis (Arabidopsis thaliana), while unloading in roots is less understood. To gain information on leaf-to-root transport, a soil-based culture system was developed to monitor root system architecture in two dimensions. Under water deficit (50% of soil water-holding capacity), total root length was strongly reduced but the depth of root foraging and the shape of the root system were less affected, likely to improve water uptake. (14)CO2 pulse-chase experiments confirmed that water deficit enhanced carbon (C) export to the roots, as suggested by the increased root-to-shoot ratio. The transcript levels of AtSWEET11 (for sugar will eventually be exported transporter), AtSWEET12, and AtSUC2 (for Suc carrier) genes, all three involved in Suc phloem loading, were significantly up-regulated in leaves of water deficit plants, in accordance with the increase in C export from the leaves to the roots. Interestingly, the transcript levels of AtSUC2 and AtSWEET11 to AtSWEET15 were also significantly higher in stressed roots, underlying the importance of Suc apoplastic unloading in Arabidopsis roots and a putative role for these Suc transporters in Suc unloading. These data demonstrate that, during water deficit, plants respond to growth limitation by allocating relatively more C to the roots to maintain an efficient root system and that a subset of Suc transporters is potentially involved in the flux of C to and in the roots. PMID:26802041

  5. A novel fluorescent assay for sucrose transporters

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    Gora Peter J

    2012-04-01

    Full Text Available Abstract Background We have developed a novel assay based on the ability of type I sucrose uptake transporters (SUTs to transport the fluorescent coumarin β-glucoside, esculin. Budding yeast (Saccharomyces cerevisiae is routinely used for the heterologous expression of SUTs and does not take up esculin. Results When type I sucrose transporters StSUT1 from potato or AtSUC2 from Arabidopsis were expressed in yeast, the cells were able to take up esculin and became brightly fluorescent. We tested a variety of incubation times, esculin concentrations, and buffer pH values and found that for these transporters, a 1 hr incubation at 0.1 to 1 mM esculin at pH 4.0 produced fluorescent cells that were easily distinguished from vector controls. Esculin uptake was assayed by several methods including fluorescence microscopy, spectrofluorometry and fluorescence-activiated cell sorting (FACS. Expression of the type II sucrose transporter OsSUT1 from rice did not result in increased esculin uptake under any conditions tested. Results were reproduced successfully in two distinct yeast strains, SEY6210 (an invertase mutant and BY4742. Conclusions The esculin uptake assay is rapid and sensitive and should be generally useful for preliminary tests of sucrose transporter function by heterologous expression in yeast. This assay is also suitable for selection of yeast showing esculin uptake activity using FACS.

  6. The Structure of Sucrose Synthase-1 from Arabidopsis thaliana and Its Functional Implications

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    Zheng, Yi; Anderson, Spencer; Zhang, Yanfeng; Garavito, R. Michael (MSU); (NWU)

    2014-10-02

    Sucrose transport is the central system for the allocation of carbon resources in vascular plants. During growth and development, plants control carbon distribution by coordinating sites of sucrose synthesis and cleavage in different plant organs and different cellular locations. Sucrose synthase, which reversibly catalyzes sucrose synthesis and cleavage, provides a direct and reversible means to regulate sucrose flux. Depending on the metabolic environment, sucrose synthase alters its cellular location to participate in cellulose, callose, and starch biosynthesis through its interactions with membranes, organelles, and cytoskeletal actin. The x-ray crystal structure of sucrose synthase isoform 1 from Arabidopsis thaliana (AtSus1) has been determined as a complex with UDP-glucose and as a complex with UDP and fructose, at 2.8- and 2.85-{angstrom} resolutions, respectively. The AtSus1 structure provides insights into sucrose catalysis and cleavage, as well as the regulation of sucrose synthase and its interactions with cellular targets.

  7. The role of sugars and sugar metabolism genes (sucrose synthase) in arabidopsis thaliana seed development

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    Odunlami, Benjamin Oladipo

    2009-01-01

    Seed development in Arabidopsis thaliana, has been studied at several levels. However, little has been done to study the role of sugar metabolism genes in seed pod development in this species. As the fertilized egg progresses to a mature seed, the sugars composition during different stages of the developing changes. These changes are related to metabolic processes in the developing seeds, but also to the activity of sucrose- converting and transporting genes, active at the interphase between ...

  8. Conserved cis-regulatory elements for DNA-binding-with-one-finger and homeo-domain-leucine-zipper transcription factors regulate companion cell-specific expression of the Arabidopsis thaliana SUCROSE TRANSPORTER 2 gene.

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    Schneidereit, Alexander; Imlau, Astrid; Sauer, Norbert

    2008-09-01

    The transition from young carbon-importing sink leaves of higher plants to mature carbon-exporting source leaves is paralleled by a complete reversal of phloem function. While sink-leaf phloem mediates the influx of reduced carbon from older source leaves and the release of this imported carbon to the sink-leaf mesophyll, source-leaf phloem catalyzes the uptake of photoassimilates into companion cells (CCs) and sieve elements (SEs) and the net carbon export from the leaf. Phloem loading in source leaves with sucrose, the main or exclusive transport form for fixed carbon in most higher plants, is catalyzed by plasma membrane-localized sucrose transporters. Consistent with the described physiological switch from sink to source, the promoter of the Arabidopsis AtSUC2 gene is active only in source-leaf CCs of Arabidopsis or of transgenic tobacco (Nicotiana tabacum). For the identification of regulatory elements involved in this companion cell-specific and source-specific gene expression, we performed detailed analyses of the AtSUC2 promoter by truncation and mutagenesis. A 126-bp promoter fragment was identified, which seems to contain these fragments and which drives AtSUC2-typical expression when combined with a 35S minimal promoter. Within this fragment, linker-scanning analyses revealed two cis-regulatory elements that were further characterized as putative binding sites for transcription factors of the DNA-binding-with-one-finger or the homeo-domain-leucine-zipper families. Similar or identical binding sites are found in other genes and in different plant species, suggesting an ancient regulatory mechanism for this important physiological switch. PMID:18551303

  9. Plant Sucrose Transporters from a Biophysical Point of View

    Institute of Scientific and Technical Information of China (English)

    Dietmar Geiger

    2011-01-01

    T The majority of higher plants use sucrose as their main mobile carbohydrate. Proton-driven sucrose transporters play a crucial role in cell-to-cell and long-distance distribution of sucrose throughout the plant. A very negative plant membrane potential and the ability of sucrose transporters to accumulate sucrose concentrations of more than 1 M indicate that plants evolved transporters with unique structural and functional features. The knowledge about the transport mechanism and structural/functional domains of these nano-machines is, however, still fragmentary. In this review,the current knowledge about the biophysical properties of plant sucrose transporters is summarized and discussed.

  10. Sucrose regulated translational control of bZip genes in Arabidopsis thaliana

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    Rahmani, F.

    2007-01-01

    Sucrose can translationally regulate the expression of bZIP11 and four other S-class bZip transcription factors in Arabidopsis thaliana. Sequence encoding 28 amino acids (SC-peptide) in the leader of the bZIP11 is sufficient to mediate sucrose induced translational control. A model proposes that suc

  11. Intracellular sucrose communicates metabolic demand to sucrose transporters in developing pea cotyledons.

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    Zhou, Yuchan; Chan, Katie; Wang, Trevor L; Hedley, Cliff L; Offler, Christina E; Patrick, John W

    2009-01-01

    Mechanistic inter-relationships in sinks between sucrose compartmentation/metabolism and phloem unloading/translocation are poorly understood. Developing grain legume seeds provide tractable experimental systems to explore this question. Metabolic demand by cotyledons is communicated to phloem unloading and ultimately import by sucrose withdrawal from the seed apoplasmic space via a turgor-homeostat mechanism. What is unknown is how metabolic demand is communicated to cotyledon sucrose transporters responsible for withdrawing sucrose from the apoplasmic space. This question was explored here using a pea rugosus mutant (rrRbRb) compromised in starch biosynthesis compared with its wild-type counterpart (RRRbRb). Sucrose influx into cotyledons was found to account for 90% of developmental variations in their absolute growth and hence starch biosynthetic rates. Furthermore, rr and RR cotyledons shared identical response surfaces, indicating that control of transporter activity was likely to be similar for both lines. In this context, sucrose influx was correlated positively with expression of a sucrose/H(+) symporter (PsSUT1) and negatively with two sucrose facilitators (PsSUF1 and PsSUF4). Sucrose influx exhibited a negative curvilinear relationship with cotyledon concentrations of sucrose and hexoses. In contrast, the impact of intracellular sugars on transporter expression was transporter dependent, with expression of PsSUT1 inhibited, PsSUF1 unaffected, and PsSUF4 enhanced by sugars. Sugar supply to, and sugar concentrations of, RR cotyledons were manipulated using in vitro pod and cotyledon culture. Collectively the results obtained showed that intracellular sucrose was the physiologically active sugar signal that communicated metabolic demand to sucrose influx and this transport function was primarily determined by PsSUT1 regulated at the transcriptional level.

  12. Overexpression of sucrose transporter gene PbSUT2 from Pyrus bretschneideri, enhances sucrose content in Solanum lycopersicum fruit.

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    Wang, Li-Fen; Qi, Xiao-Xiao; Huang, Xiao-San; Xu, Lin-Lin; Jin, Cong; Wu, Jun; Zhang, Shao-Ling

    2016-08-01

    Sucrose transporters (SUTs) belong to the major facilitator superfamily. The function of SUTs has been intensively investigated in some higher plants, whereas that in pear fruit is unknown. In this study, the cloning and functional characterization of a sucrose transporter, PbSUT2, in pear (Pyrus bretschneideri Rehd. cv. 'Yali') fruits are reported. PbSUT2 encoded a protein of 498 amino acid residues, and was localized in the plasma membrane of transformed onion epidermal cells and Arabidopsis protoplasts. Phylogenetic analysis revealed that PbSUT2 belonged to the SUT4 clade. The phenotype of overexpression of PbSUT2 tomato plants included early flowering, higher fruit quantity and lower plant height. Overexpression of PbSUT2 in transgenic tomato plants led to increases in the net photosynthetic rate in leaves and sucrose content in mature fruit compared with wild-type tomato plants, and a decrease in the contents of glucose, fructose and total soluble sugars in mature fruits. These results suggested that PbSUT2 affected sucrose content in sinks and the flowering phase during tomato plant growth and development. PMID:27105422

  13. Aspects of sucrose transport in stem parenchyma of sweet sorghum

    International Nuclear Information System (INIS)

    Sweet sorghum [Sorghum bicolor (L.) Moench] is a sucrose-storing crop with a storage tissue anatomically similar to that of sugarcane (Saccharum spp.). However, recent evidence suggests that sweet sorghum may be biochemically different from sugarcane. 14C-sucrose uptake was studied in excised tissue discs from fully-elongated internodes of Rio sweet sorghum. Washout studies gave results consistent with a 3 compartment system. After 3 hours of uptake, most of the 14C was found in the vacuole compartment, and was determined by HPLC to be sucrose. Total sucrose uptake consisted of a PCMBS-sensitive (active) and a PCMBS-insensitive (passive) component. Active sucrose uptake had a pH optimum of 4.5. Total sucrose uptake was negatively correlated with the internal sucrose content of the tissue. Fructosyl-labelled 14C-sucrose was not randomized during uptake, suggesting that sucrose cleavage is not a requirement for sucrose uptake in sweet sorghum. This data suggests that in sweet sorghum, sucrose is transported intact by a specific carrier, as opposed to the sucrose-cleavage-and-resynthesis transport system that apparently operates in sugarcane

  14. Sucrose Production Mediated by Lipid Metabolism Suppresses the Physical Interaction of Peroxisomes and Oil Bodies during Germination of Arabidopsis thaliana.

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    Cui, Songkui; Hayashi, Yasuko; Otomo, Masayoshi; Mano, Shoji; Oikawa, Kazusato; Hayashi, Makoto; Nishimura, Mikio

    2016-09-16

    Physical interaction between organelles is a flexible event and essential for cells to adapt rapidly to environmental stimuli. Germinating plants utilize oil bodies and peroxisomes to mobilize storage lipids for the generation of sucrose as the main energy source. Although membrane interaction between oil bodies and peroxisomes has been widely observed, its underlying molecular mechanism is largely unknown. Here we present genetic evidence for control of the physical interaction between oil bodies and peroxisomes. We identified alleles of the sdp1 mutant altered in oil body morphology. This mutant accumulates bigger and more oil body aggregates compared with the wild type and showed defects in lipid mobilization during germination. SUGAR DEPENDENT 1 (SDP1) encodes major triacylglycerol lipase in Arabidopsis Interestingly, sdp1 seedlings show enhanced physical interaction between oil bodies and peroxisomes compared with the wild type, whereas exogenous sucrose supplementation greatly suppresses the interaction. The same phenomenon occurs in the peroxisomal defective 1 (ped1) mutant, defective in lipid mobilization because of impaired peroxisomal β-oxidation, indicating that sucrose production is a key factor for oil body-peroxisomal dissociation. Peroxisomal dissociation and subsequent release from oil bodies is dependent on actin filaments. We also show that a peroxisomal ATP binding cassette transporter, PED3, is the potential anchor protein to the membranes of these organelles. Our results provide novel components linking lipid metabolism and oil body-peroxisome interaction whereby sucrose may act as a negative signal for the interaction of oil bodies and peroxisomes to fine-tune lipolysis. PMID:27466365

  15. Long-distance phloem transport of glucosinolates in Arabidopsis.

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    Chen, S; Petersen, B L; Olsen, C E; Schulz, A; Halkier, B A

    2001-09-01

    Glucosinolates are a large group of plant secondary metabolites found mainly in the order Capparales, which includes a large number of economically important Brassica crops and the model plant Arabidopsis. In the present study, several lines of evidence are provided for phloem transport of glucosinolates in Arabidopsis. When radiolabeled p-hydroxybenzylglucosinolate (p-OHBG) and sucrose were co-applied to the tip of detached leaves, both tracers were collected in the phloem exudates at the petioles. Long-distance transport of [(14)C]p-OHBG was investigated in wild-type and transgenic 35S::CYP79A1 plants, synthesizing high amounts of p-OHBG, which is not a natural constituent of wild-type Arabidopsis. In both wild-type and 35S::CYP79A1 plants, radiolabeled p-OHBG was rapidly transported from the application site into the whole plant and intact p-OHBG was recovered from different tissues. The pattern of distribution of the radioactivity corresponded to that expected for transport of photoassimilates such as sucrose, and was consistent with translocation in phloem following the source-sink relationship. Radiolabeled p-OHBG was shown to accumulate in the seeds of wild-type and 35S::CYP79A1 plants, where p-OHBG had been either exogenously applied or endogenously synthesized from Tyr in the leaves. p-OHBG was found in phloem exudates collected from cut petioles of leaves from both wild-type and 35S::CYP79A1 plants. Phloem exudates were shown to contain intact glucosinolates, and not desulphoglucosinolates, as the transport form. It is concluded that intact glucosinolates are readily loaded into and transported by the phloem. PMID:11553747

  16. Effects of elevated carbon dioxide and sucrose concentrations on Arabidopsis thaliana root architecture and anatomy

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    Lee-Ho, E.; Walton, L.J.; Reid, D.M.; Yeung, E.C.; Kurepin, L.V. [Calgary Univ., AB (Canada). Dept. of Biology

    2007-03-15

    Plant root growth is known to be influenced by higher levels of atmospheric carbon dioxide (CO{sub 2}). Roots of some species grown in hydroponics under elevated CO{sub 2} concentrations may be more competitive sinks for photosynthetic assimilates than roots grown under lower CO{sub 2} conditions. Root branching patterns may also be influenced by elevated CO{sub 2} concentrations. Studies have also shown that factors such as soil compaction, salinity and the availability of nitrate, phosphorous, oxygen and water also influence root growth, and the effects of higher CO{sub 2} on roots can be confounded by such environmental factors. This study evaluated the effects of elevated carbon dioxide and sucrose concentrations on Arabidopsis thaliana root growth, morphology, and architecture. Both ambient and elevated CO{sub 2} levels were used along with various sucrose concentrations. The study revealed that A. thaliana plants grown on a phytagar medium in small chambers with elevated CO{sub 2} had longer roots, more lateral root growth than plants grown in ambient CO{sub 2}. Roots in elevated CO{sub 2} were found to have wider root diameters, and more secondary growth. The addition of sucrose to the media closely resembled the effects of elevated CO{sub 2}. In addition, the increase in sucrose concentration had a bigger effect on root morphology under ambient, than elevated CO{sub 2}. Therefore, both elevated CO{sub 2} and increased sucrose concentrations promote root growth by increasing their number, length, and diameter. The dichotomy branching index (DBI) also dropped resulting in a more dichotomous branching pattern. 34 refs., 5 figs.

  17. Characteristics of Sucrose Transport through the Sucrose-Specific Porin ScrY Studied by Molecular Dynamics Simulations

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    Liping eSun

    2016-02-01

    Full Text Available Sucrose-specific porin (ScrY is a transmembrane protein that allows for the uptake of sucrose under growth-limiting conditions. The crystal structure of ScrY was resolved before by X-ray crystallography, both in its uncomplexed form and with bound sucrose. However, little is known about the molecular characteristics of the transport mechanism of ScrY. To date, there has not yet been any clear demonstration for sucrose transport through the ScrY.Here, the dynamics of the ScrY trimer embedded in a phospholipid bilayer as well as the characteristics of sucrose translocation were investigated by means of atomistic molecular dynamics (MD simulations. The potential of mean force (PMF for sucrose translocation through the pore showed two main energy barriers within the constriction region of ScrY. Energy decomposition allowed to pinpoint three aspartic acids as key residues opposing the passage of sucrose, all located within the L3 loop. Mutation of two aspartic acids to uncharged residues resulted in an accordingly modified electrostatics and decreased PMF barrier. The chosen methodology and results will aid in the design of porins with modified transport specificities.

  18. Characteristics of Sucrose Transport through the Sucrose-Specific Porin ScrY Studied by Molecular Dynamics Simulations.

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    Sun, Liping; Bertelshofer, Franziska; Greiner, Günther; Böckmann, Rainer A

    2016-01-01

    Sucrose-specific porin (ScrY) is a transmembrane protein that allows for the uptake of sucrose under growth-limiting conditions. The crystal structure of ScrY was resolved before by X-ray crystallography, both in its uncomplexed form and with bound sucrose. However, little is known about the molecular characteristics of the transport mechanism of ScrY. To date, there has not yet been any clear demonstration for sucrose transport through the ScrY. Here, the dynamics of the ScrY trimer embedded in a phospholipid bilayer as well as the characteristics of sucrose translocation were investigated by means of atomistic molecular dynamics (MD) simulations. The potential of mean force (PMF) for sucrose translocation through the pore showed two main energy barriers within the constriction region of ScrY. Energy decomposition allowed to pinpoint three aspartic acids as key residues opposing the passage of sucrose, all located within the L3 loop. Mutation of two aspartic acids to uncharged residues resulted in an accordingly modified electrostatics and decreased PMF barrier. The chosen methodology and results will aid in the design of porins with modified transport specificities. PMID:26913282

  19. Routes to the tonoplast: the sorting of tonoplast transporters in Arabidopsis mesophyll protoplasts.

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    Wolfenstetter, Susanne; Wirsching, Petra; Dotzauer, Dorina; Schneider, Sabine; Sauer, Norbert

    2012-01-01

    Vacuoles perform a multitude of functions in plant cells, including the storage of amino acids and sugars. Tonoplast-localized transporters catalyze the import and release of these molecules. The mechanisms determining the targeting of these transporters to the tonoplast are largely unknown. Using the paralogous Arabidopsis thaliana inositol transporters INT1 (tonoplast) and INT4 (plasma membrane), we performed domain swapping and mutational analyses and identified a C-terminal di-leucine motif responsible for the sorting of higher plant INT1-type transporters to the tonoplast in Arabidopsis mesophyll protoplasts. We demonstrate that this motif can reroute other proteins, such as INT4, SUCROSE TRANSPORTER2 (SUC2), or SWEET1, to the tonoplast and that the position of the motif relative to the transmembrane helix is critical. Rerouted INT4 is functionally active in the tonoplast and complements the growth phenotype of an int1 mutant. In Arabidopsis plants defective in the β-subunit of the AP-3 adaptor complex, INT1 is correctly localized to the tonoplast, while sorting of the vacuolar sucrose transporter SUC4 is blocked in cis-Golgi stacks. Moreover, we demonstrate that both INT1 and SUC4 trafficking to the tonoplast is sensitive to brefeldin A. Our data show that plants possess at least two different Golgi-dependent targeting mechanisms for newly synthesized transporters to the tonoplast.

  20. Arg188 in rice sucrose transporter OsSUT1 is crucial for substrate transport

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    Sun Ye

    2012-11-01

    Full Text Available Abstract Background Plant sucrose uptake transporters (SUTs are H+/sucrose symporters related to the major facilitator superfamily (MFS. SUTs are essential for plant growth but little is known about their transport mechanism. Recent work identified several conserved, charged amino acids within transmembrane spans (TMS in SUTs that are essential for transport activity. Here we further evaluated the role of one of these positions, R188 in the fourth TMS of OsSUT1, a type II SUT. Results The OsSUT1(R188K mutant, studied by expression in plants, yeast, and Xenopus oocytes, did not transport sucrose but showed a H+ leak that was blocked by sucrose. The H+ leak was also blocked by β-phenyl glucoside which is not translocated by OsSUT1. Replacing the corresponding Arg in type I and type III SUTs, AtSUC1(R163K and LjSUT4(R169K, respectively, also resulted in loss of sucrose transport activity. Fluorination at the glucosyl 3 and 4 positions of α-phenyl glucoside greatly decreased transport by wild type OsSUT1 but did not affect the ability to block H+ leak in the R188K mutant. Conclusion OsSUT1 R188 appears to be essential for sucrose translocation but not for substrate interaction that blocks H+ leak. Therefore, we propose that an additional binding site functions in the initial recognition of substrates. The corresponding Arg in type I and III SUTs are equally important. We propose that R188 interacts with glucosyl 3-OH and 4-OH during translocation.

  1. Differential regulation of two sucrose transporters by defoliation and light conditions in perennial ryegrass.

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    Furet, Pierre-Maxime; Berthier, Alexandre; Decau, Marie-Laure; Morvan-Bertrand, Annette; Prud'homme, Marie-Pascale; Noiraud-Romy, Nathalie; Meuriot, Frédéric

    2012-12-01

    Sucrose transport between source and sink tissues is supposed to be a key-step for an efficient regrowth of perennial rye-grass after defoliation and might be altered by light conditions. We assessed the effect of different light regimes (high vs low light applied before or after defoliation) on growth, fructans and sucrose mobilization, as well as on sucrose transporter expression during 14 days of regrowth. Our results reported that defoliation led to a mobilization of C reserves (first sucrose and then fructans), which was parallel to an induction of LpSUT1 sucrose transporter expression in source and sink tissues (i.e. leaf sheaths and elongating leaf bases, respectively) irrespective to light conditions. Light regime (high or low light) had little effects on regrowth and on C reserves mobilization during the first 48 h of regrowth after defoliation. Thereafter, low light conditions, delaying the recovery of photosynthetic capacities, had a negative effect on C reserves re-accumulation (especially sucrose). Surprisingly, high light did not enhance sucrose transporter expression. Indeed, while light conditions had no effect on LpSUT1 expression, LpSUT2 transcripts levels were enhanced for low light grown plants. These results indicate that two sucrose transporter currently identified in Lolium perenne L. are differentially regulated by light and sucrose.

  2. Membrane-Transport Systems for Sucrose in Relation to Whole-Plant Carbon Partitioning

    Institute of Scientific and Technical Information of China (English)

    Brian G. Ayre

    2011-01-01

    T Sucrose is the principal product of photosynthesis used for the distribution of assimilated carbon in plants. Transport mechanisms and efficiency influence photosynthetic productivity by relieving product inhibition and contribute to plant vigor by controlling source/sink relationships and biomass partitioning. Sucrose is synthesized in the cytoplasm and may move cell to cell through plasmodesmata or may cross membranes to be compartmentalized or exported to the apoplasm for uptake into adjacent cells. As a relatively large polar compound, sucrose requires proteins to facilitate efficient membrane transport. Transport across the tonoplast by facilitated diffusion, antiport with protons, and symport with protons have been proposed; for transport across plasma membranes, symport with protons and a mechanism resembling facilitated diffusion are evident. Despite decades of research, only symport with protons is well established at the molecular level. This review aims to integrate recent and older studies on sucrose flux across membranes with principles of whole-plant carbon partitioning.

  3. Mass spectrometry-based method to investigate the natural selectivity of sucrose as the sugar transport form for plants.

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    Yuan, Hang; Wu, Yile; Liu, Wu; Liu, Yan; Gao, Xiang; Lin, Jinming; Zhao, Yufen

    2015-04-30

    Sucrose is the carbon skeletons and energy vector for plants, which is important for plants growth. Among thousands of disaccharides in Nature, why chose sucrose for plants? In this paper, we analyzed the intrinsic structural characteristics of four sucrose isomers with different glycosidic linkage by mass spectrometry (MS) technique. Our results show that sucrose has the most labile glycosidic bond compared with other three isomers, which is helpful for releasing glucose and fructose unit. Besides, sucrose has the most stable integral structure, which is hard to dehydrate and degrade into fragments through losing one or three even four-carbon units, just as its three isomers. In other words, sucrose is more easily holds an integral structure during the transport process, whenever it is necessary, and sucrose can be cleaved into glucose and fructose easily. Besides, we also investigate the internal relationship of sucrose with K(+) by tandem mass spectrometry and viscosity measurement. The related results have shown that the K(+) can stabilize sucrose to a greater extent than the Na(+). Furthermore, under the same conditions, K(+) ions reduce the viscosity of sucrose-water system much more than Na(+). These results suggest that K(+) is a better co-transporter for sucrose. Of course, the transport of sucrose in plants is a very complicated process, which is involved in many proteins. This paper directly accounts for the basic structure feature of sucrose, and the results discovered could provide the novel insight for the answer why Nature chose sucrose for plants.

  4. Transport of sucrose, not hexose, in the phloem

    OpenAIRE

    Liu, David D.; Chao, Wesley M.; Turgeon, Robert

    2012-01-01

    Several lines of evidence indicate that glucose and fructose are essentially absent in mobile phloem sap. However, this paradigm has been called into question, especially but not entirely, with respect to species in the Ranunculaceae and Papaveraceae. In the experiments in question, phloem sap was obtained by detaching leaves and placing the cut ends of the petioles in an EDTA solution. More hexose than sucrose was detected. In the present study, these results were confirmed for four species....

  5. Inverse pH regulation of plant and fungal sucrose transporters: a mechanism to regulate competition for sucrose at the host/pathogen interface?

    Directory of Open Access Journals (Sweden)

    Kathrin Wippel

    Full Text Available BACKGROUND: Plant sucrose transporter activities were shown to respond to changes in the extracellular pH and redox status, and oxidizing compounds like glutathione (GSSG or H(2O(2 were reported to effect the subcellular targeting of these proteins. We hypothesized that changes in both parameters might be used to modulate the activities of competing sucrose transporters at a plant/pathogen interface. We, therefore, compared the effects of redox-active compounds and of extracellular pH on the sucrose transporters UmSRT1 and ZmSUT1 known to compete for extracellular sucrose in the Ustilago maydis (corn smut/Zea mays (maize pathosystem. METHODOLOGY/PRINCIPAL FINDINGS: We present functional analyses of the U. maydis sucrose transporter UmSRT1 and of the plant sucrose transporters ZmSUT1 and StSUT1 in Saccharomyces cerevisiae or in Xenopus laevis oocytes in the presence of different extracellular pH-values and redox systems, and study the possible effects of these treatments on the subcellular targeting. We observed an inverse regulation of host and pathogen sucrose transporters by changes in the apoplastic pH. Under none of the conditions analyzed, we could confirm the reported effects of redox-active compounds. CONCLUSIONS/SIGNIFICANCE: Our data suggest that changes in the extracellular pH but not of the extracellular redox status might be used to oppositely adjust the transport activities of plant and fungal sucrose transporters at the host/pathogen interface.

  6. Proton-associated sucrose transport of mammalian solute carrier family 45: an analysis in Saccharomyces cerevisiae.

    Science.gov (United States)

    Bartölke, Rabea; Heinisch, Jürgen J; Wieczorek, Helmut; Vitavska, Olga

    2014-12-01

    The members of the solute carrier 45 (SLC45) family have been implicated in the regulation of glucose homoeostasis in the brain (SLC45A1), with skin and hair pigmentation (SLC45A2), and with prostate cancer and myelination (SLC45A3). However, apart from SLC45A1, a proton-associated glucose transporter, the function of these proteins is still largely unknown, although sequence similarities to plant sucrose transporters mark them as a putative sucrose transporter family. Heterologous expression of the three members SLC45A2, SLC45A3 and SLC45A4 in Saccharomyces cerevisiae confirmed that they are indeed sucrose transporters. [(14)C]Sucrose-uptake measurements revealed intermediate transport affinities with Km values of approximately 5 mM. Transport activities were best under slightly acidic conditions and were inhibited by the protonophore carbonyl cyanide m-chlorophenylhydrazone, demonstrating an H(+)-coupled transport mechanism. Na(+), on the other hand, had no effect on sucrose transport. Competitive inhibition assays indicated a possible transport also of glucose and fructose. Real-time PCR of mouse tissues confirmed mRNA expression of SLC45A2 in eyes and skin and of SLC45A3 primarily in the prostate, but also in other tissues, whereas SLC45A4 showed a predominantly ubiquitous expression. Altogether the results provide new insights into the physiological significance of SLC45 family members and challenge existing concepts of mammalian sugar transport, as they (i) transport a disaccharide, and (ii) perform secondary active transport in a proton-dependent manner. PMID:25164149

  7. Sucrose transport and phloem unloading in peach fruit: potential role of two transporters localized in different cell types.

    Science.gov (United States)

    Zanon, Laura; Falchi, Rachele; Santi, Simonetta; Vizzotto, Giannina

    2015-06-01

    Several complex physiological processes, which include long-distance translocation in the phloem and unloading in sink tissues, govern the partitioning of sugars in economically important organs, such as peach fruit. In this study, we took advantage of a symplastic tracer, carboxyfluorescein (CF), providing evidence for an apoplastic sucrose transfer in the early (SI) and middle (SIII) phases of peach fruit development. Moreover, using a combination of in situ hybridization and laser microdissection-assisted expression analysis, three putative sucrose transporters encoding genes (PpSUT1, PpSUT2, PpSUT4) were transcriptionally analyzed to relate their expression with sucrose storage in this organ. Our study revealed that PpSUT2 and PpSUT4 are the genes predominantly expressed in fruit flesh, and the detailed analysis of their expression pattern in the different cell types enabled us to suggest a specialized role in sucrose distribution. Both PpSUTs transporters could be involved in the retrieval of sucrose lost from the symplastic continuum of the phloem and, when expressed in parenchyma cells, they could be active in the import of sucrose into sink tissues, via symport from the apoplast. An alternative hypothesis has been proposed and discussed for PpSUT4 because of its putative tonoplastic localization. Taken together, our results provide new insights into the molecular mechanisms underpinning sucrose unloading and accumulation in peach fruit.

  8. Sucrose transport and phloem unloading in peach fruit: potential role of two transporters localized in different cell types.

    Science.gov (United States)

    Zanon, Laura; Falchi, Rachele; Santi, Simonetta; Vizzotto, Giannina

    2015-06-01

    Several complex physiological processes, which include long-distance translocation in the phloem and unloading in sink tissues, govern the partitioning of sugars in economically important organs, such as peach fruit. In this study, we took advantage of a symplastic tracer, carboxyfluorescein (CF), providing evidence for an apoplastic sucrose transfer in the early (SI) and middle (SIII) phases of peach fruit development. Moreover, using a combination of in situ hybridization and laser microdissection-assisted expression analysis, three putative sucrose transporters encoding genes (PpSUT1, PpSUT2, PpSUT4) were transcriptionally analyzed to relate their expression with sucrose storage in this organ. Our study revealed that PpSUT2 and PpSUT4 are the genes predominantly expressed in fruit flesh, and the detailed analysis of their expression pattern in the different cell types enabled us to suggest a specialized role in sucrose distribution. Both PpSUTs transporters could be involved in the retrieval of sucrose lost from the symplastic continuum of the phloem and, when expressed in parenchyma cells, they could be active in the import of sucrose into sink tissues, via symport from the apoplast. An alternative hypothesis has been proposed and discussed for PpSUT4 because of its putative tonoplastic localization. Taken together, our results provide new insights into the molecular mechanisms underpinning sucrose unloading and accumulation in peach fruit. PMID:25348206

  9. Arabidopsis POLYOL TRANSPORTER5, a new member of the monosaccharide transporter-like superfamily, mediates H+-Symport of numerous substrates, including myo-inositol, glycerol, and ribose.

    Science.gov (United States)

    Klepek, Yvonne-Simone; Geiger, Dietmar; Stadler, Ruth; Klebl, Franz; Landouar-Arsivaud, Lucie; Lemoine, Rémi; Hedrich, Rainer; Sauer, Norbert

    2005-01-01

    Six genes of the Arabidopsis thaliana monosaccharide transporter-like (MST-like) superfamily share significant homology with polyol transporter genes previously identified in plants translocating polyols (mannitol or sorbitol) in their phloem (celery [Apium graveolens], common plantain [Plantago major], or sour cherry [Prunus cerasus]). The physiological role and the functional properties of this group of proteins were unclear in Arabidopsis, which translocates sucrose and small amounts of raffinose rather than polyols. Here, we describe POLYOL TRANSPORTER5 (AtPLT5), the first member of this subgroup of Arabidopsis MST-like transporters. Transient expression of an AtPLT5-green fluorescent protein fusion in plant cells and functional analyses of the AtPLT5 protein in yeast and Xenopus oocytes demonstrate that AtPLT5 is located in the plasma membrane and characterize this protein as a broad-spectrum H+-symporter for linear polyols, such as sorbitol, xylitol, erythritol, or glycerol. Unexpectedly, however, AtPLT5 catalyzes also the transport of the cyclic polyol myo-inositol and of different hexoses and pentoses, including ribose, a sugar that is not transported by any of the previously characterized plant sugar transporters. RT-PCR analyses and AtPLT5 promoter-reporter gene plants revealed that AtPLT5 is most strongly expressed in Arabidopsis roots, but also in the vascular tissue of leaves and in specific floral organs. The potential physiological role of AtPLT5 is discussed. PMID:15598803

  10. The Arabidopsis NPF3 protein is a GA transporter

    DEFF Research Database (Denmark)

    Tal, Iris; Zhang, Yi; Jørgensen, Morten Egevang;

    2016-01-01

    Gibberellins (GAs) are plant hormones that promote a wide range of developmental processes. While GA signalling is well understood, little is known about how GA is transported or how GA distribution is regulated. Here we utilize fluorescently labelled GAs (GA-Fl) to screen for Arabidopsis mutants...... deficient in GA transport. We show that the NPF3 transporter efficiently transports GA across cell membranes in vitro and GA-Fl in vivo. NPF3 is expressed in root endodermis and repressed by GA. NPF3 is targeted to the plasma membrane and subject to rapid BFA-dependent recycling. We show that abscisic acid...... (ABA), an antagonist of GA, is also transported by NPF3 in vitro. ABA promotes NPF3 expression and GA-Fl uptake in plants. On the basis of these results, we propose that GA distribution and activity in Arabidopsis is partly regulated by NPF3 acting as an influx carrier and that GA-ABA interaction may...

  11. Transcriptional coordination and abscisic acid mediated regulation of sucrose transport and sucrose-to-starch metabolism related genes during grain filling in wheat (Triticum aestivum L.).

    Science.gov (United States)

    Mukherjee, Shalini; Liu, Aihua; Deol, Kirandeep K; Kulichikhin, Konstanin; Stasolla, Claudio; Brûlé-Babel, Anita; Ayele, Belay T

    2015-11-01

    Combining physiological, molecular and biochemical approaches, this study investigated the transcriptional coordination and abscisic acid (ABA) mediated regulation of genes involved in sucrose import and its conversion to starch during grain filling in wheat. Sucrose import appears to be mediated by seed localized TaSUT1, mainly TaSUT1D, while sucrose cleavage by TaSuSy2. Temporal overlapping of the transcriptional activation of AGPL1 and AGPS1a that encode AGPase with that of the above genes suggests their significance in the synthesis of ADP-glucose; TaAGPL1A and TaAGPL1D contributing the majority of AGPL1 transcripts. ABA induced repressions of TaSUT1, TaSuSy2, TaAGPL1 and TaAGPS1a imply that ABA negatively regulates sucrose import into the endosperm and its subsequent metabolism to ADP-glucose, the substrate for starch synthesis. The formations of amyloses and amylopectin from ADP-glucose appear to be mediated by specific members of GBSS, and SS, SBE and DBE gene families, and the ABA-induced transcriptional change in most of these genes implies that ABA regulates amylose and amylopectin synthesis. The findings provide insights into the molecular mechanisms underlying the coordination and ABA mediated regulation of sucrose transport into the developing endosperm and its subsequent metabolism to starch during grain filling in wheat.

  12. Identification of mitochondrial coenzyme a transporters from maize and Arabidopsis.

    Science.gov (United States)

    Zallot, Rémi; Agrimi, Gennaro; Lerma-Ortiz, Claudia; Teresinski, Howard J; Frelin, Océane; Ellens, Kenneth W; Castegna, Alessandra; Russo, Annamaria; de Crécy-Lagard, Valérie; Mullen, Robert T; Palmieri, Ferdinando; Hanson, Andrew D

    2013-06-01

    Plants make coenzyme A (CoA) in the cytoplasm but use it for reactions in mitochondria, chloroplasts, and peroxisomes, implying that these organelles have CoA transporters. A plant peroxisomal CoA transporter is already known, but plant mitochondrial or chloroplastic CoA transporters are not. Mitochondrial CoA transporters belonging to the mitochondrial carrier family, however, have been identified in yeast (Saccharomyces cerevisiae; Leu-5p) and mammals (SLC25A42). Comparative genomic analysis indicated that angiosperms have two distinct homologs of these mitochondrial CoA transporters, whereas nonflowering plants have only one. The homologs from maize (Zea mays; GRMZM2G161299 and GRMZM2G420119) and Arabidopsis (Arabidopsis thaliana; At1g14560 and At4g26180) all complemented the growth defect of the yeast leu5Δ mitochondrial CoA carrier mutant and substantially restored its mitochondrial CoA level, confirming that these proteins have CoA transport activity. Dual-import assays with purified pea (Pisum sativum) mitochondria and chloroplasts, and subcellular localization of green fluorescent protein fusions in transiently transformed tobacco (Nicotiana tabacum) Bright Yellow-2 cells, showed that the maize and Arabidopsis proteins are targeted to mitochondria. Consistent with the ubiquitous importance of CoA, the maize and Arabidopsis mitochondrial CoA transporter genes are expressed at similar levels throughout the plant. These data show that representatives of both monocotyledons and eudicotyledons have twin, mitochondrially located mitochondrial carrier family carriers for CoA. The highly conserved nature of these carriers makes possible their reliable annotation in other angiosperm genomes. PMID:23590975

  13. Characterization, localization, and seasonal changes of the sucrose transporter FeSUT1 in the phloem of Fraxinus excelsior.

    Science.gov (United States)

    Öner-Sieben, Soner; Rappl, Christine; Sauer, Norbert; Stadler, Ruth; Lohaus, Gertrud

    2015-08-01

    Trees are generally assumed to be symplastic phloem loaders. A typical feature for most wooden species is an open minor vein structure with symplastic connections between mesophyll cells and phloem cells, which allow sucrose to move cell-to-cell through the plasmodesmata into the phloem. Fraxinus excelsior (Oleaceae) also translocates raffinose family oligosaccharides in addition to sucrose. Sucrose concentration was recently shown to be higher in the phloem sap than in the mesophyll cells. This suggests the involvement of apoplastic steps and the activity of sucrose transporters in addition to symplastic phloem-loading processes. In this study, the sucrose transporter FeSUT1 from F. excelsior was analysed. Heterologous expression in baker's yeast showed that FeSUT1 mediates the uptake of sucrose. Immunohistochemical analyses revealed that FeSUT1 was exclusively located in phloem cells of minor veins and in the transport phloem of F. excelsior. Further characterization identified these cells as sieve elements and possibly ordinary companion cells but not as intermediary cells. The localization and expression pattern point towards functions of FeSUT1 in phloem loading of sucrose as well as in sucrose retrieval. FeSUT1 is most likely responsible for the observed sucrose gradient between mesophyll and phloem. The elevated expression level of FeSUT1 indicated an increased apoplastic carbon export activity from the leaves during spring and late autumn. It is hypothesized that the importance of apoplastic loading is high under low-sucrose conditions and that the availability of two different phloem-loading mechanisms confers advantages for temperate woody species like F. excelsior. PMID:26022258

  14. Characterization, localization, and seasonal changes of the sucrose transporter FeSUT1 in the phloem of Fraxinus excelsior.

    Science.gov (United States)

    Öner-Sieben, Soner; Rappl, Christine; Sauer, Norbert; Stadler, Ruth; Lohaus, Gertrud

    2015-08-01

    Trees are generally assumed to be symplastic phloem loaders. A typical feature for most wooden species is an open minor vein structure with symplastic connections between mesophyll cells and phloem cells, which allow sucrose to move cell-to-cell through the plasmodesmata into the phloem. Fraxinus excelsior (Oleaceae) also translocates raffinose family oligosaccharides in addition to sucrose. Sucrose concentration was recently shown to be higher in the phloem sap than in the mesophyll cells. This suggests the involvement of apoplastic steps and the activity of sucrose transporters in addition to symplastic phloem-loading processes. In this study, the sucrose transporter FeSUT1 from F. excelsior was analysed. Heterologous expression in baker's yeast showed that FeSUT1 mediates the uptake of sucrose. Immunohistochemical analyses revealed that FeSUT1 was exclusively located in phloem cells of minor veins and in the transport phloem of F. excelsior. Further characterization identified these cells as sieve elements and possibly ordinary companion cells but not as intermediary cells. The localization and expression pattern point towards functions of FeSUT1 in phloem loading of sucrose as well as in sucrose retrieval. FeSUT1 is most likely responsible for the observed sucrose gradient between mesophyll and phloem. The elevated expression level of FeSUT1 indicated an increased apoplastic carbon export activity from the leaves during spring and late autumn. It is hypothesized that the importance of apoplastic loading is high under low-sucrose conditions and that the availability of two different phloem-loading mechanisms confers advantages for temperate woody species like F. excelsior.

  15. Characterization of multiple SPS knockout mutants reveals redundant functions of the four Arabidopsis sucrose phosphate synthase isoforms in plant viability, and strongly indicates that enhanced respiration and accelerated starch turnover can alleviate the blockage of sucrose biosynthesis.

    Science.gov (United States)

    Bahaji, Abdellatif; Baroja-Fernández, Edurne; Ricarte-Bermejo, Adriana; Sánchez-López, Ángela María; Muñoz, Francisco José; Romero, Jose M; Ruiz, María Teresa; Baslam, Marouane; Almagro, Goizeder; Sesma, María Teresa; Pozueta-Romero, Javier

    2015-09-01

    We characterized multiple knock-out mutants of the four Arabidopsis sucrose phosphate synthase (SPSA1, SPSA2, SPSB and SPSC) isoforms. Despite their reduced SPS activity, spsa1/spsa2, spsa1/spsb, spsa2/spsb, spsa2/spsc, spsb/spsc, spsa1/spsa2/spsb and spsa2/spsb/spsc mutants displayed wild type (WT) vegetative and reproductive morphology, and showed WT photosynthetic capacity and respiration. In contrast, growth of rosettes, flowers and siliques of the spsa1/spsc and spsa1/spsa2/spsc mutants was reduced compared with WT plants. Furthermore, these plants displayed a high dark respiration phenotype. spsa1/spsb/spsc and spsa1/spsa2/spsb/spsc seeds poorly germinated and produced aberrant and sterile plants. Leaves of all viable sps mutants, except spsa1/spsc and spsa1/spsa2/spsc, accumulated WT levels of nonstructural carbohydrates. spsa1/spsc leaves possessed high levels of metabolic intermediates and activities of enzymes of the glycolytic and tricarboxylic acid cycle pathways, and accumulated high levels of metabolic intermediates of the nocturnal starch-to-sucrose conversion process, even under continuous light conditions. Results presented in this work show that SPS is essential for plant viability, reveal redundant functions of the four SPS isoforms in processes that are important for plant growth and nonstructural carbohydrate metabolism, and strongly indicate that accelerated starch turnover and enhanced respiration can alleviate the blockage of sucrose biosynthesis in spsa1/spsc leaves.

  16. A vacuolar phosphate transporter essential for phosphate homeostasis in Arabidopsis

    OpenAIRE

    Liu, Jinlong; Yang, Lei; Luan, Mingda; Wang, Yuan; Zhang, Chi; Zhang, Bin; Shi, Jisen; Zhao, Fu-Geng; Lan, Wenzhi; Luan, Sheng

    2015-01-01

    Phosphate is an essential nutrient for plant growth, and inorganic phosphate (Pi) is stored largely in the vacuole of plant cells. Thus, vacuolar Pi maintains homeostasis of cytosolic Pi to ensure an optimal Pi supply for plants under variable Pi status in the soil. This study uncovered in Arabidopsis a vacuolar phosphate transporter, VPT1, that mediates vacuolar Pi sequestration. Lack of VPT1 caused growth defects under both low-Pi and high-Pi conditions, implicating VPT1 in plant adaptation...

  17. The Arabidopsis NPF3 protein is a GA transporter.

    Science.gov (United States)

    Tal, Iris; Zhang, Yi; Jørgensen, Morten Egevang; Pisanty, Odelia; Barbosa, Inês C R; Zourelidou, Melina; Regnault, Thomas; Crocoll, Christoph; Olsen, Carl Erik; Weinstain, Roy; Schwechheimer, Claus; Halkier, Barbara Ann; Nour-Eldin, Hussam Hassan; Estelle, Mark; Shani, Eilon

    2016-01-01

    Gibberellins (GAs) are plant hormones that promote a wide range of developmental processes. While GA signalling is well understood, little is known about how GA is transported or how GA distribution is regulated. Here we utilize fluorescently labelled GAs (GA-Fl) to screen for Arabidopsis mutants deficient in GA transport. We show that the NPF3 transporter efficiently transports GA across cell membranes in vitro and GA-Fl in vivo. NPF3 is expressed in root endodermis and repressed by GA. NPF3 is targeted to the plasma membrane and subject to rapid BFA-dependent recycling. We show that abscisic acid (ABA), an antagonist of GA, is also transported by NPF3 in vitro. ABA promotes NPF3 expression and GA-Fl uptake in plants. On the basis of these results, we propose that GA distribution and activity in Arabidopsis is partly regulated by NPF3 acting as an influx carrier and that GA-ABA interaction may occur at the level of transport. PMID:27139299

  18. Molecular analysis of the scrA and scrB genes from Klebsiella pneumoniae and plasmid pUR400 which encode the sucrose transport protein Enzyme IIScr of the phosphotransferase system and a sucrose-6-phosphate invertase

    NARCIS (Netherlands)

    Titgemeyer, F; Jahreis, K; Ebner, R; Lengeler, JW

    1996-01-01

    The Klebsiella pneumoniae genes scrA and scrB are indispensable for sucrose (Scr) utilisation. Gene scrA codes for an Enzyme IIScr (IIScr) transport protein of the phosphoenolpyruvate-dependent carbohydrate: phosphotransferase system (PTS), while scrB encodes a sucrose 6-phosphate specific invertase

  19. Molecular cloning and expression analysis of a gene for sucrose transporter from pear (Pyrus bretschneideri Rehd.) fruit.

    Science.gov (United States)

    Zhang, Huping; Zhang, Shujun; Qin, Gaihua; Wang, Lifen; Wu, Tao; Qi, Kaijie; Zhang, Shaoling

    2013-12-01

    Here we report the cloning of a sucrose transporter cDNA from pear (Pyrus bretschneideri Rehd. cv 'Yali') fruit and an analysis of the expression of the gene. A cDNA clone, designated PbSUT1 was identified as a sucrose transporter cDNA from its sequence homology at the amino acid level to sucrose transporters that have been cloned from other higher plant species. PbSUT1 potentially encoded a protein of 499 amino acid residues with a predicted molecular mass of 53.4 kDa and an isoelectric point (pI) of 9.21. Phylogenetic analysis revealed that the PbSUT1 belonged to type III SUTs and was more closely related to the MdSUT1 from apple fruit. Some major facilitator superfamily (MFS)-specific sequence motifs were found in the predicted PbSUT1 peptides, and an MFS_1 domain was located at the amino acid positions of 29-447 of the sequence. A study of gene expression along fruit development showed that PbSUT1 transcripts are present at all stages but significantly increase before fruit enlargement and during the ripening process with increasing sucrose levels. In contrast, the expression levels don't change much during the period of rapid fruit growth. This work shows that sucrose transporter may play a role in the accumulation of sugars during maturation and in maintaining the internal cellular distribution.

  20. Bayesian phylogeny of sucrose transporters: Ancient origins, differential expansion and convergent evolution in monocots and dicots

    Directory of Open Access Journals (Sweden)

    Duo ePeng

    2014-11-01

    Full Text Available Sucrose transporters (SUTs are essential for the export and efficient movement of sucrose from source leaves to sink organs in plants. The angiosperm SUT family was previously classified into three or four distinct groups, Types I, II (subgroup IIB and III, with dicot-specific Type I and monocot-specific Type IIB functioning in phloem loading. To shed light on the underlying drivers of SUT evolution, Bayesian phylogenetic inference was undertaken using 41 sequenced plant genomes, including seven basal lineages at key evolutionary junctures. Our analysis supports four phylogenetically and structurally distinct SUT subfamilies, originating from two ancient groups (AG1 and AG2 that diverged early during terrestrial colonization. In both AG1 and AG2, multiple intron acquisition events in the progenitor vascular plant established the gene structures of modern SUTs. Tonoplastic Type III and plasmalemmal Type II represent evolutionarily conserved descendants of AG1 and AG2, respectively. Type I and Type IIB were previously thought to evolve after the dicot-monocot split. We show, however, that divergence of Type I from Type III SUT predated basal angiosperms, likely associated with evolution of vascular cambium and phloem transport. Type I SUT was subsequently lost in monocots along with vascular cambium, and independent evolution of Type IIB coincided with modified monocot vasculature. Both Type I and Type IIB underwent lineage-specific expansion. In multiple unrelated taxa, the newly-derived SUTs exhibit biased expression in reproductive tissues, suggesting a functional link between phloem loading and reproductive fitness. Convergent evolution of Type I and Type IIB for SUT function in phloem loading and reproductive organs supports the idea that differential vascular development in dicots and monocots is a strong driver for SUT family evolution in angiosperms.

  1. Decrease in Leaf Sucrose Synthesis Leads to Increased Leaf Starch Turnover and Decreased RuBP-limited Photosynthesis But Not Rubisco-limited Photosynthesis in Arabidopsis Null Mutants of SPSA1

    Science.gov (United States)

    SPS (Sucrose phosphate synthase) isoforms from dicots cluster into families A, B and C. In this study, we investigated the individual effect of null mutations of each of the four SPS genes in Arabidopsis (spsa1, spsa2, spsb and spsc) on photosynthesis and carbon partitioning. Null mutants spsa1 and ...

  2. The Arabidopsis nitrate transporter AtNRT2.1 is targeted to the root plasma membrane.

    Science.gov (United States)

    Chopin, Franck; Wirth, Judith; Dorbe, Marie-France; Lejay, Laurence; Krapp, Anne; Gojon, Alain; Daniel-Vedele, Françoise

    2007-08-01

    Arabidopsis AtNRT2.1 protein is the best characterized high affinity nitrate transporter in higher plants. However, nothing is known about its sub-cellular localization. In this work, we used GFP imaging to follow the targeting of the AtNRT2.1 protein to the different cell membranes. A polyclonal antibody was also raised against a peptide derived from the AtNRT2.1 sequence. Comparison of wild type and mutant plant extracts showed that this antibody recognized specifically the AtNRT2.1 protein. Microsomal membranes were fractionated on sucrose gradients and immunological detections were performed on the different fractions. Altogether, our results demonstrate that the AtNRT2.1 protein is located in the plasma membrane of the root cells.

  3. A universal algorithm for genome-wide in silicio identification of biologically significant gene promoter putative cis-regulatory-elements; identification of new elements for reactive oxygen species and sucrose signaling in Arabidopsis.

    Science.gov (United States)

    Geisler, Matt; Kleczkowski, Leszek A; Karpinski, Stanislaw

    2006-02-01

    Short motifs of many cis-regulatory elements (CREs) can be found in the promoters of most Arabidopsis genes, and this raises the question of how their presence can confer specific regulation. We developed a universal algorithm to test the biological significance of CREs by first identifying every Arabidopsis gene with a CRE and then statistically correlating the presence or absence of the element with the gene expression profile on multiple DNA microarrays. This algorithm was successfully verified for previously characterized abscisic acid, ethylene, sucrose and drought responsive CREs in Arabidopsis, showing that the presence of these elements indeed correlates with treatment-specific gene induction. Later, we used standard motif sampling methods to identify 128 putative motifs induced by excess light, reactive oxygen species and sucrose. Our algorithm was able to filter 20 out of 128 novel CREs which significantly correlated with gene induction by either heat, reactive oxygen species and/or sucrose. The position, orientation and sequence specificity of CREs was tested in silicio by analyzing the expression of genes with naturally occurring sequence variations. In three novel CREs the forward orientation correlated with sucrose induction and the reverse orientation with sucrose suppression. The functionality of the predicted novel CREs was experimentally confirmed using Arabidopsis cell-suspension cultures transformed with short promoter fragments or artificial promoters fused with the GUS reporter gene. Our genome-wide analysis opens up new possibilities for in silicio verification of the biological significance of newly discovered CREs, and allows for subsequent selection of such CREs for experimental studies.

  4. Anthocyanin Biosynthesis Regulated by Sucrose in Arabidopsis thaliana Seedling%蔗糖调节拟南芥花青素的生物合成

    Institute of Scientific and Technical Information of China (English)

    杨少华; 王丽; 穆春; 王翔; 何静辉; 赵静尧; 王林嵩

    2011-01-01

    为了探讨糖在花青素合成过程中的调节作用,采用蔗糖和其代谢糖(葡萄糖和果糖)组合处理拟南芥幼苗.实验结果表明,60 mmol/L蔗糖处理显著提高拟南芥幼苗的花青素、还原糖含量,并上调花青素合成相关基因(CHS,FLS-1,DFR,LDOX,BANYULS)的转录,对叶绿素含量和UGT78D2基因的转录无影响;20 mmol/L葡萄糖+20 mmol/L果糖处理,对花青素、叶绿素和还原糖的含量无影响,对花青素合成相关基因转录影响不一;20 mmol/L蔗糖+20 mmol/L葡萄糖+20mmol/L果糖处理后,花青素和还原糖含量介于前两个处理之间,也上调花青素合成相关基因的转录;但和蔗糖处理组相比,上调UG778D2基因转录,下调FLS-1基因转录.在不同处理组之间,花青素含量变化和还原糖含量变化趋势相同,有可能糖在调节花青素合成的同时也调节还原糖含量.因此,蔗糖既可以通过蔗糖特异信号途径,也可以和其代谢糖通过其他途径共同调节拟南芥花青素的生物合成.%In order to investigate the sugar regulation of anthocyanin biosynthesis, the combined effects of sucrose and its metabolic product of glucose and fructose were studied in Arabidopsis thaliana seedling.The results indicated that when cultured with 60 mmol/L sucrose, the contents of anthocyanin and reductive sugars were significantly increased, and the transcription genes in anthocyanin biosynthesis were upregulated, such as chalcone synthase (CHS) , flavonol synthase-1 (FLS-1) , dihydroflavonol reductase (DFR), leucoanthocyanidin dioxygenase (LDOX), anthocyanidin reductase (BANYULS).The chlorophyll content and UDP-Glc: flavonoid 3-O-glucosyltransferase (UGT78D2) expression remained unchanged.When grown at 1∶1 mixture of 20 mmol/L glucose + 20 mmol/L fructose, no changes of anthocyanin, chlorophyll and reductive sugars were observed, whereas the expression of anthocyanin biosynthetic genes varied.In case of the treatment with 1∶ 1∶ 1 mixture of

  5. A Mitochondrial Magnesium Transporter Functions in Arabidopsis Pollen Development

    Institute of Scientific and Technical Information of China (English)

    Le-Gong Li; Lubomir N.Sokolov; Yong-Hua Yang; Dong-Ping Li; Julie Ting; Girdhar K.Pandy; Sheng Luan

    2008-01-01

    Magnesium is an abundant divalent cation in plant cells and plays a critical role in many physiological processes.We have previously described the jdentification of a 10-member Arabidopsis gene family encoding putative magnesium transport(MGT)proteins.Here,we report that a member of the MGT family,AtMGT5, functions as a dual-functional Mg-transporter that operates in a concentration-dependent manner, namely it serves as a Mg-importer at micromolar levels and facilitates the efflux in the millimolar range.The AtMGT5 protein is localized in the mitochondria,suggesting that AtMGT5 mediates Mg-trafficking between the cytosol and mitochondria.The AtMGT5 gene was exclusively expressed in anthers at early stages of flower development.Examination of two independent T-DNA insertional mutants of AtMGT5 gene demonstrated that AtMG7-5 played an essential role for pollen development and male fertility.This study suggests a critical role for Mg2+ transport between cytosol and mitochondria in male gametogenesis in plants.

  6. Organ and Tissue-specific Sucrose Transporters. Important Hubs in Gene and Metabolite Networks Regulating Carbon Use in Wood-forming Tissues of Populus

    Energy Technology Data Exchange (ETDEWEB)

    Harding, Scott A. [Univ. of Georgia, Athens, GA (United States); Tsai, Chung-Jui [Univ. of Georgia, Athens, GA (United States)

    2016-01-04

    The overall project objective was to probe the relationship between sucrose transporters and plant productivity in the biomass for biofuels woody perennial, Populus. At the time the proposal was written, sucrose transporters had already been investigated in many plant model systems, primarily with respect to the export of photosynthate sucrose from source leaves, and the uptake of sucrose in storage organs and seeds. Preliminary findings by the PI found that in Populus, sucrose transporter genes (SUTs) were well expressed in wood-forming tissues that comprise the feedstock for biofuels production. Because sucrose comprises by far the predominant form in which photosynthate is delivered from source organs to sink organs like roots and wood-forming tissues, SUTs control a gate that nominally at least could impact the allocation or partitioning of sucrose for potentially competing end uses like growth (stem biomass) and storage. In addition, water use might be conditioned by the way in which sucrose is distributed throughout the plant, and/or by the way in which sucrose is partitioned intracellularly. Several dozen transgenic lines were produced in year 1 of the project to perturb the expression ratio of multiple plasma membrane (PM) SUTs (intercellular trafficking), versus the single tonoplast membrane (TM) sucrose transporter that effectively regulates intracellular trafficking of sucrose. It was possible to obtain transgenic lines with dual SUT gene knockdown using the 35S promoter, but not the wood-specific TUA1 promoter. By the end of project year 2, a decision was made to work with the 35S plants while archiving the TUA1 plants. The PhD candidate charged with producing the transgenic lines abandoned the project during its second year, substantially contributing to the decision to operate with just the 35S lines. That student’s interests ranged more toward evolutionary topics, and a report on SUT gene evolution was published (Peng et al 2014).

  7. Putative role of the H(+)/sucrose symporter SLC45A3 as an osmolyte transporter in the kidney.

    Science.gov (United States)

    Vitavska, Olga; Edemir, Bayram; Wieczorek, Helmut

    2016-08-01

    The solute carrier family 45 a3 member (SLC45A3), known also as prostein, has been implicated with prostate cancer and the regulation of lipid metabolism in oligodendrocytes. Recently, we expressed SLC45A3 in yeast cells and characterised it as a proton-coupled sucrose symporter. However, the physiological functions of SLC45A3 were still unknown. Here, we report that SLC45A3 occurs in the kidney and is highly expressed in the medullary collecting duct (IMCD), a part of the kidney responsible for final urine concentration and faced to hyperosmotic environment. Moreover, messenger RNA (mRNA) expression of endogenous SLC45A3 in rat IMCD cells as well as in NRK52E cells increased up to four-fold under hyperosmotic conditions at 600 mOsmol/kg. Using NRK52E cells as an experimental model, we investigated the proton-coupled sugar transport and found that the uptake of sucrose or glucose was enhanced by hyperosmolarity. Down-regulation of expression by small interfering RNA (siRNA) decreased the osmotically inducible part of sucrose uptake and confirmed the involvement of SLC45A3 in this process. Furthermore, we observed an up to four-fold elevation of sucrose uptake triggered by hyperosmolarity across the apical membrane of NRK52E cells, while uptake across the basolateral membrane was not affected. Due to this finding, we conclude that SLC45A3 may occur at the luminal side of kidney epithelial cells and thus may take up solutes from the tubular fluid. Altogether, we show that SLC45A3 is a novel sugar transporter in kidney and hypothesise that the disaccharide sucrose, and probably the monosaccharides glucose and fructose, may serve as compatible osmolytes in urine. PMID:27228996

  8. Interaction of brassinosteroid functions and sucrose transporter SlSUT2 regulate the formation of arbuscular mycorrhiza

    Science.gov (United States)

    Bitterlich, Michael; Krügel, Undine; Boldt-Burisch, Katja; Franken, Philipp; Kühn, Christina

    2014-01-01

    Transgenic tomato plants with reduced expression of the sucrose transporter SlSUT2 showed higher efficiency of mycorrhization suggesting a sucrose retrieval function of SlSUT2 from the peri-arbuscular space back into the cell cytoplasm plant cytoplasm thereby limiting mycorrhiza fungal development. Sucrose uptake in colonized root cells requires efficient plasma membrane-targeting of SlSUT2 which is often retained intracellularly in vacuolar vesicles. Protein-protein interaction studies suggested a link between SISUT2 function and components of brassinosteroid biosynthesis and signaling. Indeed, the tomato DWARF mutant dx defective in BR synthesis1 showed significantly reduced mycorrhization parameters.2 The question has been raised whether the impact of brassinosteroids on mycorrhization is a general phenomenon. Here, we include a rice mutant defective in DIM1/DWARF1 involved in BR biosynthesis to investigate the effects on mycorrhization. A model is presented where brassinolides are able to impact mycorrhization by activating SUT2 internalization and inhibiting its role in sucrose retrieval. PMID:25482803

  9. A subgroup of MATE transporter genes regulates hypocotyl cell elongation in Arabidopsis.

    Science.gov (United States)

    Wang, Rui; Liu, Xiayan; Liang, Shuang; Ge, Qing; Li, Yuanfeng; Shao, Jingxia; Qi, Yafei; An, Lijun; Yu, Fei

    2015-10-01

    The growth of higher plants is under complex regulation to ensure the elaboration of developmental programmes under a changing environment. To dissect these regulatory circuits, we carried out genetic screens for Arabidopsis abnormal shoot (abs) mutants with altered shoot development. Here, we report the isolation of two dominant mutants, abs3-1D and abs4-1D, through activation tagging. Both mutants showed a 'bushy' loss of apical dominance phenotype. ABS3 and ABS4 code for two closely related putative Multidrug and Toxic Compound Extrusion (MATE) family of efflux transporters, respectively. ABS3 and ABS4, as well as two related MATE genes, ABS3-Like1 (ABS3L1) and ABS3L2, showed diverse tissue expression profiles but their gene products all localized to the late endosome/prevacuole (LE/PVC) compartment. The over-expression of these four genes individually led to the inhibition of hypocotyl cell elongation in the light. On the other hand, the quadruple knockout mutant (mateq) showed the opposite phenotype of an enhanced hypocotyl cell elongation in the light. Hypocotyl cell elongation and de-etiolation processes in the dark were also affected by the mutations of these genes. Exogenously applied sucrose attenuated the inhibition of hypocotyl elongation caused by abs3-1D and abs4-1D in the dark, and enhanced the hypocotyl elongation of mateq under prolonged dark treatment. We determined that ABS3 genetically interacts with the photoreceptor gene PHYTOCHROME B (PHYB). Our results demonstrate that ABS3 and related MATE family transporters are potential negative regulators of hypocotyl cell elongation and support a functional link between the endomembrane system, particularly the LE/PVC, and the regulation of plant cell elongation. PMID:26160579

  10. Phytotoxicity, accumulation and transport of silver nanoparticles by Arabidopsis thaliana.

    Science.gov (United States)

    Geisler-Lee, Jane; Wang, Qiang; Yao, Ying; Zhang, Wen; Geisler, Matt; Li, Kungang; Huang, Ying; Chen, Yongsheng; Kolmakov, Andrei; Ma, Xingmao

    2013-05-01

    The widespread availability of nano-enabled products in the global market may lead to the release of a substantial amount of engineered nanoparticles in the environment, which frequently display drastically different physiochemical properties than their bulk counterparts. The purpose of the study was to evaluate the impact of citrate-stabilised silver nanoparticles (AgNPs) on the plant Arabidopsis thaliana at three levels, physiological phytotoxicity, cellular accumulation and subcellular transport of AgNPs. The monodisperse AgNPs of three different sizes (20, 40 and 80 nm) aggregated into much larger sizes after mixing with quarter-strength Hoagland solution and became polydisperse. Immersion in AgNP suspension inhibited seedling root elongation and demonstrated a linear dose-response relationship within the tested concentration range. The phytotoxic effect of AgNPs could not be fully explained by the released silver ions. Plants exposed to AgNP suspensions bioaccumulated higher silver content than plants exposed to AgNO3 solutions (Ag(+) representative), indicating AgNP uptake by plants. AgNP toxicity was size and concentration dependent. AgNPs accumulated progressively in this sequence: border cells, root cap, columella and columella initials. AgNPs were apoplastically transported in the cell wall and found aggregated at plasmodesmata. In all the three levels studied, AgNP impacts differed from equivalent dosages of AgNO3.

  11. Auxin transport in an auxin-resistant mutant of arabidopsis thaliana

    Energy Technology Data Exchange (ETDEWEB)

    Lincoln, C.; Benning, C.; Estelle, M.

    1987-04-01

    The authors are studying a group of allelic recessive mutations in Arabidopsis called axr-1. Homozygous axr-1 plants are resistant to exogenously applied auxin. In addition, axr-1 mutations all confer a number of development abnormalities including an apparent reduction in apical dominance, loss of normal geotropic response, and a failure to self-fertilize due to a decrease in stamen elongation. In order to determine whether this pleiotropic phenotype is due to an alteration in auxin transport they have adapted the agar block transport assay for use in Arabidopsis stem segments. Their results indicate that as in other plant species, auxin transport is strongly polar in Arabidopsis stem segments. In addition transport is inhibited by the well characterized auxin transport inhibitor N-1-naphthylphthalamic acid and the artificial auxin 2,4-D. These results as well as the characterization of transport in axr-1 plants will be presented.

  12. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development

    OpenAIRE

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M.; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L.

    2016-01-01

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rh...

  13. A novel high-affinity sucrose transporter is required for virulence of the plant pathogen Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Ramon Wahl

    2010-02-01

    Full Text Available Plant pathogenic fungi cause massive yield losses and affect both quality and safety of food and feed produced from infected plants. The main objective of plant pathogenic fungi is to get access to the organic carbon sources of their carbon-autotrophic hosts. However, the chemical nature of the carbon source(s and the mode of uptake are largely unknown. Here, we present a novel, plasma membrane-localized sucrose transporter (Srt1 from the corn smut fungus Ustilago maydis and its characterization as a fungal virulence factor. Srt1 has an unusually high substrate affinity, is absolutely sucrose specific, and allows the direct utilization of sucrose at the plant/fungal interface without extracellular hydrolysis and, thus, without the production of extracellular monosaccharides known to elicit plant immune responses. srt1 is expressed exclusively during infection, and its deletion strongly reduces fungal virulence. This emphasizes the central role of this protein both for efficient carbon supply and for avoidance of apoplastic signals potentially recognized by the host.

  14. The Potato Sucrose Transporter StSUT1 Interacts with a DRM-Associated Protein Disulfide Isomerase

    Institute of Scientific and Technical Information of China (English)

    Undine Krügel; Hong-Xia He; Konstanze Gier; Jana Reins; Izabela Chincinska; Bernhard Grimm; Waltraud X. Schulze; Christina Kühn

    2012-01-01

    Organization of proteins into complexes is crucial for many cellular functions.Recently,the SUT1 protein was shown to form homodimeric complexes,to be associated with lipid raft-like microdomains in yeast as well as in plants and to undergo endocytosis in response to brefeldin A.We therefore aimed to identify SUT1-interacting proteins that might be involved in dimerization,endocytosis,or targeting of SUT1 to raft-like microdomains.Therefore,we identified potato membrane proteins,which are associated with the detergent-resistant membrane (DRM) fraction.Among the proteins identified,we clearly confirmed StSUT1 as part of DRM in potato source leaves.We used the yeast two-hybrid split ubiquitin system (SUS) to systematically screen for interaction between the sucrose transporter StSUT1 and other membraneassociated or soluble proteins in vivo.The SUS screen was followed by immunoprecipitation using affinity-purified StSUT1-specific peptide antibodies and mass spectrometric analysis of co-precipitated proteins.A large overlap was observed between the StSUT1-interacting proteins identified in the co-immunoprecipitation and the detergent-resistant membrane fraction.One of the SUT1-interacting proteins,a protein disulfide isomerase (PDI),interacts also with other sucrose transporter proteins.A potential role of the PDI as escort protein is discussed.

  15. PGR5-PGRL1-Dependent Cyclic Electron Transport Modulates Linear Electron Transport Rate in Arabidopsis thaliana.

    Science.gov (United States)

    Suorsa, Marjaana; Rossi, Fabio; Tadini, Luca; Labs, Mathias; Colombo, Monica; Jahns, Peter; Kater, Martin M; Leister, Dario; Finazzi, Giovanni; Aro, Eva-Mari; Barbato, Roberto; Pesaresi, Paolo

    2016-02-01

    Plants need tight regulation of photosynthetic electron transport for survival and growth under environmental and metabolic conditions. For this purpose, the linear electron transport (LET) pathway is supplemented by a number of alternative electron transfer pathways and valves. In Arabidopsis, cyclic electron transport (CET) around photosystem I (PSI), which recycles electrons from ferrodoxin to plastoquinone, is the most investigated alternative route. However, the interdependence of LET and CET and the relative importance of CET remain unclear, largely due to the difficulties in precise assessment of the contribution of CET in the presence of LET, which dominates electron flow under physiological conditions. We therefore generated Arabidopsis mutants with a minimal water-splitting activity, and thus a low rate of LET, by combining knockout mutations in PsbO1, PsbP2, PsbQ1, PsbQ2, and PsbR loci. The resulting Δ5 mutant is viable, although mature leaves contain only ∼ 20% of wild-type naturally less abundant PsbO2 protein. Δ5 plants compensate for the reduction in LET by increasing the rate of CET, and inducing a strong non-photochemical quenching (NPQ) response during dark-to-light transitions. To identify the molecular origin of such a high-capacity CET, we constructed three sextuple mutants lacking the qE component of NPQ (Δ5 npq4-1), NDH-mediated CET (Δ5 crr4-3), or PGR5-PGRL1-mediated CET (Δ5 pgr5). Their analysis revealed that PGR5-PGRL1-mediated CET plays a major role in ΔpH formation and induction of NPQ in C3 plants. Moreover, while pgr5 dies at the seedling stage under fluctuating light conditions, Δ5 pgr5 plants are able to survive, which underlines the importance of PGR5 in modulating the intersystem electron transfer. PMID:26687812

  16. Phosphoenolpyruvate-dependent phosphorylation of sucrose by Clostridium tyrobutyricum ZJU 8235: evidence for the phosphotransferase transport system.

    Science.gov (United States)

    Jiang, Ling; Cai, Jin; Wang, Jufang; Liang, Shizhong; Xu, Zhinan; Yang, Shang-Tian

    2010-01-01

    The uptake and metabolism of sucrose, the major sugar in industrial cane molasses, by Clostridium tyrobutyricum ZJU 8235 was investigated and this study provided the first definitive evidence for phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) activity in butyric acid-producing bacteria. Glucose was utilized preferentially to sucrose when both substrates were present in the medium. The PEP-dependent sucrose: PTS was induced by growing C. tyrobutyricum on sucrose (but not glucose) as the sole carbon source. Extract fractionation and PTS reconstitution experiments revealed that both soluble and membrane components were required for bioactivity. Sucrose-6-phosphate hydrolase and fructokinase activities were also detected in sucrose-grown cultures. Based on these findings, a pathway of sucrose metabolism in this organism was proposed that includes the forming of sucrose-6-phosphate via the PTS and its further degradation into glucose-6-phosphate and fructose-6-phosphate. PMID:19726178

  17. The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis

    DEFF Research Database (Denmark)

    Rautengarten, Carsten; Ebert, Berit; Moreno, Ignacio;

    2014-01-01

    that are specifically presumed to deliver the diverse array of nucleotide sugars found in plants. This study has developed a novel approach that enabled functional characterization of six bifunctional UDP-rhamnose (Rha)/UDP-galactose (Gal) transporters from Arabidopsis. An analysis of loss...

  18. Family business: the multidrug-resistance related protein (MRP) ABC transporter genes in Arabidopsis thaliana.

    Science.gov (United States)

    Kolukisaoglu, H Uner; Bovet, Lucien; Klein, Markus; Eggmann, Thomas; Geisler, Markus; Wanke, Dierk; Martinoia, Enrico; Schulz, Burkhard

    2002-11-01

    Despite the completion of the sequencing of the entire genome of Arabidopsis thaliana (L.) Heynh., the exact determination of each single gene and its function remains an open question. This is especially true for multigene families. An approach that combines analysis of genomic structure, expression data and functional genomics to ascertain the role of the members of the multidrug-resistance-related protein ( MRP) gene family, a subfamily of the ATP-binding cassette (ABC) transporters from Arabidopsis is presented. We used cDNA sequencing and alignment-based re-annotation of genomic sequences to define the exact genic structure of all known AtMRP genes. Analysis of promoter regions suggested different induction conditions even for closely related genes. Expression analysis for the entire gene family confirmed these assumptions. Phylogenetic analysis and determination of segmental duplication in the regions of AtMRP genes revealed that the evolution of the extraordinarily high number of ABC transporter genes in plants cannot solely be explained by polyploidisation during the evolution of the Arabidopsis genome. Interestingly MRP genes from Oryza sativa L. (rice; OsMRP) show very similar genomic structures to those from Arabidopsis. Screening of large populations of T-DNA-mutagenised lines of A. thaliana resulted in the isolation of AtMRP insertion mutants. This work opens the way for the defined analysis of a multigene family of important membrane transporters whose broad variety of functions expands their traditional role as cellular detoxifiers. PMID:12430019

  19. Aluminum-activated citrate and malate transporters from the MATE and ALMT families function independently to confer Arabidopsis aluminum tolerance

    Science.gov (United States)

    Aluminum (Al) activated root malate and citrate exudation play an important role in Al tolerance in many plant species. AtALMT1, an Al-activated malate transporter, is a major contributor to Arabidopsis Al tolerance. Here, we demonstrate that a second, unrelated gene, AtMATE, encodes an Arabidopsi...

  20. The Phosphate Transporter PHT4;6 Is a Determinant of Salt Tolerance that Is Localized to the Golgi Apparatus of Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Beatriz Cubero; Yuko Nakagawa; Xing-Yu jiang; Ken-Ji Miura; Fang Li; Kashchandra G.Raghothama; Ray A.Bressan; Paul M.Hasegawa; Jose M.Pardo

    2009-01-01

    Insertion mutations that disrupt the function of PHT4;6 (At5g44370) cause NaCI hypersensitivity of Arabidop-sis seedlings that is characterized by reduced growth of the primary root,enhanced lateral branching,and swelling of root tips.Mutant phenotypes were exacerbated by sucrose,but not by equiosmolar concentrations of mannitol,and atten-uated by low inorganic phosphate in the medium.Protein PHT4;6 belongs to the Major Facilitator Superfamily of per-meases that shares significant sequence similarity to mammalian type-I Pi transporters and vesicular glutamate transporters,and is a member of the PHT4 family of putative intracellular phosphate transporters of plants.PHT4;6 local-izes to the Golgi membrane and transport studies indicate that PHT4;6 facilitates the selective transport of Pi but not of chloride or inorganic anions.Phenotypic similarities with other mutants displaying root swelling suggest that PHT4;6 likely functions in protein N-glycosylation and cell wall biosynthesis,which are essential for salt tolerance.Together,our results indicate that PHT4;6 transports Pi out of the Golgi lumenal space for the re-cycling of the Pi released from glycosylation processes.

  1. Auxin distribution and transport during embryogenesis and seed germi-nation of Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Auxin distribution during embryogenesis and seed germination were studied with transgenic Arabidopsis plants expressing GUS gene driven by a synthetic DR5 promoter, an auxin responsive promoter. The results showed that GUS activity is higher in ends of hypophysis and cotyledon primordia of heart-, torpedo- and cotyledon-stage embryos, leaf tip area, lateral root primordia, root apex and cotyledon of young seedlings.And GUS accumulated in root apex of the seedlings grown on auxin transport inhibitor containing media.All these suggested that above-mentioned part of the organs and tissues have a higher level of auxin, and auxin polar transport inhibitor could cause the accumulation of auxin in root apex. And auxin transport inhibitor also resulted in aberration of Arabidopsis leaf pattern formation, root gravitropism and elongation.

  2. Photoperiodic regulation of the sucrose transporter StSUT4 affects the expression of circadian-regulated genes and ethylene production

    Directory of Open Access Journals (Sweden)

    Izabela eChincinska

    2013-02-01

    Full Text Available Several recent publications report different subcellular localisation of members of the SUT4 subfamily of sucrose transporters. The physiological function of SUT4 sucrose transporters is still not entirely clarified as down-regulation of members of the SUT4 clade had very different effects in rice, poplar and potato. Here, we provide new data on the localization and function of the Solanaceous StSUT4 protein, further elucidating involvement in the onset of flowering, tuberization and in the shade avoidance syndrome of potato plants.Induction of early flowering and tuberization in SUT4-inhibited potato plants correlates with increased sucrose export from leaves and increased sucrose and starch accumulation in terminal sink organs such as developing tubers. SUT4 does not only affect the expression of gibberellin and ethylene biosynthetic enzymes, but also the rate of ethylene synthesis in potato. In SUT4-inhibited plants, the ethylene production no longer follows a diurnal rhythm, leading to the assumption that StSUT4 controls circadian gene expression, potentially by regulating sucrose export from leaves. Furthermore, SUT4 expression affects clock-regulated genes such as StFT, StSOC1 and StCO, which might also be involved in a photoperiod-dependently controlled tuberization. A model is proposed in which StSUT4 controls a phloem-mobile signalling molecule generated in leaves which together with enhanced sucrose export affects developmental switches in apical meristems. SUT4 seems to link photoreceptor-perceived information about the light quality and day length, with phytohormone biosynthesis and the expression of circadian genes.

  3. Multidrug Resistance–like Genes of Arabidopsis Required for Auxin Transport and Auxin-Mediated Development

    Science.gov (United States)

    Noh, Bosl; Murphy, Angus S.; Spalding, Edgar P.

    2001-01-01

    Arabidopsis possesses several genes related to the multidrug resistance (MDR) genes of animals, one of which, AtMDR1, was shown to be induced by the hormone auxin. Plants having mutations in AtMDR1 or its closest relative, AtPGP1, were isolated by a reverse genetic strategy. Auxin transport activity was greatly impaired in atmdr1 and atmdr1 atpgp1 double mutant plants. Epinastic cotyledons and reduced apical dominance were mutant phenotypes consistent with the disrupted basipetal flow of auxin. The auxin transport inhibitor 1-naphthylphthalamic acid was shown to bind tightly and specifically to AtMDR1 and AtPGP1 proteins. The results indicate that these two MDR-like genes of Arabidopsis encode 1-naphthylphthalamic acid binding proteins that are required for normal auxin distribution and auxin-mediated development. PMID:11701880

  4. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development.

    Science.gov (United States)

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L

    2016-01-01

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development. PMID:27381418

  5. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development.

    Science.gov (United States)

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L

    2016-07-06

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development.

  6. Arabidopsis thaliana AtUTr7 Encodes a Golgi-Localized UDP-Glucose/UDP-Galactose Transporter that Affects Lateral Root Emergence

    Institute of Scientific and Technical Information of China (English)

    Michael Handford; Cecilia Rodríguez-Furlán; Lorena Marchant; Marcelo Segura; Daniela Gómez; Elena Alvarez-Buyll; Guang-Yan Xiong; Markus Pauly; Ariel Orellana

    2012-01-01

    Nucleotide sugar transporters (NSTs) are antiporters comprising a gene family that plays a fundamental role in the biosynthesis of complex cell wall polysaccharides and glycoproteins in plants.However,due to the limited number of related mutants that have observable phenotypes,the biological function(s) of most NSTs in cell wall biosynthesis and assembly have remained elusive.Here,we report the characterization of AtUTr7 from Arabidopsis (Arabidopsis thaliana (L.) Heynh.),which is homologous to multi-specific UDP-sugar transporters from Drosophila melanogaster,humans,and Caenorhabditis elegans.We show that AtUTr7 possesses the common structural characteristics conserved among NSTs.Using a green fluorescent protein (GFP) tagged version,we demonstrate that AtUTr7 is localized in the Golgi apparatus.We also show that AtUTr7 is widely expressed,especially in the roots and in specific floral organs.Additionally,the results of an in vitro nucleotide sugar transport assay carried out with a tobacco and a yeast expression system suggest that AtUTr7 is capable of transferring UDP-Gal and UDP-GIc,but not a range of other UDP-and GDP-sugars,into the Golgi lumen.Mutants lacking expression of AtUTr7 exhibited an early proliferation of lateral roots as well as distorted root hairs when cultivated at high sucrose concentrations.Furthermore,the distribution of homogalacturonan with a low degree of methyl esterification differed in lateral root tips of the mutant compared to wild-type plants,although additional analytical procedures revealed no further differences in the composition of the root cell walls.This evidence suggests that the transport of UDP-Gal and UDP-GIc into the Golgi under conditions of high root biomass production plays a role in lateral root and root hair development.

  7. Reproductive Toxicity and Life History Study of Silver Nanoparticle Effect, Uptake and Transport in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Jane Geisler-Lee

    2014-04-01

    Full Text Available Concerns about nanotechnology have prompted studies on how the release of these engineered nanoparticles impact our environment. Herein, the impact of 20 nm silver nanoparticles (AgNPs on the life history traits of Arabidopsis thaliana was studied in both above- and below-ground parts, at macroscopic and microscopic scales. Both gross phenotypes (in contrast to microscopic phenotypes and routes of transport and accumulation were investigated from roots to shoots. Wild type Arabidopsis growing in soil, regularly irrigated with 75 μg/L of AgNPs, did not show any obvious morphological change. However, their vegetative development was prolonged by two to three days and their reproductive growth shortened by three to four days. In addition, the germination rates of offspring decreased drastically over three generations. These findings confirmed that AgNPs induce abiotic stress and cause reproductive toxicity in Arabidopsis. To trace transport of AgNPs, this study also included an Arabidopsis reporter line genetically transformed with a green fluorescent protein and grown in an optical transparent medium with 75 μg/L AgNPs. AgNPs followed three routes: (1 At seven days after planting (DAP at S1.0 (stages defined by Boyes et al. 2001 [41], AgNPs attached to the surface of primary roots and then entered their root tips; (2 At 14 DAP at S1.04, as primary roots grew longer, AgNPs gradually moved into roots and entered new lateral root primordia and root hairs; (3 At 17 DAP at S1.06 when the Arabidopsis root system had developed multiple lateral roots, AgNPs were present in vascular tissue and throughout the whole plant from root to shoot. In some cases, if cotyledons of the Arabidopsis seedlings were immersed in melted transparent medium, then AgNPs were taken up by and accumulated in stomatal guard cells. These findings in Arabidopsis are the first to document specific routes and rates of AgNP uptake in vivo and in situ.

  8. Phytochelatin-metal(loid) transport into vacuoles shows different substrate preferences in barley and Arabidopsis.

    Science.gov (United States)

    Song, Won-Yong; Mendoza-Cózatl, David G; Lee, Youngsook; Schroeder, Julian I; Ahn, Sang-Nag; Lee, Hyun-Sook; Wicker, Thomas; Martinoia, Enrico

    2014-05-01

    Cadmium (Cd) and arsenic (As) are toxic to all living organisms, including plants and humans. In plants, Cd and As are detoxified by phytochelatins (PCs) and metal(loid)-chelating peptides and by sequestering PC-metal(loid) complexes in vacuoles. Consistent differences have been observed between As and Cd detoxification. Whereas chelation of Cd by PCs is largely sufficient to detoxify Cd, As-PC complexes must be sequestered into vacuoles to be fully detoxified. It is not clear whether this difference in detoxification pathways is ubiquitous among plants or varies across species. Here, we have conducted a PC transport study using vacuoles isolated from Arabidopsis and barley. Arabidopsis vacuoles accumulated low levels of PC2 -Cd, and vesicles from yeast cells expressing either AtABCC1 or AtABCC2 exhibited negligible PC2 -Cd transport activity compared with PC2 -As. In contrast, barley vacuoles readily accumulated comparable levels of PC2 -Cd and PC2 -As. PC transport in barley vacuoles was inhibited by vanadate, but not by ammonium, suggesting the involvement of ABC-type transporters. Interestingly, barley vacuoles exhibited enhanced PC2 transport activity when essential metal ions, such as Zn(II), Cu(II) and Mn(II), were added to the transport assay, suggesting that PCs might contribute to the homeostasis of essential metals and detoxification of non-essential toxic metal(loid)s.

  9. Gibberellins inhibit adventitious rooting in hybrid aspen and Arabidopsis by affecting auxin transport.

    Science.gov (United States)

    Mauriat, Mélanie; Petterle, Anna; Bellini, Catherine; Moritz, Thomas

    2014-05-01

    Knowledge of processes involved in adventitious rooting is important to improve both fundamental understanding of plant physiology and the propagation of numerous plants. Hybrid aspen (Populus tremula × tremuloïdes) plants overexpressing a key gibberellin (GA) biosynthesis gene (AtGA20ox1) grow rapidly but have poor rooting efficiency, which restricts their clonal propagation. Therefore, we investigated the molecular basis of adventitious rooting in Populus and the model plant Arabidopsis. The production of adventitious roots (ARs) in tree cuttings is initiated from the basal stem region, and involves the interplay of several endogenous and exogenous factors. The roles of several hormones in this process have been characterized, but the effects of GAs have not been fully investigated. Here, we show that a GA treatment negatively affects the numbers of ARs produced by wild-type hybrid aspen cuttings. Furthermore, both hybrid aspen plants and intact Arabidopsis seedlings overexpressing AtGA20ox1, PttGID1.1 or PttGID1.3 genes (with a 35S promoter) produce few ARs, although ARs develop from the basal stem region of hybrid aspen and the hypocotyl of Arabidopsis. In Arabidopsis, auxin and strigolactones are known to affect AR formation. Our data show that the inhibitory effect of GA treatment on adventitious rooting is not mediated by perturbation of the auxin signalling pathway, or of the strigolactone biosynthetic and signalling pathways. Instead, GAs appear to act by perturbing polar auxin transport, in particular auxin efflux in hybrid aspen, and both efflux and influx in Arabidopsis.

  10. A major facilitator superfamily transporter plays a dual role in polar auxin transport and drought stress tolerance in Arabidopsis.

    Science.gov (United States)

    Remy, Estelle; Cabrito, Tânia R; Baster, Pawel; Batista, Rita A; Teixeira, Miguel C; Friml, Jiri; Sá-Correia, Isabel; Duque, Paula

    2013-03-01

    Many key aspects of plant development are regulated by the polarized transport of the phytohormone auxin. Cellular auxin efflux, the rate-limiting step in this process, has been shown to rely on the coordinated action of PIN-formed (PIN) and B-type ATP binding cassette (ABCB) carriers. Here, we report that polar auxin transport in the Arabidopsis thaliana root also requires the action of a Major Facilitator Superfamily (MFS) transporter, Zinc-Induced Facilitator-Like 1 (ZIFL1). Sequencing, promoter-reporter, and fluorescent protein fusion experiments indicate that the full-length ZIFL1.1 protein and a truncated splice isoform, ZIFL1.3, localize to the tonoplast of root cells and the plasma membrane of leaf stomatal guard cells, respectively. Using reverse genetics, we show that the ZIFL1.1 transporter regulates various root auxin-related processes, while the ZIFL1.3 isoform mediates drought tolerance by regulating stomatal closure. Auxin transport and immunolocalization assays demonstrate that ZIFL1.1 indirectly modulates cellular auxin efflux during shootward auxin transport at the root tip, likely by regulating plasma membrane PIN2 abundance. Finally, heterologous expression in yeast revealed that ZIFL1.1 and ZIFL1.3 share H(+)-coupled K(+) transport activity. Thus, by determining the subcellular and tissue distribution of two isoforms, alternative splicing dictates a dual function for the ZIFL1 transporter. We propose that this MFS carrier regulates stomatal movements and polar auxin transport by modulating potassium and proton fluxes in Arabidopsis cells.

  11. Brassica oleracea MATE encodes a citrate transporter and enhances aluminum tolerance in Arabidopsis thaliana.

    Science.gov (United States)

    Wu, Xinxin; Li, Ren; Shi, Jin; Wang, Jinfang; Sun, Qianqian; Zhang, Haijun; Xing, Yanxia; Qi, Yan; Zhang, Na; Guo, Yang-Dong

    2014-08-01

    The secretion of organic acid anions from roots is an important mechanism for plant aluminum (Al) tolerance. Here we report cloning and characterizing BoMATE (KF031944), a multidrug and toxic compound extrusion (MATE) family gene from cabbage (Brassica oleracea). The expression of BoMATE was more abundant in roots than in shoots, and it was highly induced by Al treatment. The (14)C-citrate efflux experiments in oocytes demonstrated that BoMATE is a citrate transporter. Electrophysiological analysis and SIET analysis of Xenopus oocytes expressing BoMATE indicated BoMATE is activated by Al. Transient expression of BoMATE in onion epidermal cells demonstrated that it localized to the plasma membrane. Compared with the wild-type Arabidopsis, the transgenic lines constitutively overexpressing BoMATE enhanced Al tolerance and increased citrate secretion. In addition, Arabidopsis transgenic lines had a lower K(+) efflux and higher H(+) efflux, in the presence of Al, than control wild type in the distal elongation zone (DEZ). This is the first direct evidence that MATE protein is involved in the K(+) and H(+) flux in response to Al treatment. Taken together, our results show that BoMATE is an Al-induced citrate transporter and enhances aluminum tolerance in Arabidopsis thaliana.

  12. AtCCR4a and AtCCR4b are Involved in Determining the Poly(A) Length of Granule-bound starch synthase 1 Transcript and Modulating Sucrose and Starch Metabolism in Arabidopsis thaliana.

    Science.gov (United States)

    Suzuki, Yuya; Arae, Toshihiro; Green, Pamela J; Yamaguchi, Junji; Chiba, Yukako

    2015-05-01

    Removing the poly(A) tail is the first and rate-limiting step of mRNA degradation and apparently an effective step not only for modulating mRNA stability but also for translation of many eukaryotic transcripts. Carbon catabolite repressor 4 (CCR4) has been identified as a major cytoplasmic deadenylase in Saccharomyces cerevisiae. The Arabidopsis thaliana homologs of the yeast CCR4, AtCCR4a and AtCCR4b, were identified by sequence-based analysis; however, their role and physiological significance in plants remain to be elucidated. In this study, we revealed that AtCCR4a and AtCCR4b are localized to cytoplasmic mRNA processing bodies, which are specific granules consisting of many enzymes involved in mRNA turnover. Double mutants of AtCCR4a and AtCCR4b exhibited tolerance to sucrose application but not to glucose. The levels of sucrose in the seedlings of the atccr4a/4b double mutants were reduced, whereas no difference was observed in glucose levels. Further, amylose levels were slightly but significantly increased in the atccr4a/4b double mutants. Consistent with this observation, we found that the transcript encoding granule-bound starch synthase 1 (GBSS1), which is responsible for amylose synthesis, is accumulated to a higher level in the atccr4a/4b double mutant plants than in the control plants. Moreover, we revealed that GBSS1 has a longer poly(A) tail in the double mutant than in the control plant, suggesting that AtCCR4a and AtCCR4b can influence the poly(A) length of transcripts related to starch metabolism. Our results collectively suggested that AtCCR4a and AtCCR4b are involved in sucrose and starch metabolism in A. thaliana.

  13. An improved grafting technique for mature Arabidopsis plants demonstrates long-distance shoot-to-root transport of phytochelatins in Arabidopsis.

    Science.gov (United States)

    Chen, Alice; Komives, Elizabeth A; Schroeder, Julian I

    2006-05-01

    Phytochelatins (PCs) are peptides that function in heavy-metal chelation and detoxification in plants and fungi. A recent study showed that PCs have the ability to undergo long-distance transport in a root-to-shoot direction in transgenic Arabidopsis (Arabidopsis thaliana). To determine whether long-distance transport of PCs can occur in the opposite direction, from shoots to roots, the wheat (Triticum aestivum) PC synthase (TaPCS1) gene was expressed under the control of a shoot-specific promoter (CAB2) in an Arabidopsis PC-deficient mutant, cad1-3 (CAB2TaPCS1/cad1-3). Analyses demonstrated that TaPCS1 is expressed only in shoots and that CAB2TaPCS1/cad1-3 lines complement the cadmium (Cd) and arsenic metal sensitivity of cad1-3 shoots. CAB2TaPCS1/cad1-3 plants exhibited higher Cd accumulation in roots and lower Cd accumulation in shoots compared to wild type. Fluorescence HPLC coupled to mass spectrometry analyses directly detected PC2 in the roots of CAB2:TaPCS1/cad1-3 but not in cad1-3 controls, suggesting that PC2 is transported over long distances in the shoot-to-root direction. In addition, wild-type shoot tissues were grafted onto PC synthase cad1-3 atpcs2-1 double loss-of-function mutant root tissues. An Arabidopsis grafting technique for mature plants was modified to obtain an 84% success rate, significantly greater than a previous rate of approximately 11%. Fluorescence HPLC-mass spectrometry showed the presence of PC2, PC3, and PC4 in the root tissue of grafts between wild-type shoots and cad1-3 atpcs2-1 double-mutant roots, demonstrating that PCs are transported over long distances from shoots to roots in Arabidopsis.

  14. From the Soil to the Seed. Metal Transport in Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, Mary Lou [Dartmouth College, Hanover, NH (United States)

    2015-02-27

    Deficiencies of micronutrients such as Fe, Mn, and Zn commonly limit plant growth and crop yields. The long-term goals of our program are to understand how plants acquire metal micronutrients from the soil and distribute them while protecting themselves from the potential redox damage metals can cause to living tissues. Metals serve as important co-factors for photosynthesis and respiration, yet we still know very little about metal transport. Our approach combines experimental and computational tools from the physical sciences with biochemistry and molecular biology. Specifically, we combine mutant analysis with synchrotron X-ray fluorescence (SXRF) spectroscopy, a technique that allows us to image the elemental composition of living plant material in 3-D. By analyzing the phenotypes of lines carrying mutations in various metal transporters, we have identified the genes responsible for uptake of zinc from the soil as well as genes involved in loading the seeds with metal micronutrients. Several of these transporters affect the localization of metals in the seed without affecting the overall metal content. Understanding how seeds obtain and store nutrients is key to developing crops with higher agronomic and nutritional value.

  15. The Arabidopsis ATNRT2.7 nitrate transporter controls nitrate content in seeds.

    Science.gov (United States)

    Chopin, Franck; Orsel, Mathilde; Dorbe, Marie-France; Chardon, Fabien; Truong, Hoai-Nam; Miller, Anthony J; Krapp, Anne; Daniel-Vedele, Françoise

    2007-05-01

    In higher plants, nitrate is taken up by root cells where Arabidopsis thaliana NITRATE TRANSPORTER2.1 (ATNRT2.1) chiefly acts as the high-affinity nitrate uptake system. Nitrate taken up by the roots can then be translocated from the root to the leaves and the seeds. In this work, the function of the ATNRT2.7 gene, one of the seven members of the NRT2 family in Arabidopsis, was investigated. High expression of the gene was detected in reproductive organs and peaked in dry seeds. beta-Glucuronidase or green fluorescent protein reporter gene expression driven by the ATNRT2.7 promoter confirmed this organ specificity. We assessed the capacity of ATNRT2.7 to transport nitrate in Xenopus laevis oocytes or when it is expressed ectopically in mutant plants deficient in nitrate transport. We measured the impact of an ATNRT2.7 mutation and found no difference from the wild type during vegetative development. By contrast, seed nitrate content was affected by overexpression of ATNRT2.7 or a mutation in the gene. Finally, we showed that this nitrate transporter protein was localized to the vacuolar membrane. Our results demonstrate that ATNRT2.7 plays a specific role in nitrate accumulation in the seed.

  16. Cadmium and iron transport by members of a plant metal transporter family in Arabidopsis with homology to Nramp genes

    OpenAIRE

    Thomine, Sébastien; Wang, Rongchen; Ward, John M.; Crawford, Nigel M.; Schroeder, Julian I.

    2000-01-01

    Metal cation homeostasis is essential for plant nutrition and resistance to toxic heavy metals. Many plant metal transporters remain to be identified at the molecular level. In the present study, we have isolated AtNramp cDNAs from Arabidopsis and show that these genes complement the phenotype of a metal uptake deficient yeast strain, smf1. AtNramps show homology to the Nramp gene family in bacteria, yeast, plants, and animals. Expression of AtNramp cDNAs increases Cd2+ sensitivity and Cd2+ a...

  17. Endocytosis and degradation of BOR1, a boron transporter of Arabidopsis thaliana, regulated by boron availability

    OpenAIRE

    Takano, Junpei; Miwa, Kyoko; Yuan, Lixing; von Wirén, Nicolaus; Fujiwara, Toru

    2005-01-01

    Boron (B) is essential for plants but toxic when present in excess. Arabidopsis thaliana BOR1 is a B exporter for xylem loading and is essential for efficient B translocation from roots to shoots under B limitation. B translocation to shoots was enhanced under B limitation in WT but not in bor1-1 mutant plants. The enhanced translocation was suppressed upon resupply of high levels of B within several hours. Unlike a number of transporters for essential mineral nutrients, BOR1 mRNA accumulatio...

  18. Transport of the two natural auxins, indole-3-butyric acid and indole-3-acetic acid, in Arabidopsis

    Science.gov (United States)

    Rashotte, Aaron M.; Poupart, Julie; Waddell, Candace S.; Muday, Gloria K.; Brown, C. S. (Principal Investigator)

    2003-01-01

    Polar transport of the natural auxin indole-3-acetic acid (IAA) is important in a number of plant developmental processes. However, few studies have investigated the polar transport of other endogenous auxins, such as indole-3-butyric acid (IBA), in Arabidopsis. This study details the similarities and differences between IBA and IAA transport in several tissues of Arabidopsis. In the inflorescence axis, no significant IBA movement was detected, whereas IAA is transported in a basipetal direction from the meristem tip. In young seedlings, both IBA and IAA were transported only in a basipetal direction in the hypocotyl. In roots, both auxins moved in two distinct polarities and in specific tissues. The kinetics of IBA and IAA transport appear similar, with transport rates of 8 to 10 mm per hour. In addition, IBA transport, like IAA transport, is saturable at high concentrations of auxin, suggesting that IBA transport is protein mediated. Interestingly, IAA efflux inhibitors and mutations in genes encoding putative IAA transport proteins reduce IAA transport but do not alter IBA movement, suggesting that different auxin transport protein complexes are likely to mediate IBA and IAA transport. Finally, the physiological effects of IBA and IAA on hypocotyl elongation under several light conditions were examined and analyzed in the context of the differences in IBA and IAA transport. Together, these results present a detailed picture of IBA transport and provide the basis for a better understanding of the transport of these two endogenous auxins.

  19. AtMGT7: An Arabidopsis Gene Encoding a Low-Affinity Magnesium Transporter

    Institute of Scientific and Technical Information of China (English)

    Dan-Dan Mao; Lian-Fu Tian; Le-Gong Li; Jian Chen; Pei-Yi Deng; Dong-Ping Li; Sheng Luan

    2008-01-01

    Magnesium (Mg2+) is one of the essential cations in all calls. Although the Mg2+ transport mechanism has been well-documented in bacteria, less is known about Mg2+ transporters in eukaryotes. The AtMGT gene family encoding putative magnesium transport proteins had been described previously. We report here that one of the Arabidopsis MGT family members, the AtMGT7 gene, encodes two mRNAs that have resulted from alternative splicing variants, designated AtMGT7a and AtMGTTb. Interestingly, the two mRNA variants were expressed with different patterns with AtMGT7a expressing in all organs, but AtMGTTb appearing only in root and flowers. The AtMGT7a variant functionally complemented a bacterial mutant lacking Mg2+ transport capacity, whereas AtMGT7b did not. The 63Ni2+ tracer uptake analysis in the bacterial model showed that AtMGT7a mediated low-affinity transport of Mg2+. Consistent with the complementation assay result, 63Ni2+ tracer uptake analysis revealed that AtMGT7b did not transport Mg2+. This study therefore has identified from a higher plant the first low-affinity Mg2+ transporter encoded by a gana with alternatively spliced transcripts that produce proteins with distinct functions.

  20. Sucrose-mediated transcriptional regulation of sucrose symporter activity in the phloem.

    Energy Technology Data Exchange (ETDEWEB)

    Matt Vaughn Greg Harrington Daniel R Bush

    2002-08-06

    This project was based on our discovery that sucrose acts as a signaling molecule that regulates the activity of a proton-sucrose symporter in sugar beet leaf tissue. A major objective here was determining how sucrose transporter activity is being regulated. When sucrose accumulates in the phloem sucrose transport activity drops dramatically. Western blots of plasma membrane proteins isolated from sucrose treated leaves showed that the loss of sucrose transport activity was proportional to a decline in symporter abundance, demonstrating that sucrose transport is regulated by changes in the amount of BvSUT1 protein. BvSUT1 transcript levels decreased in parallel with the loss of sucrose transport activity. Nuclear run-on experiments demonstrated that BvSUT1 gene transcription was repressed significantly in nuclei from leaves fed 100 mM exogenous sucrose, showing that sucrose-dependent modulation of BvSUT1 mRNA levels is mediated by changes in transcription. To identify which secondary messenger systems might be involved in regulating symporter activity, we used a variety of pharmacological agents to probe for a role of calcium or protein phosphorylation in sucrose signaling. In a detailed analysis, only okadaic acid altered sucrose transport activity. These results suggest a protein phosphatase is involved. We hypothesized that protein kinase inhibitors would have a neutral affect or increase symporter transcription. Transpirational feeding of the protein kinase inhibitor staurosporine had no impact on sucrose transport while calphostin C, an inhibitor of protein kinase C, caused a 60% increase. These data provided good evidence that protein phosphorylation plays a central role in regulating sucrose symporter expression and sucrose transport activity. To determine whether protein phosphorylation is involved in sucrose regulation of proton-sucrose symporter activity, we pre-fed leaves with staurosporine for 4 h and then fed the treated leaves water or 100 mM sucrose

  1. Long-distance root-to-shoot transport of phytochelatins and cadmium in Arabidopsis.

    Science.gov (United States)

    Gong, Ji-Ming; Lee, David A; Schroeder, Julian I

    2003-08-19

    Phytochelatin synthases (PCS) mediate cellular heavy-metal resistance in plants, fungi, and worms. However, phytochelatins (PCs) are generally considered to function as intracellular heavy-metal detoxification mechanisms, and whether long-distance transport of PCs occurs during heavy-metal detoxification remains unknown. Here, wheat TaPCS1 cDNA expression was either targeted to Arabidopsis roots with the Arabidopsis alcohol dehydrogenase (Adh) promoter (Adh::TaPCS1/cad1-3) or ectopically expressed with the cauliflower mosaic virus 35S promoter (35S::TaPCS1/cad1-3) in the PC-deficient mutant cad1-3. Adh::TaPCS1/cad1-3 and 35S::TaPCS1/cad1-3 complemented the cadmium, mercury, and arsenic sensitivities of the cad1-3 mutant. Northern blot, RT-PCR, and Western blot analyses showed Adh promoter-driven TaPCS1 expression only in roots and thus demonstrated lack of long-distance TaPCS1 mRNA and protein transport in plants. Fluorescence HPLC analyses showed that under Cd2+ stress, no PCs were detectable in cad1-3. However, in Adh::TaPCS1/cad1-3 plants, PCs were detected in roots and in rosette leaves and stems. Inductively coupled plasma atomic emission spectrometer analyses showed that either root-specific or ectopic expression of TaPCS1 significantly enhanced long-distance Cd2+ transport into stems and rosette leaves. Unexpectedly, transgenic expression of TaPCS1 reduced Cd2+ accumulation in roots compared with cad1-3. The reduced Cd2+ accumulation in roots and enhanced root-to-shoot Cd2+ transport in transgenic plants were abrogated by l-buthionine sulfoximine. The presented findings show that (i) transgenic expression of TaPCS1 suppresses the heavy-metal sensitivity of cad1-3, (ii) PCs can be transported from roots to shoots, and (iii) transgenic expression of the TaPCS1 gene increases long-distance root-to-shoot Cd2+ transport and reduces Cd2+ accumulation in roots.

  2. Arsenic tolerance in Arabidopsis is mediated by two ABCC-type phytochelatin transporters.

    Science.gov (United States)

    Song, Won-Yong; Park, Jiyoung; Mendoza-Cózatl, David G; Suter-Grotemeyer, Marianne; Shim, Donghwan; Hörtensteiner, Stefan; Geisler, Markus; Weder, Barbara; Rea, Philip A; Rentsch, Doris; Schroeder, Julian I; Lee, Youngsook; Martinoia, Enrico

    2010-12-01

    Arsenic is an extremely toxic metalloid causing serious health problems. In Southeast Asia, aquifers providing drinking and agricultural water for tens of millions of people are contaminated with arsenic. To reduce nutritional arsenic intake through the consumption of contaminated plants, identification of the mechanisms for arsenic accumulation and detoxification in plants is a prerequisite. Phytochelatins (PCs) are glutathione-derived peptides that chelate heavy metals and metalloids such as arsenic, thereby functioning as the first step in their detoxification. Plant vacuoles act as final detoxification stores for heavy metals and arsenic. The essential PC-metal(loid) transporters that sequester toxic metal(loid)s in plant vacuoles have long been sought but remain unidentified in plants. Here we show that in the absence of two ABCC-type transporters, AtABCC1 and AtABCC2, Arabidopsis thaliana is extremely sensitive to arsenic and arsenic-based herbicides. Heterologous expression of these ABCC transporters in phytochelatin-producing Saccharomyces cerevisiae enhanced arsenic tolerance and accumulation. Furthermore, membrane vesicles isolated from these yeasts exhibited a pronounced arsenite [As(III)]-PC(2) transport activity. Vacuoles isolated from atabcc1 atabcc2 double knockout plants exhibited a very low residual As(III)-PC(2) transport activity, and interestingly, less PC was produced in mutant plants when exposed to arsenic. Overexpression of AtPCS1 and AtABCC1 resulted in plants exhibiting increased arsenic tolerance. Our findings demonstrate that AtABCC1 and AtABCC2 are the long-sought and major vacuolar PC transporters. Modulation of vacuolar PC transporters in other plants may allow engineering of plants suited either for phytoremediation or reduced accumulation of arsenic in edible organs.

  3. Sequence analysis and functional prediction of grape sucrose transporters%葡萄中蔗糖转运蛋白家族的序列分析及功能推测

    Institute of Scientific and Technical Information of China (English)

    于菲; 卢江; 张雅丽

    2011-01-01

    This study mainly focused on grape sucrose transporters (SUCs). Depending on the studies of SUCs of arabidopsis and rice, we analyzed the cis - acting element in SUCs promoter sequences, predicted their affections on mRNA transcription. We classified SUCs to three subtribes according to their amino acid sequences, and described gene structure of every subtribe. Amino acid conservation in different SUCs was calculated subsequently.This study throws light on the further functional genomics researches of SUCs and accurate control of plant metabolism by gene technology.%以葡萄中的蔗糖转运蛋白为主要研究对象,结合了功能研究较为深入的拟南芥和水稻蔗糖转运蛋白家族序列,分析讨论了这些基因在启动子区域顺式作用元件的异同,以及这些差异可能对mRNA的转录带来的影响;同时,根据蔗糖转运蛋白氨基酸序列对其家族进行了分类,分析了不同亚类蔗糖转运蛋白基因结构的特点;最后还对蔗糖转运蛋白家族中氨基酸的保守性进行了分析.这些分析将为后续蔗糖转运蛋白功能基因组学的研究以及通过基因工程技术精确调节植物代谢提供一定的依据.

  4. Brassinosteroids stimulate plant tropisms through modulation of polar auxin transport in Brassica and Arabidopsis.

    Science.gov (United States)

    Li, Li; Xu, Jian; Xu, Zhi-Hong; Xue, Hong-Wei

    2005-10-01

    Brassinosteroids (BRs) are important plant growth regulators in multiple developmental processes. Previous studies have indicated that BR treatment enhanced auxin-related responses, but the underlying mechanisms remain unknown. Using (14)C-labeled indole-3-acetic acid and Arabidopsis thaliana plants harboring an auxin-responsive reporter construct, we show that the BR brassinolide (BL) stimulates polar auxin transport capacities and modifies the distribution of endogenous auxin. In plants treated with BL or defective in BR biosynthesis or signaling, the transcription of PIN genes, which facilitate functional auxin transport in plants, was differentially regulated. In addition, BL enhanced plant tropistic responses by promoting the accumulation of the PIN2 protein from the root tip to the elongation zone and stimulating the expression and dispersed localization of ROP2 during tropistic responses. Constitutive overexpression of ROP2 results in enhanced polar accumulation of PIN2 protein in the root elongation region and increased gravitropism, which is significantly affected by latrunculin B, an inhibitor of F-actin assembly. The ROP2 dominant negative mutants (35S-ROP2-DA/DN) show delayed tropistic responses, and this delay cannot be reversed by BL addition, strongly supporting the idea that ROP2 modulates the functional localization of PIN2 through regulation of the assembly/reassembly of F-actins, thereby mediating the BR effects on polar auxin transport and tropistic responses.

  5. Getting the most sulfate from soil: Regulation of sulfate uptake transporters in Arabidopsis.

    Science.gov (United States)

    Rouached, Hatem; Secco, David; Arpat, A Bulak

    2009-06-01

    Sulfur (S) is an essential macronutrient for all living organisms. Plants require large amounts of sulfate for growth and development, and this serves as a major entry point of sulfate into the food web. Plants acquire S in its ionic form from the soil; they have evolved tightly controlled mechanisms for the regulation of sulfate uptake in response to its external and internal availability. In the model plant Arabidopsis thaliana, the first key step in sulfate uptake is presumed to be carried out exclusively by only two high-affinity sulfate transporters: SULTR1;1 and SULTR1;2. A better understanding of the mode of regulation for these two transporters is crucial because they constitute the first determinative step in balancing sulfate in respect to its supply and demand. Here, we review the recent progress achieved in our comprehension of (i) mechanisms that regulate these two high-affinity sulfate transporters at the transcriptional and post-transcriptional levels, and (ii) their structure-function relationship. Such progress is important to enable biotechnological and agronomic strategies aimed at enhancing sulfate uptake and improving crop yield in S-deficient soils. PMID:19375816

  6. The Vacuolar Manganese Transporter MTP8 Determines Tolerance to Iron Deficiency-Induced Chlorosis in Arabidopsis.

    Science.gov (United States)

    Eroglu, Seckin; Meier, Bastian; von Wirén, Nicolaus; Peiter, Edgar

    2016-02-01

    Iron (Fe) deficiency is a widespread nutritional disorder on calcareous soils. To identify genes involved in the Fe deficiency response, Arabidopsis (Arabidopsis thaliana) transfer DNA insertion lines were screened on a high-pH medium with low Fe availability. This approach identified METAL TOLERANCE PROTEIN8 (MTP8), a member of the Cation Diffusion Facilitator family, as a critical determinant for the tolerance to Fe deficiency-induced chlorosis, also on soil substrate. Subcellular localization to the tonoplast, complementation of a manganese (Mn)-sensitive Saccharomyces cerevisiae yeast strain, and Mn sensitivity of mtp8 knockout mutants characterized the protein as a vacuolar Mn transporter suitable to prevent plant cells from Mn toxicity. MTP8 expression was strongly induced on low-Fe as well as high-Mn medium, which were both strictly dependent on the transcription factor FIT, indicating that high-Mn stress induces Fe deficiency. mtp8 mutants were only hypersensitive to Fe deficiency when Mn was present in the medium, which further suggested an Mn-specific role of MTP8 during Fe limitation. Under those conditions, mtp8 mutants not only translocated more Mn to the shoot than did wild-type plants but suffered in particular from critically low Fe concentrations and, hence, Fe chlorosis, although the transcriptional Fe deficiency response was up-regulated more strongly in mtp8. The diminished uptake of Fe from Mn-containing low-Fe medium by mtp8 mutants was caused by an impaired ability to boost the ferric chelate reductase activity, which is an essential process in Fe acquisition. These findings provide a mechanistic explanation for the long-known interference of Mn in Fe nutrition and define the molecular processes by which plants alleviate this antagonism. PMID:26668333

  7. Potassium Transporter KUP7 Is Involved in K(+) Acquisition and Translocation in Arabidopsis Root under K(+)-Limited Conditions.

    Science.gov (United States)

    Han, Min; Wu, Wei; Wu, Wei-Hua; Wang, Yi

    2016-03-01

    Potassium (K(+)) is one of the essential macronutrients for plant growth and development. K(+) uptake from environment and K(+) translocation in plants are conducted by K(+) channels and transporters. In this study, we demonstrated that KT/HAK/KUP transporter KUP7 plays crucial roles in K(+) uptake and translocation in Arabidopsis root. The kup7 mutant exhibited a sensitive phenotype on low-K(+) medium, whose leaves showed chlorosis symptoms compared with wild-type plants. Loss of function of KUP7 led to a reduction of K(+) uptake rate and K(+) content in xylem sap under K(+)-deficient conditions. Thus, the K(+) content in kup7 shoot was significantly reduced under low-K(+) conditions. Localization analysis revealed that KUP7 was predominantly targeted to the plasma membrane. The complementation assay in yeast suggested that KUP7 could mediate K(+) transport. In addition, phosphorylation on S80, S719, and S721 was important for KUP7 activity. KUP7 was ubiquitously expressed in many organs/tissues, and showed a higher expression level in Arabidopsis root. Together, our data demonstrated that KUP7 is crucial for K(+) uptake in Arabidopsis root and might be also involved in K(+) transport into xylem sap, affecting K(+) translocation from root toward shoot, especially under K(+)-limited conditions. PMID:26851373

  8. Transport of sucrose-modified nanoscale zero-valent iron in saturated porous media: role of media size, injection rate and input concentration.

    Science.gov (United States)

    Li, Hui; Zhao, Yong-sheng; Han, Zhan-tao; Hong, Mei

    2015-01-01

    The growing use of nanoscale zero-valent iron (NZVI) in the remediation of contaminated groundwater raises concerns regarding its transport in aquifers. Laboratory-scale sand-packed column experiments were conducted with bare and sucrose-modified NZVI (SM-NZVI) to improve our understanding of the transport of the nanoparticles in saturated porous media, as well as the role of media size, suspension injection rate and concentration on the nanoparticle behavior. As the main indicative parameters, the normalized effluent concentration was measured and the deposition rate coefficient (k) was calculated for different simulated conditions. Overall, compared to the high retention of bare NZVI in the saturated silica column, SM-NZVI suspension could travel through the coarse sand column easily. However, the transport of SM-NZVI particles was not very satisfactory in a smaller size granular matrix especially in fine silica sand. Furthermore, the value of k regularly decreased with the increasing injection rate of suspension but increased with suspension concentration, which could reflect the role of these factors in the SM-NZVI travel process. The calculation of k-value at the tests condition adequately described the experimental results from the point of deposition dynamics, which meant the assumption of first-order deposition kinetics for the transport of NZVI particles was reasonable and feasible. PMID:26524436

  9. Overexpression of OsWRKY72 gene interferes in the abscisic acid signal and auxin transport pathway of Arabidopsis

    Indian Academy of Sciences (India)

    Song Yu; Chen Ligang; Zhang Liping; Yu Diqiu

    2010-09-01

    Through activating specific transcriptional programmes, plants can launch resistance mechanisms to stressful environments and acquire a new equilibrium between development and defence. To screen the rice WRKY transcription factor which functions in abiotic stress tolerance and modulates the abscisic acid (ABA) response, we generated a whole array of 35S-OsWRKY transgenic Arabidopsis. In this study, we report that 35S-OsWRKY72 transgenic Arabidopsis, whose seed germination was retarded under normal conditions, emerged more sensitive to mannitol, NaCl, ABA stresses and sugar starvation than vector plants. Meanwhile, 35S-OsWRKY72 transgenic Arabidopsis displayed early flowering, reduced apical dominance, lost high temperature-induced hypocotyl elongation response, and enhanced gravitropism response, which were similar to the auxin-related gene mutants aux1, axr1 and bud1. Further, semi-quantitative RT-PCR showed that the expression patterns of three auxin-related genes AUX1, AXR1 and BUD1 were significantly altered in rosette leaves and inflorescences of 35S-OsWRKY72 plants compared with control Arabidopsis, and two ABA-related genes ABA2 and ABI4 were induced in 35S-OsWRKY72 seedlings. In addition, northern blot analysis indicated that, in rice, OsWRKY72 was inducible by polyethylene glycol (PEG), NaCl, naphthalene acetic acid (NAA), ABA and 42°C, similar to its orthologue AtWRKY75 in Arabidopsis, implying that these two WRKY genes might be required for multiple physiological processes in their plants. Together, these results suggest that OsWRKY72 interferes in the signal cross-talk between the ABA signal and auxin transport pathway in transgenic Arabidopsis.

  10. Structure-Function Relationship of a Plant NCS1 Member – Homology Modeling and Mutagenesis Identified Residues Critical for Substrate Specificity of PLUTO, a Nucleobase Transporter from Arabidopsis

    OpenAIRE

    Sandra Witz; Pankaj Panwar; Markus Schober; Johannes Deppe; Farhan Ahmad Pasha; Joanne Lemieux, M; Torsten Möhlmann

    2014-01-01

    Plastidic uracil salvage is essential for plant growth and development. So far, PLUTO, the plastidic nucleobase transporter from Arabidopsis thaliana is the only known uracil importer at the inner plastidic membrane which represents the permeability barrier of this organelle. We present the first homology model of PLUTO, the sole plant NCS1 member from Arabidopsis based on the crystal structure of the benzyl hydantoin transporter MHP1 from Microbacterium liquefaciens and validated by molecula...

  11. Allyl Isothiocyanate Inhibits Actin-Dependent Intracellular Transport in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Bjørnar Sporsheim

    2015-12-01

    Full Text Available Volatile allyl isothiocyanate (AITC derives from the biodegradation of the glucosinolate sinigrin and has been associated with growth inhibition in several plants, including the model plant Arabidopsis thaliana. However, the underlying cellular mechanisms of this feature remain scarcely investigated in plants. In this study, we present evidence of an AITC-induced inhibition of actin-dependent intracellular transport in A. thaliana. A transgenic line of A. thaliana expressing yellow fluorescent protein (YFP-tagged actin filaments was used to show attenuation of actin filament movement by AITC. This appeared gradually in a time- and dose-dependent manner and resulted in actin filaments appearing close to static. Further, we employed four transgenic lines with YFP-fusion proteins labeling the Golgi apparatus, endoplasmic reticulum (ER, vacuoles and peroxisomes to demonstrate an AITC-induced inhibition of actin-dependent intracellular transport of or, in these structures, consistent with the decline in actin filament movement. Furthermore, the morphologies of actin filaments, ER and vacuoles appeared aberrant following AITC-exposure. However, AITC-treated seedlings of all transgenic lines tested displayed morphologies and intracellular movements similar to that of the corresponding untreated and control-treated plants, following overnight incubation in an AITC-absent environment, indicating that AITC-induced decline in actin-related movements is a reversible process. These findings provide novel insights into the cellular events in plant cells following exposure to AITC, which may further expose clues to the physiological significance of the glucosinolate-myrosinase system.

  12. PIN6 auxin transporter at endoplasmic reticulum and plasma membrane mediates auxin homeostasis and organogenesis in Arabidopsis.

    Science.gov (United States)

    Simon, Sibu; Skůpa, Petr; Viaene, Tom; Zwiewka, Marta; Tejos, Ricardo; Klíma, Petr; Čarná, Mária; Rolčík, Jakub; De Rycke, Riet; Moreno, Ignacio; Dobrev, Petre I; Orellana, Ariel; Zažímalová, Eva; Friml, Jiří

    2016-07-01

    Plant development mediated by the phytohormone auxin depends on tightly controlled cellular auxin levels at its target tissue that are largely established by intercellular and intracellular auxin transport mediated by PIN auxin transporters. Among the eight members of the Arabidopsis PIN family, PIN6 is the least characterized candidate. In this study we generated functional, fluorescent protein-tagged PIN6 proteins and performed comprehensive analysis of their subcellular localization and also performed a detailed functional characterization of PIN6 and its developmental roles. The localization study of PIN6 revealed a dual localization at the plasma membrane (PM) and endoplasmic reticulum (ER). Transport and metabolic profiling assays in cultured cells and Arabidopsis strongly suggest that PIN6 mediates both auxin transport across the PM and intracellular auxin homeostasis, including the regulation of free auxin and auxin conjugates levels. As evidenced by the loss- and gain-of-function analysis, the complex function of PIN6 in auxin transport and homeostasis is required for auxin distribution during lateral and adventitious root organogenesis and for progression of these developmental processes. These results illustrate a unique position of PIN6 within the family of PIN auxin transporters and further add complexity to the developmentally crucial process of auxin transport. PMID:27240710

  13. Mutations in the Arabidopsis Peroxisomal ABC Transporter COMATOSE Allow Differentiation between Multiple Functions In Planta: Insights from an Allelic Series

    OpenAIRE

    Dietrich, Daniela; Schmuths, Heike; Lousa, Carine De Marcos; Baldwin, Jocelyn M.; Baldwin, Stephen A.; Baker, Alison; Theodoulou, Frederica L; Holdsworth, Michael J

    2009-01-01

    COMATOSE (CTS), the Arabidopsis homologue of human Adrenoleukodystrophy protein (ALDP), is required for import of substrates for peroxisomal β-oxidation. A new allelic series and a homology model based on the bacterial ABC transporter, Sav1866, provide novel insights into structure-function relations of ABC subfamily D proteins. In contrast to ALDP, where the majority of mutations result in protein absence from the peroxisomal membrane, all CTS mutants produced stable protein. Mutation of con...

  14. Identification of a Stelar-Localized Transport Protein That Facilitates Root-to-Shoot Transfer of Chloride in Arabidopsis

    KAUST Repository

    Li, Bo

    2015-12-11

    Under saline conditions, higher plants restrict the accumulation of chloride ions (Cl–) in the shoot by regulating their transfer from the root symplast into the xylem-associated apoplast. To identify molecular mechanisms underpinning this phenomenon, we undertook a transcriptional screen of salt stressed Arabidopsis (Arabidopsis thaliana) roots. Microarrays, quantitative RT-PCR, and promoter-GUS fusions identified a candidate gene involved in Cl– xylem loading from the Nitrate transporter 1/Peptide Transporter family (NPF2.4). This gene was highly expressed in the root stele compared to the cortex, and its expression decreased after exposure to NaCl or abscisic acid. NPF2.4 fused to fluorescent proteins, expressed either transiently or stably, was targeted to the plasma membrane. Electrophysiological analysis of NPF2.4 in Xenopus laevis oocytes suggested that NPF2.4 catalyzed passive Cl– efflux out of cells and was much less permeable to NO3−. Shoot Cl– accumulation was decreased following NPF2.4 artificial microRNA knockdown, whereas it was increased by overexpression of NPF2.4. Taken together, these results suggest that NPF2.4 is involved in long-distance transport of Cl– in plants, playing a role in the loading and the regulation of Cl– loading into the xylem of Arabidopsis roots during salinity stress.

  15. Sucrose nonfermenting AMPK-related kinase (SNARK) mediates contraction-stimulated glucose transport in mouse skeletal muscle

    DEFF Research Database (Denmark)

    Koh, Ho-Jin; Toyoda, Taro; Fujii, Nobuharu;

    2010-01-01

    . Whole-body SNARK heterozygotic knockout mice also had impaired contraction-stimulated glucose transport in skeletal muscle, and knockdown of SNARK in C2C12 muscle cells impaired sorbitol-stimulated glucose transport. SNARK is activated by muscle contraction and is a unique mediator of contraction...

  16. Modelling the dynamics of polar auxin transport in inflorescence stems of Arabidopsis thaliana.

    Science.gov (United States)

    Boot, Kees J M; Hille, Sander C; Libbenga, Kees R; Peletier, Lambertus A; van Spronsen, Paulina C; van Duijn, Bert; Offringa, Remko

    2016-02-01

    The polar transport of the plant hormone auxin has been the subject of many studies, several involving mathematical modelling. Unfortunately, most of these models have not been experimentally verified. Here we present experimental measurements of long-distance polar auxin transport (PAT) in segments of inflorescence stems of Arabidopsis thaliana together with a descriptive mathematical model that was developed from these data. It is based on a general advection-diffusion equation for auxin density, as suggested by the chemiosmotic theory, but is extended to incorporate both immobilization of auxin and exchange with the surrounding tissue of cells involved in PAT, in order to account for crucial observations. We found that development of the present model assisted effectively in the analysis of experimental observations. As an example, we discuss the analysis of a quadruple mutant for all four AUX1/LAX1-LAX3 influx carriers genes. We found a drastic change in the parameters governing the exchange of PAT channels with the surrounding tissue, whereas the velocity was still of the order of magnitude of the wild type. In addition, the steady-state flux of auxin through the PAT system of the mutant did not exhibit a saturable component, as we found for the wild type, suggesting that the import carriers are responsible for the saturable component in the wild type. In the accompanying Supplementary data available at JXB online, we describe in more detail the data-driven development of the model, review and derive predictions from a mathematical model of the chemiosmotic theory, and explore relationships between parameters in our model and processes and parameters at the cellular level. PMID:26531101

  17. ESKIMO1 disruption in Arabidopsis alters vascular tissue and impairs water transport.

    Directory of Open Access Journals (Sweden)

    Valérie Lefebvre

    Full Text Available Water economy in agricultural practices is an issue that is being addressed through studies aimed at understanding both plant water-use efficiency (WUE, i.e. biomass produced per water consumed, and responses to water shortage. In the model species Arabidopsis thaliana, the ESKIMO1 (ESK1 gene has been described as involved in freezing, cold and salt tolerance as well as in water economy: esk1 mutants have very low evapo-transpiration rates and high water-use efficiency. In order to establish ESK1 function, detailed characterization of esk1 mutants has been carried out. The stress hormone ABA (abscisic acid was present at high levels in esk1 compared to wild type, nevertheless, the weak water loss of esk1 was independent of stomata closure through ABA biosynthesis, as combining mutant in this pathway with esk1 led to additive phenotypes. Measurement of root hydraulic conductivity suggests that the esk1 vegetative apparatus suffers water deficit due to a defect in water transport. ESK1 promoter-driven reporter gene expression was observed in xylem and fibers, the vascular tissue responsible for the transport of water and mineral nutrients from the soil to the shoots, via the roots. Moreover, in cross sections of hypocotyls, roots and stems, esk1 xylem vessels were collapsed. Finally, using Fourier-Transform Infrared (FTIR spectroscopy, severe chemical modifications of xylem cell wall composition were highlighted in the esk1 mutants. Taken together our findings show that ESK1 is necessary for the production of functional xylem vessels, through its implication in the laying down of secondary cell wall components.

  18. Modelling the dynamics of polar auxin transport in inflorescence stems of Arabidopsis thaliana

    Science.gov (United States)

    Boot, Kees J.M.; Hille, Sander C.; Libbenga, Kees R.; Peletier, Lambertus A.; van Spronsen, Paulina C.; van Duijn, Bert; Offringa, Remko

    2016-01-01

    The polar transport of the plant hormone auxin has been the subject of many studies, several involving mathematical modelling. Unfortunately, most of these models have not been experimentally verified. Here we present experimental measurements of long-distance polar auxin transport (PAT) in segments of inflorescence stems of Arabidopsis thaliana together with a descriptive mathematical model that was developed from these data. It is based on a general advection–diffusion equation for auxin density, as suggested by the chemiosmotic theory, but is extended to incorporate both immobilization of auxin and exchange with the surrounding tissue of cells involved in PAT, in order to account for crucial observations. We found that development of the present model assisted effectively in the analysis of experimental observations. As an example, we discuss the analysis of a quadruple mutant for all four AUX1/LAX1–LAX3 influx carriers genes. We found a drastic change in the parameters governing the exchange of PAT channels with the surrounding tissue, whereas the velocity was still of the order of magnitude of the wild type. In addition, the steady-state flux of auxin through the PAT system of the mutant did not exhibit a saturable component, as we found for the wild type, suggesting that the import carriers are responsible for the saturable component in the wild type. In the accompanying Supplementary data available at JXB online, we describe in more detail the data-driven development of the model, review and derive predictions from a mathematical model of the chemiosmotic theory, and explore relationships between parameters in our model and processes and parameters at the cellular level. PMID:26531101

  19. 拟南芥MicroRNA828负调控蔗糖诱导的花青素合成%MicroRNA828 Negatively Regulates Sucrose-Induced Anthocyanin Biosynthesis in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    谢烨; 孙毅; 李淡宁; 黄继荣

    2013-01-01

    花青素生物合成途径及转录调控因子虽然已基本被阐明,但其调控机理仍在日益更新.本研究利用蔗糖诱导花青素合成的表型,建立了一种筛选拟南芥花青素代谢突变体的方法.我们从T-DNA插入突变体库中筛选出一株花青素合成过量突变体,基因克隆结果表明是由MicroRNA828 (miR828)的功能缺失所致.进一步研究发现miR828过表达植株中蔗糖诱导的花青素积累较野生型减少,这与敲除miR828的靶基因TAS4导致花青素积累比野生型高的结果一致,表明miR828负调控花青素合成.miR828在各组织中表达量很低,但其表达受到蔗糖诱导.在讨论中,我们提出了miR828调控蔗糖诱导花青素合成的模型.%Anthocyanins displaying from red, blue to purple give plants a colorful world. They play an important role in pollination, seed dispersal, and stress resistance. Although the anthocyanin biosynthetic pathway and the transcription factors have been well-documented, regulatory mechanisms underlying anthocyanin biosynthesis are not fully understood. In this study, we established a system to screen mutants with high accumulation of anthocyanin in Arabidopsis thaliana, and provided new evidence that small RNA is involved in anthocyanin biosynthesis. Using the phenomenon of sugar-induced anthocyanin biosynthesis, we obtained a mutant accumulated a higher level of anthocyanin compared with the wild type (WT). TAIL-PCR analysis revealed that the phenotype was resulted from the loss-of-function microRNA828 (miR828). Consistently, anthocyanin content was reduced in miR828 overexpressors under sucrose treatment. In addition, knockout of TAS4, the target of miR828, also led to higher accumulation of anthocyanin in sugar-treated seedlings compared with WT. These results indicate that miR828 negatively regulates anthocyanin biosynthesis. Further analysis demonstrated that the expression level of miR828 was quite low in various tissues, but was

  20. Splicing factor SR34b mutation reduces cadmium tolerance in Arabidopsis by regulating iron-regulated transporter 1 gene

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Wentao; Du, Bojing; Liu, Di; Qi, Xiaoting, E-mail: qixiaoting@cnu.edu.cn

    2014-12-12

    Highlights: • Arabidopsis splicing factor SR34b gene is cadmium-inducible. • SR34b T-DNA insertion mutant is sensitive to cadmium due to high cadmium uptake. • SR34b is a regulator of cadmium transporter IRT1 at the posttranscription level. • These results highlight the roles of splicing factors in cadmium tolerance of plant. - Abstract: Serine/arginine-rich (SR) proteins are important splicing factors. However, the biological functions of plant SR proteins remain unclear especially in abiotic stresses. Cadmium (Cd) is a non-essential element that negatively affects plant growth and development. In this study, we provided clear evidence for SR gene involved in Cd tolerance in planta. Systemic expression analysis of 17 Arabidopsis SR genes revealed that SR34b is the only SR gene upregulated by Cd, suggesting its potential roles in Arabidopsis Cd tolerance. Consistent with this, a SR34b T-DNA insertion mutant (sr34b) was moderately sensitive to Cd, which had higher Cd{sup 2+} uptake rate and accumulated Cd in greater amounts than wild-type. This was due to the altered expression of iron-regulated transporter 1 (IRT1) gene in sr34b mutant. Under normal growth conditions, IRT1 mRNAs highly accumulated in sr34b mutant, which was a result of increased stability of IRT1 mRNA. Under Cd stress, however, sr34b mutant plants had a splicing defect in IRT1 gene, thus reducing the IRT1 mRNA accumulation. Despite of this, sr34b mutant plants still constitutively expressed IRT1 proteins under Cd stress, thereby resulting in Cd stress-sensitive phenotype. We therefore propose the essential roles of SR34b in posttranscriptional regulation of IRT1 expression and identify it as a regulator of Arabidopsis Cd tolerance.

  1. Splicing factor SR34b mutation reduces cadmium tolerance in Arabidopsis by regulating iron-regulated transporter 1 gene

    International Nuclear Information System (INIS)

    Highlights: • Arabidopsis splicing factor SR34b gene is cadmium-inducible. • SR34b T-DNA insertion mutant is sensitive to cadmium due to high cadmium uptake. • SR34b is a regulator of cadmium transporter IRT1 at the posttranscription level. • These results highlight the roles of splicing factors in cadmium tolerance of plant. - Abstract: Serine/arginine-rich (SR) proteins are important splicing factors. However, the biological functions of plant SR proteins remain unclear especially in abiotic stresses. Cadmium (Cd) is a non-essential element that negatively affects plant growth and development. In this study, we provided clear evidence for SR gene involved in Cd tolerance in planta. Systemic expression analysis of 17 Arabidopsis SR genes revealed that SR34b is the only SR gene upregulated by Cd, suggesting its potential roles in Arabidopsis Cd tolerance. Consistent with this, a SR34b T-DNA insertion mutant (sr34b) was moderately sensitive to Cd, which had higher Cd2+ uptake rate and accumulated Cd in greater amounts than wild-type. This was due to the altered expression of iron-regulated transporter 1 (IRT1) gene in sr34b mutant. Under normal growth conditions, IRT1 mRNAs highly accumulated in sr34b mutant, which was a result of increased stability of IRT1 mRNA. Under Cd stress, however, sr34b mutant plants had a splicing defect in IRT1 gene, thus reducing the IRT1 mRNA accumulation. Despite of this, sr34b mutant plants still constitutively expressed IRT1 proteins under Cd stress, thereby resulting in Cd stress-sensitive phenotype. We therefore propose the essential roles of SR34b in posttranscriptional regulation of IRT1 expression and identify it as a regulator of Arabidopsis Cd tolerance

  2. Iron Sucrose Injection

    Science.gov (United States)

    Iron sucrose injection is used treat iron-deficiency anemia (a lower than normal number of red blood cells due ... may cause the kidneys to stop working). Iron sucrose injection is in a class of medications called ...

  3. Mutations in exocyst complex subunit SEC6 gene impaired polar auxin transport and PIN protein recycling in Arabidopsis primary root.

    Science.gov (United States)

    Tan, Xiaoyun; Feng, Yihong; Liu, Yulong; Bao, Yiqun

    2016-09-01

    Polar auxin transport, which is critical for land plant pattern formation and directional growth, is largely depended on asymmetric distribution of PIN proteins at the plasma membrane (PM). Endocytosis and recycling processes play important roles in regulating PIN protein distribution and abundance at the PM. Two subunits (SEC8, EXO70A1) of exocyst, an octameric vesicle-tethering complex, have been reported to be involved in PIN protein recycling in Arabidopsis. However, the function of exocyst complex in PIN protein recycling and polar auxin transport remains incompletely understood. In this study, we utilized two SEC6 down-regulation mutants (PRsec6-1 and PRsec6-2) to investigate the role of exocyst subunit SEC6 in the primary root development, polar auxin transport and PIN proteins recycling. We found that in PRsec6 mutants: 1. Primary root growth was retarded, and lateral root initiation were compromised. 2. Primary roots were sensitive to exogenous auxin 1-napthalene acetic acid (NAA) but not 2,4-dichlorophenoxy (2.4-D). 3. Recycling of PIN1 and PIN2 proteins from the Brefeldin A (BFA) compartment to the PM was delayed. 4. Vesicles accumulated in the primary root tip cells, especially accumulated in the cytosol closed to the PM. These results further demonstrated that the exocyst complex plays an important role in PIN protein recycling and polar auxin transport in Arabidopsis primary root.

  4. Mutations in exocyst complex subunit SEC6 gene impaired polar auxin transport and PIN protein recycling in Arabidopsis primary root.

    Science.gov (United States)

    Tan, Xiaoyun; Feng, Yihong; Liu, Yulong; Bao, Yiqun

    2016-09-01

    Polar auxin transport, which is critical for land plant pattern formation and directional growth, is largely depended on asymmetric distribution of PIN proteins at the plasma membrane (PM). Endocytosis and recycling processes play important roles in regulating PIN protein distribution and abundance at the PM. Two subunits (SEC8, EXO70A1) of exocyst, an octameric vesicle-tethering complex, have been reported to be involved in PIN protein recycling in Arabidopsis. However, the function of exocyst complex in PIN protein recycling and polar auxin transport remains incompletely understood. In this study, we utilized two SEC6 down-regulation mutants (PRsec6-1 and PRsec6-2) to investigate the role of exocyst subunit SEC6 in the primary root development, polar auxin transport and PIN proteins recycling. We found that in PRsec6 mutants: 1. Primary root growth was retarded, and lateral root initiation were compromised. 2. Primary roots were sensitive to exogenous auxin 1-napthalene acetic acid (NAA) but not 2,4-dichlorophenoxy (2.4-D). 3. Recycling of PIN1 and PIN2 proteins from the Brefeldin A (BFA) compartment to the PM was delayed. 4. Vesicles accumulated in the primary root tip cells, especially accumulated in the cytosol closed to the PM. These results further demonstrated that the exocyst complex plays an important role in PIN protein recycling and polar auxin transport in Arabidopsis primary root. PMID:27457987

  5. The arabidopsis thaliana AGRAVITROPIC 1 gene encodes a component of the polar-auxin-transport efflux carrier

    Science.gov (United States)

    Chen, R.; Hilson, P.; Sedbrook, J.; Rosen, E.; Caspar, T.; Masson, P. H.

    1998-01-01

    Auxins are plant hormones that mediate many aspects of plant growth and development. In higher plants, auxins are polarly transported from sites of synthesis in the shoot apex to their sites of action in the basal regions of shoots and in roots. Polar auxin transport is an important aspect of auxin functions and is mediated by cellular influx and efflux carriers. Little is known about the molecular identity of its regulatory component, the efflux carrier [Estelle, M. (1996) Current Biol. 6, 1589-1591]. Here we show that mutations in the Arabidopsis thaliana AGRAVITROPIC 1 (AGR1) gene involved in root gravitropism confer increased root-growth sensitivity to auxin and decreased sensitivity to ethylene and an auxin transport inhibitor, and cause retention of exogenously added auxin in root tip cells. We used positional cloning to show that AGR1 encodes a putative transmembrane protein whose amino acid sequence shares homologies with bacterial transporters. When expressed in Saccharomyces cerevisiae, AGR1 promotes an increased efflux of radiolabeled IAA from the cells and confers increased resistance to fluoro-IAA, a toxic IAA-derived compound. AGR1 transcripts were localized to the root distal elongation zone, a region undergoing a curvature response upon gravistimulation. We have identified several AGR1-related genes in Arabidopsis, suggesting a global role of this gene family in the control of auxin-regulated growth and developmental processes.

  6. Expression of the Znt1 zinc transporter from the metal hyperaccumulator noccaea caerulescens confers enhanced zinc and cadmium tolerance and accumulation to arabidopsis thaliana

    NARCIS (Netherlands)

    Lin, Ya Fen; Hassan, Zeshan; Talukdar, S.; Schat, Henk; Aarts, Mark G.M.

    2016-01-01

    Prompt regulation of transition metal transporters is crucial for plant zinc homeostasis. NcZNT1 is one of such transporters, found in the metal hyperaccumulator Brassicaceae species Noccaea caerulescens. It is orthologous to AtZIP4 from Arabidopsis thaliana, an important actor in Zn homeostasis.

  7. Arabidopsis copper transport protein COPT2 participates in the cross talk between iron deficiency responses and low-phosphate signaling.

    Science.gov (United States)

    Perea-García, Ana; Garcia-Molina, Antoni; Andrés-Colás, Nuria; Vera-Sirera, Francisco; Pérez-Amador, Miguel A; Puig, Sergi; Peñarrubia, Lola

    2013-05-01

    Copper and iron are essential micronutrients for most living organisms because they participate as cofactors in biological processes, including respiration, photosynthesis, and oxidative stress protection. In many eukaryotic organisms, including yeast (Saccharomyces cerevisiae) and mammals, copper and iron homeostases are highly interconnected; yet, such interdependence is not well established in higher plants. Here, we propose that COPT2, a high-affinity copper transport protein, functions under copper and iron deficiencies in Arabidopsis (Arabidopsis thaliana). COPT2 is a plasma membrane protein that functions in copper acquisition and distribution. Characterization of the COPT2 expression pattern indicates a synergic response to copper and iron limitation in roots. We characterized a knockout of COPT2, copt2-1, that leads to increased resistance to simultaneous copper and iron deficiencies, measured as reduced leaf chlorosis and improved maintenance of the photosynthetic apparatus. We propose that COPT2 could play a dual role under iron deficiency. First, COPT2 participates in the attenuation of copper deficiency responses driven by iron limitation, possibly to minimize further iron consumption. Second, global expression analyses of copt2-1 versus wild-type Arabidopsis plants indicate that low-phosphate responses increase in the mutant. These results open up new biotechnological approaches to fight iron deficiency in crops.

  8. Large-Scale Public Transcriptomic Data Mining Reveals a Tight Connection between the Transport of Nitrogen and Other Transport Processes in Arabidopsis.

    Science.gov (United States)

    He, Fei; Karve, Abhijit A; Maslov, Sergei; Babst, Benjamin A

    2016-01-01

    Movement of nitrogen to the plant tissues where it is needed for growth is an important contribution to nitrogen use efficiency. However, we have very limited knowledge about the mechanisms of nitrogen transport. Loading of nitrogen into the xylem and/or phloem by transporter proteins is likely important, but there are several families of genes that encode transporters of nitrogenous molecules (collectively referred to as N transporters here), each comprised of many gene members. In this study, we leveraged publicly available microarray data of Arabidopsis to investigate the gene networks of N transporters to elucidate their possible biological roles. First, we showed that tissue-specificity of nitrogen (N) transporters was well reflected among the public microarray data. Then, we built coexpression networks of N transporters, which showed relationships between N transporters and particular aspects of plant metabolism, such as phenylpropanoid biosynthesis and carbohydrate metabolism. Furthermore, genes associated with several biological pathways were found to be tightly coexpressed with N transporters in different tissues. Our coexpression networks provide information at the systems-level that will serve as a resource for future investigation of nitrogen transport systems in plants, including candidate gene clusters that may work together in related biological roles. PMID:27563305

  9. Constitutive overexpression of the sucrose transporter SoSUT1 in potato plants increases arbuscular mycorrhiza fungal root colonization under high, but not under low, soil phosphorus availability.

    Science.gov (United States)

    Gabriel-Neumann, Elke; Neumann, Günter; Leggewie, Georg; George, Eckhard

    2011-06-15

    The sucrose transporter SUT1 functions in phloem loading of photoassimilates in solanaceous plant species. In the present study, wildtype and transgenic potato plants with either constitutive overexpression or antisense inhibition of SUT1 were grown under high or low phosphorus (P) fertilization levels in the presence or absence of the arbuscular mycorrhizal (AM) fungus Glomus intraradices. At a low soil P fertilization level, the extent of AM fungal root colonization was not different among the genotypes. In all plants, the AM symbiosis contributed significantly to P uptake under these conditions. In response to a high soil P fertilization level, all genotypes showed a decrease in AM fungal root colonization, indicating that the expression level of SUT1 does not constitute a major mechanism of control over AM development in response to the soil P availability. However, plants with overexpression of SUT1 showed a higher extent of AM fungal root colonization compared with the other genotypes when the soil P availability was high. Whether an increased symbiotic C supply, alterations in the phytohormonal balance, or a decreased synthesis of antimicrobial compounds was the major cause for this effect requires further investigation. In plants with impaired phloem loading, a low C status of plant sink tissues did apparently not negatively affect plant C supply to the AM symbiosis. It is possible that, at least during vegetative and early generative growth, source rather than sink tissues exert control over amounts of C supplied to AM fungi. PMID:21382646

  10. Wax and cutin mutants of Arabidopsis: Quantitative characterization of the cuticular transport barrier in relation to chemical composition.

    Science.gov (United States)

    Sadler, Christina; Schroll, Bettina; Zeisler, Viktoria; Waßmann, Friedrich; Franke, Rochus; Schreiber, Lukas

    2016-09-01

    Using (14)C-labeled epoxiconazole as a tracer, cuticular permeability of Arabidopsis thaliana leaves was quantitatively measured in order to compare different wax and cutin mutants (wax2, cut1, cer5, att1, bdg, shn3 and shn1) to the corresponding wild types (Col-0 and Ws). Mutants were characterized by decreases or increases in wax and/or cutin amounts. Permeances [ms(-1)] of Arabidopsis cuticles either increased in the mutants compared to wild type or were not affected. Thus, genetic changes in wax and cutin biosynthesis in some of the investigated Arabidopsis mutants obviously impaired the coordinated cutin and wax deposition at the outer leaf epidermal cell wall. As a consequence, barrier properties of cuticles were significantly decreased. However, increasing cutin and wax amounts by genetic modifications, did not automatically lead to improved cuticular barrier properties. As an alternative approach to the radioactive transport assay, changes in chlorophyll fluorescence were monitored after foliar application of metribuzine, an herbicide inhibiting electron transport in chloroplasts. Since both, half-times of photosynthesis inhibition as well as times of complete inhibition, in fact correlated with (14)C-epoxiconazole permeances, different rates of decline of photosynthetic yield between mutants and wild type must be a function of foliar uptake of the herbicide across the cuticle. Thus, monitoring changes in chlorophyll fluorescence, instead of conducting radioactive transport assays, represents an easy-to-handle and fast alternative evaluating cuticular barrier properties of different genotypes. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner.

  11. Sucrose and Related Oligosaccharides

    Science.gov (United States)

    Eggleston, Gillian

    Sucrose (α-D-glucopyranosyl-(1↔2)-β-D-fructofuranoside) is the most common low-molecular-weight sugar found in the plant kingdom. It is ubiquitously known as common table sugar and primarily produced industrially from sugarcane (Saccharum officinarum) and sugar beet (Beta vulgaris); the basics of the industrial manufacture of sucrose are outlined in this chapter. Commercial sucrose has a very high purity (> 99.9%) making it one of the purest organic substances produced on an industrial scale. Value-addition to sucrose via chemical and biotechnological reactions is becoming more important for the diversification of the sugar industry to maintain the industries' competitiveness in a world increasingly turning to a bio-based economy. The basis for the chemical reactivity of sucrose is the eight hydroxyl groups present on the molecule, although, sucrose chemical reactivity is regarded as difficult. Increasing use of enzymatic biotechnological techniques to derivatize sucrose is expected, to add special functionalities to sucrose products like biodegradability, biocompatibility, and non-toxicity. Analysis of sucrose by colorimetric, enzymatic, oxidation-reduction and chromatography methods are discussed. Oligosaccharides related to sucrose are outlined in detail and include sucrose-based plant, honey and in vitro oligosaccharides.

  12. Specific and efficient targeting of cyanobacterial bicarbonate transporters to the inner envelope membrane of chloroplasts in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Susumu eUehara

    2016-02-01

    Full Text Available Installation of cyanobacterial bicarbonate transporters to the inner envelope membrane (IEM of chloroplasts in C3 plants has been thought to improve photosynthetic performance. However, the method to deliver cyanobacterial bicarbonate transporters to the chloroplast IEM remains to be established. In this study, we provide evidence that the cyanobacterial bicarbonate transporters, BicA and SbtA, can be specifically installed into the chloroplast IEM using the chloroplast IEM targeting signal in conjunction with the transit peptide. We fused the transit peptide and the mature portion of Cor413im1, whose targeting mechanism to the IEM has been characterized in detail, to either BicA or SbtA isolated from Synechocystis sp. PCC6803. Among the seven chimeric constructs tested, we confirmed that four chimeric bicarbonate transporters, designated as BicAI, BicAII, SbtAII, and SbtAIII, were expressed in Arabidopsis. Furthermore, these chimeric transporters were specifically targeted to the chloroplast IEM. They were also resistant to alkaline extraction but can be solubilized by Triton X-100, indicating that they are integral membrane proteins in the chloroplast IEM. One of the transporters, BicA, could reside in the chloroplast IEM even after removal of the IEM targeting signal. Taken together, our results indicate that the addition of IEM targeting signal, as well as the transit peptide, to bicarbonate transporters allows us to efficiently target nuclear-encoded chimeric bicarbonate transporters to the chloroplast IEM.

  13. Separating the roles of acropetal and basipetal auxin transport on gravitropism with mutations in two Arabidopsis multidrug resistance-like ABC transporter genes.

    Science.gov (United States)

    Lewis, Daniel R; Miller, Nathan D; Splitt, Bessie L; Wu, Guosheng; Spalding, Edgar P

    2007-06-01

    Two Arabidopsis thaliana ABC transporter genes linked to auxin transport by various previous results were studied in a reverse-genetic fashion. Mutations in Multidrug Resistance-Like1 (MDR1) reduced acropetal auxin transport in roots by 80% without affecting basipetal transport. Conversely, mutations in MDR4 blocked 50% of basipetal transport without affecting acropetal transport. Developmental and auxin distribution phenotypes associated with these altered auxin flows were studied with a high-resolution morphometric system and confocal microscopy, respectively. Vertically grown mdr1 roots produced positive and negative curvatures threefold greater than the wild type, possibly due to abnormal auxin distribution observed in the elongation zone. However, upon 90 degrees reorientation, mdr1 gravitropism was inseparable from the wild type. Thus, acropetal auxin transport maintains straight growth but contributes surprisingly little to gravitropism. Conversely, vertically maintained mdr4 roots grew as straight as the wild type, but their gravitropism was enhanced. Upon reorientation, curvature in this mutant developed faster, was distributed more basally, and produced a greater total angle than the wild type. An amplified auxin asymmetry may explain the mdr4 hypertropism. Double mutant analysis indicated that the two auxin transport streams are more independent than interdependent. The hypothesis that flavanols regulate MDR-dependent auxin transport was supported by the epistatic relationship of mdr4 to the tt4 phenylpropanoid pathway mutation.

  14. Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and 2: fructose and xylitol/H+ symporters in pollen and young xylem cells

    OpenAIRE

    Klepek, Yvonne-Simone; Volke, Melanie; Konrad, Kai R.; Wippel, Kathrin; Hoth, Stefan; Hedrich, Rainer; Sauer, Norbert

    2009-01-01

    The genome of Arabidopsis thaliana contains six genes, AtPMT1 to AtPMT6 (Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTER 1–6), which form a distinct subfamily within the large family of more than 50 monosaccharide transporter-like (MST-like) genes. So far, only AtPMT5 [formerly named AtPLT5 (At3g18830)] has been characterized and was shown to be a plasma membrane-localized H+-symporter with broad substrate specificity. The characterization of AtPMT1 (At2g16120) and AtPMT2 (At2g16130), ...

  15. The ABCG transporter PEC1/ABCG32 is required for the formation of the developing leaf cuticle in Arabidopsis.

    Science.gov (United States)

    Fabre, Guillaume; Garroum, Imène; Mazurek, Sylwester; Daraspe, Jean; Mucciolo, Antonio; Sankar, Martial; Humbel, Bruno M; Nawrath, Christiane

    2016-01-01

    The cuticle is an essential diffusion barrier on aerial surfaces of land plants whose structural component is the polyester cutin. The PERMEABLE CUTICLE1/ABCG32 (PEC1) transporter is involved in plant cuticle formation in Arabidopsis. The gpat6 pec1 and gpat4 gapt8 pec1 double and triple mutants are characterized. Their PEC1-specific contributions to aliphatic cutin composition and cuticle formation during plant development are revealed by gas chromatography/mass spectrometry and Fourier-transform infrared spectroscopy. The composition of cutin changes during rosette leaf expansion in Arabidopsis. C16:0 monomers are in higher abundance in expanding than in fully expanded leaves. The atypical cutin monomer C18:2 dicarboxylic acid is more prominent in fully expanded leaves. Findings point to differences in the regulation of several pathways of cutin precursor synthesis. PEC1 plays an essential role during expansion of the rosette leaf cuticle. The reduction of C16 monomers in the pec1 mutant during leaf expansion is unlikely to cause permeability of the leaf cuticle because the gpat6 mutant with even fewer C16:0 monomers forms a functional rosette leaf cuticle at all stages of development. PEC1/ABCG32 transport activity affects cutin composition and cuticle structure in a specific and non-redundant fashion.

  16. Identification of sucrose binding, membrane proteins using a photolyzable sucrose analog. [P. saccharophila

    Energy Technology Data Exchange (ETDEWEB)

    Ripp, K.G.; Liu, D.F.; Viitanen, P.; Hitz, W.D.

    1986-04-01

    The sucrose derivative 6'-deoxy-6'-(2-hydroxy-4-azido)benzamidosucrose (6'-HABS) was prepared from sucrose (via 6'-deoxy-6'-aminosucrose) and 4-amino-salicylic acid. 6'-HABS is a competitive inhibitor of sucrose influx into protoplasts from developing soybean cotyledons and of sucrose binding to membranes from the bacteria P. saccharophila. The Ki for inhibition in the soybean protoplasts was 75..mu..M. 6'-Deoxy-6'-(2-hydroxy-3-/sup 125/Iodo-4-azido)benzamidosucrose was prepared by lactoperoxidase iodination of 6'-HABS. Upon photolysis in the presence of membranes from P saccharophila, label from the photoprobe is incorporated into a sucrose inducible polypeptide of mass 84 KD in SDS-PAGE. The polypeptide is protected from labeling by the inclusion of sucrose in the photolysis mixture. Photolysis conditions which lead to specific labeling of the sucrose protectable polypeptide in bacterial membranes also give sucrose protectable labeling of a 66 KD polypeptide in microsomal preparations made from developing soybeans. The possibility that this is a sucrose transporting protein is being tested.

  17. AtHKT1;1 mediates nernstian sodium channel transport properties in Arabidopsis root stelar cells.

    Directory of Open Access Journals (Sweden)

    Shaowu Xue

    Full Text Available The Arabidopsis AtHKT1;1 protein was identified as a sodium (Na⁺ transporter by heterologous expression in Xenopus laevis oocytes and Saccharomyces cerevisiae. However, direct comparative in vivo electrophysiological analyses of a plant HKT transporter in wild-type and hkt loss-of-function mutants has not yet been reported and it has been recently argued that heterologous expression systems may alter properties of plant transporters, including HKT transporters. In this report, we analyze several key functions of AtHKT1;1-mediated ion currents in their native root stelar cells, including Na⁺ and K⁺ conductances, AtHKT1;1-mediated outward currents, and shifts in reversal potentials in the presence of defined intracellular and extracellular salt concentrations. Enhancer trap Arabidopsis plants with GFP-labeled root stelar cells were used to investigate AtHKT1;1-dependent ion transport properties using patch clamp electrophysiology in wild-type and athkt1;1 mutant plants. AtHKT1;1-dependent currents were carried by sodium ions and these currents were not observed in athkt1;1 mutant stelar cells. However, K⁺ currents in wild-type and athkt1;1 root stelar cell protoplasts were indistinguishable correlating with the Na⁺ over K⁺ selectivity of AtHKT1;1-mediated transport. Moreover, AtHKT1;1-mediated currents did not show a strong voltage dependence in vivo. Unexpectedly, removal of extracellular Na⁺ caused a reduction in AtHKT1;1-mediated outward currents in Columbia root stelar cells and Xenopus oocytes, indicating a role for external Na⁺ in regulation of AtHKT1;1 activity. Shifting the NaCl gradient in root stelar cells showed a Nernstian shift in the reversal potential providing biophysical evidence for the model that AtHKT1;1 mediates passive Na⁺ channel transport properties.

  18. Studies on the role of heavy-metal transporting P-type ATPase family genes on zinc (Zn) transport and accumulation in Arabidopsis thaliana

    Energy Technology Data Exchange (ETDEWEB)

    Bagavathiannan, M.V. [Manitoba Univ., Winnipeg, MB (Canada). Dept. of Plant Science; Broadley, M.R.; Donnelly, S.J.; Smith, R.J.; Mills, V. [Nottingham Univ., Loughborough, Leicestershire (United Kingdom). School of Biosciences, Plant Sciences Division

    2006-07-01

    Although zinc (Zn) is an essential plant mineral nutrient for normal crop growth, excess amounts can cause environmental contamination problems. Higher amounts of Zn are added into soils every year through effluents from tanning industries and other sources such as sewage treatment plants, metal inputs from rivers and the atmosphere. Studies have shown that specific metal tolerances exist at the cellular level in plants, indicating that specific adaptations to metal ions occur in cells as well as in the whole plant. This paper described the mechanisms that plants develop in order to tolerate heavy metals and showed that transporter genes play a key role in uptake and sequestration of heavy metals in plant systems. Since metal ion transporting genes are also involved in transport and homeostasis of heavy metals in plants, this study examined the role of the metal ion transporting gene family members in Zn transport and tolerance in plant systems. Among the metal ion transporting gene families, P-type ATPase gene family members are considered to be efficient in metal transport in the model plant Arabidopsis thaliana. They form a diverse superfamily of transporters which carry a range of essential and potentially toxic metals across cellular membranes. Genetic-screening experiments were performed in which 3 SALK lines with known disruption in the target gene were studied physiologically and at the molecular level to determine their role in heavy-metal transportation and accumulation. The study showed that one of the family lines may have altered Zn tolerance and uptake characteristics. Ongoing research continues to examine the characteristics of this line. 27 refs., 1 tab., 8 figs.

  19. Uncovering Arabidopsis membrane protein interactome enriched in transporters using mating-based split ubiquitin assays and classification models

    Directory of Open Access Journals (Sweden)

    Jin eChen

    2012-06-01

    Full Text Available High-throughput data are a double-edged sword; for the benefit of large amount of data, there is an associated cost of noise. To increase reliability and scalability of high-throughput protein interaction data generation, we tested the efficacy of classification to enrich potential protein-protein interactions (pPPIs. We applied this method to identify interactions among Arabidopsis membrane proteins enriched in transporters. We validated our method with multiple retests. Classification improved the quality of the ensuing interaction network and was effective in reducing the search space and increasing true positive rate. The final network of 541 interactions among 239 proteins (of which 179 are transporters is the first protein interaction network enriched in membrane transporters reported for any organism. This network has similar topological attributes to other published protein interaction networks. It also extends and fills gaps in currently available biological networks in plants and allows building a number of hypotheses about processes and mechanisms involving signal-transduction and transport systems.

  20. Glucose-1-phosphate transport into protoplasts and chloroplasts from leaves of Arabidopsis.

    Science.gov (United States)

    Fettke, Joerg; Malinova, Irina; Albrecht, Tanja; Hejazi, Mahdi; Steup, Martin

    2011-04-01

    Almost all glucosyl transfer reactions rely on glucose-1-phosphate (Glc-1-P) that either immediately acts as glucosyl donor or as substrate for the synthesis of the more widely used Glc dinucleotides, ADPglucose or UDPglucose. In this communication, we have analyzed two Glc-1-P-related processes: the carbon flux from externally supplied Glc-1-P to starch by either mesophyll protoplasts or intact chloroplasts from Arabidopsis (Arabidopsis thaliana). When intact protoplasts or chloroplasts are incubated with [U-(14)C]Glc-1-P, starch is rapidly labeled. Incorporation into starch is unaffected by the addition of unlabeled Glc-6-P or Glc, indicating a selective flux from Glc-1-P to starch. However, illuminated protoplasts incorporate less (14)C into starch when unlabeled bicarbonate is supplied in addition to the (14)C-labeled Glc-1-P. Mesophyll protoplasts incubated with [U-(14)C]Glc-1-P incorporate (14)C into the plastidial pool of adenosine diphosphoglucose. Protoplasts prepared from leaves of mutants of Arabidopsis that lack either the plastidial phosphorylase or the phosphoglucomutase isozyme incorporate (14)C derived from external Glc-1-P into starch, but incorporation into starch is insignificant when protoplasts from a mutant possessing a highly reduced ADPglucose pyrophosphorylase activity are studied. Thus, the path of assimilatory starch biosynthesis initiated by extraplastidial Glc-1-P leads to the plastidial pool of adenosine diphosphoglucose, and at this intermediate it is fused with the Calvin cycle-driven route. Mutants lacking the plastidial phosphoglucomutase contain a small yet significant amount of transitory starch.

  1. Arabidopsis SOI33/AtENT8 Gene Encodes a Putative Equilibrative Nucleoside Transporter That Is Involved in Cytokinin Transport In Planta

    Institute of Scientific and Technical Information of China (English)

    Jiaqiang SUN; Naoya HIROSE; Xingchun WANG; Pei WEN; Li XUE; Hitoshi SAKAKIBARA; Jianru ZUO

    2005-01-01

    The plant phytohormone cytokinin plays an important role in many facets of plant growth and development by regulating cell division and differentiation. Recent studies have shed significant light into the mechanisms of cytokinin metabolism and signaling. However, little is known about how the hormone is transported in planta, although it has been proposed that the hormone is presumably transported in nucleoside-conjugated forms. Here, we report the identification and characterization of cytokinin transport ers in Arabidopsis. We previously reported that a gain-of-function mutation in the PGA22/AtIPT8 gene caused overproduction of cytokinins in planta. In an effort to screen for suppressor of pga22/atipt8 (soi) mutants, we identified a mutant soi33-1. Molecular and genetic analyses indicated that SOI33 encodes a putative equilibrative nucleoside transporter (ENT), previously designated as AtENT8. Members of this small gene family are presumed to be involved in the transport of nucleosides in eukaryotic cells. Under conditions of nitrogen starvation, loss-of-function mutations in SOI33/AtENT8 or in a related gene AtENT3 cause a reduced sensitivity to the nucleoside-type cytokinins isopentenyladenine riboside (iPR) and trans zeatin riboside (tZR), but display a normal response to the free base-type cytokinins isopentenyladenine (iP) and trans-zeatin (tZ). Conversely, overexpression of SOI33/AtENT8 renders transgenic plants hyper sensitive to iPR but not to iP. An in planta measurement experiment indicated that uptake efficiency of 3H labeled iPR was reduced more than 40% in soi33 and atent3 mutants. However, a mutation inAtENT1 had no substantial effect on the cytokinin response and iPR uptake efficiency. Our results suggest that SOI33/ AtENT8 and AtENT3 are involved in the transport of nucleoside-type cytokinins in Arabidopsis.

  2. Functional Characterization of AtSP1, A Sucrose-Responsive Receptor-Like Kinase

    Institute of Scientific and Technical Information of China (English)

    Xuna Wu; Kerstin Zander; Eugenia Maximova; Gerhard Obermeyer; Waltraud Schulze

    2012-01-01

    Sucrose is the main product of photosynthesis and an important metabolite.It functions as signaling molecule,also potentially being involved in the regulation of growth and development in plants.AtSP1 is one of putative RLKs identified in sucrose resupply after starvation experiment and hypothesized to be involved in sucrose-related regulatory processes (Niittyl(a) et al.,2007).The aim of this work was to characterize the effect of AtSP1 and its phosphorylation site Ser744 on sucrose induced phosphorylation cascade.Therefore,proteins which changed their phosphorylation status upon sucrose supply were compared in WT,the sp1 mutant,complementation lines (35S::AtSP1-GFP/sp1),and two mutagenesis lines (35S::AtSP1S744A-GFP/sp1 and 35S::AtSP1S744D-GFP/sp1).Proteins with altered phosphorylation response in the mutant lines are candidates for direct or indirect substrates of AtSP1.To test this,Arabidopsis seedlings were cultured for 24 h without sucrose in JPL medium,and either sucrose or mannitol was resupplied for 3 min.The treatment time of 3 min was chosen as the maximum phosphorylation change of AtSP1 occurred after 3 min (Niittyl(a) et al.,2007).Seedlings were then harvested.The soluble proteins and microsome fraction were isolated and identified via LC-MS/MS.The phosphorylated amino acids were identified by MAXQUANT and quantified based on normalized ion intensities.The quantified phosphoproteins in five genotypes were compared to find candidate proteins which could be regulated by AtSP1 in response to sucrose resupply after starvation.The analysis of the short term effects regulated by AtSP1 provided new insights about sucrose-induced regulatory processes.In total,93 phosphoproteins were identified,42 of which showed a significant lower phosphorylation in the sp1 mutant.These results indicated that when expression levels of AtSP1 were severely reduced,multiple other proteins were impaired in their phosphorylation upon sucrose reply,linking them potentially with At

  3. Sucrose secreted by the engineered cyanobacterium and its fermentability

    Science.gov (United States)

    Duan, Yangkai; Luo, Quan; Liang, Feiyan; Lu, Xuefeng

    2016-10-01

    The unicellular cyanobacterium, Synechococcus elongatus PCC 7942 (Syn7942), synthesizes sucrose as the only compatible solute under salt stress. A series of engineered Syn7942 strains for sucrose production were constructed. The overexpression of the native sps (encoding a natively fused protein of sucrose phosphate synthase SPS and sucrose phosphate phosphatase SPP) in Syn7942 wild type caused a 93% improvement of sucrose productivity. The strain FL130 co-overexpressing sps and cscB (encoding a sucrose transporter) exhibited a 74% higher extracellular sucrose production than that overexpressing cscB only. Both results showed the significant improvement of sucrose productivity by the double functional protein SPS-SPP. Afterwards, FL130 was cultivated under a modified condition, and the cell-free culture medium containing 1.5 g L-1 sucrose was pre-treated with an acid hydrolysis technique. Cultivated with the neutralized hydrolysates as the starting media, two widely used microorganisms, Escherichia coli and Saccharomyces cerevisiae, showed a comparable growth with that in the control media supplemented with glucose. These results clearly demonstrated that the cell-free culture of sucrose-secreting cyanobacteria can be applied as starting media in microbial cultivation.

  4. Structure-function relationship of a plant NCS1 member - Homology modeling and mutagenesis identified residues critical for substrate specificity of PLUTO, a nucleobase transporter from arabidopsis

    KAUST Repository

    Witz, Sandra

    2014-03-12

    Plastidic uracil salvage is essential for plant growth and development. So far, PLUTO, the plastidic nucleobase transporter from Arabidopsis thaliana is the only known uracil importer at the inner plastidic membrane which represents the permeability barrier of this organelle. We present the first homology model of PLUTO, the sole plant NCS1 member from Arabidopsis based on the crystal structure of the benzyl hydantoin transporter MHP1 from Microbacterium liquefaciens and validated by molecular dynamics simulations. Polar side chains of residues Glu-227 and backbones of Val-145, Gly-147 and Thr-425 are proposed to form the binding site for the three PLUTO substrates uracil, adenine and guanine. Mutational analysis and competition studies identified Glu-227 as an important residue for uracil and to a lesser extent for guanine transport. A differential response in substrate transport was apparent with PLUTO double mutants E227Q G147Q and E227Q T425A, both of which most strongly affected adenine transport, and in V145A G147Q, which markedly affected guanine transport. These differences could be explained by docking studies, showing that uracil and guanine exhibit a similar binding mode whereas adenine binds deep into the catalytic pocket of PLUTO. Furthermore, competition studies confirmed these results. The present study defines the molecular determinants for PLUTO substrate binding and demonstrates key differences in structure-function relations between PLUTO and other NCS1 family members. 2014 Witz et al.

  5. Structure-function relationship of a plant NCS1 member--homology modeling and mutagenesis identified residues critical for substrate specificity of PLUTO, a nucleobase transporter from Arabidopsis.

    Science.gov (United States)

    Witz, Sandra; Panwar, Pankaj; Schober, Markus; Deppe, Johannes; Pasha, Farhan Ahmad; Lemieux, M Joanne; Möhlmann, Torsten

    2014-01-01

    Plastidic uracil salvage is essential for plant growth and development. So far, PLUTO, the plastidic nucleobase transporter from Arabidopsis thaliana is the only known uracil importer at the inner plastidic membrane which represents the permeability barrier of this organelle. We present the first homology model of PLUTO, the sole plant NCS1 member from Arabidopsis based on the crystal structure of the benzyl hydantoin transporter MHP1 from Microbacterium liquefaciens and validated by molecular dynamics simulations. Polar side chains of residues Glu-227 and backbones of Val-145, Gly-147 and Thr-425 are proposed to form the binding site for the three PLUTO substrates uracil, adenine and guanine. Mutational analysis and competition studies identified Glu-227 as an important residue for uracil and to a lesser extent for guanine transport. A differential response in substrate transport was apparent with PLUTO double mutants E227Q G147Q and E227Q T425A, both of which most strongly affected adenine transport, and in V145A G147Q, which markedly affected guanine transport. These differences could be explained by docking studies, showing that uracil and guanine exhibit a similar binding mode whereas adenine binds deep into the catalytic pocket of PLUTO. Furthermore, competition studies confirmed these results. The present study defines the molecular determinants for PLUTO substrate binding and demonstrates key differences in structure-function relations between PLUTO and other NCS1 family members. PMID:24621654

  6. Structure-function relationship of a plant NCS1 member--homology modeling and mutagenesis identified residues critical for substrate specificity of PLUTO, a nucleobase transporter from Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Sandra Witz

    Full Text Available Plastidic uracil salvage is essential for plant growth and development. So far, PLUTO, the plastidic nucleobase transporter from Arabidopsis thaliana is the only known uracil importer at the inner plastidic membrane which represents the permeability barrier of this organelle. We present the first homology model of PLUTO, the sole plant NCS1 member from Arabidopsis based on the crystal structure of the benzyl hydantoin transporter MHP1 from Microbacterium liquefaciens and validated by molecular dynamics simulations. Polar side chains of residues Glu-227 and backbones of Val-145, Gly-147 and Thr-425 are proposed to form the binding site for the three PLUTO substrates uracil, adenine and guanine. Mutational analysis and competition studies identified Glu-227 as an important residue for uracil and to a lesser extent for guanine transport. A differential response in substrate transport was apparent with PLUTO double mutants E227Q G147Q and E227Q T425A, both of which most strongly affected adenine transport, and in V145A G147Q, which markedly affected guanine transport. These differences could be explained by docking studies, showing that uracil and guanine exhibit a similar binding mode whereas adenine binds deep into the catalytic pocket of PLUTO. Furthermore, competition studies confirmed these results. The present study defines the molecular determinants for PLUTO substrate binding and demonstrates key differences in structure-function relations between PLUTO and other NCS1 family members.

  7. Transport of antimony salts by Arabidopsis thaliana protoplasts over-expressing the human multidrug resistance-associated protein 1 (MRP1/ABCC1).

    Science.gov (United States)

    Gayet, Landry; Picault, Nathalie; Cazalé, Anne-Claire; Beyly, Audrey; Lucas, Philippe; Jacquet, Hélène; Suso, Henri-Pierre; Vavasseur, Alain; Peltier, Gilles; Forestier, Cyrille

    2006-12-22

    ABC transporters from the multidrug resistance-associated protein (MRP) subfamily are glutathione S-conjugate pumps exhibiting a broad substrate specificity illustrated by numerous xenobiotics, such as anticancer drugs, herbicides, pesticides and heavy metals. The engineering of MRP transporters into plants might be interesting either to reduce the quantity of xenobiotics taken up by the plant in the context of "safe-food" strategies or, conversely, in the development of phytoremediation strategies in which xenobiotics are sequestered in the vacuolar compartment. In this report, we obtained Arabidopsis transgenic plants overexpressing human MRP1. In these plants, expression of MRP1 did not increase plant resistance to antimony salts (Sb(III)), a classical glutathione-conjugate substrate of MRP1. However, the transporter was fully translated in roots and shoots, and targeted to the plasma membrane. In order to investigate the functionality of MRP1 in Arabidopsis, mesophyll cell protoplasts (MCPs) were isolated from transgenic plants and transport activities were measured by using calcein or Sb(III) as substrates. Expression of MRP1 at the plasma membrane was correlated with an increase in the MCPs resistance to Sb(III) and a limitation of the metalloid content in the protoplasts due to an improvement in Sb(III) efflux. Moreover, Sb(III) transport was sensitive to classical inhibitors of the human MRP1, such as MK571 or glibenclamide. These results demonstrate that a human ABC transporter can be functionally introduced in Arabidopsis, which might be useful, with the help of stronger promoters, to reduce the accumulation of xenobiotics in plants, such as heavy metals from multi-contaminated soils.

  8. Sucrose Metabolism in Plastids

    NARCIS (Netherlands)

    Gerrits, N.; Turk, S.C.H.J.; Dun, van K.P.M.; Hulleman, H.D.; Visser, R.G.F.; Weisbeek, P.J.; Smeekens, S.C.M.

    2001-01-01

    The question whether sucrose (Suc) is present inside plastids has been long debated. Low Suc levels were reported to be present inside isolated chloroplasts, but these were argued to be artifacts of the isolation procedures used. We have introduced Suc-metabolizing enzymes in plastids and our experi

  9. A chimeric protein of aluminum-activated malate transporter generated from wheat and Arabidopsis shows enhanced response to trivalent cations.

    Science.gov (United States)

    Sasaki, Takayuki; Tsuchiya, Yoshiyuki; Ariyoshi, Michiyo; Ryan, Peter R; Yamamoto, Yoko

    2016-07-01

    TaALMT1 from wheat (Triticum aestivum) and AtALMT1 from Arabidopsis thaliana encode aluminum (Al)-activated malate transporters, which confer acid-soil tolerance by releasing malate from roots. Chimeric proteins from TaALMT1 and AtALMT1 (Ta::At, At::Ta) were previously analyzed in Xenopus laevis oocytes. Those studies showed that Al could activate malate efflux from the Ta::At chimera but not from At::Ta. Here, functions of TaALMT1, AtALMT1 and the chimeric protein Ta::At were compared in cultured tobacco BY-2 cells. We focused on the sensitivity and specificity of their activation by trivalent cations. The activation of malate efflux by Al was at least two-fold greater in the chimera than the native proteins. All proteins were also activated by lanthanides (erbium, ytterbium, gadolinium, and lanthanum), but the chimera again released more malate than TaALMT1 or AtALMT1. In Xenopus oocytes, Al, ytterbium, and erbium activated inward currents from the native TaALMT1 and the chimeric protein, but gadolinium only activated currents from the chimera. Lanthanum inhibited currents from both proteins. These results demonstrated that function of the chimera protein was altered compared to the native proteins and was more responsive to a range of trivalent cations when expressed in plant cells. PMID:27039280

  10. Complete Sucrose Metabolism Requires Fructose Phosphotransferase Activity in Corynebacterium glutamicum To Ensure Phosphorylation of Liberated Fructose

    OpenAIRE

    Dominguez, H.; Lindley, N. D.

    1996-01-01

    Sucrose uptake by Corynebacterium glutamicum involves a phosphoenolpyruvate-dependent sucrose phosphotransferase (PTS), but in the absence of fructokinase, further metabolism of the liberated fructose requires efflux of the fructose and reassimilation via the fructose PTS. Mutant strains lacking detectable fructose-transporting PTS activity accumulated fructose extracellularly but consumed sucrose at rates comparable to those of the wild-type strain.

  11. Rapid hyperosmotic-induced Ca2+ responses in Arabidopsis thaliana exhibit sensory potentiation and involvement of plastidial KEA transporters.

    Science.gov (United States)

    Stephan, Aaron B; Kunz, Hans-Henning; Yang, Eric; Schroeder, Julian I

    2016-08-30

    Plants experience hyperosmotic stress when faced with saline soils and possibly with drought stress, but it is currently unclear how plant roots perceive this stress in an environment of dynamic water availabilities. Hyperosmotic stress induces a rapid rise in intracellular Ca(2+) concentrations ([Ca(2+)]i) in plants, and this Ca(2+) response may reflect the activities of osmo-sensory components. Here, we find in the reference plant Arabidopsis thaliana that the rapid hyperosmotic-induced Ca(2+) response exhibited enhanced response magnitudes after preexposure to an intermediate hyperosmotic stress. We term this phenomenon "osmo-sensory potentiation." The initial sensing and potentiation occurred in intact plants as well as in roots. Having established a quantitative understanding of wild-type responses, we investigated effects of pharmacological inhibitors and candidate channel/transporter mutants. Quintuple mechano-sensitive channels of small conductance-like (MSL) plasma membrane-targeted channel mutants as well as double mid1-complementing activity (MCA) channel mutants did not affect the response. Interestingly, however, double mutations in the plastid K(+) exchange antiporter (KEA) transporters kea1kea2 and a single mutation that does not visibly affect chloroplast structure, kea3, impaired the rapid hyperosmotic-induced Ca(2+) responses. These mutations did not significantly affect sensory potentiation of the response. These findings suggest that plastids may play an important role in early steps mediating the response to hyperosmotic stimuli. Together, these findings demonstrate that the plant osmo-sensory components necessary to generate rapid osmotic-induced Ca(2+) responses remain responsive under varying osmolarities, endowing plants with the ability to perceive the dynamic intensities of water limitation imposed by osmotic stress. PMID:27528686

  12. Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and 2: fructose and xylitol/H+ symporters in pollen and young xylem cells

    Science.gov (United States)

    Klepek, Yvonne-Simone; Konrad, Kai R.; Wippel, Kathrin; Hoth, Stefan; Hedrich, Rainer; Sauer, Norbert

    2010-01-01

    The genome of Arabidopsis thaliana contains six genes, AtPMT1 to AtPMT6 (Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTER 1–6), which form a distinct subfamily within the large family of more than 50 monosaccharide transporter-like (MST-like) genes. So far, only AtPMT5 [formerly named AtPLT5 (At3g18830)] has been characterized and was shown to be a plasma membrane-localized H+-symporter with broad substrate specificity. The characterization of AtPMT1 (At2g16120) and AtPMT2 (At2g16130), two other, almost identical, members of this transporter subfamily, are presented here. Expression of the AtPMT1 and AtPMT2 cDNAs in baker's yeast (Saccharomyces cerevisiae) revealed that these proteins catalyse the energy-dependent, high-capacity transport of fructose and xylitol, and the transport of several other compounds with lower rates. Expression of their cRNAs in Xenopus laevis oocytes showed that both proteins are voltage-dependent and catalyse the symport of their substrates with protons. Fusions of AtPMT1 or AtPMT2 with the green fluorescent protein (GFP) localized to Arabidopsis plasma membranes. Analyses of reporter genes performed with AtPMT1 or AtPMT2 promoter sequences showed expression in mature (AtPMT2) or germinating (AtPMT1) pollen grains, as well as in growing pollen tubes, hydathodes, and young xylem cells (both genes). The expression was confirmed with an anti-AtPMT1/AtPMT2 antiserum (αAtPMT1/2) raised against peptides conserved in AtPMT1 and AtPMT2. The physiological roles of the proteins are discussed and related to plant cell wall modifications. PMID:19969532

  13. Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and 2: fructose and xylitol/H+ symporters in pollen and young xylem cells.

    Science.gov (United States)

    Klepek, Yvonne-Simone; Volke, Melanie; Konrad, Kai R; Wippel, Kathrin; Hoth, Stefan; Hedrich, Rainer; Sauer, Norbert

    2010-01-01

    The genome of Arabidopsis thaliana contains six genes, AtPMT1 to AtPMT6 (Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTER 1-6), which form a distinct subfamily within the large family of more than 50 monosaccharide transporter-like (MST-like) genes. So far, only AtPMT5 [formerly named AtPLT5 (At3g18830)] has been characterized and was shown to be a plasma membrane-localized H(+)-symporter with broad substrate specificity. The characterization of AtPMT1 (At2g16120) and AtPMT2 (At2g16130), two other, almost identical, members of this transporter subfamily, are presented here. Expression of the AtPMT1 and AtPMT2 cDNAs in baker's yeast (Saccharomyces cerevisiae) revealed that these proteins catalyse the energy-dependent, high-capacity transport of fructose and xylitol, and the transport of several other compounds with lower rates. Expression of their cRNAs in Xenopus laevis oocytes showed that both proteins are voltage-dependent and catalyse the symport of their substrates with protons. Fusions of AtPMT1 or AtPMT2 with the green fluorescent protein (GFP) localized to Arabidopsis plasma membranes. Analyses of reporter genes performed with AtPMT1 or AtPMT2 promoter sequences showed expression in mature (AtPMT2) or germinating (AtPMT1) pollen grains, as well as in growing pollen tubes, hydathodes, and young xylem cells (both genes). The expression was confirmed with an anti-AtPMT1/AtPMT2 antiserum (alphaAtPMT1/2) raised against peptides conserved in AtPMT1 and AtPMT2. The physiological roles of the proteins are discussed and related to plant cell wall modifications. PMID:19969532

  14. Evidence for several cysteine transport mechanisms in the mitochondrial membranes of Arabidopsis thaliana.

    Science.gov (United States)

    Lee, Chun Pong; Wirtz, Markus; Hell, Rüdiger

    2014-01-01

    Cysteine is essential for many mitochondrial processes in plants, including translation, iron-sulfur cluster biogenesis and cyanide detoxification. Its biosynthesis is carried out by serine acetyltransferase (SAT) and O-acetylserine (thiol) lyase (OAS-TL) which can be found in the cytosol, plastids and mitochondria. Mutants lacking one compartment-specific OAS-TL isoform show viable phenotypes, leading to the hypothesis that the organellar membranes are permeable to substrates and products of the cysteine biosynthetic pathway. In this report, we show that exogenouslly supplied [(35)S]cysteine accumulates in the mitochondrial fraction and is taken up into isolated mitochondria for in organello protein synthesis. Analysis of cysteine uptake by isolated mitochondria and mitoplasts indicates that cysteine is transported by multiple facilitated mechanisms that operate in a concentration gradient-dependent manner. In addition, cysteine uptake is dependent mainly on the ΔpH across the inner membrane. The rates of mitochondrial cysteine transport can be mildly altered by specific metabolites in the cyanide detoxification-linked sulfide oxidation, but not by most substrates and products of the cysteine biosynthetic pathway. Based on these results, we propose that the transport of cysteine plays a pivotal role in regulating cellular cysteine biosynthesis as well as modulating the availability of sulfur for mitochondrial metabolism.

  15. Assessment of Sugar Components and Genes Involved in the Regulation of Sucrose Accumulation in Peach Fruit.

    Science.gov (United States)

    Vimolmangkang, Sornkanok; Zheng, Hongyu; Peng, Qian; Jiang, Quan; Wang, Huiliang; Fang, Ting; Liao, Liao; Wang, Lu; He, Huaping; Han, Yuepeng

    2016-09-01

    Soluble sugar contents in mature fruits of 45 peach accessions were quantified using gas chromatography analysis. Sucrose is the predominant sugar in mature fruit, followed by glucose and fructose, which have similar concentrations. Overall, sucrose metabolism and accumulation are crucial determinants of sugar content in peach fruit, and there is a wide range of sucrose concentrations among peach genotypes. To understand the mechanisms regulating sucrose accumulation in peach fruit, expression profiles of genes involved in sucrose metabolism and transport were compared among four genotypes. Two sucrose-cleaving enzyme genes (SUS4 and NINV8), one gene involved in sucrose resynthesis (SPS3), and three sugar transporter genes (SUT2, SUT4, and TMT2) were prevalently expressed in peach fruit, and their expression levels are significantly correlated with sucrose accumulation. In contrast, the VAINV genes responsible for sucrose cleavage in the vacuole were weakly expressed in mature fruit, suggesting that the sucrose-cleaving reaction is not active in the vacuole of sink cells of mature peach fruit. This study suggests that sucrose accumulation in peach fruit involves the coordinated interaction of genes related to sucrose cleavage, resynthesis, and transport, which could be helpful for future peach breeding. PMID:27537219

  16. Sugar (sucrose) holograms

    Science.gov (United States)

    Ponce-Lee, E. L.; Olivares-Pérez, A.; Fuentes-Tapia, I.

    2004-06-01

    Computer holograms made with sugar crystals are reported. This material is well known as a good sweetener; the sugar from sugar cane or sugar beet (sucrose). These sweetener can be applied as honey "water and diluted sugar" easily on any substrate such as plastics or glasses without critical conditions for developed process. This step corresponds only to the cured sucrose as a photopolymer process. The maximum absorption spectra is localized at UV region λ=240 nm. We record with lithographic techniques some gratings, showing a good diffraction efficiency around 45%. This material has good resolution to make diffraction gratings. These properties are attractive because they open the possibility to make phase holograms on candies. Mainly the phase modulation is by refraction index.

  17. Aluminium toxicity targets PIN2 in Arabidopsis root apices: Effects on PIN2 endocytosis, vesicular recycling,and polar auxin transport

    Institute of Scientific and Technical Information of China (English)

    SHEN Hong; HOU NingYan; Markus SCHLICHT; WAN YingLang; Stefano MANCUSO; Frantisek BALUSKA

    2008-01-01

    The most obvious symptom of AI toxicity is the inhibition of root growth.However,the mechanism of AI-inhibiting root growth remains to be elucidated.In this study,auxin transport and vesicle movement of an auxin-efflux carrier (PIN2) were investigated in Arabidopsis roots in response to AI stress.Results indicated that AI inhibited the apical transport of auxin in root tips of Arabidopsis significantly.The severe inhibition was localized in the cells of transition zone,where the concentration of auxin was only 34% that of the control.Brefeldin A (BFA),an inhibitor of vesicle transport,induced the dot-like structure of PIN2 vesicle significantly.Al decreased the size of dot-like structure of PIN2 vesicles.Re-sults of real-time RT-PCR and Western-blotting analysis showed that Al increased the transcript level of PIN2 and the accumulation of PIN2 protein in horizontal direction of plasma membrane,but decreased its distribution in endosomes,suggesting that AI inhibited the transport of PIN2 vesicles from plasma membrane to endosomes.Results of cytoskeleton-depolymering drugs indicated that it was via the pathway of disruption of actin microfilaments that AI inhibited the transport of PIN2 vesicles.Exposed to AI stress,the cells of elongation zone had less AI uptake and less transport frequency of vesicles than cells of transition zone.Taken together,our results suggested that AI inhibited root growth mainly by modulating the transport of PIN2 vesicles between plasma membrane and endosomes,thus block-ing auxin transport and root growth.

  18. Arabidopsis TT19 Functions as a Carrier to Transport Anthocyanin from the Cytosol to Tonoplasts

    Institute of Scientific and Technical Information of China (English)

    Yi Sun; Hong Li; Ji-Rong Huang

    2012-01-01

    Anthocyanins are synthesized in the cytosolic surface of the endoplasmic reticulum (ER) but dominantly accumulate in the vacuole.Little is known about how anthocyanins are transported from the ER to the vacuole.Here,we provide evidence supporting that Transparent Testa 19 (TT19),a glutathione S-transferase (GST),functions as a carrier to transport cyanidin and/or anthocyanins to the tonoplast.We identified a novel tt19 mutant (tt19-7),which barely accumulates anthocyanins but produces a 36% higher level of flavonol than the wild-type (WT),from ethyl methanesulfonate mutagenized seeds.Expressing TT19-fused green fluorescence protein (GFP) in tt19-7 rescues the mutant phenotype in defective anthocyanin biosynthesis,indicating that TT19-GFP is functional.We further showed that TT19-GFP is localized not only in the cytoplasm and nuclei,but also on the tonoplast.The membrane localization of TT19-GFP was further ascertained by immunoblot analysis.In vitro assay showed that the purified recombinant TT19 increases water solubility of cyanidin (Cya) and cyanidin-3-O-glycoside (C3G).Compared with C3G,Cya can dramatically quench the intrinsic tryptophan fluorescence of TT19 to much lower levels,indicating a higher affinity of TT19 to Cya than to C3G.Isothermal titration calorimetry analysis also confirmed physical interaction between TT19 and C3G.Taken together,our data reveal molecular mechanism underlying TT19-mediated anthocyanin transportation.

  19. Single-particle analysis reveals shutoff control of the Arabidopsis ammonium transporter AMT1;3 by clustering and internalization.

    Science.gov (United States)

    Wang, Qinli; Zhao, Yuanyuan; Luo, Wangxi; Li, Ruili; He, Qihua; Fang, Xiaohong; Michele, Roberto De; Ast, Cindy; von Wirén, Nicolaus; Lin, Jinxing

    2013-08-01

    Ammonium is a preferred source of nitrogen for plants but is toxic at high levels. Plant ammonium transporters (AMTs) play an essential role in NH4(+) uptake, but the mechanism by which AMTs are regulated remains unclear. To study how AMTs are regulated in the presence of ammonium, we used variable-angle total internal reflection fluorescence microscopy and fluorescence cross-correlation spectroscopy for single-particle fluorescence imaging of EGFP-tagged AMT1;3 on the plasma membrane of Arabidopsis root cells at various ammonium levels. We demonstrated that AMT1;3-EGFP dynamically appeared and disappeared on the plasma membrane as moving fluorescent spots in low oligomeric states under N-deprived and N-sufficient conditions. Under external high-ammonium stress, however, AMT1;3-EGFPs were found to amass into clusters, which were then internalized into the cytoplasm. A similar phenomenon also occurred in the glutamine synthetase mutant gln1;2 background. Single-particle analysis of AMT1;3-EGFPs in the clathrin heavy chain 2 mutant (chc2 mutant) and Flotllin1 artificial microRNA (Flot1 amiRNA) backgrounds, together with chemical inhibitor treatments, demonstrated that the endocytosis of AMT1;3 clusters induced by high-ammonium stress could occur mainly through clathrin-mediated endocytic pathways, but the contribution of microdomain-associated endocytic pathway cannot be excluded in the internalization. Our results revealed that the clustering and endocytosis of AMT1;3 provides an effective mechanism by which plant cells can avoid accumulation of toxic levels of ammonium by eliminating active AMT1;3 from the plasma membrane.

  20. Sucrose metabolism gene families and their biological functions.

    Science.gov (United States)

    Jiang, Shu-Ye; Chi, Yun-Hua; Wang, Ji-Zhou; Zhou, Jun-Xia; Cheng, Yan-Song; Zhang, Bao-Lan; Ma, Ali; Vanitha, Jeevanandam; Ramachandran, Srinivasan

    2015-11-30

    Sucrose, as the main product of photosynthesis, plays crucial roles in plant development. Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions. We focused on four sucrose metabolism related gene families including sucrose synthase, sucrose phosphate synthase, sucrose phosphate phosphatase and UDP-glucose pyrophosphorylase. These gene families exhibited different expansion and evolutionary history as their host genomes experienced differentiated rates of the whole genome duplication, tandem and segmental duplication, or mobile element mediated gene gain and loss. They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion. However, we have detected recent positive selection during intra-species divergence. Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses. Our studies uncovered the molecular mechanisms of gene expansion and evolution and also provided new insight into the role of positive selection in intra-species divergence. Overexpression data revealed novel biological functions of these genes in flowering time control and seed germination under normal and stress conditions.

  1. Proof of concept for a novel functional screening system for plant sucrose effluxers

    Directory of Open Access Journals (Sweden)

    Yuchan Zhou

    2014-07-01

    Full Text Available Membrane transporters play pivotal roles in facilitating sucrose transport in plants and their activities have been shown to impact plant growth rates and crop yield.  In contrast to the well-defined mechanism of sucrose influx across plasma membranes, less is known about sucrose efflux mechanisms and the membrane proteins supporting this function.  A major impediment blocking progress in this key area of plant science is the absence of a functional screening system for genes encoding sucrose effluxers.  Here we report a novel yeast system for screening sucrose effluxers based on sucrose release from yeast cells genetically modified to synthesize, but not to metabolize, sucrose.  Inhibiting sucrose metabolism was achieved using yeast strains, SEY 6210 and YSL4-6, carrying mutations in genes encoding invertase and maltase, respectively.  Genes encoding essential components of sucrose biosynthesis, sucrose phosphate synthase (SPS and sucrose phosphate phosphatase (SPP, were used to transform the two yeast hosts to make strains SuPy (from SEY6210 and Ysu (from YSL4-6.  Cultures of SuPy15 cells were found to be capable of synthesizing sucrose when supplied with various compounds as the sole carbon source, including non-fermentable sugars and non-sugar substrates.  A proof of concept of the screening system was demonstrated by transforming SuPy15 with sucrose transporter genes known to encode plasma membrane proteins that mediate sucrose efflux.  The robustness of the yeast SuPy15 system as a novel platform to screen putative plant sucrose effluxers is discussed.

  2. Synthesis of insecticidal sucrose esters

    Institute of Scientific and Technical Information of China (English)

    Song Zi-juan; Li Shu-jun; Chen Xi; Liu Li-mei; Song Zhan-qian

    2006-01-01

    Some synthetic sucrose esters (SE) are a relatively new class of insecticidal compounds produced by reacting sugars with fatty acids, which are safe for the environment. Especially, sucrose esters composed of C6-C12 fatty acids have desirable insecticidal properties against many soft-bodied arthropod pests. In our study, sucrose octanoate which has the highest activity against a range of arthropod species was synthesized by a trans-esterification method and proved its insecticidal property. Under the condition of a homogeneous liquid, sucrose octanoate was prepared by reacting ethyl octanoate with sucrose at reduced pressure; the yield was 79.11%. Sucrose octanoate synthesized was identified and its property analyzed by IR, TLC and spectrophotometric analysis. It was shown that the ratio of monoester to polyester in sucrose octanoate was 1.48:1. The insecticidal activity of the synthetic sucrose octanoate was evaluated at a concentration of 4 and 8 mg·mL-1. The mortality of first-instar larvae ofLymantria dispar from its contact toxicity was 72.5% after 36 hours, the revision insect reduced rate of Aphis glycines reached above 80% at 4 and 8 mg·mL-1 after being treated for 5 days. Since the SE products are nontoxic to humans and higher animals, fully biodegradable and hydrolyzed to readily metabolizable sucrose and fatty acid, they are not harmful to crops and appear to be good insecticide candidates.

  3. Switching the mode of sucrose utilization by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Miletti Luiz C

    2008-02-01

    Full Text Available Abstract Background Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. Results We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Conclusion Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by

  4. Sucrose accumulation in mature sweet melon fruits

    International Nuclear Information System (INIS)

    Mesocarp tissue from sucrose-accumulating sweet melon (Cucumis melo cv. Galia) showed sucrose synthase activity (ca 1 nkat/gfw) while soluble acid invertase and sucrose phosphate synthase activities were not observed. Sucrose uptake into mesocarp discs was linear with sucrose concentration (1-500 mM) and unaffected by PCMBS and CCCP. Sucrose compartmentation into the vacuole also increased linearly with sucrose concentration as indicated by compartmental efflux kinetics. Mesocarp discs incubated in 14C-fructose + UDP-glu synthesized 14C-sucrose and efflux kinetics indicated that the 14C-sucrose was compartmentalized. These data support the hypothesis that two mechanisms are involved in sucrose accumulation in sweet melon: (1) compartmentation of intact sucrose and (2) synthesis of sucrose via sucrose synthase and subsequent compartmentation in the vacuole

  5. MzPIP2;1: An Aquaporin Involved in Radial Water Movement in Both Water Uptake and Transportation, Altered the Drought and Salt Tolerance of Transgenic Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Lin Wang

    Full Text Available Plants are unavoidably subjected to various abiotic stressors, including high salinity, drought and low temperature, which results in water deficit and even death. Water uptake and transportation play a critical role in response to these stresses. Many aquaporin proteins, localized at different tissues, function in various transmembrane water movements. We targeted at the key aquaporin in charge of both water uptake in roots and radial water transportation from vascular tissues through the whole plant.The MzPIP2;1 gene encoding a plasma membrane intrinsic protein was cloned from salt-tolerant apple rootstock Malus zumi Mats. The GUS gene was driven by MzPIP2;1 promoter in transgenic Arabidopsis. It indicated that MzPIP2;1 might function in the epidermal and vascular cells of roots, parenchyma cells around vessels through the stems and vascular tissues of leaves. The ectopically expressed MzPIP2;1 conferred the transgenic Arabidopsis plants enhanced tolerance to slight salt and drought stresses, but sensitive to moderate salt stress, which was indicated by root length, lateral root number, fresh weight and K+/Na+ ratio. In addition, the possible key cis-elements in response to salt, drought and cold stresses were isolated by the promoter deletion experiment.The MzPIP2;1 protein, as a PIP2 aquaporins subgroup member, involved in radial water movement, controls water absorption and usage efficiency and alters transgenic plants drought and salt tolerance.

  6. The transparent testa4 mutation prevents flavonoid synthesis and alters auxin transport and the response of Arabidopsis roots to gravity and light.

    Science.gov (United States)

    Buer, Charles S; Muday, Gloria K

    2004-05-01

    We examined whether flavonoids act as endogenous auxin transport regulators during gravity vector and light intensity changes in Arabidopsis thaliana roots. Flavonoid deficient transparent testa4 [tt4(2YY6)] seedlings had elevated root basipetal auxin transport compared with the wild type, consistent with the absence of a negative auxin transport regulator. The tt4(2YY6) roots had delayed gravitropism that was chemically complemented with a flavonoid intermediate. Flavonoid accumulation was found in wild-type columella cells, the site of gravity perception, and in epidermal and cortical cells, the site of differential growth, but flavonoid accumulation was absent in tt4(2YY6) roots. Flavonoid accumulation was higher in gravity-stimulated root tips as compared with vertical controls, with maximum differences coinciding with the timing of gravitropic bending, and was located in epidermal cells. Exogenous indole-3-acetic acid (IAA) also elevated flavonoid accumulation, suggesting that flavonoid changes in response to gravity might be partly as a result of changing IAA distribution. Acropetal IAA transport was also elevated in roots of tt4(2YY6). Flavonoid synthesis was repressed in the dark, as were differences in root acropetal transport in tt4(2YY6). These results are consistent with light- and gravity-induced flavonoid stimulation that alters auxin transport in roots and dependent physiological processes, including gravitropic bending and root development.

  7. Vacuolar invertase regulates elongation of Arabidopsis thaliana roots as revealed by QTL and mutant analysis.

    NARCIS (Netherlands)

    Sergeeva, L.I.; Keurentjes, J.J.B.; Bentsink, L.; Vonk, J.; Plas, van der L.H.W.; Koornneef, M.; Vreugdenhil, D.

    2006-01-01

    The possible role of the sucrose-splitting enzymes sucrose synthase and invertase in elongating roots and hypocotyls of Arabidopsis was tested by using a combination of histochemical methods and quantitative trait locus (QTL) analysis. Lengths of roots and hypocotyls correlated better with invertase

  8. Copper-induced alteration in sucrose partitioning and its relationship to the root growth of two Elsholtzia haichowensis Sun populations.

    Science.gov (United States)

    Li, Min-Jing; Xiong, Zhi-Ting; Liu, Hui; Kuo, Yi-Ming; Tong, Lei

    2016-10-01

    Hydroponic culture was used to comparatively investigate the copper (Cu)-induced alteration to sucrose metabolism and biomass allocation in two Elsholtzia haichowensis Sun populations with one from a Cu-contaminated site (CS) and the other from a non-contaminated site (NCS). Experimental results revealed that biomass allocation preferred roots over shoots in CS population, and shoots over roots in NCS population under Cu exposure. The difference in biomass allocation was correlated with the difference in sucrose partitioning between the two populations. Cu treatment (45 μM) significantly decreased leaf sucrose content and increased root sucrose content in CS population as a result of the increased activities of leaf sucrose synthesis enzymes (sucrose phosphate synthetase and sucrose synthase) and root sucrose cleavage enzyme (vacuolar invertase), which led to increased sucrose transport from leaves to roots. In contrast, higher Cu treatment increased sucrose content in leaves and decreased sucrose content in roots in NCS population as a result of the decreased activities of root sucrose cleavage enzymes (vacuolar and cell wall invertases) that led to less sucrose transport from leaves to roots. These results provide important insights into carbon resource partitioning and biomass allocation strategies in metallophytes and are beneficial for the implementation of phytoremediation techniques. PMID:27153457

  9. HY5 regulates nitrite reductase 1 (NIR1) and ammonium transporter1;2 (AMT1;2) in Arabidopsis seedlings.

    Science.gov (United States)

    Huang, Lifen; Zhang, Hongcheng; Zhang, Huiyong; Deng, Xing Wang; Wei, Ning

    2015-09-01

    HY5 (Long Hypocotyles 5) is a key transcription factor in Arabidopsis thaliana that has a pivotal role in seedling development. Soil nitrogen is an essential macronutrient, and its uptake, assimilation and metabolism are influenced by nutrient availability and by lights. To understand the role of HY5 in nitrogen assimilation pathways, we examined the phenotype as well as the expression of selected nitrogen assimilation-related genes in hy5 mutant grown under various nitrogen limiting and nitrogen sufficient conditions, or different light conditions. We report that HY5 positively regulates nitrite reductase gene NIR1 and negatively regulates the ammonium transporter gene AMT1;2 under all nitrogen and light conditions tested, while it affects several other genes in a nitrogen supply-dependent manner. HY5 is not required for light induction of NIR1, AMT1;2 and NIA genes, but it is necessary for high level expression of NIR1 and NIA under optimal nutrient and light conditions. In addition, nitrogen deficiency exacerbates the abnormal root system of hy5. Together, our results suggest that HY5 exhibits the growth-promoting activity only when sufficient nutrients, including lights, are provided, and that HY5 has a complex involvement in nitrogen acquisition and metabolism in Arabidopsis seedlings. PMID:26259199

  10. Vacuolar invertase regulates elongation of Arabidopsis thaliana roots as revealed by QTL and mutant analysis

    OpenAIRE

    Sergeeva, L.I.; Keurentjes, J. J. B.; Bentsink, L.; Vonk, J.; Plas, van der, M..; Koornneef, M; Vreugdenhil, D.

    2006-01-01

    The possible role of the sucrose-splitting enzymes sucrose synthase and invertase in elongating roots and hypocotyls of Arabidopsis was tested by using a combination of histochemical methods and quantitative trait locus (QTL) analysis. Lengths of roots and hypocotyls correlated better with invertase activities than with sucrose synthase activities. The highest correlations were observed with activities in the elongating zones of roots. The genetic basis of these correlations was studied by us...

  11. Arabidopsis CDS blastp result: AK064342 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064342 002-107-H07 At5g58270.1 mitochondrial half-ABC transporter (STA1) identical to half...-molecule ABC transporter ATM3 GI:9964121 from [Arabidopsis thaliana]; almost identical to mitochondrial half...-ABC transporter STA1 GI:9187883 from [Arabidopsis thaliana]; identical to cDNA mitochondrial half-ABC transporter (STA1 gene)GI:9187882 0.0 ...

  12. Arabidopsis CDS blastp result: AK287662 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287662 J065112L10 At5g58270.1 68418.m07295 mitochondrial half-ABC transporter (STA1) identical to half...-molecule ABC transporter ATM3 GI:9964121 from [Arabidopsis thaliana]; almost identical to mitochondrial half...-ABC transporter STA1 GI:9187883 from [Arabidopsis thaliana]; identical to cDNA mitochondrial half-ABC transporter (STA1 gene)GI:9187882 1e-65 ...

  13. Arabidopsis CDS blastp result: AK242094 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242094 J075142E09 At5g58270.1 68418.m07295 mitochondrial half-ABC transporter (STA1) identical to half...-molecule ABC transporter ATM3 GI:9964121 from [Arabidopsis thaliana]; almost identical to mitochondrial half...-ABC transporter STA1 GI:9187883 from [Arabidopsis thaliana]; identical to cDNA mitochondrial half-ABC transporter (STA1 gene)GI:9187882 2e-33 ...

  14. Arabidopsis CDS blastp result: AK102879 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102879 J033112G11 At5g58270.1 mitochondrial half-ABC transporter (STA1) identical to half...-molecule ABC transporter ATM3 GI:9964121 from [Arabidopsis thaliana]; almost identical to mitochondrial half...-ABC transporter STA1 GI:9187883 from [Arabidopsis thaliana]; identical to cDNA mitochondrial half-ABC transporter (STA1 gene)GI:9187882 1e-122 ...

  15. Arabidopsis CDS blastp result: AK287488 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287488 J043029O04 At5g58270.1 68418.m07295 mitochondrial half-ABC transporter (STA1) identical to half...-molecule ABC transporter ATM3 GI:9964121 from [Arabidopsis thaliana]; almost identical to mitochondrial half...-ABC transporter STA1 GI:9187883 from [Arabidopsis thaliana]; identical to cDNA mitochondrial half-ABC transporter (STA1 gene)GI:9187882 4e-27 ...

  16. 27 CFR 21.131 - Sucrose octaacetate.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Sucrose octaacetate. 21....131 Sucrose octaacetate. (a) Sucrose octaacetate is an organic acetylation product occurring as a.... Sucrose octaacetate 98 percent minimum by weight when determined by the following procedure: Transfer...

  17. 21 CFR 172.869 - Sucrose oligoesters.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sucrose oligoesters. 172.869 Section 172.869 Food... Multipurpose Additives § 172.869 Sucrose oligoesters. Sucrose oligoesters, as identified in this section, may be safely used in accordance with the following conditions: (a) Sucrose oligoesters consist...

  18. Sucrose release from polysaccharide gels.

    Science.gov (United States)

    Nishinari, Katsuyoshi; Fang, Yapeng

    2016-05-18

    Sucrose release from polysaccharide gels has been studied extensively because it is expected to be useful in understanding flavour release from solid foods and to find a new processing method which produces more palatable and healthier foods. We provide an overview of the release of sucrose and other sugars from gels of agar and related polysaccharides. The addition of sucrose to agar solutions leads to the increase in transparency of the resulting gels and the decrease in syneresis, which is attributed to the decrease in mesh size in gels. The syneresis occurring in the quiescent condition and fluid release induced by compression is discussed. The relationship between the sugar release and the structural, rheological and thermal properties of gels is also discussed. Finally, the future research direction is proposed.

  19. A novel sucrose/H+ symport system and an intracellular sucrase in Leishmania donovani.

    Science.gov (United States)

    Singh, Arpita; Mandal, Debjani

    2011-07-01

    The flagellated form of pathogenic parasitic protozoa Leishmania, resides in the alimentary tract of its sandfly vector, where sucrose serves as a major nutrient source. In this study we report the presence of a sucrose transport system in Leishmania donovani promastigotes. The kinetics of sucrose uptake in promastigotes are biphasic in nature with both high affinity K(m) (K(m) of ∼ 75 μM) and low affinity K(m) (K(m)∼ 1.38 mM) components. By contrast the virulent amastigotes take up sucrose via a low affinity process with a K(m) of 2.5mM. The transport of sucrose into promastigotes leads to rapid intracellular acidification, as indicated by changes in the fluorescence of the pH indicator 2',7'-bis-(2-carboxyethyl)-5-(6) Carboxyfluorescein (BCECF). In experiments with right side-out plasma membrane vesicles derived from L. donovani promastigotes, an artificial pH gradient was able to drive the active accumulation of sucrose. These data are consistent with the operation of a H(+)-sucrose symporter. The symporter was shown to be independent of Na(+) and to be insensitive to cytochalasin B, to the flavonoid phloretin and to the Na(+)/K(+) ATPase inhibitor ouabain. However, the protonophore carbonylcyanide P- (trifluromethoxy) phenylhydrazone (FCCP) and a number of thiol reagents caused significant inhibition of sucrose uptake. Evidence was also obtained for the presence of a stable intracellular pool of the sucrose splitting enzyme, sucrase, in promastigote stage parasites. The results are consistent with the hypothesis that L. donovani promastigotes take up sucrose via a novel H(+)-sucrose symport system and that, on entering the cell, the sucrose is hydrolysed to its component monosaccharides by an intracellular sucrase, thereby providing an energy source for the parasites. PMID:21515279

  20. A Bacterial Glucanotransferase Can Replace the Complex Maltose Metabolism Required for Starch to Sucrose Conversion in Leaves at Night

    DEFF Research Database (Denmark)

    Ruzanski, Christian; Smirnova, Julia; Rejzek, Martin;

    2013-01-01

    Controlled conversion of leaf starch to sucrose at night is essential for the normal growth of Arabidopsis. The conversion involves the cytosolic metabolism of maltose to hexose phosphates via an unusual, multidomain protein with 4-glucanotransferase activity, DPE2, believed to transfer glucosyl...

  1. Cadmium-inducible expression of the ABC-type transporter AtABCC3 increases phytochelatin-mediated cadmium tolerance in Arabidopsis.

    Science.gov (United States)

    Brunetti, Patrizia; Zanella, Letizia; De Paolis, Angelo; Di Litta, Davide; Cecchetti, Valentina; Falasca, Giuseppina; Barbieri, Maurizio; Altamura, Maria Maddalena; Costantino, Paolo; Cardarelli, Maura

    2015-07-01

    The heavy metal cadmium (Cd) is a widespread environmental contaminant with harmful effects on living cells. In plants, phytochelatin (PC)-dependent Cd detoxification requires that PC-Cd complexes are transported into vacuoles. Here, it is shown that Arabidopsis thaliana seedlings defective in the ABCC transporter AtABCC3 (abcc3) have an increased sensitivity to different Cd concentrations, and that seedlings overexpressing AtABCC3 (AtABCC3ox) have an increased Cd tolerance. The cellular distribution of Cd was analysed in protoplasts from abcc3 mutants and AtABCC3 overexpressors grown in the presence of Cd, by means of the Cd-specific fluorochromes 5-nitrobenzothiazole coumarin (BTC-5N) and Leadmium™ Green AM dye. This analysis revealed that Cd is mostly localized in the cytosol of abcc3 mutant protoplasts whereas there is an increase in vacuolar Cd in protoplasts from AtABCC3ox plants. Overexpression of AtABCC3 in cad1-3 mutant seedlings defective in PC production and in plants treated with l-buthionine sulphoximine (BSO), an inhibitor of PC biosynthesis, had no effect on Cd tolerance, suggesting that AtABCC3 acts via PCs. In addition, overexpression of AtABCC3 in atabcc1 atabcc2 mutant seedlings defective in the Cd transporters AtABCC1 and AtABCC2 complements the Cd sensitivity of double mutants, but not in the presence of BSO. Accordingly, the level of AtABCC3 transcript in wild type seedlings was lower than that of AtABCC1 and AtABCC2 in the absence of Cd but higher after Cd exposure, and even higher in atabcc1 atabcc2 mutants. The results point to AtABCC3 as a transporter of PC-Cd complexes, and suggest that its activity is regulated by Cd and is co-ordinated with the activity of AtABCC1/AtABCC2.

  2. Effects of molybdenum deficiency and defects in molybdate transporter MOT1 on transcript accumulation and nitrogen/sulphur metabolism in Arabidopsis thaliana.

    Science.gov (United States)

    Ide, Yoko; Kusano, Miyako; Oikawa, Akira; Fukushima, Atsushi; Tomatsu, Hajime; Saito, Kazuki; Hirai, Masami Yokota; Fujiwara, Toru

    2011-02-01

    Molybdenum (Mo) is a micronutrient essential for plant growth, as several key enzymes of plant metabolic pathways contain Mo cofactor in their catalytic centres. Mo-containing oxidoreductases include nitrate reductase, sulphite oxidase, xanthine dehydrogenase, and aldehyde oxidase. These are involved in nitrate assimilation, sulphite detoxification, purine metabolism or the synthesis of abscisic acid, auxin and glucosinolates in plants. To understand the effects of Mo deficiency and a mutation in a molybdate transporter, MOT1, on nitrogen and sulphur metabolism in Arabidopsis thaliana, transcript and metabolite profiling of the mutant lacking MOT1 was conducted in the presence or absence of Mo. Transcriptome analysis revealed that Mo deficiency had impacts on genes involved in metabolisms, transport, stress responses, and signal transductions. The transcript level of a nitrate reductase NR1 was highly induced under Mo deficiency in mot1-1. The metabolite profiles were analysed further by using gas chromatography time-of-flight mass spectrometry, capillary electrophoresis time-of-flight mass spectrometry, and ultra high performance liquid chromatography. The levels of amino acids, sugars, organic acids, and purine metabolites were altered significantly in the Mo-deficient plants. These results are the first investigation of the global effect of Mo nutrition and MOT1 on plant gene expressions and metabolism. PMID:21131548

  3. CHX14 is a plasma membrane K-efflux transporter that regulates K+ redistribution in "Arabidopsis thaliana"

    Science.gov (United States)

    Potassium (K(+)) is essential for plant growth and development, yet the molecular identity of many K(+) transporters remains elusive. Here we characterized cation/H(+) exchanger (CHX) 14 as a plasma membrane K(+) transporter. "CHX14" expression was induced by elevated K(+) and histochemical analysis...

  4. Sucrose compared with artificial sweeteners

    DEFF Research Database (Denmark)

    Sørensen, Lone Brinkmann; Vasilaras, Tatjana H; Astrup, Arne;

    2014-01-01

    There is a lack of appetite studies in free-living subjects supplying the habitual diet with either sucrose or artificially sweetened beverages and foods. Furthermore, the focus of artificial sweeteners has only been on the energy intake (EI) side of the energy-balance equation. The data are from...

  5. The Arabidopsis nitrate transporter NRT2.5 plays a role in nitrate acquisition and remobilization in nitrogen-starved plants.

    Science.gov (United States)

    Lezhneva, Lina; Kiba, Takatoshi; Feria-Bourrellier, Ana-Belen; Lafouge, Florence; Boutet-Mercey, Stéphanie; Zoufan, Parzhak; Sakakibara, Hitoshi; Daniel-Vedele, Françoise; Krapp, Anne

    2014-10-01

    Nitrogen is a key mineral nutrient playing a crucial role in plant growth and development. Understanding the mechanisms of nitrate uptake from the soil and distribution through the plant in response to nitrogen starvation is an important step on the way to improve nitrogen uptake and utilization efficiency for better growth and productivity of plants, and to prevent negative effects of nitrogen fertilizers on the environment and human health. In this study, we show that Arabidopsis NITRATE TRANSPORTER 2.5 (NRT2.5) is a plasma membrane-localized high-affinity nitrate transporter playing an essential role in adult plants under severe nitrogen starvation. NRT2.5 expression is induced under nitrogen starvation and NRT2.5 becomes the most abundant transcript amongst the seven NRT2 family members in shoots and roots of adult plants after long-term starvation. GUS reporter analyses showed that NRT2.5 is expressed in the epidermis and the cortex of roots at the root hair zone and in minor veins of mature leaves. Reduction of NRT2.5 expression resulted in a decrease in high-affinity nitrate uptake without impacting low-affinity uptake. In the background of the high-affinity nitrate transporter mutant nrt2.4, an nrt2.5 mutation reduced nitrate levels in the phloem of N-starved plants further than in the single nrt2.4 mutants. Growth analyses of multiple mutants between NRT2.1, NRT2.2, NRT2.4, and NRT2.5 revealed that NRT2.5 is required to support growth of nitrogen-starved adult plants by ensuring the efficient uptake of nitrate collectively with NRT2.1, NRT2.2 and NRT2.4 and by taking part in nitrate loading into the phloem during nitrate remobilization. PMID:25065551

  6. The impact of the absence of aliphatic glucosinolates on water transport under salt stress in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Mcarmen eMartinez-Ballesta

    2015-07-01

    Full Text Available Members of the Brassicaceae are known for their contents of nutrients and health-promoting phytochemicals, including glucosinolatesExposure to salinity increases the levels of several of these compounds, but their role in abiotic stress response is unclear. The effect of aliphatic glucosinolates on plant water balance and growth under salt stress, involving aquaporins, was investigated by means of Arabidopsis thaliana mutants impaired in aliphatic glucosinolate biosynthesis, which is controlled by two transcription factors: Myb28 and Myb29. The double mutant myb28myb29, completely lacking aliphatic glucosinolates, was compared to wild type Col-0 (WT and the single mutant myb28. A greater reduction in the hydraulic conductivity of myb28myb29 was observed under salt stress, when compared to the WT and myb28; this correlated with the abundance of both PIP1 and PIP2 aquaporin subfamilies. Also, changes in root architecture in response to salinity were genotype dependent. Treatment with NaCl altered glucosinolates biosynthesis in a similar way in WT and the single mutant and differently in the double mutant. The results indicate that short-chain aliphatic glucosinolates may contribute to water saving under salt stress

  7. Arabidopsis ABCB21 is a facultative auxin importer/exporter regulated by cytoplasmic auxin concentration.

    Science.gov (United States)

    Kamimoto, Yoshihisa; Terasaka, Kazuyoshi; Hamamoto, Masafumi; Takanashi, Kojiro; Fukuda, Shoju; Shitan, Nobukazu; Sugiyama, Akifumi; Suzuki, Hideyuki; Shibata, Daisuke; Wang, Bangjun; Pollmann, Stephan; Geisler, Markus; Yazaki, Kazufumi

    2012-12-01

    The phytohormone auxin is critical for plant growth and many developmental processes. Members of the P-glycoprotein (PGP/ABCB) subfamily of ATP-binding cassette (ABC) transporters have been shown to function in the polar movement of auxin by transporting auxin over the plasma membrane in both monocots and dicots. Here, we characterize a new Arabidopsis member of the ABCB subfamily, ABCB21/PGP21, a close homolog of ABCB4, for which conflicting transport directionalities have been reported. ABCB21 is strongly expressed in the abaxial side of cotyledons and in junctions of lateral organs in the aerial part, whereas in roots it is specifically expressed in pericycle cells. Membrane fractionation by sucrose density gradient centrifugation followed by Western blot showed that ABCB21 is a plasma membrane-localized ABC transporter. A transport assay with Arabidopsis protoplasts suggested that ABCB21 was involved in IAA transport in an outward direction, while naphthalene acetic acid (NAA) was a less preferable substrate for ABCB21. Further functional analysis of ABCB21 using yeast import and export assays showed that ABCB21 mediates the 1-N-naphthylphthalamic acid (NPA)-sensitive translocation of auxin in an inward direction when the cytoplasmic IAA concentration is low, whereas this transporter mediates outward transport under high internal IAA. An increase in the cytoplasmic IAA concentration by pre-loading of IAA into yeast cells abolished the IAA uptake activity by ABCB21 as well as ABCB4. These findings suggest that ABCB21 functions as a facultative importer/exporter controlling auxin concentrations in plant cells.

  8. 21 CFR 184.1854 - Sucrose.

    Science.gov (United States)

    2010-04-01

    ... Substances Affirmed as GRAS § 184.1854 Sucrose. (a) Sucrose (C12H22O11, CAS Reg. No. 57-50-11-1) sugar, cane sugar, or beet sugar is the chemical β-D-fructofuranosyl-α-D-glucopyranoside. Sucrose is obtained by... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sucrose. 184.1854 Section 184.1854 Food and...

  9. Arabidopsis CDS blastp result: AK288065 [KOME

    Lifescience Database Archive (English)

    Full Text Available al to sulfate tansporter Sultr1;3 [Arabidopsis thaliana] GI:10716805; contains Pfam profile PF00916: Sulfate... transporter family; contains Pfam profile PF01740: STAS domain; contains TIGRfam profile TIGR00815: sulfate permease 1e-145 ...

  10. The signal transducer NPH3 integrates the phototropin1 photosensor with PIN2-based polar auxin transport in Arabidopsis root phototropism.

    Science.gov (United States)

    Wan, Yinglang; Jasik, Jan; Wang, Li; Hao, Huaiqing; Volkmann, Dieter; Menzel, Diedrik; Mancuso, Stefano; Baluška, František; Lin, Jinxing

    2012-02-01

    Under blue light (BL) illumination, Arabidopsis thaliana roots grow away from the light source, showing a negative phototropic response. However, the mechanism of root phototropism is still unclear. Using a noninvasive microelectrode system, we showed that the BL sensor phototropin1 (phot1), the signal transducer NONPHOTOTROPIC HYPOCOTYL3 (NPH3), and the auxin efflux transporter PIN2 were essential for BL-induced auxin flux in the root apex transition zone. We also found that PIN2-green fluorescent protein (GFP) localized to vacuole-like compartments (VLCs) in dark-grown root epidermal and cortical cells, and phot1/NPH3 mediated a BL-initiated pathway that caused PIN2 redistribution to the plasma membrane. When dark-grown roots were exposed to brefeldin A (BFA), PIN2-GFP remained in VLCs in darkness, and BL caused PIN2-GFP disappearance from VLCs and induced PIN2-GFP-FM4-64 colocalization within enlarged compartments. In the nph3 mutant, both dark and BL BFA treatments caused the disappearance of PIN2-GFP from VLCs. However, in the phot1 mutant, PIN2-GFP remained within VLCs under both dark and BL BFA treatments, suggesting that phot1 and NPH3 play different roles in PIN2 localization. In conclusion, BL-induced root phototropism is based on the phot1/NPH3 signaling pathway, which stimulates the shootward auxin flux by modifying the subcellular targeting of PIN2 in the root apex transition zone. PMID:22374399

  11. Suppression of NDA-type alternative mitochondrial NAD(P)H dehydrogenases in arabidopsis thaliana modifies growth and metabolism, but not high light stimulation of mitochondrial electron transport.

    Science.gov (United States)

    Wallström, Sabá V; Florez-Sarasa, Igor; Araújo, Wagner L; Escobar, Matthew A; Geisler, Daniela A; Aidemark, Mari; Lager, Ida; Fernie, Alisdair R; Ribas-Carbó, Miquel; Rasmusson, Allan G

    2014-05-01

    The plant respiratory chain contains several pathways which bypass the energy-conserving electron transport complexes I, III and IV. These energy bypasses, including type II NAD(P)H dehydrogenases and the alternative oxidase (AOX), may have a role in redox stabilization and regulation, but current evidence is inconclusive. Using RNA interference, we generated Arabidopsis thaliana plants simultaneously suppressing the type II NAD(P)H dehydrogenase genes NDA1 and NDA2. Leaf mitochondria contained substantially reduced levels of both proteins. In sterile culture in the light, the transgenic lines displayed a slow growth phenotype, which was more severe when the complex I inhibitor rotenone was present. Slower growth was also observed in soil. In rosette leaves, a higher NAD(P)H/NAD(P)⁺ ratio and elevated levels of lactate relative to sugars and citric acid cycle metabolites were observed. However, photosynthetic performance was unaffected and microarray analyses indicated few transcriptional changes. A high light treatment increased AOX1a mRNA levels, in vivo AOX and cytochrome oxidase activities, and levels of citric acid cycle intermediates and hexoses in all genotypes. However, NDA-suppressing plants deviated from the wild type merely by having higher levels of several amino acids. These results suggest that NDA suppression restricts citric acid cycle reactions, inducing a shift towards increased levels of fermentation products, but do not support a direct association between photosynthesis and NDA proteins.

  12. Transcriptome Analysis of Sucrose Metabolism during Bulb Swelling and Development in Onion (Allium cepa L.)

    Science.gov (United States)

    Zhang, Chunsha; Zhang, Hongwei; Zhan, Zongxiang; Liu, Bingjiang; Chen, Zhentai; Liang, Yi

    2016-01-01

    Allium cepa L. is a widely cultivated and economically significant vegetable crop worldwide, with beneficial dietary and health-related properties, but its sucrose metabolism is still poorly understood. To analyze sucrose metabolism during bulb swelling, and the development of sweet taste in onion, a global transcriptome profile of onion bulbs was undertaken at three different developmental stages, using RNA-seq. A total of 79,376 unigenes, with a mean length of 678 bp, was obtained. In total, 7% of annotated Clusters of Orthologous Groups (COG) were involved in carbohydrate transport and metabolism. In the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, “starch and sucrose metabolism” (147, 2.40%) constituted the primary metabolism pathway in the integrated library. The expression of sucrose transporter genes was greatest during the early-swelling stage, suggesting that sucrose transporters (SUTs) participated in sucrose metabolism mainly at an early stage of bulb development. A gene-expression analysis of the key enzymes of sucrose metabolism suggested that sucrose synthase, cell wall invertase, and invertase were all likely to participate in the hydrolysis of sucrose, generating glucose, and fructose. In addition, trehalose was hydrolyzed to two molecules of glucose by trehalase. From 15 to 40 days after swelling (DAS), both the glucose and fructose contents of bulbs increased, whereas the sucrose content decreased. The growth rate between 15 and 30 DAS was slower than that between 30 and 40 DAS, suggesting that the latter was a period of rapid expansion. The dataset generated by our transcriptome profiling will provide valuable information for further research. PMID:27713754

  13. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    Science.gov (United States)

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar.

  14. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    Science.gov (United States)

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar. PMID:25873679

  15. Generation of boron-deficiency-tolerant tomato by overexpressing an Arabidopsis thaliana borate transporter AtBOR1

    Directory of Open Access Journals (Sweden)

    Shimpei eUraguchi

    2014-04-01

    Full Text Available Nutrient deficiency in soil poses a widespread agricultural problem. Boron (B is an essential micronutrient in plants, and its deficiency causes defects in both vegetative and reproductive growth in various crops in the field. In Arabidopsis thaliana, increased expression of a major borate transporter gene AtBOR1 or boric acid channel gene AtNIP5;1 improves plant growth under B-deficient conditions. In this study, we examined whether high expression of a borate transporter gene increases B accumulation in shoots and improves the growth of tomato plant, a model of fruit-bearing crops, under B-deficient conditions. We established three independent transgenic tomato plants lines expressing AtBOR1 using Agrobacterium-mediated transformation of tomato (Solanum lycopersicum L. cv. Micro-Tom. Reverse transcription-polymerase chain reaction (RT-PCR analysis confirmed that two lines (Line 1 and Line 2 more strongly expressed AtBOR1 than Line 3. Wild-type plants and the transgenic plants were grown hydroponically under B-sufficient and B-deficient conditions. Wild-type and Line 3 (weakly expressing transgenic line showed a defect in shoot growth under B-deficient conditions, especially in the development of new leaves. However, seedlings of Line 1 and Line 2, the transgenic lines showing strong AtBOR1 expression, did not show the B-deficiency phenotype in newly developing leaves. In agreement with this phenotype, shoot biomass under low-B conditions was higher in the strongly expressing AtBOR1 line. B concentrations in leaves or fruits were also higher in Line 2 and Line 1. The present study demonstrates that strong expression of AtBOR1 improved growth in tomato under B-deficient conditions.

  16. Magnetostructural study of iron sucrose

    International Nuclear Information System (INIS)

    Magnetic and structural analyses have been performed on an iron sucrose complex used as a haematinic agent. The system contains two-line ferrihydrite particles of about 5 nm that are superparamagnetic above approximately 50 K. The observed low-temperature magnetic dynamics of this compound is closer to simple models than in the case of other iron-containing drugs for intravenous use like iron dextran

  17. Expression of the ZNT1 Zinc Transporter from the Metal Hyperaccumulator Noccaea caerulescens Confers Enhanced Zinc and Cadmium Tolerance and Accumulation to Arabidopsis thaliana.

    Science.gov (United States)

    Lin, Ya-Fen; Hassan, Zeshan; Talukdar, Sangita; Schat, Henk; Aarts, Mark G M

    2016-01-01

    Prompt regulation of transition metal transporters is crucial for plant zinc homeostasis. NcZNT1 is one of such transporters, found in the metal hyperaccumulator Brassicaceae species Noccaea caerulescens. It is orthologous to AtZIP4 from Arabidopsis thaliana, an important actor in Zn homeostasis. We examined if the NcZNT1 function contributes to the metal hyperaccumulation of N. caerulescens. NcZNT1 was found to be a plasma-membrane located metal transporter. Constitutive overexpression of NcZNT1 in A. thaliana conferred enhanced tolerance to exposure to excess Zn and Cd supply, as well as increased accumulation of Zn and Cd and induction of the Fe deficiency response, when compared to non-transformed wild-type plants. Promoters of both genes were induced by Zn deficiency in roots and shoots of A. thaliana. In A. thaliana, the AtZIP4 and NcZNT1 promoters were mainly active in cortex, endodermis and pericycle cells under Zn deficient conditions. In N. caerulescens, the promoters were active in the same tissues, though the activity of the NcZNT1 promoter was higher and not limited to Zn deficient conditions. Common cis elements were identified in both promoters by 5' deletion analysis. These correspond to the previously determined Zinc Deficiency Responsive Elements found in A. thaliana to interact with two redundantly acting transcription factors, bZIP19 and bZIP23, controlling the Zn deficiency response. In conclusion, these results suggest that NcZNT1 is an important factor in contributing to Zn and Cd hyperaccumulation in N. caerulescens. Differences in cis- and trans-regulators are likely to account for the differences in expression between A. thaliana and N. caerulescens. The high, constitutive NcZNT1 expression in the stele of N. caerulescens roots implicates its involvement in long distance root-to-shoot metal transport by maintaining a Zn/Cd influx into cells responsible for xylem loading. PMID:26930473

  18. Expression of the ZNT1 Zinc Transporter from the Metal Hyperaccumulator Noccaea caerulescens Confers Enhanced Zinc and Cadmium Tolerance and Accumulation to Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Ya-Fen Lin

    Full Text Available Prompt regulation of transition metal transporters is crucial for plant zinc homeostasis. NcZNT1 is one of such transporters, found in the metal hyperaccumulator Brassicaceae species Noccaea caerulescens. It is orthologous to AtZIP4 from Arabidopsis thaliana, an important actor in Zn homeostasis. We examined if the NcZNT1 function contributes to the metal hyperaccumulation of N. caerulescens. NcZNT1 was found to be a plasma-membrane located metal transporter. Constitutive overexpression of NcZNT1 in A. thaliana conferred enhanced tolerance to exposure to excess Zn and Cd supply, as well as increased accumulation of Zn and Cd and induction of the Fe deficiency response, when compared to non-transformed wild-type plants. Promoters of both genes were induced by Zn deficiency in roots and shoots of A. thaliana. In A. thaliana, the AtZIP4 and NcZNT1 promoters were mainly active in cortex, endodermis and pericycle cells under Zn deficient conditions. In N. caerulescens, the promoters were active in the same tissues, though the activity of the NcZNT1 promoter was higher and not limited to Zn deficient conditions. Common cis elements were identified in both promoters by 5' deletion analysis. These correspond to the previously determined Zinc Deficiency Responsive Elements found in A. thaliana to interact with two redundantly acting transcription factors, bZIP19 and bZIP23, controlling the Zn deficiency response. In conclusion, these results suggest that NcZNT1 is an important factor in contributing to Zn and Cd hyperaccumulation in N. caerulescens. Differences in cis- and trans-regulators are likely to account for the differences in expression between A. thaliana and N. caerulescens. The high, constitutive NcZNT1 expression in the stele of N. caerulescens roots implicates its involvement in long distance root-to-shoot metal transport by maintaining a Zn/Cd influx into cells responsible for xylem loading.

  19. Identification of a crucial histidine involved in metal transport activity in the Arabidopsis cation/H(+) exchanger CAX1

    Science.gov (United States)

    In plants, yeast and bacteria, cation/H(+) exchangers (CAXs), have been shown to translocate Ca(2+) and other metals. The best characterized of these related transporters is the plant vacuolar-localized CAX1. We used site-directed mutagenesis to assess the impact of altering the seven histidine re...

  20. Arabidopsis phosphatidylinositol monophosphate 5-kinase 2 is involved in root gravitropism through regulation of polar auxin transport by affecting the cycling of PIN proteins.

    Science.gov (United States)

    Mei, Yu; Jia, Wen-Jing; Chu, Yu-Jia; Xue, Hong-Wei

    2012-03-01

    Phosphatidylinositol monophosphate 5-kinase (PIP5K) catalyzes the synthesis of PI-4,5-bisphosphate (PtdIns(4,5)P(2)) by phosphorylation of PI-4-phosphate at the 5 position of the inositol ring, and is involved in regulating multiple developmental processes and stress responses. We here report on the functional characterization of Arabidopsis PIP5K2, which is expressed during lateral root initiation and elongation, and whose expression is enhanced by exogenous auxin. The knockout mutant pip5k2 shows reduced lateral root formation, which could be recovered with exogenous auxin, and interestingly, delayed root gravity response that could not be recovered with exogenous auxin. Crossing with the DR5-GUS marker line and measurement of free IAA content confirmed the reduced auxin accumulation in pip5k2. In addition, analysis using the membrane-selective dye FM4-64 revealed the decelerated vesicle trafficking caused by PtdIns(4,5)P(2) reduction, which hence results in suppressed cycling of PIN proteins (PIN2 and 3), and delayed redistribution of PIN2 and auxin under gravistimulation in pip5k2 roots. On the contrary, PtdIns(4,5)P(2) significantly enhanced the vesicle trafficking and cycling of PIN proteins. These results demonstrate that PIP5K2 is involved in regulating lateral root formation and root gravity response, and reveal a critical role of PIP5K2/PtdIns(4,5)P(2) in root development through regulation of PIN proteins, providing direct evidence of crosstalk between the phosphatidylinositol signaling pathway and auxin response, and new insights into the control of polar auxin transport. PMID:21894193

  1. Arabidopsis phosphatidylinositol monophosphate 5-kinase 2 is involved in root gravitropism through regulation of polar auxin transport by affecting the cycling of PIN proteins

    Institute of Scientific and Technical Information of China (English)

    Yu Mei; Wen-Jing Jia; Yu-Jia Chu; Hong-Wei Xue

    2012-01-01

    Phosphatidylinositol monophosphate 5-kinase(PIP5K)catalyzes the synthesis of PI-4,5-bisphosphate(PtdIns(4,5)P2)by phosphorylation of PI-4-phosphate at the 5 position of the inositol ring,and is involved in regulating multiple developmental processes and stress responses.We here report on the functional characterization of Arabidopsis PIP5K2,which is expressed during lateral root initiation and elongation,and whose expression is enhanced by exogenous auxin.The knockout mutant pip5k2 shows reduced lateral root formation,which could be recovered with exogenous auxin,and interestingly,delayed root gravity response that could not be recovered with exogenous auxin.Crossing with the DR5-GUS marker line and measurement of free IAA content confirmed the reduced auxin accumulation in pip5k2.In addition,analysis using the membrane-selective dye FM4-64 revealed the decelerated vesicle trafficking caused by PtdIns(4,5)P2 reduction,which hence results in suppressed cycling of PIN proteins(PIN2 and 3),and delayed redistribution of PIN2 and auxin under gravistimulation in pipSk2 roots.On the contrary,PtdIns(4,5)P2 significantly enhanced the vesicle trafficking and cycling of PIN proteins.These results demonstrate that PIP5K2 is involved in regulating lateral root formation and root gravity response,and reveal a critical role of PIP5K2/Ptdlns(4,5)P2 in root development through regulation of PIN proteins,providing direct evidence of crosstalk between the phosphatidylinositol signaling pathway and auxin response,and new insights into the control of polar auxin transport.

  2. Sucrose induces vesicle accumulation and autophagy.

    Science.gov (United States)

    Higuchi, Takahiro; Nishikawa, Jun; Inoue, Hiroko

    2015-04-01

    It has been shown that the treatment of mammalian cells with sucrose leads to vacuole accumulation associated with lysosomes and upregulation of lysosomal enzyme expression and activity. Autophagy is an evolutionarily conserved homeostatic process by which cells deliver cytoplasmic material for degradation into lysosomes, thus it is probable that sucrose affects the autophagic activity. The role of sucrose in autophagy is unknown; however, another disaccharide, trehalose has been shown to induce autophagy. In the current study, we used mouse embryonic fibroblasts to investigate whether sucrose induces autophagy and whether vesicle formation is associated with autophagy. The results showed that sucrose induces autophagy while being accumulated within the endosomes/lysosomes. These vesicles were swollen and packed within the cytoplasm. Furthermore, trehalose and the trisaccharide raffinose, which are not hydrolyzed in mammalian cells, increased the rate of vesicles accumulation and LC3-II level (a protein marker of autophagy). However, fructose and maltose did not show the same effects. The correlation between the two processes, vesicle accumulation and autophagy induction, was confirmed by treatment of cells with sucrose plus invertase, or maltose plus acarbose-the α-glucosidase inhibitor-and by sucrose deprivation. Results also showed that vesicle accumulation was not affected by autophagy inhibition. Therefore, the data suggest that sucrose-induced autophagy through accumulation of sucrose-containing vesicles is caused by the absence of hydrolysis enzymes.

  3. Sucrose induces vesicle accumulation and autophagy.

    Science.gov (United States)

    Higuchi, Takahiro; Nishikawa, Jun; Inoue, Hiroko

    2015-04-01

    It has been shown that the treatment of mammalian cells with sucrose leads to vacuole accumulation associated with lysosomes and upregulation of lysosomal enzyme expression and activity. Autophagy is an evolutionarily conserved homeostatic process by which cells deliver cytoplasmic material for degradation into lysosomes, thus it is probable that sucrose affects the autophagic activity. The role of sucrose in autophagy is unknown; however, another disaccharide, trehalose has been shown to induce autophagy. In the current study, we used mouse embryonic fibroblasts to investigate whether sucrose induces autophagy and whether vesicle formation is associated with autophagy. The results showed that sucrose induces autophagy while being accumulated within the endosomes/lysosomes. These vesicles were swollen and packed within the cytoplasm. Furthermore, trehalose and the trisaccharide raffinose, which are not hydrolyzed in mammalian cells, increased the rate of vesicles accumulation and LC3-II level (a protein marker of autophagy). However, fructose and maltose did not show the same effects. The correlation between the two processes, vesicle accumulation and autophagy induction, was confirmed by treatment of cells with sucrose plus invertase, or maltose plus acarbose-the α-glucosidase inhibitor-and by sucrose deprivation. Results also showed that vesicle accumulation was not affected by autophagy inhibition. Therefore, the data suggest that sucrose-induced autophagy through accumulation of sucrose-containing vesicles is caused by the absence of hydrolysis enzymes. PMID:25389129

  4. Sucrose accumulation in mature sweet melon fruits. [Cucumis melo

    Energy Technology Data Exchange (ETDEWEB)

    Schaffer, A.A.; Aloni, B.

    1987-04-01

    Mesocarp tissue from sucrose-accumulating sweet melon (Cucumis melo cv. Galia) showed sucrose synthase activity (ca 1 nkat/gfw) while soluble acid invertase and sucrose phosphate synthase activities were not observed. Sucrose uptake into mesocarp discs was linear with sucrose concentration (1-500 mM) and unaffected by PCMBS and CCCP. Sucrose compartmentation into the vacuole also increased linearly with sucrose concentration as indicated by compartmental efflux kinetics. Mesocarp discs incubated in /sup 14/C-fructose + UDP-glu synthesized /sup 14/C-sucrose and efflux kinetics indicated that the /sup 14/C-sucrose was compartmentalized. These data support the hypothesis that two mechanisms are involved in sucrose accumulation in sweet melon: (1) compartmentation of intact sucrose and (2) synthesis of sucrose via sucrose synthase and subsequent compartmentation in the vacuole.

  5. Sugarcane genes associated with sucrose content

    Directory of Open Access Journals (Sweden)

    Vincentz Michel GA

    2009-03-01

    Full Text Available Abstract Background - Sucrose content is a highly desirable trait in sugarcane as the worldwide demand for cost-effective biofuels surges. Sugarcane cultivars differ in their capacity to accumulate sucrose and breeding programs routinely perform crosses to identify genotypes able to produce more sucrose. Sucrose content in the mature internodes reach around 20% of the culms dry weight. Genotypes in the populations reflect their genetic program and may display contrasting growth, development, and physiology, all of which affect carbohydrate metabolism. Few studies have profiled gene expression related to sugarcane's sugar content. The identification of signal transduction components and transcription factors that might regulate sugar accumulation is highly desirable if we are to improve this characteristic of sugarcane plants. Results - We have evaluated thirty genotypes that have different Brix (sugar levels and identified genes differentially expressed in internodes using cDNA microarrays. These genes were compared to existing gene expression data for sugarcane plants subjected to diverse stress and hormone treatments. The comparisons revealed a strong overlap between the drought and sucrose-content datasets and a limited overlap with ABA signaling. Genes associated with sucrose content were extensively validated by qRT-PCR, which highlighted several protein kinases and transcription factors that are likely to be regulators of sucrose accumulation. The data also indicate that aquaporins, as well as lignin biosynthesis and cell wall metabolism genes, are strongly related to sucrose accumulation. Moreover, sucrose-associated genes were shown to be directly responsive to short term sucrose stimuli, confirming their role in sugar-related pathways. Conclusion - Gene expression analysis of sugarcane populations contrasting for sucrose content indicated a possible overlap with drought and cell wall metabolism processes and suggested signaling and

  6. Omics analysis of high-energy Arabidopsis thaliana

    OpenAIRE

    Liang, Chao; 梁超

    2014-01-01

    Arabidopsis thaliana purple acid phosphatase 2 (AtPAP2) is a phosphatase dually targeted to both chloroplasts and mitochondria. Overexpression (OE) of AtPAP2 in Arabidopsis thaliana was reported to speed up plant growth and promote flowering, seed yield and biomass at maturity in a previous study. Under long-day (16 hours light at 22°C / 8 hours dark at 18°C) growth conditions, the leaves of 20-day-old OE lines contained significant higher sucrose and glucose than the wild-type (WT) plants, r...

  7. 21 CFR 172.859 - Sucrose fatty acid esters.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sucrose fatty acid esters. 172.859 Section 172.859... CONSUMPTION Multipurpose Additives § 172.859 Sucrose fatty acid esters. Sucrose fatty acid esters identified...) Sucrose fatty acid esters are the mono-, di-, and tri-esters of sucrose with fatty acids and are...

  8. Monogalactosyldiacylglycerol synthesis in the outer envelope membrane of chloroplasts is required for enhanced growth under sucrose supplementation

    Directory of Open Access Journals (Sweden)

    Masato eMurakawa

    2014-06-01

    Full Text Available Plant galactolipid synthesis on the outer envelope membranes of chloroplasts is an important biosynthetic pathway for sustained growth under conditions of phosphate (Pi depletion. During Pi starvation, the amount of digalactosyldiacylglycerol (DGDG is increased to substitute for the phospholipids that are degraded for supplying Pi. An increase in DGDG concentration depends on an adequate supply of monogalactosyldiacylglycerol (MGDG, which is a substrate for DGDG synthesis and is synthesized by a type-B MGDG synthase, MGD3. Recently, sucrose was suggested to be a global regulator of plant responses to Pi starvation. Thus, we analyzed expression levels of several genes involved in lipid remodeling during Pi starvation in Arabidopsis thaliana and found that the abundance of MGD3 mRNA increased when sucrose was exogenously supplied to the growth medium. Sucrose supplementation retarded the growth of the Arabidopsis MGD3 knockout mutant mgd3 but enhanced the growth of transgenic Arabidopsis plants overexpressing MGD3 compared with wild type, indicating the involvement of MGD3 in plant growth under sucrose-replete conditions. Although most features such as chlorophyll content, photosynthetic activity, and Pi content were comparable between wild-type and the transgenic plants overexpressing MGD3, sucrose content in shoot tissues decreased and incorporation of exogenously supplied carbon to DGDG was enhanced in the MGD3-overexpressing plants compared with wild type. Our results suggest that MGD3 plays an important role in supplying DGDG as a component of extraplastidial membranes to support enhanced plant growth under conditions of carbon excess.

  9. Arabidopsis CDS blastp result: AK287911 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287911 J065213B08 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 3e-85 ...

  10. Arabidopsis CDS blastp result: AK318551 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK318551 J075138M12 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 4e-27 ...

  11. Arabidopsis CDS blastp result: AK241823 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241823 J065212G21 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 1e-150 ...

  12. Arabidopsis CDS blastp result: AK243378 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243378 J100063A13 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 5e-18 ...

  13. Arabidopsis CDS blastp result: AK288351 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288351 J090024C17 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 2e-24 ...

  14. Arabidopsis CDS blastp result: AK242252 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242252 J075182G16 At1g12110.1 68414.m01402 nitrate/chlorate transporter (NRT1.1) ...(CHL1) identical to nitrate/chlorate transporter SP:Q05085 from [Arabidopsis thaliana]; contains Pfam profile: PF00854 POT family 6e-88 ...

  15. Arabidopsis CDS blastp result: AK241728 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241728 J065199H08 At1g50310.1 68414.m05640 monosaccharide transporter (STP9) iden...tical to monosaccharide transporter STP9 protein [Arabidopsis thaliana] GI:15487254; contains Pfam profile PF00083: major facilitator superfamily protein 3e-36 ...

  16. Arabidopsis CDS blastp result: AK240645 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240645 J023003B03 At1g50310.1 68414.m05640 monosaccharide transporter (STP9) iden...tical to monosaccharide transporter STP9 protein [Arabidopsis thaliana] GI:15487254; contains Pfam profile PF00083: major facilitator superfamily protein 1e-17 ...

  17. Arabidopsis CDS blastp result: AK243302 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243302 J100054J17 At1g50310.1 68414.m05640 monosaccharide transporter (STP9) iden...tical to monosaccharide transporter STP9 protein [Arabidopsis thaliana] GI:15487254; contains Pfam profile PF00083: major facilitator superfamily protein 4e-82 ...

  18. Arabidopsis CDS blastp result: AK241015 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241015 J065054A13 At1g50310.1 68414.m05640 monosaccharide transporter (STP9) iden...tical to monosaccharide transporter STP9 protein [Arabidopsis thaliana] GI:15487254; contains Pfam profile PF00083: major facilitator superfamily protein 8e-37 ...

  19. Arabidopsis CDS blastp result: AK288091 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288091 J075184D14 At1g50310.1 68414.m05640 monosaccharide transporter (STP9) iden...tical to monosaccharide transporter STP9 protein [Arabidopsis thaliana] GI:15487254; contains Pfam profile PF00083: major facilitator superfamily protein 4e-29 ...

  20. IDENTIFICATION AND CHARACTERIZATION OF THE SUCROSE SYNTHASE 2 GENE (Sus2 IN DURUM WHEAT

    Directory of Open Access Journals (Sweden)

    Mariateresa eVolpicella

    2016-03-01

    Full Text Available Sucrose transport is the central system for the allocation of carbon resources in vascular plants. Sucrose synthase, which reversibly catalyzes sucrose synthesis and cleavage, represents a key enzyme in the control of the flow of carbon into starch biosynthesis. In the present study the genomic identification and characterization of the Sus2-2A and Sus2-2B genes coding for sucrose synthase in durum wheat (cultivars Ciccio and Svevo is reported. The genes were analyzed for their expression in different tissues and at different seed maturation stages, in four tetraploid wheat genotypes (Svevo, Ciccio, Primadur and 5-BIL42. The activity of the encoded proteins was evaluated by specific activity assays on endosperm extracts and their structure established by modelling approaches. The combined results of SUS2 expression and activity levels were then considered in the light of their possible involvement in starch yield.

  1. Sucrose- and Fructose-Specific Effects on the Transcriptome of Streptococcus mutans, as Determined by RNA Sequencing.

    Science.gov (United States)

    Zeng, Lin; Burne, Robert A

    2015-10-16

    Recent genome-scale studies have begun to establish the scope and magnitude of the impacts of carbohydrate source and availability on the regulation of gene expression in bacteria. The effects of sugars on gene expression are particularly profound in a group of lactic acid bacteria that rely almost entirely on their saccharolytic activities for energy production and growth. For Streptococcus mutans, the major etiologic agent of human dental caries, sucrose is the carbohydrate that contributes in the most significant manner to establishment, persistence, and virulence of the organism. However, because this organism produces multiple extracellular sucrolytic enzymes that can release hexoses from sucrose, it has not been possible to study the specific effects of sucrose transport and metabolism on gene expression in the absence of carbohydrates that by themselves can elicit catabolite repression and induce expression of multiple genes. By employing RNA deep-sequencing (RNA-Seq) technology and mutants that lacked particular sucrose-metabolizing enzymes, we compared the transcriptomes of S. mutans bacteria growing on glucose, fructose, or sucrose as the sole carbohydrate source. The results provide a variety of new insights into the impact of sucrose transport and metabolism by S. mutans, including the likely expulsion of fructose after sucrose internalization and hydrolysis, and identify a set of genes that are differentially regulated by sucrose versus fructose. The findings significantly enhance our understanding of the genetics and physiology of this cariogenic pathogen.

  2. Sucrose- and Fructose-Specific Effects on the Transcriptome of Streptococcus mutans, as Determined by RNA Sequencing.

    Science.gov (United States)

    Zeng, Lin; Burne, Robert A

    2016-01-01

    Recent genome-scale studies have begun to establish the scope and magnitude of the impacts of carbohydrate source and availability on the regulation of gene expression in bacteria. The effects of sugars on gene expression are particularly profound in a group of lactic acid bacteria that rely almost entirely on their saccharolytic activities for energy production and growth. For Streptococcus mutans, the major etiologic agent of human dental caries, sucrose is the carbohydrate that contributes in the most significant manner to establishment, persistence, and virulence of the organism. However, because this organism produces multiple extracellular sucrolytic enzymes that can release hexoses from sucrose, it has not been possible to study the specific effects of sucrose transport and metabolism on gene expression in the absence of carbohydrates that by themselves can elicit catabolite repression and induce expression of multiple genes. By employing RNA deep-sequencing (RNA-Seq) technology and mutants that lacked particular sucrose-metabolizing enzymes, we compared the transcriptomes of S. mutans bacteria growing on glucose, fructose, or sucrose as the sole carbohydrate source. The results provide a variety of new insights into the impact of sucrose transport and metabolism by S. mutans, including the likely expulsion of fructose after sucrose internalization and hydrolysis, and identify a set of genes that are differentially regulated by sucrose versus fructose. The findings significantly enhance our understanding of the genetics and physiology of this cariogenic pathogen. PMID:26475108

  3. [Sucrose reward promotes rats' motivation for cocaine].

    Science.gov (United States)

    Li, Yan-Qing; LE, Qiu-Min; Yu, Xiang-Chen; Ma, Lan; Wang, Fei-Fei

    2016-06-25

    Caloric diet, such as fat and sugar intake, has rewarding effects, and has been indicated to affect the responses to addictive substances in animal experiments. However, the possible association between sucrose reward and the motivation for addictive drugs remains to be elucidated. Thus, we carried out behavioral tests after sucrose self-administration training to determine the effects of sucrose experience on rats' motivation for cocaine, locomotor sensitivity to cocaine, basal locomotor activity, anxiety level, and associative learning ability. The sucrose-experienced (sucrose) group exhibited higher lever press, cocaine infusion and break point, as well as upshift of cocaine dose-response curve in cocaine self-administration test, as compared with the control (chow) group. Additionally, despite similar locomotor activity in open field test and comparable score in cocaine-induced conditioned place preference, the sucrose group showed higher cocaine-induced locomotor sensitivity as compared with the chow group. The anxiety level and the performance in vocal-cue induced fear memory were similar between these two groups in elevated plus maze and fear conditioning tests, respectively. Taken together, our work indicates that sucrose experience promotes the rats' motivation for cocaine. PMID:27350195

  4. Assessment of preconscious sucrose perception using EEG

    DEFF Research Database (Denmark)

    Rotvel, Camilla Arndal; Møller, Stine; Nielsen, Rene R.;

    to the brain cortex. The method complements sensory panel assessment by providing insight to pre-conscious taste perception. In the empirical study the subject was stimulated with an aqueous sucrose solution at two concentrations: 1 mL 0.1g/L sucrose (below detection threshold) and 100g/L sucrose, respectively...... is known to be involved in sensory integration. The proposed method demonstrates promising results in assessing pre-conscious taste perception, suggesting its viability complementing conventional taste panels....

  5. Structural development of sucrose-sweetened and sucrose-free sponge cakes during baking.

    Science.gov (United States)

    Baeva, Marianna Rousseva; Terzieva, Vesselina Velichkova; Panchev, Ivan Nedelchev

    2003-06-01

    The influence of sucrose, wheat starch and sorbitol upon the heat- and mass-exchanging processes forming the structure of sponge cake was studied. Under the influence of wheat starch and sorbitol the structure of the sucrose-free sponge cake was formed at more uniform total moisture release. This process was done at lower temperatures and smoother change of the sponge cake height with respect to the sucrose-sweetened sponge cake. The porous and steady structure of both cakes was finally formed at identical time--between 18th and 19th minute, at the applied conditions for baking of each batter (metal pan with diameter 15.4 cm and depth 6.2 cm containing 300 g of batter and placed in an electric oven "Rahovetz-02", Bulgaria for 30 min at 180 degrees C). The water-losses at the end of baking (10.30% and 10.40% for the sucrose-sweetened cake and sucrose-free cake, respectively) and the final temperatures reached in the crumb central layers (96.6 degrees C and 96.3 degrees C for the sucrose-sweetened cake and sucrose-free cake, respectively) during baking of both samples were not statistically different. The addition of wheat starch and sorbitol in sucrose-free sponge cake lead to the statistically different values for the porosity (76.15% and 72.98%) and the volume (1014.17 cm3 and 984.25 cm3) of the sucrose-sweetened and sucrose-free sponge cakes, respectively. As a result, the sucrose-free sponge cake formed during baking had a more homogeneous and finer microstructure with respect to that ofthe sucrose-sweetened one. PMID:12866615

  6. Effect of High Temperature on Sucrose Content and Sucrose Cleaving Enzyme Activity in Rice Grain During the Filling Stage

    Institute of Scientific and Technical Information of China (English)

    LI Tian; LIU Qi-hua; Ryu OHSUGI; Tohru YAMAGISHI; Haruto SASAKI

    2006-01-01

    Dynamic changes of sucrose, fructose, glucose contents and differences in activities of sucrose synthase, vacuolar invertase, and cell wall bound invertase in rice grain after flowering stage were studied under natural and high temperatures by using two japonica rice varieties Koshihikari and Sasanishiki. In rice grains, the sucrose synthase activity was higher than that of invertase, which was significantly correlated with starch accumulation rate, indicating that the sucrose synthase played an important role in sucrose degradation and starch synthesis. Under high temperature, the significant increase in grain sucrose content without any increase in fructose and glucose contents, suggested that the high temperature treatment enhanced sucrose accumulation, while diminished sucrose degradation in rice grains. Compared with the control plants, the decrease in activities of sucrose synthase, vacuolar invertase, and cell wall bound invertase with high temperature treated plants indicated that the deceleration of sucrose-degradation was related to the decrease in activities of sucrose synthase and invertase.

  7. Asparagine Metabolic Pathways in Arabidopsis.

    Science.gov (United States)

    Gaufichon, Laure; Rothstein, Steven J; Suzuki, Akira

    2016-04-01

    Inorganic nitrogen in the form of ammonium is assimilated into asparagine via multiple steps involving glutamine synthetase (GS), glutamate synthase (GOGAT), aspartate aminotransferase (AspAT) and asparagine synthetase (AS) in Arabidopsis. The asparagine amide group is liberated by the reaction catalyzed by asparaginase (ASPG) and also the amino group of asparagine is released by asparagine aminotransferase (AsnAT) for use in the biosynthesis of amino acids. Asparagine plays a primary role in nitrogen recycling, storage and transport in developing and germinating seeds, as well as in vegetative and senescence organs. A small multigene family encodes isoenzymes of each step of asparagine metabolism in Arabidopsis, except for asparagine aminotransferase encoded by a single gene. The aim of this study is to highlight the structure of the genes and encoded enzyme proteins involved in asparagine metabolic pathways; the regulation and role of different isogenes; and kinetic and physiological properties of encoded enzymes in different tissues and developmental stages. PMID:26628609

  8. Coordinated post-translational responses of aquaporins to abiotic and nutritional stimuli in Arabidopsis roots.

    Science.gov (United States)

    di Pietro, Magali; Vialaret, Jérôme; Li, Guo-Wei; Hem, Sonia; Prado, Karine; Rossignol, Michel; Maurel, Christophe; Santoni, Véronique

    2013-12-01

    In plants, aquaporins play a crucial role in regulating root water transport in response to environmental and physiological cues. Controls achieved at the post-translational level are thought to be of critical importance for regulating aquaporin function. To investigate the general molecular mechanisms involved, we performed, using the model species Arabidopsis, a comprehensive proteomic analysis of root aquaporins in a large set of physiological contexts. We identified nine physiological treatments that modulate root hydraulics in time frames of minutes (NO and H2O2 treatments), hours (mannitol and NaCl treatments, exposure to darkness and reversal with sucrose, phosphate supply to phosphate-starved roots), or days (phosphate or nitrogen starvation). All treatments induced inhibition of root water transport except for sucrose supply to dark-grown plants and phosphate resupply to phosphate-starved plants, which had opposing effects. Using a robust label-free quantitative proteomic methodology, we identified 12 of 13 plasma membrane intrinsic protein (PIP) aquaporin isoforms, 4 of the 10 tonoplast intrinsic protein isoforms, and a diversity of post-translational modifications including phosphorylation, methylation, deamidation, and acetylation. A total of 55 aquaporin peptides displayed significant changes after treatments and enabled the identification of specific and as yet unknown patterns of response to stimuli. The data show that the regulation of PIP and tonoplast intrinsic protein abundance was involved in response to a few treatments (i.e. NaCl, NO, and nitrate starvation), whereas changes in the phosphorylation status of PIP aquaporins were positively correlated to changes in root hydraulic conductivity in the whole set of treatments. The identification of in vivo deamidated forms of aquaporins and their stimulus-induced changes in abundance may reflect a new mechanism of aquaporin regulation. The overall work provides deep insights into the in vivo post

  9. Transcriptomic analysis of Arabidopsis overexpressing flowering locus T driven by a meristem-specific promoter that induces early flowering

    Science.gov (United States)

    Duplat-Bermúdez, L.; Ruiz-Medrano, R.; Landsman, D.; Mariño-Ramírez, L.; Xoconostle-Cázares, B.

    2016-01-01

    Here we analyzed in leaves the effect of FT overexpression driven by meristem-specific KNAT1 gene homolog of Arabidopsis thaliana (Lincoln et al., 1994; Long et al., 1996) on the transcriptomic response during plant development. Our results demonstrated that meristematic FT overexpression generates a phenotype with an early flowering independent of photoperiod when compared with wild type (WT) plants. Arabidopsis FT-overexpressor lines (AtFTOE) did not show significant differences compared with WT lines neither in leaf number nor in rosette diameter up to day 21, when AtFTOE flowered. After this period AtFTOE plants started flower production and no new rosette leaves were produced. Additionally, WT plants continued on vegetative stage up to day 40, producing 12–14 rosette leaves before flowering. Transcriptomic analysis of rosette leaves studied by sequencing Illumina RNA-seq allowed us to determine the differential expression in mature leaf rosette of 3652 genes, being 626 of them up-regulated and 3026 down-regulated. Overexpressed genes related with flowering showed up-regulated transcription factors such as MADS-box that are known as flowering markers in meristem and which overexpression has been related with meristem identity preservation and the transition from vegetative to floral stage. Genes related with sugar transport have shown a higher demand of monosaccharides derived from the hydrolysis of sucrose to glucose and probably fructose, which can also be influenced by reproductive stage of AtFTOE plants. PMID:27154816

  10. Arabidopsis phosphatidylinositol monophosphate 5-kinase 2 is involved in root gravitropism through regulation of polar auxin transport by affecting the cycling of PIN proteins

    OpenAIRE

    Mei, Yu; Jia, Wen-Jing; Chu, Yu-Jia; Xue, Hong-Wei

    2011-01-01

    Phosphatidylinositol monophosphate 5-kinase (PIP5K) catalyzes the synthesis of PI-4,5-bisphosphate (PtdIns(4,5)P2) by phosphorylation of PI-4-phosphate at the 5 position of the inositol ring, and is involved in regulating multiple developmental processes and stress responses. We here report on the functional characterization of Arabidopsis PIP5K2, which is expressed during lateral root initiation and elongation, and whose expression is enhanced by exogenous auxin. The knockout mutant pip5k...

  11. A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells.

    Science.gov (United States)

    Huber, S C; Akazawa, T

    1986-08-01

    Enzymes of sucrose degradation and glycolysis in cultured sycamore (Acer pseudoplatanus L.) cells were assayed and characterized in crude extracts and after partial purification, in an attempt to identify pathways for sucrose catabolism. Desalted cell extracts contained similar activities (20-40 nanomoles per milligram protein per minute) of sucrose synthase, neutral invertase, glucokinase, fructokinase, phosphofructokinase, and UDPglucose pyrophosphorylase (assayed with 2 micromolar pyrophosphate (PPi). PPi-linked phosphofructokinase activity was virtually dependent upon fructose 2,6-bisphosphate, and the maximum activity exceeded that of ATP-linked phosphofructokinase. Hexokinase activity, with glucose as substrate, was highly specific for ATP, whereas fructokinase activity was relatively nonspecific. At 1 millimolar nucleoside triphosphate, fructokinase activity decreased in the order: UTP > ATP > CTP > GTP. We propose two pathways for sucrose degradation. One involves invertase action, followed by classical glycolysis of hexose sugars, and the other is a novel pathway initiated by sucrose synthase. The K(m) for sucrose of sucrose synthase was severalfold lower than that of neutral invertase (15 versus 65 millimolar), which may determine carbon partitioning between the two pathways. The sucrose synthase pathway proposed involves cycling of uridylates and PPi. UDPglucose pyrophosphorylase, which is shown to be an effective ;PPi-scavenger,' would consume PPi and form UTP. The UTP could be then utilized in the UTP-linked fructokinase reaction, thereby forming UDP for sucrose synthase. The source of PPi is postulated to arise from the back reaction of PPi-linked phosphofructokinase. Sycamore cells contained a substantial endogenous pool of PPi (about 3 nanomoles per gram fresh weight, roughly 1/10 the amount of ATP in these cells), and sufficient fructose 2,6-bisphosphate (0.09 nanomole per gram fresh weight) to activate the PPi-linked phosphofructokinase. Possible

  12. Shoot-supplied ammonium targets the root auxin influx carrier AUX1 and inhibits lateral root emergence in Arabidopsis

    KAUST Repository

    Li, Baohai

    2011-03-24

    Deposition of ammonium (NH4 +) from the atmosphere is a substantial environmental problem. While toxicity resulting from root exposure to NH4 + is well studied, little is known about how shoot-supplied ammonium (SSA) affects root growth. In this study, we show that SSA significantly affects lateral root (LR) development. We show that SSA inhibits lateral root primordium (LRP) emergence, but not LRP initiation, resulting in significantly impaired LR number. We show that the inhibition is independent of abscisic acid (ABA) signalling and sucrose uptake in shoots but relates to the auxin response in roots. Expression analyses of an auxin-responsive reporter, DR5:GUS, and direct assays of auxin transport demonstrated that SSA inhibits root acropetal (rootward) auxin transport while not affecting basipetal (shootward) transport or auxin sensitivity of root cells. Mutant analyses indicated that the auxin influx carrier AUX1, but not the auxin efflux carriers PIN-FORMED (PIN)1 or PIN2, is required for this inhibition of LRP emergence and the observed auxin response. We found that AUX1 expression was modulated by SSA in vascular tissues rather than LR cap cells in roots. Taken together, our results suggest that SSA inhibits LRP emergence in Arabidopsis by interfering with AUX1-dependent auxin transport from shoot to root. © 2011 Blackwell Publishing Ltd.

  13. 过量表达蔗糖转运蛋白基因增强转基因小麦的耐旱性%Overexpression of Sucrose Transporter (TaSUT1A) Improves Drought Tolerance in Transgenic Wheat

    Institute of Scientific and Technical Information of China (English)

    胡梦芸; 李辉; 庞建周; 刘茜; 张颖君; 孙丽静

    2015-01-01

    [Objective]Drought is one of the most important constraints resulting in large yield losses and limiting the average yield increase of wheat in China. The aim of this study is to develop and select stable TaSUT1A transgenic wheat lines with resistance to drought.[Method]The open-reading-frame sequence of TaSUT1A was synthesized and used to construct the gene transformation vector pUBI::cas-TaSUT1A, in which TaSUT1A gene was driven by maize ubiquitin promoter and should be highly expressed in monocot plants. Particle bombardment method was used to introduce TaSUT1A into wheat cultivar Kenong 199. After bialaphos screening, DNA PCR and RT-PCR methods were used to detect the presence and transcript levels of TaSUT1A in the transgenic wheat plants of T0-T4 generations. Subsequently, the resistance to water stress of transgenic wheat plants and non-transgenic wheat Kenong 199 at young seedling-stage was evaluated. Further, their determination of physiologic index related to abiotic stress was finished and the potential of improving abiotic stress tolerance in plant was elucidated. [Result]The results indicated that the introduced TaSUT1A gene was stably inherited, and 220%PEG treatment could significantly induce the expression of TaSUT1A in root and leaf tissues of three transgenic wheat lines. The average germination rate of transgenic wheat lines was 85.53%, which was more higher than those of non-transgenic controls, and the average celoeptile and primary root length significantly higher than the non-transgenic plants. The results showed that the transgenic wheat significantly enhanced seed germination in 20%PEG treatment, as indicated by enhancement of the growth of celoeptile and primary root. Further, three pure lines with higher TaSUT1A expression were selected to elucidate physiologic index related to abiotic stress. In detail, in 20% PEG treatment, the transgenic lines overexpressing TaSUT1A increased sucrose and total soluble sugar content in leaf and root

  14. Invertase and sucrose synthase activities in coffee plants sprayed with sucrose solution

    Directory of Open Access Journals (Sweden)

    Silva José Carlos da

    2003-01-01

    Full Text Available One management practice of which the efficiency has not yet been scientifically tested is spraying coffee plants with diluted sucrose solutions as a source of carbon for the plant. This paper evaluates the effect of foliar spraying with sugar on the endogenous level of carbohydrates and on the activities of invertase and sucrose synthase in coffee (Coffea arabica L. seedlings with reduced (low and high (normal levels of carbon reserve. The concentrations used were 0.5 and 1.0% sucrose, and water as a control. The use of sucrose at 1.0% caused an increase in the concentration of total soluble sugars in depauperate plants, as well as increased the activity of the following enzymes: cell wall and vacuole acid invertase, neutral cytosol invertase and sucrose synthase. In plants with high level of carbon reserve, no increments in total soluble sugar levels or in enzymatic activity were observed. Regardless of treatments or plants physiological state, no differences in transpiration or stomatal conductance were observed, demonstrating the stomatal control of transpiration. Photosynthesis was stimulated with the use of 0.5 and 1.0 % sucrose only in depauperate plants. Coffee seedling spraying with sucrose is only efficient for depauperate plants, at the concentration of 1.0%.

  15. Differential expression analysis of boron transporters and some stress-related genes in response to 24-epibrassinolide and boron by semi-quantitative RT-PCR in Arabidopsis thaliana (L. Heynh

    Directory of Open Access Journals (Sweden)

    Surgun Yonca

    2016-01-01

    Full Text Available Plant steroidal hormones, brassinosteroids (BRs, promote plant developmental processes and enhance tolerance to several abiotic stresses including high boron (B stress. To examine the possible role of BR in high B-induced stress at the transcriptional level, we investigated the response of B transporter genes (BOR1-4, high B-induced genes (MATE, Hsp-like, BR-induced genes (Hsp70-4, Hsp90-1 and other stress-related genes (LTI/COR78, LEA4-5 upon exogenous treatments of 24-epibrassinolide (EBL on Arabidopsis thaliana (L. Heynh exposed to high concentrations of boric acid (BA using semi-quantitative RT-PCR. BA treatments led to down regulation of BOR1 and BOR3 genes in leaf and root tissues and higher concentration of EBL further decreased expression of these genes in roots. The expression of high B-induced genes was observed to be upregulated by 1 μM EBL treatment under high B stress in both tissues of the seedlings. The upregulation of BR-induced genes were clearly evident in root tissues co-treated with 1 μM EBL and BA as compared to BA alone. Higher concentration of EBL was found to be more effective in increasing expression of LTI/COR78 gene in root and LEA4-5 gene in shoot tissues. To our knowledge, this is the first report how exogenous application of EBL modulates high B stress responses at molecular level in model plant Arabidopsis thaliana.

  16. Identification of drought-response genes and a study of their expression during sucrose accumulation and water deficit in sugarcane culms

    Directory of Open Access Journals (Sweden)

    Xu Jingsheng

    2011-01-01

    Full Text Available Abstract Background The ability of sugarcane to accumulate high concentrations of sucrose in its culm requires adaptation to maintain cellular function under the high solute load. We have investigated the expression of 51 genes implicated in abiotic stress to determine their expression in the context of sucrose accumulation by studying mature and immature culm internodes of a high sucrose accumulating sugarcane cultivar. Using a sub-set of eight genes, expression was examined in mature internode tissues of sugarcane cultivars as well as ancestral and more widely related species with a range of sucrose contents. Expression of these genes was also analysed in internode tissue from a high sucrose cultivar undergoing water deficit stress to compare effects of sucrose accumulation and water deficit. Results A sub-set of stress-related genes that are potentially associated with sucrose accumulation in sugarcane culms was identified through correlation analysis, and these included genes encoding enzymes involved in amino acid metabolism, a sugar transporter and a transcription factor. Subsequent analysis of the expression of these stress-response genes in sugarcane plants that were under water deficit stress revealed a different transcriptional profile to that which correlated with sucrose accumulation. For example, genes with homology to late embryogenesis abundant-related proteins and dehydrin were strongly induced under water deficit but this did not correlate with sucrose content. The expression of genes encoding proline biosynthesis was associated with both sucrose accumulation and water deficit, but amino acid analysis indicated that proline was negatively correlated with sucrose concentration, and whilst total amino acid concentrations increased about seven-fold under water deficit, the relatively low concentration of proline suggested that it had no osmoprotectant role in sugarcane culms. Conclusions The results show that while there was a

  17. Arabidopsis type I proton-pumping pyrophosphatase expresses strongly in phloem, where it is required for pyrophosphate metabolism and photosynthate partitioning.

    Science.gov (United States)

    Pizzio, Gaston A; Paez-Valencia, Julio; Khadilkar, Aswad S; Regmi, Kamesh; Patron-Soberano, Araceli; Zhang, Shangji; Sanchez-Lares, Jonathan; Furstenau, Tara; Li, Jisheng; Sanchez-Gomez, Concepcion; Valencia-Mayoral, Pedro; Yadav, Umesh P; Ayre, Brian G; Gaxiola, Roberto A

    2015-04-01

    Phloem loading is a critical process in plant physiology. The potential of regulating the translocation of photoassimilates from source to sink tissues represents an opportunity to increase crop yield. Pyrophosphate homeostasis is crucial for normal phloem function in apoplasmic loaders. The involvement of Arabidopsis (Arabidopsis thaliana) type I proton-pumping pyrophosphatase (AVP1) in phloem loading was analyzed at genetic, histochemical, and physiological levels. A transcriptional AVP1 promoter::GUS fusion revealed phloem activity in source leaves. Ubiquitous AVP1 overexpression (35S::AVP1 cassette) enhanced shoot biomass, photoassimilate production and transport, rhizosphere acidification, and expression of sugar-induced root ion transporter genes (POTASSIUM TRANSPORTER2 [KUP2], NITRATE TRANSPORTER2.1 [NRT2.1], NRT2.4, and PHOSPHATE TRANSPORTER1.4 [PHT1.4]). Phloem-specific AVP1 overexpression (Commelina Yellow Mottle Virus promoter [pCOYMV]::AVP1) elicited similar phenotypes. By contrast, phloem-specific AVP1 knockdown (pCoYMV::RNAiAVP1) resulted in stunted seedlings in sucrose-deprived medium. We also present a promoter mutant avp1-2 (SALK046492) with a 70% reduction of expression that did not show severe growth impairment. Interestingly, AVP1 protein in this mutant is prominent in the phloem. Moreover, expression of an Escherichia coli-soluble pyrophosphatase in the phloem (pCoYMV::pyrophosphatase) of avp1-2 plants resulted in severe dwarf phenotype and abnormal leaf morphology. We conclude that the Proton-Pumping Pyrophosphatase AVP1 localized at the plasma membrane of the sieve element-companion cell complexes functions as a synthase, and that this activity is critical for the maintenance of pyrophosphate homeostasis required for phloem function. PMID:25681328

  18. Arabidopsis Type I Proton-Pumping Pyrophosphatase Expresses Strongly in Phloem, Where It Is Required for Pyrophosphate Metabolism and Photosynthate Partitioning1[OPEN

    Science.gov (United States)

    Pizzio, Gaston A.; Paez-Valencia, Julio; Khadilkar, Aswad S.; Regmi, Kamesh; Patron-Soberano, Araceli; Zhang, Shangji; Sanchez-Lares, Jonathan; Furstenau, Tara; Li, Jisheng; Sanchez-Gomez, Concepcion; Valencia-Mayoral, Pedro; Yadav, Umesh P.; Ayre, Brian G.; Gaxiola, Roberto A.

    2015-01-01

    Phloem loading is a critical process in plant physiology. The potential of regulating the translocation of photoassimilates from source to sink tissues represents an opportunity to increase crop yield. Pyrophosphate homeostasis is crucial for normal phloem function in apoplasmic loaders. The involvement of Arabidopsis (Arabidopsis thaliana) type I proton-pumping pyrophosphatase (AVP1) in phloem loading was analyzed at genetic, histochemical, and physiological levels. A transcriptional AVP1 promoter::GUS fusion revealed phloem activity in source leaves. Ubiquitous AVP1 overexpression (35S::AVP1 cassette) enhanced shoot biomass, photoassimilate production and transport, rhizosphere acidification, and expression of sugar-induced root ion transporter genes (POTASSIUM TRANSPORTER2 [KUP2], NITRATE TRANSPORTER2.1 [NRT2.1], NRT2.4, and PHOSPHATE TRANSPORTER1.4 [PHT1.4]). Phloem-specific AVP1 overexpression (Commelina Yellow Mottle Virus promoter [pCOYMV]::AVP1) elicited similar phenotypes. By contrast, phloem-specific AVP1 knockdown (pCoYMV::RNAiAVP1) resulted in stunted seedlings in sucrose-deprived medium. We also present a promoter mutant avp1-2 (SALK046492) with a 70% reduction of expression that did not show severe growth impairment. Interestingly, AVP1 protein in this mutant is prominent in the phloem. Moreover, expression of an Escherichia coli-soluble pyrophosphatase in the phloem (pCoYMV::pyrophosphatase) of avp1-2 plants resulted in severe dwarf phenotype and abnormal leaf morphology. We conclude that the Proton-Pumping Pyrophosphatase AVP1 localized at the plasma membrane of the sieve element-companion cell complexes functions as a synthase, and that this activity is critical for the maintenance of pyrophosphate homeostasis required for phloem function. PMID:25681328

  19. A phloem-specific sucrose-H+ symporter from Plantago major L. supports the model of apoplastic phloem loading.

    Science.gov (United States)

    Gahrtz, M; Stolz, J; Sauer, N

    1994-11-01

    In this paper the cloning of a full-length cDNA clone encoding the PmSUC2 sucrose-H+ symporter from Plantago major is described. This plant allows the simple preparation of vascular bundles from the basal regions of fully developed source leaves and thus a separation of vascular and non-vascular tissue. A cDNA library was constructed from poly(A)+ RNA isolated from vascular bundles and used for the subsequent cloning of cDNAs. The respective mRNA is specifically expressed in the vascular bundles as shown on Northern blots of total RNA from vascular and non-vascular tissues. The PmSUC2 protein has 12 putative transmembrane helices and is highly homologous to other plant sucrose transporters. Substrate specificity and energy dependence of the transporter encoded by this cDNA were determined by expression in baker's yeast Saccharomyces cerevisiae. The PmSUC2 protein catalyses the transport of sucrose into transgenic yeast cells. Invertase null mutants of yeast expressing PmSUC2 accumulate sucrose more than 200-fold. This transport was sensitive to uncouplers or SH-group inhibitors. Plasma membranes from yeast cells expressing the PmSUC2 protein were purified and fused to proteoliposomes containing cytochrome-c-oxidase. In this system sucrose is accumulated only when proton motive force is generated, indicating that PmSUC2 is a sucrose-H+ symporter. The apparent molecular weight of the PmSUC2 protein is 35 kDa on 10% SDS-polyacrylamide gels. The presented data strongly support the theory of phloem loading from the apoplastic space by a sucrose-H+ symporter.

  20. Effect of sucrose on adventitious root regeneration in apple

    NARCIS (Netherlands)

    Calamar, A.; Klerk, de G.J.M.

    2002-01-01

    We have examined the effect of sucrose on adventitious root formation in apple microcuttings and in 1-mm stem slices cut from apple microcuttings. The sucrose concentration influenced the number of adventitious roots, but at a broad range of sucrose concentrations (1¿9%) the effect was small. In add

  1. Metabolism of sucrose during storage in intact sorghum stalk

    International Nuclear Information System (INIS)

    Grasses of the Andropogoneae, such as sorghum (Sorghum bicolor [L.] Moench) and sugarcane (Saccharum spp.), accumulate large amounts of harvestable sucrose in the stalk. Sucrose is translocated from the leaves, yet the metabolic fate of sucrose up to and during storage in the stalk is uncertain. This study determined whether sucrose is necessarily routed through inversion and hexose-phosphate isomerization for storage in cells of the intact stalk. Sorghum stalks were dual-infused with [U-14C]sucrose and [fructose-1-3H(N)]sucrose during stages of sucrose accumulation. Unperturbed tissue was used for subsequent sugar analyses. Solutes in cell extracts were separated sequentially through enzymatic conversion and ion-exchange removal of the reaction products. The hexose moieties of sucrose were analyzed separately. A mean of 95% of recovered 14C was in sucrose. In sucrose, 46% of 14C and 77% of 3H were recovered in the fructose moiety. The significant retention of asymmetry in tritium labeling of sucrose indicates that a cycle of cleavage and resynthesis did not dominate sucrose accumulation in cells of the sorghum stalk. (author)

  2. Sham-feeding response of rats to Polycose and sucrose.

    Science.gov (United States)

    Nissenbaum, J W; Sclafani, A

    1987-01-01

    Adult female rats were fitted with gastric fistulas and maintained at 85% of their ad lib body weight. Their real-feeding (fistula closed) and sham-feeding (fistula open) responses to polysaccharide (Polycose) and sucrose solutions were measured during 30 min/day one-bottle tests. The rats consumed similar amounts of a 1% Polycose solution during real- and sham-feeding tests, but their sham-intakes of 4%, 16% and 32% Polycose solutions greatly exceeded their real-intakes of these solutions. Similar results were obtained with sucrose solutions. The rats sham-fed more Polycose than sucrose at the 1% and 4% concentrations, while their sham-intakes of the 16% and 32% Polycose and sucrose solutions were comparable. In subsequent two-solution sham-feeding tests, the rats preferred 1% Polycose to 1% sucrose, but preferred sucrose to Polycose at 4%, 16% and 32% concentrations. These preference results indicate that rats find Polycose more palatable than sucrose at low concentrations, but sucrose more palatable at high concentrations. In addition, the findings that the rats preferred 4% sucrose to 4% Polycose in the two-bottle test, but sham-fed more 4% Polycose than 4% sucrose in the one-bottle tests, suggest that sucrose is more "orally-satiating" than is Polycose. These results provide further evidence for qualitative differences in the tastes of sucrose and polysaccharide. They also indicate that the amount of solution sham-fed does not necessarily reflect the palatability of the solution.

  3. New insights into carbon transport and incorporation to wood

    OpenAIRE

    Mahboubi, Amir

    2015-01-01

    Wood formation in trees requires carbon import from the photosynthetic tissues. In several tree species the majority of this carbon is derived from sucrose transported in the phloem. This thesis describes experimental work on the mechanism of radial sucrose transport from phloem to developing wood and subsequent incorporation of carbon into wood polymers. I investigated the role of active sucrose transport during secondary cell wall formation in hybrid aspen (Populus tremula x tremuloides...

  4. Sucrose Inversion An Experiment on Heterogeneous Catalysis

    OpenAIRE

    Adélio Mendes; Magalhães, Fernão D.; Luis M. Madeira

    2003-01-01

    llustration of heterogeneous catalysis concepts in laboratory courses is not usually simple or economical. For our undergraduate senior lab course we have developed an environmentally friendly experiment dealing with several aspects of heterogeneous catalysis, having in mind the use of readily available and relatively inexpensive equipment, and chemicals on a compact setup, which students can safely operate. The experiment deals with the acid-catalyzed sucrose inversion, performed in packed b...

  5. Withanolides and Sucrose Esters from Physalis neomexicana.

    Science.gov (United States)

    Cao, Cong-Mei; Wu, Xiaoqing; Kindscher, Kelly; Xu, Liang; Timmermann, Barbara N

    2015-10-23

    Four withanolides (1-4) and two sucrose esters (5, 6) were isolated from the aerial parts of Physalis neomexicana. The structures of 1-6 were elucidated through a variety of spectroscopic techniques. Cytotoxicity studies of the isolates revealed that 2 inhibited human breast cancer cell lines (MDA-MB-231 and MCF-7) with IC50 values of 1.7 and 6.3 μM, respectively.

  6. Crystallization inhibition of an amorphous sucrose system using raffinose

    Institute of Scientific and Technical Information of China (English)

    LEINEN K.M.; LABUZA T.P.

    2006-01-01

    The shelf life of pure amorphous sucrose systems, such as cotton candy, can be very short. Previous studies have shown that amorphous sucrose systems held above the glass transition temperature will collapse and crystallize. One study,however, showed that adding a small percent of another type of sugar, such as trehalose, to sucrose can extend the shelf life of the amorphous system by slowing crystallization. This study explores the hypothesis that raffinose increases the stability of an amorphous sucrose system. Cotton candy at 5 wt% raffinose and 95 wt% sucrose was made and stored at room temperature and three different relative humidities (%RH) 11%RH, 33%RH, and 43%RH. XRD patterns, and glass transition temperatures were obtained to determine the stability as a function of %RH. The data collected showed that raffinose slows sucrose crystallization in a low moisture amorphous state above the glass transition temperature and therefore improves the stability of amorphous sucrose systems.

  7. STUDY ON THE SYNTHESIS AND PHYSICOCHEMICAL PROPERTIES OF SUCROSE POLYESTER

    Institute of Scientific and Technical Information of China (English)

    LIU Zhongdong; LIU Huihua; LUO Peng; LIU Peng; Xiao Fugang

    2006-01-01

    Orthogonal test was used to evaluate the effects of synthetic such as temperature (120~140 ℃), reaction time (4-6) and substrate molar ratio of methyl oleate to sucrose (8:1-12:1) on the percent quantity conversion to sucrose polyester. Sucrose polyester was synthesized by a solvent-free one-stage interesterification. The optimum reaction conditions are as follows: methyl oleate/sucrose = 10∶1 (mol/mol); reaction temperature is 140 ℃;yield reaches 88.15%, and the degree of esterification (DE) is over 7 in the conditions. Thin layer chromatography (TLC), column chromatography (CC), High-performance liquid chromatography (HPLC) were used to analyze the product, the results show that the percent of sucrose polyoleate is over 70% in the product. The physicochemical properties of sucrose polyesters were compared with cooking oil. The results show that the qualities of sucrose polyesters are all up to the triglyceride.

  8. Protein-based microhydraulic transport for controllable actuation

    Science.gov (United States)

    Sundaresan, Vishnu Baba; Leo, Donald J.

    2006-03-01

    Plants have the ability to develop large mechanical force from chemical energy available with bio-fuels. The energy released by the cleavage of a terminal phosphate ion during the hydrolysis of a bio- fuel assists the transport of ions and fluids in cellular homeostasis. Materials that develop pressure and hence strain similar to the response of plants to an external stimuli are classified as nastic materials. Calculations for controlled actuation of an active material inspired by biological transport mechanism demonstrated the feasibility of developing such a material with actuation energy densities on the order of 100 kJ/m 3. The mathematical model for a simplified proof of concept actuator referred to as micro hydraulic actuator uses ion transporters extracted from plants reconstituted on a synthetic bilayer lipid membrane (BLM). Thermodynamic model of the concept actuator predicted the ability to develop 5 percent normalized deformation in thickness of the micro- hydraulic actuator. Controlled fluid transport through AtSUT4 (Proton-sucrose co-transporter from Arabidopsis thaliana) reconstituted on a 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-[Phospho-L- Serine] (Sodium Salt) (POPS), 1-Palmitoyl-2-Oleoyl-sn-Glycero-3- Phosphoethanolamine (POPE) BLM on a porous lead silicate glass plate (50μm with 61μm pitch) was driven by proton gradient. Bulk fluid flux of 1.2 μl/min was observed for each microliter of AtSUT4 transporter suspension (16.6 mg/ml in pH7.0 medium) reconstituted on the BLM. The flux rate is observed to be dependent on the concentration of sucrose present in pH4 buffer. Flux rate of 10 μl/min is observed for 5 mM sucrose in the first 10 minutes. The observed flux scales linearly with BLM area and the amount of proteins reconstituted on the lipid membrane. This article details the next step in the development of the micro hydraulic actuator - fluid transport driven by exergonic Adenosine triphosphate (ATP) hydrolysis reaction in the presence of ATP

  9. Comparison of freezing tolerance, compatible solutes and polyamines in geographically diverse collections of Thellungiella sp. and Arabidopsis thaliana accessions

    Directory of Open Access Journals (Sweden)

    Lee Yang

    2012-08-01

    Full Text Available Abstract Background Thellungiella has been proposed as an extremophile alternative to Arabidopsis to investigate environmental stress tolerance. However, Arabidopsis accessions show large natural variation in their freezing tolerance and here the tolerance ranges of collections of accessions in the two species were compared. Results Leaf freezing tolerance of 16 Thellungiella accessions was assessed with an electrolyte leakage assay before and after 14 days of cold acclimation at 4°C. Soluble sugars (glucose, fructose, sucrose, raffinose and free polyamines (putrescine, spermidine, spermine were quantified by HPLC, proline photometrically. The ranges in nonacclimated freezing tolerance completely overlapped between Arabidopsis and Thellungiella. After cold acclimation, some Thellungiella accessions were more freezing tolerant than any Arabidopsis accessions. Acclimated freezing tolerance was correlated with sucrose levels in both species, but raffinose accumulation was lower in Thellungiella and only correlated with freezing tolerance in Arabidopsis. The reverse was true for leaf proline contents. Polyamine levels were generally similar between the species. Only spermine content was higher in nonacclimated Thellungiella plants, but decreased during acclimation and was negatively correlated with freezing tolerance. Conclusion Thellungiella is not an extremophile with regard to freezing tolerance, but some accessions significantly expand the range present in Arabidopsis. The metabolite data indicate different metabolic adaptation strategies between the species.

  10. 蔗糖转运蛋白VvSUC11和VvSUC12累加作用对提高转基因甜菜含糖量的影响%Improvement of the Saccharinity by Additive Action of Sucrose Transporters VvSUC11 and VvSUC12 in Transgenic Sugar Beet

    Institute of Scientific and Technical Information of China (English)

    闫甜甜; 郭新勇; 向本春; 祝建波

    2014-01-01

    an important sugar crop in the world. The sucrose content in roots is the key determinant of sugar beet quality. In order to study the improvement of saccharinity of sugar beet by two sucrose transporter genes from grape, VvSUC11 and VvSUC12, were introduced into sugar beet root using plant genetic engineering. The work will lay a foundation for using sucrose transporter genes to increase crop yield and quality. [Method] A bivalent root specific expression vector called pCAMBIA2301-SP1-VvSUC11-SP2-VvSUC12 containing grape sucrose transporters VvSUC11 and VvSUC12 genes was constructed and transformed into sugar beet variety KWS-9103 by Agrobacterium tumefaciens mediated transformation. Then PCR and RT-PCR analyses were used to confirm the integration and expression of the two genes in transgenic sugar beets. The leaf characters, physiological targets, tuberous root weight and saccharinity of transgenic and control plants were measured after transplanting to the field.[Result]Thirteen transgenic sugar beet lines were obtained by PCR and RT-PCR analyses. Results indicated that the average number of leaf width, petiole length and leaf number of transgenic plants were 19.31 cm, 29.17 cm and 38.33, which increased obviously by 31.81%, 16.61% and, 17.04%, respectively, the average number of leaf length were 25.16 cm, which increased insignificantly by 1.68%. However, there were no significant difference in the content of Chlorophyll a(1.31 mg·g-1), Chlorophyll b (0.562 mg·g-1) and total Chlorophyll(1.87 mg·g-1) between transgenic and control plants. Compared to control, soluble sugar content (14.59 mg·g-1) of leaf was reduced markedly by 13.04%, but soluble sugar content (21.90 mg·g-1) of petiole was only increased by 3.36%. The average root weight and sugar content of individual plants were 3.067 kg and 183.2 g·kg-1, respectively. Compared with control plants, the root weight of transgenic plants increased by 5.91%, while saccharinity increased significantly by 9

  11. A re-assessment of sucrose signaling involved in cluster-root formation and function in phosphate-deficient white lupin (Lupinus albus).

    Science.gov (United States)

    Wang, Zhengrui; Shen, Jianbo; Ludewig, Uwe; Neumann, Günter

    2015-07-01

    Apart from substrate functions, a signaling role of sucrose in root growth regulation is well established. This raised the question whether sucrose signals might also be involved in formation of cluster-roots (CRs) under phosphate (Pi) limitation, mediating exudation of phosphorus (P)-mobilizing root exudates, e.g. in Lupinus albus and members of the Proteaceae. Earlier studies demonstrated that CR formation in L. albus was mimicked to some extent by external application of high sucrose concentrations (25 mM) in the presence of extremely high P supply (1-10 mM), usually suppressing CR formation. In this study, we re-addressed this question using an axenic hydroponic culture system with normal P supply (0.1 mM) and a range of sucrose applications (0.25-25 mM). The 2.5 mM sucrose concentration was comparable with internal sucrose levels in the zone of CR initiation in first-order laterals of P-deficient plants (3.4 mM) and induced the same CR morphology. Similar to earlier studies, high sucrose concentrations (25 mM) resulted in root thickening and inhibition of root elongation, associated with a 10-fold increase of the internal sucrose level. The sucrose analog palatinose and a combination of glucose/fructose failed to stimulate CR formation under P-sufficient conditions, demonstrating a signal function of sucrose and excluding osmotic or carbon source effects. In contrast to earlier findings, sucrose was able to induce CR formation but had no effect on CR functioning with respect to citrate exudation, in vitro activity and expression of genes encoding phosphoenolpyruvate carboxylase, secretory acid phosphatase and MATE transporters, mediating P-mobilizing functions of CRs.

  12. Structural rearrangements of sucrose phosphorylase from Bifidobacterium adolescentis during sucrose conversion

    DEFF Research Database (Denmark)

    Mirza, Osman; Henriksen, Lars Skov; Sprogøe, Desiree;

    2006-01-01

    in other related sucrose-acting enzymes. Wild-type BiSP was also crystallized in the presence of sucrose. In the dimeric structure, a covalent glucosyl intermediate was formed in one molecule of the BiSP dimer, and after hydrolysis of the glucosyl intermediate, a beta-D-glucose product complex was formed...... in the other molecule. Although the overall structure of the BiSP-glucosyl intermediate complex is similar to that of the BiSP(E232Q)-sucrose complex, the glucose complex discloses major differences in loop conformations. Two loops (residues 336-344 and 132-137) in the proximity of the active site move up...... to 16 and 4 A, respectively. On the basis of these findings, we have suggested a reaction cycle that takes into account the large movements in the active-site entrance loops....

  13. Sucrose esters increase drug penetration, but do not inhibit p-glycoprotein in caco-2 intestinal epithelial cells.

    Science.gov (United States)

    Kiss, Lóránd; Hellinger, Éva; Pilbat, Ana-Maria; Kittel, Ágnes; Török, Zsolt; Füredi, András; Szakács, Gergely; Veszelka, Szilvia; Sipos, Péter; Ózsvári, Béla; Puskás, László G; Vastag, Monika; Szabó-Révész, Piroska; Deli, Mária A

    2014-10-01

    Sucrose fatty acid esters are increasingly used as excipients in pharmaceutical products, but few data are available on their toxicity profile, mode of action, and efficacy on intestinal epithelial models. Three water-soluble sucrose esters, palmitate (P-1695), myristate (M-1695), laurate (D-1216), and two reference absorption enhancers, Tween 80 and Cremophor RH40, were tested on Caco-2 cells. Caco-2 monolayers formed a good barrier as reflected by high transepithelial resistance and positive immunostaining for junctional proteins claudin-1, ZO-1, and β-catenin. Sucrose esters in nontoxic concentrations significantly reduced resistance and impedance, and increased permeability for atenolol, fluorescein, vinblastine, and rhodamine 123 in Caco-2 monolayers. No visible opening of the tight junctions was induced by sucrose esters assessed by immunohistochemistry and electron microscopy, but some alterations were seen in the structure of filamentous actin microfilaments. Sucrose esters fluidized the plasma membrane and enhanced the accumulation of efflux transporter ligands rhodamine 123 and calcein AM in epithelial cells, but did not inhibit the P-glycoprotein (P-gp)-mediated calcein AM accumulation in MES-SA/Dx5 cell line. These data indicate that in addition to their dissolution-increasing properties sucrose esters can enhance drug permeability through both the transcellular and paracellular routes without inhibiting P-gp.

  14. Tissue- and isoform-specific phytochrome regulation of light-dependent anthocyanin accumulation in Arabidopsis thaliana

    OpenAIRE

    Warnasooriya, Sankalpi N.; Porter, Katie J.; Montgomery, Beronda L

    2011-01-01

    Phytochromes regulate light- and sucrose-dependent anthocyanin synthesis and accumulation in many plants. Mesophyll-specific phyA alone has been linked to the regulation of anthocyanin accumulation in response to far-red light in Arabidopsis thaliana. However, multiple mesophyll-localized phytochromes were implicated in the photoregulation of anthocyanin accumulation in red-light conditions. Here, we report a role for mesophyll-specific phyA in blue-light-dependent regulation of anthocyanin l...

  15. Sucrose and Warmth for Analgesia in Healthy Newborns: An RCT

    Science.gov (United States)

    Garza, Elizabeth; Zageris, Danielle; Heilman, Keri J.; Porges, Stephen W.

    2015-01-01

    BACKGROUND AND OBJECTIVE: Increasing data suggest that neonatal pain has long-term consequences. Nonpharmacologic techniques (sucrose taste, pacifier suckling, breastfeeding) are effective and now widely used to combat minor neonatal pain. This study examined the analgesic effect of sucrose combined with radiant warmth compared with the taste of sucrose alone during a painful procedure in healthy full-term newborns. METHODS: A randomized, controlled trial included 29 healthy, full-term newborns born at the University of Chicago Hospital. Both groups of infants were given 1.0 mL of 25% sucrose solution 2 minutes before the vaccination, and 1 group additionally was given radiant warmth from an infant warmer before the vaccination. We assessed pain by comparing differences in cry, grimace, heart rate variability (ie, respiratory sinus arrhythmia), and heart rate between the groups. RESULTS: The sucrose plus warmer group cried and grimaced for 50% less time after the vaccination than the sucrose alone group (P < .05, respectively). The sucrose plus warmer group had lower heart rate and heart rate variability (ie, respiratory sinus arrhythmia) responses compared with the sucrose alone group (P < .01), reflecting a greater ability to physiologically regulate in response to the painful vaccination. CONCLUSIONS: The combination of sucrose and radiant warmth is an effective analgesic in newborns and reduces pain better than sucrose alone. The ready availability of this practical nonpharmacologic technique has the potential to reduce the burden of newborn pain. PMID:25687147

  16. Leaves Of Cut Rose Flower Convert Exogenously Applied Glucose To Sucrose And Translocate It To Petals

    Directory of Open Access Journals (Sweden)

    Horibe Takanori

    2014-12-01

    Full Text Available To understand the role that the leaves play in the translocation of soluble carbohydrates in cut rose flowers, we first evaluated the effect of leaf removal on flower quality and the sugar content in petals. Cut rose flowers with leaves had higher soluble sugar content in petals compared with cut flower without leaves. Next, we treated cut flowers with radioactive glucose to clarify translocation routes of exogenously applied sugar. There was no significant difference between the specific radioactivity of sucrose and glucose in leaves, but specific radioactivity of sucrose in petals was much higher than that of glucose. These results suggested that most of the exogenously applied glucose first moved to the leaves, where it was converted into sucrose and then the synthesised sucrose was translocated to the petals. Our results showed that the leaves of cut rose flowers play an important role in the metabolism and transportation of exogenously applied soluble carbohydrates toward the petals, thus contributing to sustaining the post-harvest quality.

  17. Comprehensive Mutational Analysis of Sucrose-Metabolizing Pathways in Streptococcus mutans Reveals Novel Roles for the Sucrose Phosphotransferase System Permease

    Science.gov (United States)

    Zeng, Lin

    2013-01-01

    Sucrose is perhaps the most efficient carbohydrate for the promotion of dental caries in humans, and the primary caries pathogen Streptococcus mutans encodes multiple enzymes involved in the metabolism of this disaccharide. Here, we engineered a series of mutants lacking individual or combinations of sucrolytic pathways to understand the control of sucrose catabolism and to determine whether as-yet-undisclosed pathways for sucrose utilization were present in S. mutans. Growth phenotypes indicated that gtfBCD (encoding glucan exopolysaccharide synthases), ftf (encoding the fructan exopolysaccharide synthase), and the scrAB pathway (sugar-phosphotransferase system [PTS] permease and sucrose-6-PO4 hydrolase) constitute the majority of the sucrose-catabolizing activity; however, mutations in any one of these genes alone did not affect planktonic growth on sucrose. The multiple-sugar metabolism pathway (msm) contributed minimally to growth on sucrose. Notably, a mutant lacking gtfBC, which cannot produce water-insoluble glucan, displayed improved planktonic growth on sucrose. Meanwhile, loss of scrA led to growth stimulation on fructooligosaccharides, due in large part to increased expression of the fruAB (fructanase) operon. Using the LevQRST four-component signal transduction system as a model for carbohydrate-dependent gene expression in strains lacking extracellular sucrases, a PlevD-cat (EIIALev) reporter was activated by pulsing with sucrose. Interestingly, ScrA was required for activation of levD expression by sucrose through components of the LevQRST complex, but not for activation by the cognate LevQRST sugars fructose or mannose. Sucrose-dependent catabolite repression was also evident in strains containing an intact sucrose PTS. Collectively, these results reveal a novel regulatory circuitry for the control of sucrose catabolism, with a central role for ScrA. PMID:23222725

  18. Control of Enzyme IIscr and Sucrose-6-Phosphate Hydrolase Activities in Streptococcus mutans by Transcriptional Repressor ScrR Binding to the cis-Active Determinants of the scr Regulon

    OpenAIRE

    Wang, Bing; Kuramitsu, Howard K.

    2003-01-01

    In Streptococcus mutans, enzyme IIscr and sucrose-6-phosphate hydrolase are two important enzymes in the transport and metabolism of dietary sucrose. The scr regulon of S. mutans is composed of three genes, scrA and scrB, which code for enzyme IIscr and sucrose-6-phosphate hydrolase, respectively, and scrR, which codes for a GalR-LacI-type transcription regulator. It was previously shown that expression of both scrA and scrB is similarly induced by sucrose. Mutation in the scrR gene resulted ...

  19. Kinetics of sucrose crystallization in whey protein films.

    Science.gov (United States)

    Dangaran, Kirsten L; Krochta, John M

    2006-09-20

    The kinetics of sucrose crystallization in whey protein isolate (WPI) films was studied at 25 degrees C in four different relative humidity environments: 23, 33, 44, and 53%. The effects of protein matrix, crystallization inhibitors, and storage environment on the rate constants of sucrose crystallization were determined using the Avrami model of crystallization. It was found that a cross-linked, denatured whey protein (WP) matrix more effectively hindered sucrose crystallization than a protein matrix of native WP. The crystallization inhibitors tested were lactose, raffinose, modified starch (Purity 69), and polyvinylpyrrolidone (Plasdone C15). Raffinose and modified starch were determined to be the more effective inhibitors of sucrose crystallization. At lower relative humidities (23, 33, and 44%), the cross-linked protein matrix played a more important role in sucrose crystallization than the inhibitors. As relative humidity increased (53%), the crystallization inhibitors were more central to controlling sucrose crystallization in WPI films.

  20. Effect of salt on the response of birds to sucrose

    Science.gov (United States)

    Rogers, J.G.; Maller, O.

    1973-01-01

    The preference of male red-winged blackbirds for solutions of sucrose and sucrose with 0.03 M sodium chloride was tested, using a two-bottle choice test. Preliminary experiments demonstrated that the birds were indifferent to 0.03 M NaCl in water. Both control and experimental animals exhibited indifference to the solutions at the lowest concentration and aversion at the highest. The data suggest that the added sodium chloride makes the sucrose stimulus more discriminable.

  1. STP10 encodes a high-affinity monosaccharide transporter and is induced under low-glucose conditions in pollen tubes of Arabidopsis.

    Science.gov (United States)

    Rottmann, Theresa; Zierer, Wolfgang; Subert, Christa; Sauer, Norbert; Stadler, Ruth

    2016-04-01

    Pollen tubes are fast growing, photosynthetically inactive cells. Their energy demand is covered by specific transport proteins in the plasma membrane that mediate the uptake of sugars. Here we report on the functional characterization of AtSTP10, a previously uncharacterized member of the SUGAR TRANSPORT PROTEIN family. Heterologous expression of STP10 cDNA in yeast revealed that the encoded protein catalyses the high-affinity uptake of glucose, galactose and mannose. The transporter is sensitive to uncouplers of transmembrane proton gradients, indicating that the protein acts as a hexose-H(+)symporter. Analyses of STP10 mRNA and STP10 promoter-reporter gene studies revealed a sink-specific expression pattern of STP10 in primordia of lateral roots and in pollen tubes. This restriction to sink organs is mediated by intragenic regions of STP10 qPCR analyses with cDNA of in vitro grown pollen tubes showed that STP10 expression was down-regulated in the presence of 50mM glucose. However, in pollen tubes of glucose-insensitive plants, which lack the glucose sensor hexokinase1 (HXK1), no glucose-induced down-regulation of STP10 expression was detected. A stp10T-DNA insertion line developed normally, which may point towards functional redundancy. The data presented in this paper indicate that a high-affinity glucose uptake system is induced in growing pollen tubes under low glucose conditions and that this regulation may occur through the hexokinase pathway. PMID:26893494

  2. Sucrose metabolism in halotolerant methanotroph Methylomicrobium alcaliphilum 20Z.

    Science.gov (United States)

    But, Sergey Y; Khmelenina, Valentina N; Reshetnikov, Alexander S; Mustakhimov, Ildar I; Kalyuzhnaya, Marina G; Trotsenko, Yuri A

    2015-04-01

    Sucrose accumulation has been observed in some methylotrophic bacteria utilizing methane, methanol, or methylated amines as a carbon and energy source. In this work, we have investigated the biochemical pathways for sucrose metabolism in the model halotolerant methanotroph Methylomicrobium alcaliphilum 20Z. The genes encoding sucrose-phosphate synthase (Sps), sucrose-phosphate phosphatase (Spp), fructokinase (FruK), and amylosucrase (Ams) were co-transcribed and displayed similar expression levels. Functional Spp and Ams were purified after heterologous expression in Escherichia coli. Recombinant Spp exhibited high affinity for sucrose-6-phosphate and stayed active at very high levels of sucrose (K i  = 1.0 ± 0.6 M). The recombinant amylosucrase obeyed the classical Michaelis-Menten kinetics in the reactions of sucrose hydrolysis and transglycosylation. As a result, the complete metabolic network for sucrose biosynthesis and re-utilization in the non-phototrophic organism was reconstructed for the first time. Comparative genomic studies revealed analogous gene clusters in various Proteobacteria, thus indicating that the ability to produce and metabolize sucrose is widespread among prokaryotes.

  3. Sucrose and KF quenching system for solution phase parallel synthesis.

    Science.gov (United States)

    Chavan, Sunil; Watpade, Rahul; Toche, Raghunath

    2016-01-01

    The KF, sucrose (table sugar) exploited as quenching system in solution phase parallel synthesis. Excess of electrophiles were covalently trapped with hydroxyl functionality of sucrose and due to polar nature of sucrose derivative was solubilize in water. Potassium fluoride used to convert various excess electrophilic reagents such as acid chlorides, sulfonyl chlorides, isocyanates to corresponding fluorides, which are less susceptible for hydrolysis and subsequently sucrose traps these fluorides and dissolves them in water thus removing them from reaction mixture. Various excess electrophilic reagents such as acid chlorides, sulfonyl chlorides, and isocyanates were quenched successfully to give pure products in excellent yields. PMID:27462506

  4. Exogenous sucrose supply changes sugar metabolism and reduces photosynthesis of sugarcane through the down-regulation of Rubisco abundance and activity.

    Science.gov (United States)

    Lobo, Ana Karla Moreira; de Oliveira Martins, Marcio; Lima Neto, Milton Costa; Machado, Eduardo Caruso; Ribeiro, Rafael Vasconcelos; Silveira, Joaquim Albenisio Gomes

    2015-05-01

    Photosynthetic modulation by sugars has been known for many years, but the biochemical and molecular comprehension of this process is lacking. We studied how the exogenous sucrose supplied to leaves could affect sugar metabolism in leaf, sheath and stalk and inhibit photosynthesis in four-month old sugarcane plants. Exogenous sucrose 50mM sprayed on attached leaves strongly impaired the net CO2 assimilation (PN) and decreased the instantaneous carboxylation efficiency (PN/Ci), suggesting that the impairment in photosynthesis was caused by biochemical restrictions. The photosystem II activity was also affected by excess sucrose as indicated by the reduction in the apparent electron transport rate, effective quantum yield and increase in non-photochemical quenching. In leaf segments, sucrose accumulation was related to increases in the activities of soluble acid and neutral invertases, sucrose synthase and sucrose phosphate synthase, whereas the contents of fructose increased and glucose slightly decreased. Changes in the activities of sucrose hydrolyzing and synthesizing enzymes in leaf, sheath and stalk and sugar profile in intact plants were not enough to identify which sugar(s) or enzyme(s) were directly involved in photosynthesis modulation. However, exogenous sucrose was able to trigger down-regulation in the Rubisco abundance, activation state and enzymatic activity. Despite the fact that PN/Ci had been notably decreased by sucrose, in vitro activity and abundance of PEPCase did not change, suggesting an in vivo modulation of this enzyme. The data reveal that sucrose and/or other derivative sugars in leaves inhibited sugarcane photosynthesis by down-regulation of Rubisco synthesis and activity. Our data also suggest that sugar modulation was not exerted by a feedback mechanism induced by the accumulation of sugars in immature sugarcane stalk.

  5. Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes

    Science.gov (United States)

    Winter, H.; Huber, S. C.; Brown, C. S. (Principal Investigator)

    2000-01-01

    Sucrose (Suc) plays a central role in plant growth and development. It is a major end product of photosynthesis and functions as a primary transport sugar and in some cases as a direct or indirect regulator of gene expression. Research during the last 2 decades has identified the pathways involved and which enzymes contribute to the control of flux. Availability of metabolites for Suc synthesis and 'demand' for products of sucrose degradation are important factors, but this review specifically focuses on the biosynthetic enzyme sucrose-phosphate synthase (SPS), and the degradative enzymes, sucrose synthase (SuSy), and the invertases. Recent progress has included the cloning of genes encoding these enzymes and the elucidation of posttranslational regulatory mechanisms. Protein phosphorylation is emerging as an important mechanism controlling SPS activity in response to various environmental and endogenous signals. In terms of Suc degradation, invertase-catalyzed hydrolysis generally has been associated with cell expansion, whereas SuSy-catalyzed metabolism has been linked with biosynthetic processes (e.g., cell wall or storage products). Recent results indicate that SuSy may be localized in multiple cellular compartments: (1) as a soluble enzyme in the cytosol (as traditionally assumed); (2) associated with the plasma membrane; and (3) associated with the actin cytoskeleton. Phosphorylation of SuSy has been shown to occur and may be one of the factors controlling localization of the enzyme. The purpose of this review is to summarize some of the recent developments relating to regulation of activity and localization of key enzymes involved in sucrose metabolism in plants.

  6. Compartmentation of sucrose during radial transfer in mature sorghum culm

    Directory of Open Access Journals (Sweden)

    Vietor Donald M

    2007-06-01

    Full Text Available Abstract Background The sucrose that accumulates in the culm of sorghum (Sorghum bicolor (L. Moench and other large tropical andropogonoid grasses can be of commercial value, and can buffer assimilate supply during development. Previous study conducted with intact plants showed that sucrose can be radially transferred to the intracellular compartment of mature ripening sorghum internode without being hydrolysed. In this study, culm-infused radiolabelled sucrose was traced between cellular compartments and among related metabolites to determine if the compartmental path of sucrose during radial transfer in culm tissue was symplasmic or included an apoplasmic step. This transfer path was evaluated for elongating and ripening culm tissue of intact plants of two semidwarf grain sorghums. The metabolic path in elongating internode tissue was also evaluated. Results On the day after culm infusion of the tracer sucrose, the specific radioactivity of sucrose recovered from the intracellular compartment of growing axillary-branch tissue was greater (nearly twice than that in the free space, indicating that sucrose was preferentially transferred through symplasmic routes. In contrast, the sucrose specific radioactivity in the intracellular compartment of the mature (ripening culm tissue was probably less (about 3/4's than that in free space indicating that sucrose was preferentially transferred through routes that included an apoplasmic step. In growing internodes of the axillary branch of sorghum, the tritium label initially provided in the fructose moiety of sucrose molecules was largely (81% recovered in the fructose moiety, indicating that a large portion of sucrose molecules is not hydrolysed and resynthesized during radial transfer. Conclusion During radial transfer of sucrose in ripening internodes of intact sorghum plants, much of the sucrose is transferred intact (without hydrolysis and resynthesis and primarily through a path that includes an

  7. Effects of Soil Salinity on Sucrose Metabolism in Cotton Fiber.

    Science.gov (United States)

    Peng, Jun; Zhang, Lei; Liu, Jingran; Luo, Junyu; Zhao, Xinhua; Dong, Helin; Ma, Yan; Sui, Ning; Zhou, Zhiguo; Meng, Yali

    2016-01-01

    Cotton (Gosspium hirsutum L.) is classified as a salt tolerant crop. However, its yield and fiber quality are negatively affected by soil salinity. Studies on the enzymatic differences in sucrose metabolism under different soil salinity levels are lacking. Therefore, field experiments, using two cotton cultivars, CCRI-79 (salt-tolerant) and Simian 3 (salt-sensitive), were conducted in 2013 and 2014 at three different salinity levels (1.15 dS m-1 [low soil salinity], 6.00 dS m-1 [medium soil salinity], and 11.46 dS m-1 [high soil salinity]). The objective was to elucidate the effects of soil salinity on sucrose content and the activity of key enzymes that are related to sucrose metabolism in cotton fiber. Results showed that as the soil salinity increased, cellulose content, sucrose content, and sucrose transformation rate declined; the decreases in cellulose content and sucrose transformation rate caused by the increase in soil salinity were more in Simian 3 than those in CCRI-79. With increase in soil salinity, activities of sucrose metabolism enzymes sucrose phophate synthase (SPS), acidic invertase, and alkaline invertase were decreased, whereas sucrose synthase (SuSy) activity increased. However, the changes displayed in the SuSy and SPS activities in response to increase in soil salinity were different and the differences were large between the two cotton cultivars. These results illustrated that suppressed cellulose synthesis and sucrose metabolism under high soil salinity were mainly due to the change in SPS, SuSy, and invertase activities, and the difference in cellulose synthesis and sucrose metabolism in fiber for the two cotton cultivars in response to soil salinity was determined mainly by both SuSy and SPS activities. PMID:27227773

  8. Effects of Soil Salinity on Sucrose Metabolism in Cotton Fiber.

    Directory of Open Access Journals (Sweden)

    Jun Peng

    Full Text Available Cotton (Gosspium hirsutum L. is classified as a salt tolerant crop. However, its yield and fiber quality are negatively affected by soil salinity. Studies on the enzymatic differences in sucrose metabolism under different soil salinity levels are lacking. Therefore, field experiments, using two cotton cultivars, CCRI-79 (salt-tolerant and Simian 3 (salt-sensitive, were conducted in 2013 and 2014 at three different salinity levels (1.15 dS m-1 [low soil salinity], 6.00 dS m-1 [medium soil salinity], and 11.46 dS m-1 [high soil salinity]. The objective was to elucidate the effects of soil salinity on sucrose content and the activity of key enzymes that are related to sucrose metabolism in cotton fiber. Results showed that as the soil salinity increased, cellulose content, sucrose content, and sucrose transformation rate declined; the decreases in cellulose content and sucrose transformation rate caused by the increase in soil salinity were more in Simian 3 than those in CCRI-79. With increase in soil salinity, activities of sucrose metabolism enzymes sucrose phophate synthase (SPS, acidic invertase, and alkaline invertase were decreased, whereas sucrose synthase (SuSy activity increased. However, the changes displayed in the SuSy and SPS activities in response to increase in soil salinity were different and the differences were large between the two cotton cultivars. These results illustrated that suppressed cellulose synthesis and sucrose metabolism under high soil salinity were mainly due to the change in SPS, SuSy, and invertase activities, and the difference in cellulose synthesis and sucrose metabolism in fiber for the two cotton cultivars in response to soil salinity was determined mainly by both SuSy and SPS activities.

  9. Leaves of the Arabidopsis maltose exporter1 mutant exhibit a metabolic profile with features of cold acclimation in the warm.

    Directory of Open Access Journals (Sweden)

    Sarah J Purdy

    Full Text Available BACKGROUND: Arabidopsis plants accumulate maltose from starch breakdown during cold acclimation. The Arabidopsis mutant, maltose excess1-1, accumulates large amounts of maltose in the plastid even in the warm, due to a deficient plastid envelope maltose transporter. We therefore investigated whether the elevated maltose level in mex1-1 in the warm could result in changes in metabolism and physiology typical of WT plants grown in the cold. PRINCIPAL FINDINGS: Grown at 21 °C, mex1-1 plants were much smaller, with fewer leaves, and elevated carbohydrates and amino acids compared to WT. However, after transfer to 4 °C the total soluble sugar pool and amino acid concentration was in equal abundance in both genotypes, although the most abundant sugar in mex1-1 was still maltose whereas sucrose was in greatest abundance in WT. The chlorophyll a/b ratio in WT was much lower in the cold than in the warm, but in mex1-1 it was low in both warm and cold. After prolonged growth at 4 °C, the shoot biomass, rosette diameter and number of leaves at bolting were similar in mex1-1 and WT. CONCLUSIONS: The mex1-1 mutation in warm-grown plants confers aspects of cold acclimation, including elevated levels of sugars and amino acids and low chlorophyll a/b ratio. This may in turn compromise growth of mex1-1 in the warm relative to WT. We suggest that elevated maltose in the plastid could be responsible for key aspects of cold acclimation.

  10. Sucrose and Saccharomyces cerevisiae: a relationship most sweet.

    Science.gov (United States)

    Marques, Wesley Leoricy; Raghavendran, Vijayendran; Stambuk, Boris Ugarte; Gombert, Andreas Karoly

    2016-02-01

    Sucrose is an abundant, readily available and inexpensive substrate for industrial biotechnology processes and its use is demonstrated with much success in the production of fuel ethanol in Brazil. Saccharomyces cerevisiae, which naturally evolved to efficiently consume sugars such as sucrose, is one of the most important cell factories due to its robustness, stress tolerance, genetic accessibility, simple nutrient requirements and long history as an industrial workhorse. This minireview is focused on sucrose metabolism in S. cerevisiae, a rather unexplored subject in the scientific literature. An analysis of sucrose availability in nature and yeast sugar metabolism was performed, in order to understand the molecular background that makes S. cerevisiae consume this sugar efficiently. A historical overview on the use of sucrose and S. cerevisiae by humans is also presented considering sugarcane and sugarbeet as the main sources of this carbohydrate. Physiological aspects of sucrose consumption are compared with those concerning other economically relevant sugars. Also, metabolic engineering efforts to alter sucrose catabolism are presented in a chronological manner. In spite of its extensive use in yeast-based industries, a lot of basic and applied research on sucrose metabolism is imperative, mainly in fields such as genetics, physiology and metabolic engineering.

  11. Functionality of Inulin as a Sucrose Replacer in Cookie Baking

    Science.gov (United States)

    Inulin was evaluated as a sucrose replacer for healthy cookie production with benefits of low glycemic impact and prebiotic soluble fiber. Sucrose (as a reference) and three inulin products of different concentrations (as soluble fibers) were used to explore the effects of sugar-replacer type on so...

  12. Citrate increases glass transition temperature of vitrified sucrose preparations

    NARCIS (Netherlands)

    Kets, E.P.W.; Lipelaar, P.J.; Hoekstra, F.A.; Vromans, H.

    2004-01-01

    The aim of this study was to investigate the effect of sodium citrate on the properties of dried amorphous sucrose glasses. Addition of sodium citrate to a sucrose solution followed by freeze-drying or convective drying resulted in a glass transition temperature (T-g) that was higher than the well-s

  13. Probing Osmotic Effects on Invertase with l-(−)-Sucrose

    OpenAIRE

    Seo, Seung-Kee; Wei, Alexander

    2008-01-01

    l-(−)-Sucrose‡ was efficiently synthesized using intramolecular aglycon delivery and used to elucidate osmotic effects on the activity of invertase, which catalyzes the hydrolysis of d-(+)-sucrose. The osmotic effect imposed by l-sucrose was responsible for more than 30% of the activity loss ascribed otherwise to “substrate inhibition.”

  14. ROP3 GTPase contributes to polar auxin transport and auxin responses and is important for embryogenesis and seedling growth in Arabidopsis.

    Science.gov (United States)

    Huang, Jia-bao; Liu, Huili; Chen, Min; Li, Xiaojuan; Wang, Mingyan; Yang, Yali; Wang, Chunling; Huang, Jiaqing; Liu, Guolan; Liu, Yuting; Xu, Jian; Cheung, Alice Y; Tao, Li-zhen

    2014-09-01

    ROP GTPases are crucial for the establishment of cell polarity and for controlling responses to hormones and environmental signals in plants. In this work, we show that ROP3 plays important roles in embryo development and auxin-dependent plant growth. Loss-of-function and dominant-negative (DN) mutations in ROP3 induced a spectrum of similar defects starting with altered cell division patterning during early embryogenesis to postembryonic auxin-regulated growth and developmental responses. These resulted in distorted embryo development, defective organ formation, retarded root gravitropism, and reduced auxin-dependent hypocotyl elongation. Our results showed that the expression of AUXIN RESPONSE FACTOR5/MONOPTEROS and root master regulators PLETHORA1 (PLT1) and PLT2 was reduced in DN-rop3 mutant embryos, accounting for some of the observed patterning defects. ROP3 mutations also altered polar localization of auxin efflux proteins (PINs) at the plasma membrane (PM), thus disrupting auxin maxima in the root. Notably, ROP3 is induced by auxin and prominently detected in root stele cells, an expression pattern similar to those of several stele-enriched PINs. Our results demonstrate that ROP3 is important for maintaining the polarity of PIN proteins at the PM, which in turn ensures polar auxin transport and distribution, thereby controlling plant patterning and auxin-regulated responses.

  15. The Arabidopsis thaliana RNA Editing FactorSLO2, which Affects the Mitochondrial ElectronTransport Chain, Participates in Multiple Stressand Hormone Resoonses

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Recently, we reported that the novel mitochondrial RNA editing factor SLO2 is essential for mitochondrialelectron transport, and vital for plant growth through regulation of carbon and energy metabolism. Here, we show thatmutation in SL02 causes hypersensitivity to ABA and insensitivity to ethylene, suggesting a link with stress responses.Indeed, slo2 mutants are hypersensitive to salt and osmotic stress during the germination stage, while adult plantsshow increased drought and salt tolerance. Moreover, slo2 mutants are more susceptible to Botrytis cinerea infection.An increased expression of nuclear-encoded stress-responsive genes, as well as mitochondrial-encoded NAD genes ofcomplex I and genes of the alternative respiratory pathway, was observed in slo2 mutants, further enhanced by ABAtreatment. In addition, H202 accumulation and altered amino acid levels were recorded in slo2 mutants. We conclude thatSLO2 is required for plant sensitivity to ABA, ethylene, biotic, and abiotic stress. Although two stress-related RNA editingfactors were reported very recently, this study demonstrates a unique role of SLO2, and further supports a link betweenmitochondrial RNA editing events and stress response.

  16. Sucrose controlled translation mediated by a sequence conserved peptide in Arabidopsis thaliana

    NARCIS (Netherlands)

    Lastdrager, Jeroen

    2015-01-01

    The sessile nature of plants demands adaptation to a variety of external stimuli. Sugars have well established roles in steering growth and developmental processes, and sugar levels are a good proxy for the energy status in plants. Molecular sugar signaling networks perceive the availability of suga

  17. Sucrose mimics the light induction of Arabidopsis nitrate reductase gene transcription

    DEFF Research Database (Denmark)

    Cheng, Chi-Lien; Acedo, Gregoria N; Kristensen, Michael;

    1992-01-01

    Nitrate reductase, the first enzyme in nitrate assimilation, is located at the crossroad of two energy-consuming pathways: nitrate assimilation and carbon fixation. Light, which regulates the expression of many higher-plant carbon fixation genes, also regulates nitrate reductase gene expression. ...

  18. Sucrose utilization by Zymomonas mobilis: formation of a levan

    Science.gov (United States)

    Dawes, E. A.; Ribbons, D. W.; Rees, D. A.

    1966-01-01

    1. Molar growth-yield coefficients of Zymomonas mobilis for glucose, fructose, glucose plus fructose, and sucrose are reported. Yield coefficients for sucrose are appreciably lower than those for the equivalent concentrations of glucose plus fructose. 2. Only 2·6% of [U-14C]glucose supplied in the growth medium is incorporated into cell substance by Z. mobilis utilizing glucose as the energy source. 3. During growth on sucrose a levan is formed. It has been characterized and shown to resemble other bacterial levans. 4. Levan formation from sucrose could be demonstrated with both washed cell suspensions and cell extracts of Z. mobilis. 5. Sucrose phosphorylase could not be demonstrated in extracts of the organism. PMID:4287843

  19. Sucrose behenate as a crystallization enhancer for soft fats.

    Science.gov (United States)

    Domingues, Maria Aliciane Fontenele; da Silva, Thaís Lomonaco Teodoro; Ribeiro, Ana Paula Badan; Chiu, Ming Chih; Gonçalves, Lireny Aparecida Guaraldo

    2016-02-01

    The addition of sucrose behenate for the modification of the physical properties of soft fats, such as soybean oil-based interesterified fat, refined palm oil, and palm mid fraction was studied. The addition of sucrose behenate was verified to affect the crystalline network of fats, changing the hardness and solids profile. The isothermal crystallization behaviors of the fat blends with 1% sucrose behenate were analyzed at 20 and 25 °C. Temperature had a greater effect on the speed of crystallization (k) than the presence of the emulsifier. Sucrose behenate did, however, influence the crystallization mechanism, with changes observed in the Avrami exponent (n). These changes were also observed in the microstructure of the fats. Changes in the polymorphic behavior were observed with the addition of sucrose behenate, such as a possible delay in the α → β transition for interesterified fat, and the initial formation of the β polymorph in palm oil.

  20. Sucrose behenate as a crystallization enhancer for soft fats.

    Science.gov (United States)

    Domingues, Maria Aliciane Fontenele; da Silva, Thaís Lomonaco Teodoro; Ribeiro, Ana Paula Badan; Chiu, Ming Chih; Gonçalves, Lireny Aparecida Guaraldo

    2016-02-01

    The addition of sucrose behenate for the modification of the physical properties of soft fats, such as soybean oil-based interesterified fat, refined palm oil, and palm mid fraction was studied. The addition of sucrose behenate was verified to affect the crystalline network of fats, changing the hardness and solids profile. The isothermal crystallization behaviors of the fat blends with 1% sucrose behenate were analyzed at 20 and 25 °C. Temperature had a greater effect on the speed of crystallization (k) than the presence of the emulsifier. Sucrose behenate did, however, influence the crystallization mechanism, with changes observed in the Avrami exponent (n). These changes were also observed in the microstructure of the fats. Changes in the polymorphic behavior were observed with the addition of sucrose behenate, such as a possible delay in the α → β transition for interesterified fat, and the initial formation of the β polymorph in palm oil. PMID:26304437

  1. Analysis and protease-catalysed synthesis of sucrose alkanoate regioisomers

    DEFF Research Database (Denmark)

    Lie, Aleksander

    2014-01-01

    laurate in DMF using serine proteases and a metalloprotease. A broad range of elution strategies for the chromatographic analysis of sucrose alkanoate regioisomers was systematically investigated using design of experiments strategies and statistical and multivariate analysis and modelling. Efficiency......The aims of the presented research were to develop quantifiable methods for reversed-phase high-performance liquid chromatography analysis of sucrose alkanoate regioisomers and to investigate the activity and regioisomeric distribution in the biocatalytic esterification of sucrose with vinyl...... evaluation of the elution strategies, in terms of the resolution metric general resolution deviation for multiple peaks (RDm) and analysis time, showed that the best results were offered by step-down gradient elution for sucrose caprate and isocratic elution with increased flow for sucrose laurate. Step...

  2. Acid Phosphatase Activity May Affect the Tuber Swelling by Partially Regulating Sucrose-mediated Sugar Resorption in Potato

    Institute of Scientific and Technical Information of China (English)

    Da-Yong Wang; Yong Lian; De-Wei Zhu

    2008-01-01

    APase activity is involved in regulating many physiological and developmental events by affecting the resorption process.In this study, we investigate the role of APase activity in tuber development in potato. APase activities were mainly localized in cytoplasm, gaps among cells and stroma of amyloplasts of parenchyma cells at the stage of tuber swelling. AP1, encoding a putative APase, was also highly expressed in swelling tubers and a low level of expression was observed in elongated stolons and matured tubers. Inhibition of APase activity by applying Brefeldin A, an inhibitor of APase production and secretion, significantly suppressed the tuber swelling and moderately affected the stolon elongation and the tuberization frequency. During tuber development, sucrose serves as the main soluble sugar for long-distance transportation and resorption. Moreover, Inhibition of APase activity by Brefeldin A markedly reduced the sucrose content in tubers and further decreased the starch accumulation, suggesting that the function of APase in regulating the tuber swelling might be at least artially mediated by the sugar resorption. Exogenous sucrose treatments further indicate the important role of sucrose-mediated sugar resorption in tuber swelling. These results suggest that the APase activity might affect the tuber swelling by partially regulating the sucrose-mediated sugar resorption.

  3. 78 FR 66743 - Draft Guidance for Industry on Bioequivalence Recommendations for Iron Sucrose; Availability

    Science.gov (United States)

    2013-11-06

    ... Recommendations for Iron Sucrose; Availability AGENCY: Food and Drug Administration, HHS. ACTION: Notice. SUMMARY... entitled ``Bioequivalence Recommendations for Iron Sucrose.'' The recommendations provide specific guidance... iron sucrose injection. The draft guidance is a revised version of a previously issued draft...

  4. Homologous electron transport components fail to increase fatty acid hydroxylation in transgenic Arabidopsis thaliana [v2; ref status: indexed, http://f1000r.es/2a3

    Directory of Open Access Journals (Sweden)

    Laura L. Wayne

    2013-11-01

    Full Text Available Ricinoleic acid, a hydroxylated fatty acid (HFA present in castor (Ricinus communis seeds, is an important industrial commodity used in products ranging from inks and paints to polymers and fuels. However, due to the deadly toxin ricin and allergens also present in castor, it would be advantageous to produce ricinoleic acid in a different agricultural crop. Unfortunately, repeated efforts at heterologous expression of the castor fatty acid hydroxylase (RcFAH12 in the model plant Arabidopsis thaliana have produced only 17-19% HFA in the seed triacylglycerols (TAG, whereas castor seeds accumulate up to 90% ricinoleic acid in the endosperm TAG. RcFAH12 requires an electron supply from NADH:cytochrome b5 reductase (CBR1 and cytochrome b5 (Cb5 to synthesize ricinoleic acid. Previously, our laboratory found a mutation in the Arabidopsis CBR1 gene, cbr1-1, that caused an 85% decrease in HFA levels in the RcFAH12 Arabidopsis line. These results raise the possibility that electron supply to the heterologous RcFAH12 may limit the production of HFA. Therefore, we hypothesized that by heterologously expressing RcCb5, the reductant supply to RcFAH12 would be improved and lead to increased HFA accumulation in Arabidopsis seeds. Contrary to this proposal, heterologous expression of the top three RcCb5 candidates did not increase HFA accumulation. Furthermore, coexpression of RcCBR1 and RcCb5 in RcFAH12 Arabidopsis also did not increase in HFA levels compared to the parental lines. These results demonstrate that the Arabidopsis electron transfer system is supplying sufficient reductant to RcFAH12 and that there must be other bottlenecks limiting the accumulation of HFA.

  5. Suppressor Screens in Arabidopsis.

    Science.gov (United States)

    Li, Xin; Zhang, Yuelin

    2016-01-01

    Genetic screens have proven to be a useful tool in the dissection of biological processes in plants. Specifically, suppressor screens have been widely used to study signal transduction pathways. Here we provide a detailed protocol for ethyl methanesulfonate (EMS) mutagenesis used in our suppressor screens in Arabidopsis and discuss the basic principles behind suppressor screen design and downstream analyses. PMID:26577776

  6. Characterization of Sucrose Thin Films for Biomedical Applications

    Directory of Open Access Journals (Sweden)

    S. L. Iconaru

    2011-01-01

    Full Text Available Sucrose is a natural osmolyte accumulated in the cells of organisms as they adapt to environmental stress. In vitro sucrose increases protein stability and forces partially unfolded structures to refold. Thin films of sucrose (C12H22O11 were deposited on thin cut glass substrates by the thermal evaporation technique (P∼10−5 torr. Characteristics of thin films were put into evidence by Fourier Transform Infrared Spectroscopy (FTIR, X-ray Photoelectron Spectroscopy (XPS, scanning electron microscopy (SEM, and differential thermal analysis and thermal gravimetric analysis (TG/DTA. The experimental results confirm a uniform deposition of an adherent layer. In this paper we present a part of the characteristics of sucrose thin films deposited on glass in medium vacuum conditions, as a part of a culture medium for osteoblast cells. Osteoblast cells were used to determine proliferation, viability, and cytotoxicity interactions with sucrose powder and sucrose thin films. The osteoblast cells have been provided from the American Type Culture Collection (ATCC Centre. The outcome of this study demonstrated the effectiveness of sucrose thin films as a possible nontoxic agent for biomedical applications.

  7. Sucrose and IQ induced mutations in rat colon by independent

    DEFF Research Database (Denmark)

    Hansen, Max; Hald, M. T.; Autrup, H.;

    2004-01-01

    Sucrose-rich diets have repeatedly been observed to have co-carcinogenic actions in colon and liver of rats and to increase the number of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) induced aberrant crypt foci in rat colon. To investigate a possible interaction between sucrose and IQ...... on the genotoxicity in rat liver and colon, we gave Big Blue rats(TM) a diet containing sucrose (0%, 3.45% or 13.4% w/w) and/or IQ (70 ppm) for a period of 3 weeks. Sucrose and IQ increased the mutation frequency in the colon. The effect of combined treatments with IQ and sucrose on the mutation frequencies...... was additive indicating that sucrose and IQ act independently. This was supported by the mutation spectra where sucrose expands the background mutations in the colon, whereas IQ, in other studies, more specifically has induced G:C --> T:A transversions. In the liver IQ increased the mutation frequency, whereas...

  8. Functional Relationship between Sucrose and a Cariogenic Biofilm Formation.

    Directory of Open Access Journals (Sweden)

    Jian-Na Cai

    Full Text Available Sucrose is an important dietary factor in cariogenic biofilm formation and subsequent initiation of dental caries. This study investigated the functional relationships between sucrose concentration and Streptococcus mutans adherence and biofilm formation. Changes in morphological characteristics of the biofilms with increasing sucrose concentration were also evaluated. S. mutans biofilms were formed on saliva-coated hydroxyapatite discs in culture medium containing 0, 0.05, 0.1, 0.5, 1, 2, 5, 10, 20, or 40% (w/v sucrose. The adherence (in 4-hour biofilms and biofilm composition (in 46-hour biofilms of the biofilms were analyzed using microbiological, biochemical, laser scanning confocal fluorescence microscopic, and scanning electron microscopic methods. To determine the relationships, 2nd order polynomial curve fitting was performed. In this study, the influence of sucrose on bacterial adhesion, biofilm composition (dry weight, bacterial counts, and water-insoluble extracellular polysaccharide (EPS content, and acidogenicity followed a 2nd order polynomial curve with concentration dependence, and the maximum effective concentrations (MECs of sucrose ranged from 0.45 to 2.4%. The bacterial and EPS bio-volume and thickness in the biofilms also gradually increased and then decreased as sucrose concentration increased. Furthermore, the size and shape of the micro-colonies of the biofilms depended on the sucrose concentration. Around the MECs, the micro-colonies were bigger and more homogeneous than those at 0 and 40%, and were surrounded by enough EPSs to support their structure. These results suggest that the relationship between sucrose concentration and cariogenic biofilm formation in the oral cavity could be described by a functional relationship.

  9. Effects of high-sucrose feeding on insulin resistance and hemodynamic responses to insulin in spontaneously hypertensive rats.

    Science.gov (United States)

    Mélançon, Sébastien; Bachelard, Hélène; Badeau, Mylène; Bourgoin, Frédéric; Pitre, Maryse; Larivière, Richard; Nadeau, André

    2006-06-01

    This study was designed to investigate the effects of a sucrose diet on vascular and metabolic actions of insulin in spontaneously hypertensive rats (SHR). Male SHR were randomized to receive a sucrose or regular chow diet for 4 wk. Age-matched, chow-fed Wistar-Kyoto (WKY) rats were used as normotensive control. In a first series of experiments, the three groups of rats had pulsed Doppler flow probes and intravascular catheters implanted to determine blood pressure, heart rate, and blood flows. Insulin sensitivity was assessed during a euglycemic hyperinsulinemic clamp performed in conscious rats. In a second series of experiments, new groups of rats were used to examine glucose transport activity in isolated muscles and to determine endothelial nitric oxide synthase (eNOS) protein expression in muscles and endothelin content in vascular tissues. Sucrose feeding was shown to markedly enhance the pressor response to insulin and its hindquarter vasoconstrictor effect when compared with chow-fed SHR. A reduction in eNOS protein content in muscle, but no change in vascular endothelin-1 protein, was noted in sucrose-fed SHR when compared with WKY rats, but these changes were not different from those noted in chow-fed SHR. Similar reductions in insulin-stimulated glucose transport were observed in soleus muscles from both groups of SHR when compared with WKY rats. In extensor digitorum longus muscles, a significant reduction in insulin-stimulated glucose transport was only seen in sucrose-fed rats when compared with the other two groups. Environmental factors, that is, high intake of simple sugars, could possibly potentiate the genetic predisposition in SHR to endothelial dysfunction and insulin resistance.

  10. Cytokinin-Regulated Sucrose Metabolism in Stay-Green Wheat Phenotype

    Science.gov (United States)

    Wang, Wenqiang; Hao, Qunqun; Tian, Fengxia; Li, Qinxue; Wang, Wei

    2016-01-01

    A wheat stay-green mutant, tasg1, was observed to exhibit significantly delayed senescence in the late developmental stage. The photosynthetic capacity of the flag leaf was greater in tasg1 than in wild type (WT) plants. In addition, the grain volume of tasg1 was significantly higher than that of WT at the early filling stage. The content of various cytokinins (CKs) in the grain was significantly higher in tasg1 than in WT and was accompanied by an upregulated expression of some cell cycle-related genes. Examination of the metabolism of soluble sugars in tasg1 and WT revealed that the concentrations of glucose (Glu), fructose (Fru), and sucrose (Suc) were higher in the flag leaves and grains of tasg1 than in WT plants. The activities of sucrose-phosphate synthase (SPS), sucrose synthase (SuSy), and cell wall invertase (CW-invertase) were higher in tasg1, suggesting an altered metabolism and transport of soluble sugars. Furthermore, when tasg1 was treated with the CK inhibitor lovastatin, the activity of invertase was inhibited and was associated with premature senescence phenotype. However, the activity of invertase was partially recovered in tasg1 when treated with 6-benzylaminopurine (BAP). The trend of change in the concentrations of Glu, Fru, and Suc was similar to that of invertase. Our results suggest that CKs might regulate the stay-green phenotype of tasg1 by regulating the invertase activity involved in Suc remobilization. PMID:27580166

  11. Cytokinin-Regulated Sucrose Metabolism in Stay-Green Wheat Phenotype.

    Science.gov (United States)

    Wang, Wenqiang; Hao, Qunqun; Tian, Fengxia; Li, Qinxue; Wang, Wei

    2016-01-01

    A wheat stay-green mutant, tasg1, was observed to exhibit significantly delayed senescence in the late developmental stage. The photosynthetic capacity of the flag leaf was greater in tasg1 than in wild type (WT) plants. In addition, the grain volume of tasg1 was significantly higher than that of WT at the early filling stage. The content of various cytokinins (CKs) in the grain was significantly higher in tasg1 than in WT and was accompanied by an upregulated expression of some cell cycle-related genes. Examination of the metabolism of soluble sugars in tasg1 and WT revealed that the concentrations of glucose (Glu), fructose (Fru), and sucrose (Suc) were higher in the flag leaves and grains of tasg1 than in WT plants. The activities of sucrose-phosphate synthase (SPS), sucrose synthase (SuSy), and cell wall invertase (CW-invertase) were higher in tasg1, suggesting an altered metabolism and transport of soluble sugars. Furthermore, when tasg1 was treated with the CK inhibitor lovastatin, the activity of invertase was inhibited and was associated with premature senescence phenotype. However, the activity of invertase was partially recovered in tasg1 when treated with 6-benzylaminopurine (BAP). The trend of change in the concentrations of Glu, Fru, and Suc was similar to that of invertase. Our results suggest that CKs might regulate the stay-green phenotype of tasg1 by regulating the invertase activity involved in Suc remobilization. PMID:27580166

  12. Arbuscular mycorrhiza symbiosis induces a major transcriptional reprogramming of the potato SWEET sugar transporter family

    Directory of Open Access Journals (Sweden)

    Jasmin eManck-Götzenberger

    2016-04-01

    Full Text Available Biotrophic microbes feeding on plants must obtain carbon from their hosts without killing the cells. The symbiotic arbuscular mycorrhizal (AM fungi colonizing plant roots do so by inducing major transcriptional changes in the host that ultimately also reprogram the whole carbon partitioning of the plant. AM fungi obtain carbohydrates from the root cortex apoplast, in particular from the periarbuscular space that surrounds arbuscules. However, the mechanisms by which cortical cells export sugars into the apoplast for fungal nutrition are unknown. Recently a novel type of sugar transporter, the SWEET, able to perform not only uptake but also efflux from cells was identified. Plant SWEETs have been shown to be involved in the feeding of pathogenic microbes and are, therefore, good candidates to play a similar role in symbiotic associations. Here we have carried out the first phylogenetic and expression analyses of the potato SWEET family and investigated its role during mycorrhiza symbiosis. The potato genome contains 35 SWEETs that cluster into the same four clades defined in Arabidopsis. Colonization of potato roots by the AM fungus Rhizophagus irregularis imposes major transcriptional rewiring of the SWEET family involving, only in roots, changes in 22 of the 35 members. None of the SWEETs showed mycorrhiza-exclusive induction and most of the twelve induced genes belong to the putative hexose transporters of clade I and II, while only two are putative sucrose transporters from clade III. In contrast, most of the repressed transcripts (10 corresponded to clade III SWEETs. Promoter-reporter assays for three of the induced genes, each from one cluster, showed re-localization of expression to arbuscule-containing cells, supporting a role for SWEETs in the supply of sugars at biotrophic interfaces. The complex transcriptional regulation of SWEETs in roots in response to AM fungal colonization supports a model in which symplastic sucrose in cortical

  13. Arbuscular mycorrhiza Symbiosis Induces a Major Transcriptional Reprogramming of the Potato SWEET Sugar Transporter Family.

    Science.gov (United States)

    Manck-Götzenberger, Jasmin; Requena, Natalia

    2016-01-01

    Biotrophic microbes feeding on plants must obtain carbon from their hosts without killing the cells. The symbiotic Arbuscular mycorrhizal (AM) fungi colonizing plant roots do so by inducing major transcriptional changes in the host that ultimately also reprogram the whole carbon partitioning of the plant. AM fungi obtain carbohydrates from the root cortex apoplast, in particular from the periarbuscular space that surrounds arbuscules. However, the mechanisms by which cortical cells export sugars into the apoplast for fungal nutrition are unknown. Recently a novel type of sugar transporter, the SWEET, able to perform not only uptake but also efflux from cells was identified. Plant SWEETs have been shown to be involved in the feeding of pathogenic microbes and are, therefore, good candidates to play a similar role in symbiotic associations. Here we have carried out the first phylogenetic and expression analyses of the potato SWEET family and investigated its role during mycorrhiza symbiosis. The potato genome contains 35 SWEETs that cluster into the same four clades defined in Arabidopsis. Colonization of potato roots by the AM fungus Rhizophagus irregularis imposes major transcriptional rewiring of the SWEET family involving, only in roots, changes in 22 of the 35 members. None of the SWEETs showed mycorrhiza-exclusive induction and most of the 12 induced genes belong to the putative hexose transporters of clade I and II, while only two are putative sucrose transporters from clade III. In contrast, most of the repressed transcripts (10) corresponded to clade III SWEETs. Promoter-reporter assays for three of the induced genes, each from one cluster, showed re-localization of expression to arbuscule-containing cells, supporting a role for SWEETs in the supply of sugars at biotrophic interfaces. The complex transcriptional regulation of SWEETs in roots in response to AM fungal colonization supports a model in which symplastic sucrose in cortical cells could be cleaved

  14. Dynamics of Streptococcus mutans transcriptome in response to starch and sucrose during biofilm development.

    Directory of Open Access Journals (Sweden)

    Marlise I Klein

    Full Text Available The combination of sucrose and starch in the presence of surface-adsorbed salivary α-amylase and bacterial glucosyltransferases increase the formation of a structurally and metabolically distinctive biofilm by Streptococcus mutans. This host-pathogen-diet interaction may modulate the formation of pathogenic biofilms related to dental caries disease. We conducted a comprehensive study to further investigate the influence of the dietary carbohydrates on S. mutans-transcriptome at distinct stages of biofilm development using whole genomic profiling with a new computational tool (MDV for data mining. S. mutans UA159 biofilms were formed on amylase-active saliva coated hydroxyapatite discs in the presence of various concentrations of sucrose alone (ranging from 0.25 to 5% w/v or in combination with starch (0.5 to 1% w/v. Overall, the presence of sucrose and starch (suc+st influenced the dynamics of S. mutans transcriptome (vs. sucrose alone, which may be associated with gradual digestion of starch by surface-adsorbed amylase. At 21 h of biofilm formation, most of the differentially expressed genes were related to sugar metabolism, such as upregulation of genes involved in maltose/maltotriose uptake and glycogen synthesis. In addition, the groEL/groES chaperones were induced in the suc+st-biofilm, indicating that presence of starch hydrolysates may cause environmental stress. In contrast, at 30 h of biofilm development, multiple genes associated with sugar uptake/transport (e.g. maltose, two-component systems, fermentation/glycolysis and iron transport were differentially expressed in suc+st-biofilms (vs. sucrose-biofilms. Interestingly, lytT (bacteria autolysis was upregulated, which was correlated with presence of extracellular DNA in the matrix of suc+st-biofilms. Specific genes related to carbohydrate uptake and glycogen metabolism were detected in suc+st-biofilms in more than one time point, indicating an association between presence of starch

  15. Sucrose Improves Insecticide Activity Against Drosophila suzukii (Diptera: Drosophilidae).

    Science.gov (United States)

    Cowles, Richard S; Rodriguez-Saona, Cesar; Holdcraft, Robert; Loeb, Gregory M; Elsensohn, Johanna E; Hesler, Steven P

    2015-04-01

    The addition of sucrose to insecticides targeting spotted wing drosophila, Drosophila suzukii (Matsumura), enhanced lethality in laboratory, semifield, and field tests. In the laboratory, 0.1% sucrose added to a spray solution enhanced spotted wing drosophila feeding. Flies died 120 min earlier when exposed to spinosad residues at label rates enhanced with sucrose. Added sucrose reduced the LC50 for dried acetamiprid residues from 82 to 41 ppm in the spray solution. Laboratory bioassays of spotted wing drosophila mortality followed exposure to grape and blueberry foliage and/or fruit sprayed and aged in the field. On grape foliage, the addition of 2.4 g/liter of sugar with insecticide sprays resulted in an 11 and 6% increase of spotted wing drosophila mortality at 1 and 2 d exposures to residues, respectively, averaged over seven insecticides with three concentrations. In a separate experiment, spinetoram and cyantraniliprole reduced by 95-100% the larval infestation of blueberries, relative to the untreated control, 7 d after application at labeled rates when applied with 1.2 g/liter sucrose in a spray mixture, irrespective of rainfall; without sucrose infestation was reduced by 46-91%. Adding sugar to the organically acceptable spinosyn, Entrust, reduced larval infestation of strawberries by >50% relative to without sugar for five of the six sample dates during a season-long field trial. In a small-plot field test with blueberries, weekly applications in alternating sprays of sucrose plus reduced-risk insecticides, spinetoram or acetamiprid, reduced larval infestation relative to the untreated control by 76%; alternating bifenthrin and phosmet (without sucrose) reduced infestation by 65%.

  16. Effect of sucrose-containing snacks on blood glucose control.

    Science.gov (United States)

    Wise, J E; Keim, K S; Huisinga, J L; Willmann, P A

    1989-06-01

    To determine whether ingestion of sucrose-containing snacks would affect blood glucose (BG) control, 16 subjects with insulin-dependent diabetes mellitus participated in a 5-day double-blind study at a diabetes camp. Eight subjects in the sucrose group ate sucrose-sweetened snacks twice a day, and 8 subjects in the control group ingested snacks that were sweetened with aspartame. The percentage of total daily calories derived from added sucrose was 7% for the sucrose group and 1% for the control group. Metabolic control was assessed by daily capillary BG measurements obtained before meals and the bedtime snack and by determination of serum fructosamine (F) concentrations on arrival at camp (day 0) and after 5 days on the study protocol (day 5). No significant difference was seen between the groups on day 0 (sucrose group [mean +/- SD]: BG 9.9 +/- 3.6 mM, F 3.54 +/- 0.38 mM; control group: BG 9.1 +/- 2.8 mM, F 3.74 +/- 0.71 mM) or day 5 (sucrose group: BG 8.8 +/- 2.6 mM, F 2.94 +/- 0.32 mM; control group: BG 7.4 +/- 2.8 mM, F 2.92 +/- 0.59 mM). We conclude that ingestion of sucrose, added to snacks in an amount up to 7% of total energy intake, does not adversely affect short-term BG control. PMID:2659302

  17. Arabidopsis in Wageningen

    OpenAIRE

    Koornneef, M

    2013-01-01

    Arabidopsis thaliana is the plant species that in the past 25 years has developed into the major model species in plant biology research. This was due to its properties such as short generation time, its small genome and its easiness to be transformed. Wageningen University has played an important role in the development of this model, based on interdisciplinary collaborations using genetics as a major tool to investigate aspects of physiology, development, plant-microbe interactions and evol...

  18. Effects of Soil Salinity on Sucrose Metabolism in Cotton Leaves.

    Science.gov (United States)

    Peng, Jun; Liu, Jingran; Zhang, Lei; Luo, Junyu; Dong, Helin; Ma, Yan; Zhao, Xinhua; Chen, Binglin; Sui, Ning; Zhou, Zhiguo; Meng, Yali

    2016-01-01

    This study investigated sucrose metabolism of the youngest fully expanded main-stem leaf (MSL) and the subtending leaf of cotton (Gossypium hirsutum L.) boll (LSCB) of salt-tolerant (CCRI-79) and salt-sensitive (Simian 3) cultivars and its relationship to boll weight under low, medium and high soil salinity stress in Dafeng, China, in 2013 and 2014. The results showed that with increased soil salinity, 1) both the chlorophyll content and net photosynthetic rate (Pn) decreased, while the internal CO2 concentration firstly declined, and then increased in the MSL and LSCB; 2) carbohydrate contents in the MSL reduced significantly, while sucrose and starch contents in the LSCB increased, as did the activities of sucrose phosphate synthase (SPS) and sucrose synthase (SuSy) in both the MSL and LSCB; 3) but invertase activity in both the MSL and LSCB did not change significantly. Our study also showed that the LSCB was more sensitive to soil salinity than was the MSL. Of the measured physiological indices, higher SPS activity, mainly controlled by sps3, may contribute to adaption of the LSCB to soil salinity stress because SPS is beneficial for efficiently sucrose synthesis, reduction of cellular osmotic potential and combined actions of Pn, and sucrose transformation rate and SPS may contribute to the reduction in boll weight under soil salinity stress. PMID:27228029

  19. Effects of Soil Salinity on Sucrose Metabolism in Cotton Leaves.

    Directory of Open Access Journals (Sweden)

    Jun Peng

    Full Text Available This study investigated sucrose metabolism of the youngest fully expanded main-stem leaf (MSL and the subtending leaf of cotton (Gossypium hirsutum L. boll (LSCB of salt-tolerant (CCRI-79 and salt-sensitive (Simian 3 cultivars and its relationship to boll weight under low, medium and high soil salinity stress in Dafeng, China, in 2013 and 2014. The results showed that with increased soil salinity, 1 both the chlorophyll content and net photosynthetic rate (Pn decreased, while the internal CO2 concentration firstly declined, and then increased in the MSL and LSCB; 2 carbohydrate contents in the MSL reduced significantly, while sucrose and starch contents in the LSCB increased, as did the activities of sucrose phosphate synthase (SPS and sucrose synthase (SuSy in both the MSL and LSCB; 3 but invertase activity in both the MSL and LSCB did not change significantly. Our study also showed that the LSCB was more sensitive to soil salinity than was the MSL. Of the measured physiological indices, higher SPS activity, mainly controlled by sps3, may contribute to adaption of the LSCB to soil salinity stress because SPS is beneficial for efficiently sucrose synthesis, reduction of cellular osmotic potential and combined actions of Pn, and sucrose transformation rate and SPS may contribute to the reduction in boll weight under soil salinity stress.

  20. Enzymatic Synthesis of Sucrose Polyester as Food Emulsifier Compound

    Directory of Open Access Journals (Sweden)

    Sri Handayani

    2012-12-01

    Full Text Available Sucrose polyester (SPE is a carbohydrate ester compound that has diverse functions, from surfactant to low-calorie food products. Sucrose fatty acid ester with the degree of substitution 1-3 can be used as emulsifier in foods and cosmetics. The enzymatic synthesis of sucrose polyesters can be carried out using lipase in organic solvent and contain small amount of water. In these studies sucrose esters were synthesized by esterification reaction between sucrose with fatty acids from coconut and palm oil using Candida rugosa lipase in n-hexane. Optimization esterification reaction carried out for parameters of incubation time, temperature, and the ratio of the substrate. The optimum incubation time is at 18 hours for coconut oil and 12 hours palm oil, the optimum temperature is 30 oC for coconut and palm oil, and the mole ratio of fatty acid to sucrose is 40:1 for coconut oil and 64:1 for palm oil. Esterification products were characterized by FT-IR. The FT-IR spectrum showed the ester bond was formed as indicated by the wave number 1739.79/cm. Esterification products have 2 substitution degrees.

  1. Effect of troglitazone on vascular and glucose metabolic actions of insulin in high-sucrose-fed rats.

    Science.gov (United States)

    Santuré, Marta; Pitre, Maryse; Nadeau, André; Bachelard, Hélène

    2003-08-01

    In rats, diets high in simple sugar induce insulin resistance and alter vascular reactivity. The present study was designed to evaluate the effects of 5 weeks treatment with troglitazone on insulin sensitivity, regional hemodynamics, and vascular responses to insulin in chow-fed and high-sucrose-fed rats. Male rats were randomly divided in 4 groups to receive a regular chow diet in the absence (group 1) or presence of troglitazone (0.2% in food; group 2), or a sucrose-enriched diet in the absence (group 3) or presence of troglitazone (group 4) for 5 weeks. The rats were instrumented with Doppler flow probes and intravascular catheters to determine blood pressure, heart rate, and regional blood flows. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Glucose transport activity was examined in isolated muscles. Sucrose feeding was found to induce insulin resistance and to impair the insulin-mediated skeletal muscle vasodilation. Treatment with troglitazone was found to increase whole-body insulin sensitivity in sucrose- and chow-fed rats, but had no effect on skeletal muscle glucose transport activity measured in isolated muscles from both dietary groups. Changes in regional hemodynamics were observed in both dietary cohorts treated with troglitazone, and the hindquarter vasoconstrictor response to insulin noted in sucrose-fed rats was abolished by the treatment. The vascular effects of troglitazone, and its insulin-related attenuating effects on contractile tone, could have contributed, in part, to improve insulin action on peripheral glucose disposal, presumably by improving blood flow distribution and glucose delivery.

  2. Melatonin induces the transcripts of CBF/DREB1s and their involvement in both abiotic and biotic stresses in Arabidopsis.

    Science.gov (United States)

    Shi, Haitao; Qian, Yongqiang; Tan, Dun-Xian; Reiter, Russel J; He, Chaozu

    2015-10-01

    Melatonin (N-acetyl-5-methoxytryptamine) is a naturally occurring small molecule that acts as an important secondary messenger in plant stress responses. However, the mechanism underlying the melatonin-mediated signaling pathway in plant stress responses has not been established. C-repeat-binding factors (CBFs)/Drought response element Binding 1 factors (DREB1s) encode transcription factors that play important roles in plant stress responses. This study has determined that endogenous melatonin and transcripts level of CBFs (AtCBF1, AtCBF2, and AtCBF3) in Arabidopsis leaves were significantly induced by salt, drought, and cold stresses and by pathogen Pseudomonas syringe pv. tomato (Pst) DC3000 infection. Moreover, both exogenous melatonin treatment and overexpression of CBFs conferred enhanced resistance to both abiotic and biotic stresses in Arabidopsis. Notably, AtCBFs and exogenous melatonin treatment positively regulated the mRNA expression of several stress-responsive genes (COR15A, RD22, and KIN1) and accumulation of soluble sugars content such as sucrose in Arabidopsis under control and stress conditions. Additionally, exogenous sucrose also conferred improved resistance to both abiotic and biotic stresses in Arabidopsis. Taken together, this study indicates that AtCBFs confer enhanced resistance to both abiotic and biotic stresses, and AtCBF-mediated signaling pathway and sugar accumulation may be involved in melatonin-mediated stress response in Arabidopsis, at least partially.

  3. Melatonin induces the transcripts of CBF/DREB1s and their involvement in both abiotic and biotic stresses in Arabidopsis.

    Science.gov (United States)

    Shi, Haitao; Qian, Yongqiang; Tan, Dun-Xian; Reiter, Russel J; He, Chaozu

    2015-10-01

    Melatonin (N-acetyl-5-methoxytryptamine) is a naturally occurring small molecule that acts as an important secondary messenger in plant stress responses. However, the mechanism underlying the melatonin-mediated signaling pathway in plant stress responses has not been established. C-repeat-binding factors (CBFs)/Drought response element Binding 1 factors (DREB1s) encode transcription factors that play important roles in plant stress responses. This study has determined that endogenous melatonin and transcripts level of CBFs (AtCBF1, AtCBF2, and AtCBF3) in Arabidopsis leaves were significantly induced by salt, drought, and cold stresses and by pathogen Pseudomonas syringe pv. tomato (Pst) DC3000 infection. Moreover, both exogenous melatonin treatment and overexpression of CBFs conferred enhanced resistance to both abiotic and biotic stresses in Arabidopsis. Notably, AtCBFs and exogenous melatonin treatment positively regulated the mRNA expression of several stress-responsive genes (COR15A, RD22, and KIN1) and accumulation of soluble sugars content such as sucrose in Arabidopsis under control and stress conditions. Additionally, exogenous sucrose also conferred improved resistance to both abiotic and biotic stresses in Arabidopsis. Taken together, this study indicates that AtCBFs confer enhanced resistance to both abiotic and biotic stresses, and AtCBF-mediated signaling pathway and sugar accumulation may be involved in melatonin-mediated stress response in Arabidopsis, at least partially. PMID:26182834

  4. Induction of insulin resistance by high-sucrose feeding does not raise mean arterial blood pressure but impairs haemodynamic responses to insulin in rats.

    Science.gov (United States)

    Santuré, Marta; Pitre, Maryse; Marette, André; Deshaies, Yves; Lemieux, Christian; Larivière, Richard; Nadeau, André; Bachelard, Hélène

    2002-09-01

    1. This study was undertaken to further investigate the effects of a sucrose-enriched diet on vascular function and insulin sensitivity in rats. 2. Male Sprague-Dawley rats were randomized to receive a sucrose- or regular rat chow-diet for 4 weeks. A first group of sucrose- and chow-fed rats was instrumented with pulsed Doppler flow probes and intravascular catheters to determine blood pressure, heart rate, regional blood flows and insulin sensitivity in conscious rats. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Glucose transport activity was examined in isolated muscles by using the glucose analogue [(3)H]-2-deoxy-D-glucose. A second group of sucrose- and chow-fed rats was used to obtain information regarding nitric oxide synthase (NOS) isozymes protein expression in muscles, and determine endothelin content in vascular tissues isolated from both dietary groups. 3. Sucrose feeding was found to induce insulin resistance, but had no effect on resting blood pressure, heart rate, or regional haemodynamics. This insulin resistance was accompanied by alteration in the vascular responses to insulin. Insulin-mediated skeletal muscle vasodilation was impaired, whereas the mesenteric vasoconstrictor response was potentiated in sucrose-fed rats. A reduction in eNOS protein content in muscle and an increase in vascular endothelin peptide were noted in these animals. Moreover, a reduction in insulin-simulated glucose transport activity was also noted in muscles isolated from sucrose-fed rats. 4. Together these data suggest that a cluster of metabolic and haemodynamic abnormalities occur in response to the intake of simple sugars in rats.

  5. 生长素极性运输PIN基因在拟南芥和荠菜不同组织表达的定量分析%Quantitative analysis of expression of auxin polar transport PIN genes in different tissues of Arabidopsis thaliana and Capsella bursa-pastoris

    Institute of Scientific and Technical Information of China (English)

    朱占伟; 彭彦; 赵燕; 胡清云; 张学文

    2013-01-01

    In order to understand the morphogenesis difference of Arabidopsis thaliana and Capsella bursa-pastoris and elucidate the difference of auxin distribution,fluorescence quantitative PCR (qPCR) method with the β-actin gene as internal reference was used to quantify the expression levels of PIN1,PIN3,PIN7 in the roots,stems,leaves,flowers of the two kinds of plants.The results show that the PIN1 expression in all Arabidopsis thaliana organs is higher than they are in the Capsella organs.The expressions of PIN3 in the stem and flower of Arabidopsis thaliana were 3 and 10 times higher than those in the corresponding organs of Capsella bursa-pastoris.PIN7 expression is also higher in Arabidopsis thaliana than it is in the corresponding organs of Capsella with the expression of PIN7 in the leaf of Aradidopsis thaliana more than 10 times higher than those in the leaf of Capsella bursa-pastoris.PIN3 expression is higher in all tissues of Capsella bursa-pastoris than it is in the Arabidopsis thaliana.The expressions of PIN3 in the stem and flower ofArabidopsis thaliana were 3 and 10 times higher than those in the corresponding organs ofCapsella bursa-pastoris.The differential expression of auxin polar transport protein related genes may be the direct reason causing the morphology difference in the two plants.%为了解拟南芥和荠菜形态差异与其生长素极性分布的关系,采用荧光定量PCR,以β-actin为内参基因,对拟南芥和荠菜的根、茎、叶、花等组织中与生长素极性运输相关的PIN1、PIN3、PIN7基因的表达进行定量分析.结果表明:拟南芥各组织中PIN1的表达量都高于荠菜各组织中PIN1的表达量,拟南芥的茎和花中PIN1的表达量分别比荠菜茎和花中PIN1表达量高3倍和10倍;拟南芥各组织中PIN7的表达量也高于荠菜各组织的PIN7表达量,其叶片中PIN7的表达量比荠菜叶片中PIN7的表达量高10倍以上;荠菜各组织中PIN3的表达都高

  6. Effect of carbon/nitrogen ratio on carbohydrate metabolism and light energy dissipation mechanisms in Arabidopsis thaliana.

    Science.gov (United States)

    Huarancca Reyes, Thais; Scartazza, Andrea; Lu, Yu; Yamaguchi, Junji; Guglielminetti, Lorenzo

    2016-08-01

    Carbon (C) and nitrogen (N) nutrient sources are essential elements for metabolism, and their availability must be tightly coordinated for the optimal growth and development in plants. Plants are able to sense and respond to different C/N conditions via specific partitioning of C and N sources and the regulation of a complex cellular metabolic activity. We studied how the interaction between C and N signaling could affect carbohydrate metabolism, soluble sugar levels, photochemical efficiency of photosystem II (PSII) and the ability to drive the excess energy in Arabidopsis seedlings under moderated and disrupted C/N-nutrient conditions. Invertase and sucrose synthase activities were markedly affected by C/N-nutrient status depending on the phosphorylation status, suggesting that these enzymes may necessarily be modulated by their direct phosphorylation or phosphorylation of proteins that form complex with them in response to C/N stress. In addition, the enzymatic activity of these enzymes was also correlated with the amount of sugars, which not only act as substrate but also as signaling compounds. Analysis of chlorophyll fluorescence in plants under disrupted C/N condition suggested a reduction of electron transport rate at PSII level associated with a higher capacity for non-radiative energy dissipation in comparison with plants under moderated C/N condition. In conclusion, the tight coordination between C and N not only affects the carbohydrates metabolism and their concentration within plant tissues, but also the partitioning of the excitation energy at PSII level between radiative (electron transport) and non-radiative (heat) dissipation pathways. PMID:27108206

  7. An Arabidopsis callose synthase

    DEFF Research Database (Denmark)

    Ostergaard, Lars; Petersen, Morten; Mattsson, Ole;

    2002-01-01

    in the Arabidopsis mpk4 mutant which exhibits systemic acquired resistance (SAR), elevated beta-1,3-glucan synthase activity, and increased callose levels. In addition, AtGsl5 is a likely target of salicylic acid (SA)-dependent SAR, since AtGsl5 mRNA accumulation is induced by SA in wild-type plants, while...... expression of the nahG salicylate hydroxylase reduces AtGsl5 mRNA levels in the mpk4 mutant. These results indicate that AtGsl5 is likely involved in callose synthesis in flowering tissues and in the mpk4 mutant....

  8. Textural and sensory properties of spreads with sucrose and maltitol

    Directory of Open Access Journals (Sweden)

    Šoronja-Simović Dragana M.

    2012-01-01

    Full Text Available Spreads are confectionery products based on sugar, vegetable fat, cocoa powder, milk powder and other ingredients. Basic properties of these products are good spreadability in wide temperature range (from ambience to fridge temperature, rich creamy chocolate taste, and homogenous smooth structure without oil phase migration. Undesirable attribute of these products is their relatively high energy value (2300 kJ/100 g. In recent years, cocoa cream products with reduced energy values have become very popular among consumers and today they are present in the assortment of many confectionery manufacturers. One way to produce spreads with reduced energy value is the replacement of sugar (sucrose with adequate sweetener. Maltitol is a low-energy poliol capable to qualitatively and quantitatively replace sucrose. Cocoa spreads with maltitol and with the combination of maltitol and sucrose (produced at different temperatures and mixer rotation speeds have similar texture and rheological properties compared to the spreads with sucrose. The spreads with maltitol have about 15% lower energy value in comparison to the same product with sucrose.

  9. Adverse effects of sucrose-rich diets on uraemic rats.

    Science.gov (United States)

    Laouari, D; Kleinknecht, C; Burtin, M; Hinglais, N; Lacour, B; Landais, P; Broyer, M

    1990-01-01

    The nature of carbohydrate may affect the tolerance and progression of uraemia. The effects of three diets differing only in their carbohydrate source: namely corn starch (C), glucose (G) or sucrose (S) were examined. Study 1 examined the effects of the three carbohydrate diets on unilaterally nephrectomised control rats and severely uraemic rats. The three carbohydrates produced similar nutritional effects in uninephrectomised rats, whereas sucrose rapidly induced anorexia, stunting and slightly accelerated renal damage in uraemia. Study 2 examined the long-term effects of the three carbohydrates in moderate uraemia under conditions of high and identical carbohydrate intakes. Hyperphagic Zucker uraemic rats (F rats) received a daily allotment of each diet plus pure carbohydrate. Lean uraemic rats (L rats) received the same dietary allotment without the carbohydrate supplement. The F rats fed sucrose showed greater morbidity and mortality but little renal deterioration. Their plasma triglycerides increased dramatically. The L rats fed sucrose had the greatest urinary protein, the least creatinine clearance and the most severe renal damage. Thus, sucrose-rich but not glucose-rich diets have two adverse effects in uraemia: a deterioration in nutritional status, perhaps related to abnormal fructose utilisation, and a long-term effect on the kidney, resulting in accelerated renal deterioration.

  10. Analysis of mRNA translation states in Arabidopsis over the diurnal cycle by polysome microarray.

    Science.gov (United States)

    Missra, Anamika; von Arnim, Albrecht G

    2014-01-01

    Gene regulation at the level of translation occurs in response to environmental perturbation and is increasingly recognized as a factor affecting plant development. Despite extensive knowledge of transcriptional control, very little is known about translational regulation of genes in response to the daily light/dark cycles. Here we describe the experimental layout designed to address how the translation states of genes change at various times during a diurnal cycle in Arabidopsis thaliana seedlings. We have adopted a strategy combining sucrose-gradient profiling of ribosomes and high-throughput microarray analysis of the ribosome-associated mRNA to investigate the translational landscape of the Arabidopsis genome. This is a powerful technique that can be easily extended to study translation regulation in different genetic backgrounds and under various environmental conditions. PMID:24792050

  11. The overexpression of the pine transcription factor PpDof5 in Arabidopsis leads to increased lignin content and affects carbon and nitrogen metabolism.

    Science.gov (United States)

    Rueda-López, Marina; Cañas, Rafael A; Canales, Javier; Cánovas, Francisco M; Ávila, Concepción

    2015-12-01

    PpDof 5 is a regulator of the expression of glutamine synthetase (GS; EC 6.3.1.2) genes in photosynthetic and non-photosynthetic tissues of maritime pine. We have used Arabidopsis thaliana as a model system to study PpDof 5 function in planta, generating transgenic lines overexpressing the pine transcription factor. The overexpression of PpDof 5 resulted in a substantial increase of lignin content with a simultaneous regulation of carbon and nitrogen key genes. In addition, partitioning in carbon and nitrogen compounds was spread via various secondary metabolic pathways. These results suggest pleiotropic effects of PpDof 5 expression on various metabolic pathways of carbon and nitrogen metabolism. Plants overexpressing PpDof 5 exhibited upregulation of genes encoding enzymes for sucrose and starch biosynthesis, with a parallel increase in the content of soluble sugars. When the plants were grown under nitrate as the sole nitrogen source, they exhibited a significant regulation of the expression of genes involved mainly in signaling, but similar growth rates to wild-type plants. However, plants grown under ammonium exhibited major induction of the expression of photosynthetic genes and differential expression of ammonium and nitrate transporters. All these data suggest that in addition to controlling ammonium assimilation, PpDof 5 could be also involved in the regulation of other pathways in carbon and nitrogen metabolism in pine trees. PMID:26333592

  12. Taste preference thresholds for Polycose, maltose, and sucrose in rats.

    Science.gov (United States)

    Sclafani, A; Nissenbaum, J W

    1987-01-01

    The taste preference thresholds of adult female rats for polysaccharide (Polycose), maltose, and sucrose were compared. The nondeprived animals were given 24-hr two-bottle preference tests (saccharide solution vs. water) and, starting at 0.008%, the saccharide concentration was increased daily. The rats first preferred the Polycose solution to water at 0.01% (0.0001 M), the maltose solution to water at 0.09% (0.0025 M), and the sucrose solution to water at 0.09% (0.0026 M). Thus, on a molar basis the rats' Polycose threshold was 25 to 26 times lower than their maltose and sucrose threshold. It was postulated that the low taste threshold for polysaccharides allows the rat to detect starch which, unlike sugar, is very low in solubility.

  13. Abscisic acid and sucrose regulate tomato and strawberry fruit ripening through the abscisic acid-stress-ripening transcription factor.

    Science.gov (United States)

    Jia, Haifeng; Jiu, Songtao; Zhang, Cheng; Wang, Chen; Tariq, Pervaiz; Liu, Zhongjie; Wang, Baoju; Cui, Liwen; Fang, Jinggui

    2016-10-01

    Although great progress has been made towards understanding the role of abscisic acid (ABA) and sucrose in fruit ripening, the mechanisms underlying the ABA and sucrose signalling pathways remain elusive. In this study, transcription factor ABA-stress-ripening (ASR), which is involved in the transduction of ABA and sucrose signalling pathways, was isolated and analysed in the nonclimacteric fruit, strawberry and the climacteric fruit, tomato. We have identified four ASR isoforms in tomato and one in strawberry. All ASR sequences contained the ABA stress- and ripening-induced proteins and water-deficit stress-induced proteins (ABA/WDS) domain and all ASR transcripts showed increased expression during fruit development. The expression of the ASR gene was influenced not only by sucrose and ABA, but also by jasmonic acid (JA) and indole-3-acetic acid (IAA), and these four factors were correlated with each other during fruit development. ASR bound the hexose transporter (HT) promoter, which contained a sugar box that activated downstream gene expression. Overexpression of the ASR gene promoted fruit softening and ripening, whereas RNA interference delayed fruit ripening, as well as affected fruit physiological changes. Change in ASR gene expression influenced the expression of several ripening-related genes such as CHS, CHI, F3H, DFR, ANS, UFGT, PG, PL, EXP1/2, XET16, Cel1/2 and PME. Taken together, this study may provide new evidence on the important role of ASR in cross-signalling between ABA and sucrose to regulate tomato and strawberry fruit ripening. The findings of this study also provide new insights into the regulatory mechanism underlying fruit development.

  14. Abscisic acid and sucrose regulate tomato and strawberry fruit ripening through the abscisic acid-stress-ripening transcription factor.

    Science.gov (United States)

    Jia, Haifeng; Jiu, Songtao; Zhang, Cheng; Wang, Chen; Tariq, Pervaiz; Liu, Zhongjie; Wang, Baoju; Cui, Liwen; Fang, Jinggui

    2016-10-01

    Although great progress has been made towards understanding the role of abscisic acid (ABA) and sucrose in fruit ripening, the mechanisms underlying the ABA and sucrose signalling pathways remain elusive. In this study, transcription factor ABA-stress-ripening (ASR), which is involved in the transduction of ABA and sucrose signalling pathways, was isolated and analysed in the nonclimacteric fruit, strawberry and the climacteric fruit, tomato. We have identified four ASR isoforms in tomato and one in strawberry. All ASR sequences contained the ABA stress- and ripening-induced proteins and water-deficit stress-induced proteins (ABA/WDS) domain and all ASR transcripts showed increased expression during fruit development. The expression of the ASR gene was influenced not only by sucrose and ABA, but also by jasmonic acid (JA) and indole-3-acetic acid (IAA), and these four factors were correlated with each other during fruit development. ASR bound the hexose transporter (HT) promoter, which contained a sugar box that activated downstream gene expression. Overexpression of the ASR gene promoted fruit softening and ripening, whereas RNA interference delayed fruit ripening, as well as affected fruit physiological changes. Change in ASR gene expression influenced the expression of several ripening-related genes such as CHS, CHI, F3H, DFR, ANS, UFGT, PG, PL, EXP1/2, XET16, Cel1/2 and PME. Taken together, this study may provide new evidence on the important role of ASR in cross-signalling between ABA and sucrose to regulate tomato and strawberry fruit ripening. The findings of this study also provide new insights into the regulatory mechanism underlying fruit development. PMID:27005823

  15. Amperometric biosensors for determination of glucose, maltose, and sucrose

    Science.gov (United States)

    Zawicki, Ignacy; Filipiak, Marian; Jarzyna, Marta; Laskowska, Janina

    1995-06-01

    In the presented paper there are reported some results of the author's research on membranes containing glucose oxidase (GOx), enzymes hydrolyzing maltose and sucrose and on biosensors equipped with these membranes. The results relate to ways of extending the linear range of glucose sensors, influence of composition of the membranes on levels of the output signals of maltose and sucrose (saccharose) sensors, temperature dependence of the sensor's response and on disturbing effects of glucose in the sample on accuracy of determination of the disaccharides.

  16. Method for converting sucrose to .beta.-D-glucose

    Science.gov (United States)

    Simmons, Blake A.; Volponi, Joanne V.; Ingersoll, David; Walker, Andrew

    2009-07-07

    Disclosed is an apparatus and method for continuously converting sucrose to .beta.-D-glucose. The method comprises a three-stage enzymatic reactor in which an aqueous solution of sucrose is first converted into a solution of fructose and .alpha.-D-glucose by passing it through a porous, packed column containing an inert media on which invertase is immobilized. This solution is then sent through a second packed column containing glucose isomerase and finally a third packed column containing mutarotase. Solution temperature and pH are adjusted to maximize glucose output.

  17. Anharmonicity and hydrogen bonding in electrooptic sucrose crystal

    Science.gov (United States)

    Szostak, M. M.; Giermańska, J.

    1990-03-01

    The polarized absorption spectra of the sucrose crystal in the 5300 - 7300 cm -1 region have been measured. The assignments of all the eight OH stretching overtones are proposed and their mechanical anharmonicities are estimated. The discrepancies from the oriented gas model (OGM) in the observed relative band intensities, especially of the -CH vibrations, are assumed to be connected with vibronic couplings enhanced by the helical arrangement of molecules joined by hydrogen bondings. It seems that this kind of interactions might be important for the second harmonic generation (SHG) by the sucrose crystal.

  18. Novel Unsaturated Sucrose Ethers and Their Application as Monomers

    Directory of Open Access Journals (Sweden)

    Maria T. Barros

    2007-03-01

    Full Text Available Novel unsaturated ethers were synthesised in good yields starting from sucrose,using a two-step mild and efficient procedure based on the Gassman method, whichconsists in forming a vinyl group by the elimination of ethanol from mixed acetals withtrimethylsilyl trifluoromethanesulfonate in the presence of alkyl amines. Mixed acetals arereadily obtained from the corresponding alcohols and ethyl vinyl ether, using an acidiccatalyst. Conventional etherification involving a primary halide was also examined. Themonomers thus obtained were successfully polymerised by a free radical mechanism,yielding unbranched linear and soluble polymers with pending sucrose moieties, and someof their physical properties were determined.

  19. Sucrose: A Prospering and Sustainable Organic Raw Material

    Science.gov (United States)

    Peters, Siegfried; Rose, Thomas; Moser, Matthias

    Sucrose (α-d-glucopyranosyl-(1→2)-β-d-fructofuranoside) is an inexpensive chemical produced by sugar cane and sugar beet cultivation. Chemical and/or biochemical transformations convert it into highly valuable synthetic intermediates such as 5-hydroxymethylfurfural (HMF), bioethylene, 1,2-propylene glycol and levulinic acid. Sucrose can also be converted into biodegradable polymers such as polyesters and polyurethanes, as well as into novel carbohydrates such as isomaltulose, trehalulose, inulin, levan, Neo-amylose, and dextran, highly valuable additives for food and cosmetics and materials for separation and purification technologies.

  20. Arabidopsis thaliana as Bioindicator of Fungal VOCs in Indoor Air

    Science.gov (United States)

    Hung, Richard; Yin, Guohua; Klich, Maren A.; Grimm, Casey; Bennett, Joan W.

    2016-01-01

    In this paper, we demonstrate the ability of Arabidopsis thaliana to detect different mixtures of volatile organic compounds (VOCs) emitted by the common indoor fungus, Aspergillus versicolor, and demonstrate the potential usage of the plant as a bioindicator to monitor fungal VOCs in indoor air. We evaluated the volatile production of Aspergillus versicolor strains SRRC 108 (NRRL 3449) and SRRC 2559 (ATCC 32662) grown on nutrient rich fungal medium, and grown under conditions to mimic the substrate encountered in the built environment where fungi would typically grow indoors (moist wallboard and ceiling tiles). Using headspace solid phase microextraction/gas chromatography-mass spectrometry, we analyzed VOC profiles of the two strains. The most abundant compound produced by both strains on all three media was 1-octen-3-ol. Strain SRRC 2559 made several terpenes not detected from strain SRRC 108. Using a split-plate bioassay, we grew Arabidopsis thaliana in a shared atmosphere with VOCs from the two strains of Aspergillus versicolor grown on yeast extract sucrose medium. The VOCs emitted by SRRC 2559 had an adverse impact on seed germination and plant growth. Chemical standards of individual VOCs from the Aspergillus versicolor mixture (2-methyl-1-butanol, 3-methyl-1-butanol, 1-octen-3-ol, limonene, and β-farnesene), and β-caryophyllene were tested one by one in seed germination and vegetative plant growth assays. The most inhibitory compound to both seed germination and plant growth was 1-octen-3-ol. Our data suggest that Arabidopsis is a useful model for monitoring indoor air quality as it is sensitive to naturally emitted fungal volatile mixtures as well as to chemical standards of individual compounds, and it exhibits relatively quick concentration- and duration-dependent responses.

  1. Distribution of sucrose around the mouth and its clearance after a sucrose mouthrinse or consumption of three different foods.

    Science.gov (United States)

    Macpherson, L M; Dawes, C

    1994-01-01

    The distribution of sucrose in whole saliva and in saliva from seven different regions of the mouth was determined in 10 subjects over the 10-min period following the chewing of a doughnut, sucking on a mint candy, the drinking of orange juice, or use of a 10% sucrose mouthrinse. With all products, the sucrose was distributed non-uniformly, with particularly low concentrations on the lingual surfaces of the lower incisors and the facial surfaces of the upper molars. Clearance was also most rapid from these sites. Since the depth and duration of a Stephan curve in dental plaque is influenced by the sugar concentration to which the plaque is exposed, the results, together with previous results on salivary film velocity in different regions of the mouth, help to provide an explanation for the site-specificity of smooth-surface caries and of supragingival calculus deposition. PMID:8033187

  2. The effect of sucrose on unfrozen water and syneresis of acidified sodium caseinate-xanthan gels.

    Science.gov (United States)

    Braga, A L M; Cunha, R L

    2005-07-01

    The influence of the ingredients of acidified Na caseinate-xanthan-sucrose gels on thermophysical properties and syneresis of the gels was studied. Sucrose concentration affected all of the gel equilibrium properties and the rate of syneresis. The positive effect of sucrose on syneresis and unfrozen water (UFW) values was attributed to different effects. The amount of UFW was governed mainly by the colligative properties of sucrose whereas the equilibrium syneresis behaviour was associated with the changes in network dynamics caused by the kosmotropic properties of sucrose. The latter could enhance xanthan-sucrose association or favour xanthan-protein interactions.

  3. Transgenic Arabidopsis Gene Expression System

    Science.gov (United States)

    Ferl, Robert; Paul, Anna-Lisa

    2009-01-01

    The Transgenic Arabidopsis Gene Expression System (TAGES) investigation is one in a pair of investigations that use the Advanced Biological Research System (ABRS) facility. TAGES uses Arabidopsis thaliana, thale cress, with sensor promoter-reporter gene constructs that render the plants as biomonitors (an organism used to determine the quality of the surrounding environment) of their environment using real-time nondestructive Green Fluorescent Protein (GFP) imagery and traditional postflight analyses.

  4. Maternal chocolate and sucrose soft drink intake induces hepatic steatosis in rat offspring associated with altered lipid gene expression profile

    DEFF Research Database (Denmark)

    Kjærgaard, Maj; Nilsson, C.; Rosendal, A.;

    2014-01-01

    to decrease. Litter size reduction in offspring from high-fat/high-sucrose-fed dams further increased body weight and adiposity, and up-regulated genes involved in hepatic mitochondrial lipid oxidation and VLDL transport compared with all other groups. Litter size reduction did not have any impact on body...... until weaning, giving four dietary groups. Results: At postnatal day 1, offspring from high-fat/high-sucrose-fed dams were heavier and had increased hepatic triglycerides (TG), hepatic glycogen, blood glucose and plasma insulin compared with offspring from chow-fed dams. Hepatic genes involved in lipid...... weight gain and adiposity in offspring born to chow-fed dams. Conclusion: Our results suggest that supplementation of chocolate and soft drink during gestation and lactation contributes to early onset of hepatic steatosis associated with changes in hepatic gene expression and lipid handling. © 2013...

  5. Trichoderma volatiles effecting Arabidopsis

    DEFF Research Database (Denmark)

    Ramadan, Metwaly; Gigolashvili, Tamara; Grosskinsky, Dominik Kilian;

    2015-01-01

    Trichoderma species are present in many ecosystems and some strains have the ability to reduce the severity of plant diseases by activating various defense pathways via specific biologically active signaling molecules. Hence we investigated the effects of low molecular weight volatile compounds...... of Trichoderma asperellum IsmT5 on Arabidopsis thaliana. During co-cultivation of T. asperellum IsmT5 without physical contact to A. thaliana we observed smaller but vital and robust plants. The exposed plants exhibit increased trichome numbers, accumulation of defense-related compounds such as H2O2, anthocyanin......, camalexin, and increased expression of defense-related genes. We conclude that A. thaliana perceives the Trichoderma volatiles as stress compounds and subsequently initiates multilayered adaptations including activation of signaling cascades to withstand this environmental influence. The prominent headspace...

  6. Co-overexpressing a plasma membrane and a vacuolar membrane sodium/proton antiporter significantly improves salt tolerance in transgenic Arabidopsis plants.

    Science.gov (United States)

    The Arabidopsis gene AtNHX1 encodes a vacuolar membrane bound sodium/proton (Sodium/Hydrogen) antiporter that transports sodium into the vacuole and exports hydrogen into the cytoplasm. The Arabidopsis gene SOS1 encodes a plasma membrane bound sodium/hydrogen antiporter that exports sodium to the ex...

  7. Enzymatic synthesis and NMR studies of acylated sucrose acetates

    NARCIS (Netherlands)

    Steverink-De Zoete, M.C.; Kneepkens, M.F.M.; Waard, de P.; Woudenberg-van Oosterom, M.; Gotlieb, K.F.; Slaghek, T.

    1999-01-01

    The lipase-catalyzed esterification of partially acetylated sucrose has been studied. It was shown that the chemical acetylation increased the reaction rate of the subsequent enzymatic acylation. Thus it was possible to perform the enzymatic acylation in the absence of solvents while underivatized s

  8. Sucrose Responsiveness, Learning Success, and Task Specialization in Ants

    Science.gov (United States)

    Perez, Margot; Rolland, Uther; Giurfa,, Martin; d'Ettorre, Patrizia

    2013-01-01

    Social insects possess remarkable learning capabilities, which are crucial for their ecological success. They also exhibit interindividual differences in responsiveness to environmental stimuli, which underlie task specialization and division of labor. Here we investigated for the first time the relationships between sucrose responsiveness,…

  9. Structural analyses of sucrose laurate regioisomers by mass spectrometry techniques

    DEFF Research Database (Denmark)

    Lie, Aleksander; Stensballe, Allan; Pedersen, Lars Haastrup

    2015-01-01

    6- And 6′-O-lauroyl sucrose were isolated and analyzed by matrix-assisted laser desorption/ionisation (MALDI) time-of-flight (TOF) mass spectrometry (MS), Orbitrap high-resolution (HR) MS, and electrospray-ionization (ESI) tandem mass spectrometry (MS/MS). The analyses aimed to explore...

  10. Characterization of Ion Contents and Metabolic Responses to Salt Stress of Different Arabidopsis AtHKT1;1 Genotypes and Their Parental Strains

    Institute of Scientific and Technical Information of China (English)

    Camilla B.Hill; Deepa Jha; Antony Bacic; Mark Tester; Ute Roessner

    2013-01-01

    Plants employ several strategies to maintain cellular ion homeostasis under salinity stress,including mediating ion fluxes by transmembrane transport proteins and adjusting osmotic pressure by accumulating osmolytes.The HKT (high-affinity potassium transporter) gene family comprises Na+ and Na+/K+ transporters in diverse plant species,with HKT1;1 as the only member in Arabidopsis thaliana.Cell-type-specific overexpression of AtHKT1;1 has been shown to prevent shoot Na+ overaccumulation under salinity stress.Here,we analyzed a broad range of metabolites and elements in shoots and roots of different AtHKT1;1 genotypes and their parental strains before and after salinity stress,revealing a reciprocal relationship of metabolite differences between an AtHKT1;1 knockout line (hktl;1) and the AtHKT1;1 overexpressing lines (E2586 UASGAL4:HKT1;1 and J2731*UASGAL4:HKT1;1).Although levels of root sugars were increased after salt stress in both AtHKT1;1 overexpressing lines,E2586 UASGAL4:HKT1;1 showed higher accumulation of the osmoprotectants trehalose,gentiobiose,and melibiose,whereas J2731*UASGAL4:HKT1;1 showed higher levels of sucrose and raffinose,compared with their parental lines,respectively.In contrast,the knockout line hktl;1 showed strong increases in the levels of the tricarboxylic acid (TCA) cycle intermediates in the shoots after salt treatment.This coincided with a significant depletion of sugars,suggesting that there is an increased rate of carbon influx into the TCA cycle at a constant rate of C-efflux from the cycle,which might be needed to support plant survival during salt stress.Using correlation analysis,we identified associations between the Na+ content and several sugars,suggesting that regulation of sugar metabolism is important in plant responses to salinity stress.

  11. Measuring whole plant CO2 exchange with the environment reveals opposing effects of the gin2-1 mutation in shoots and roots of Arabidopsis thaliana.

    Science.gov (United States)

    Brauner, Katrin; Stutz, Simon; Paul, Martin; Heyer, Arnd G

    2015-01-01

    Using a cuvette for simultaneous measurement of net photosynthesis in above ground plant organs and root respiration we investigated the effect of reduced leaf glucokinase activity on plant carbon balance. The gin2-1 mutant of Arabidopsis thaliana is characterized by a 50% reduction of glucokinase activity in the shoot, while activity in roots is about fivefold higher and similar to wild type plants. High levels of sucrose accumulating in leaves during the light period correlated with elevated root respiration in gin2-1. Despite substantial respiratory losses in roots, growth retardation was moderate, probably because photosynthetic carbon fixation was simultaneously elevated in gin2-1. Our data indicate that futile cycling of sucrose in shoots exerts a reduction on net CO2 gain, but this is over-compensated by the prevention of exaggerated root respiration resulting from high sucrose concentration in leaf tissue.

  12. New insight into the catalytic properties of rice sucrose synthase.

    Science.gov (United States)

    Huang, Yu-Chiao; Hsiang, Erh-Chieh; Yang, Chien-Chih; Wang, Ai-Yu

    2016-01-01

    Sucrose synthase (SuS), which catalyzes the reversible conversion of sucrose and uridine diphosphate (UDP) into fructose and UDP-glucose, is a key enzyme in sucrose metabolism in higher plants. SuS belongs to family 4 of the glycosyltransferases (GT4) and contains an E-X7-E motif that is conserved in members of GT4 and two other GT families. To gain insight into the roles of this motif in rice sucrose synthase 3 (RSuS3), the two conserved glutamate residues (E678 and E686) in this motif and a phenylalanine residue (F680) that resides between the two glutamate residues were changed by site-directed mutagenesis. All mutant proteins maintained their tetrameric conformation. The mutants E686D and F680Y retained partial enzymatic activity and the mutants E678D, E678Q, F680S, and E686Q were inactive. Substrate binding assays indicated that UDP and fructose, respectively, were the leading substrates in the sucrose degradation and synthesis reactions of RSuS3. Mutations on E678, F680, and E686 affected the binding of fructose, but not of UDP. The results indicated that E678, F680, and E686 in the E-X7-E motif of RSuS3 are essential for the activity of the enzyme and the sequential binding of substrates. The sequential binding of the substrates implied that the reaction catalyzed by RSuS can be controlled by the availability of fructose and UDP, depending on the metabolic status of a tissue.

  13. Sucrose Diffusion in Decellularized Heart Valves for Freeze-Drying.

    Science.gov (United States)

    Wang, Shangping; Oldenhof, Harriëtte; Goecke, Tobias; Ramm, Robert; Harder, Michael; Haverich, Axel; Hilfiker, Andres; Wolkers, Willem Frederik

    2015-09-01

    Decellularized heart valves can be used as starter matrix implants for heart valve replacement therapies in terms of guided tissue regeneration. Decellularized matrices ideally need to be long-term storable to assure off-the-shelf availability. Freeze-drying is an attractive preservation method, allowing storage at room temperature in a dried state. However, the two inherent processing steps, freezing and drying, can cause severe damage to extracellular matrix (ECM) proteins and the overall tissue histoarchitecture and thus impair biomechanical characteristics of resulting matrices. Freeze-drying therefore requires a lyoprotective agent that stabilizes endogenous structural proteins during both substeps and that forms a protective glassy state at room temperature. To estimate incubation times needed to infiltrate decellularized heart valves with the lyoprotectant sucrose, temperature-dependent diffusion studies were done using Fourier transform infrared spectroscopy. Glycerol, a cryoprotective agent, was studied for comparison. Diffusion of both protectants was found to exhibit Arrhenius behavior. The activation energies of sucrose and glycerol diffusion were found to be 15.9 and 37.7 kJ·mol(-1), respectively. It was estimated that 4 h of incubation at 37°C is sufficient to infiltrate heart valves with sucrose before freeze-drying. Application of a 5% sucrose solution was shown to stabilize acellular valve scaffolds during freeze-drying. Such freeze-dried tissues, however, displayed pores, which were attributed to ice crystal damage, whereas vacuum-dried scaffolds in comparison revealed no pores after drying and rehydration. Exposure to a hygroscopic sucrose solution (80%) before freeze-drying was shown to be an effective method to diminish pore formation in freeze-dried ECMs: matrix structures closely resembled those of control samples that were not freeze-dried. Heart valve matrices were shown to be in a glassy state after drying, suggesting that they can

  14. Sucrose substitutes affect the cariogenic potential of Streptococcus mutans biofilms.

    Science.gov (United States)

    Durso, S C; Vieira, L M; Cruz, J N S; Azevedo, C S; Rodrigues, P H; Simionato, M R L

    2014-01-01

    Streptococcus mutans is considered the primary etiologic agent of dental caries and contributes significantly to the virulence of dental plaque, especially in the presence of sucrose. To avoid the role of sucrose on the virulence factors of S. mutans, sugar substitutes are commonly consumed because they lead to lower or no production of acids and interfere with biofilm formation. This study aimed to investigate the contribution of sugar substitutes in the cariogenic potential of S. mutans biofilms. Thus, in the presence of sucrose, glucose, sucralose and sorbitol, the biofilm mass was quantified up to 96 h, the pH of the spent culture media was measured, the expression of biofilm-related genes was determined, and demineralization challenge experiments were conduct in enamel fragments. The presence of sugars or sugar substitutes profoundly affected the expression of spaP, gtfB, gtfC, gbpB, ftf, vicR and vicX in either biofilm or planktonic cells. The substitution of sucrose induced a down-regulation of most genes involved in sucrose-dependent colonization in biofilm cells. When the ratio between the expression of biofilm and planktonic cells was considered, most of those genes were down-regulated in biofilm cells in the presence of sugars and up-regulated in the presence of sugar substitutes. However, sucralose but not sorbitol fulfilled the purpose of reducing the cariogenic potential of the diet since it induced the biofilm formation with the lowest biomass, did not change the pH of the medium and led to the lowest lesion depth in the cariogenic challenge.

  15. In vivo measurements of sulcal plaque pH in rats after topical applications of xylitol, sorbitol, glucose, sucrose, and sucrose plus 53 mM sodium fluoride.

    Science.gov (United States)

    Firestone, A R; Navia, J M

    1986-01-01

    In two series of experiments, Sprague-Dawley-derived rats were infected orally with cariogenic micro-organisms and fed caries-promoting diets. By means of an antimony electrode, resting pH values were measured in the mesial sulcus of the maxillary left first molar. 100 or 200 microL of the test solutions were applied, and the change in pH (delta pH) was recorded for three min. Test solutions were: (a) 10% sucrose, 10% glucose, 10% sorbitol, or 10% xylitol; (b) 0%, 10%, 20%, or 40% sucrose; (c) 0%, 3%, 7%, or 10% sucrose; and (d) 10% sucrose, 10% sucrose + 53 mmol/L NaF (1000 ppmF-), or 10% sucrose + 53 mmol/L NaCl. Experimental design was a 4 X 4 Latin square (a, b, c) or a cross-over design (d). Solutions of sucrose and glucose gave significantly greater decreases in pH than did sorbitol or xylitol. pH fall was maximal for 10% sucrose and significantly less for 40% sucrose during the three-minute experimental period. For sucrose solutions ranging in concentration from 3 to 10%, pH fall was highest after application of 10% sucrose when plaque was previously rinsed with water, but this pH fall did not differ significantly from that obtained using a 7% sucrose solution. Adding 1000 ppmF- to a 10% sucrose solution caused an increase in pH. Rinsing the teeth to remove saliva resulted in significantly lower resting pH values. The results of these experiments are in agreement with the results of human plaque pH measurements. PMID:3455697

  16. Healing effect of Sodium-Sucrose-Octasulfate and EGF on epithelial corneal abrasions in rabbits

    DEFF Research Database (Denmark)

    Johansen, Sven; Heegaard, Steffen; Bjerrum, Kirsten Birgitte;

    1998-01-01

    ophthalmology, Sjögrens syndrome, cornea, abrasion, epidermal growth factor, sodium-sucrose-octasulfate......ophthalmology, Sjögrens syndrome, cornea, abrasion, epidermal growth factor, sodium-sucrose-octasulfate...

  17. The Path of Carbon in Photosynthesis XIX. The Identification of Sucrose Phosphate in Sugar Beet Leaves

    Science.gov (United States)

    Buchanan, J. G.

    1952-09-01

    The recognition and characterization of a sucrose phosphate as an intermediate in sucrose by synthesis by green plants is described. A tentative structure for this phosphate is proposed and its mode of formation suggested.

  18. The Malus domestica sugar transporter gene family: identifications based on genome and expression profiling related to the accumulation of fruit sugars

    Directory of Open Access Journals (Sweden)

    Xiaoyu eWei

    2014-11-01

    Full Text Available In plants, sugar transporters are involved not only in long-distance transport, but also in sugar accumulations in sink cells. To identify members of sugar transporter gene families and to analyze their function in fruit sugar accumulation, we conducted a phylogenetic analysis of the Malus domestica genome. Expression profiling was performed with shoot tips, mature leaves, and developed fruit of ‘Gala’ apple. Genes for sugar alcohol (including 17 sorbitol transporters, sucrose, and monosaccharide transporters, plus SWEET genes, were selected as candidates in 31, 9, 50, and 27 loci, respectively, of the genome. The monosaccharide transporter family appears to include five subfamilies (30 MdHTs, 8 MdEDR6s, 5 MdTMTs, 3 MdvGTs, and 4 MdpGLTs. Phylogenetic analysis of the protein sequences indicated that orthologs exist among Malus, Vitis, and Arabidopsis. Investigations of transcripts revealed that 68 candidate transporters are expressed in apple, albeit to different extents. Here, we discuss their possible roles based on the relationship between their levels of expression and sugar concentrations. The high accumulation of fructose in apple fruit is possibly linked to the coordination and cooperation between MdTMT1/2 and MdEDR6. By contrast, these fruits show low MdSWEET4.1 expression and a high flux of fructose produced from sorbitol. Our study provides an exhaustive survey of sugar transporter genes and demonstrates that sugar transporter gene families in M. domestica are comparable to those in other species. Expression profiling of these transporters will likely contribute to improving our understanding of their physiological functions in fruit formation and the development of sweetness properties.

  19. Structural insights into the cubic-hexagonal phase transition kinetics of monoolein modulated by sucrose solutions

    OpenAIRE

    Reese, Caleb W.; Strango, Zachariah I.; Dell, Zachary R.; Tristram-Nagle, Stephanie; Harper, Paul E.

    2015-01-01

    Using DSC (differential scanning calorimetry), we measure the kinetics of the cubic-HII phase transition of monoolein in bulk sucrose solutions. We find that the transition temperature is dramatically lowered, with each 1 mol/kg of sucrose concentration dropping the transition by 20 °C. The kinetics of this transition also slow greatly with increasing sucrose concentration. For low sucrose concentrations, the kinetics are asymmetric, with the cooling (HII-cubic) transition taking twice as lon...

  20. AKINβ1 is Involved in the Regulation of Nitrogen Metabolism and Sugar Signaling in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    XiaoFang Li; YuJu Li; YingHui An; LiJun Xiong; XingHua Shao; Yang Wang; Yue Sun

    2009-01-01

    Sucrose non-fermenting-1-related protein kinase 1 (SnRK1) has been located at the heart of the control of metabolism and development in plants. The active SnRK1 form is usually a heterotrimeric complex. Subcellular localization and specific target of the SnRK1 kinase are regulated by specific beta subunits. In Arabidopsis, there are at least seven genes encoding beta subunits, of which the regulatory functions are not yet clear. Here, we tried to study the function of one beta subunit, AKINβ1. It showed that AKINβ1 expression was dramatically induced by ammonia nitrate but not potassium nitrate, and the investigation of AKINβ1 transgenic Arabidopsis and T-DNA insertion lines showed that AKINβ1 negatively regulated the activity of nitrate ruductase and was positively involved in sugar repression in early seedling development. Meanwhile AKINβ1 expression was reduced upon sugar treatment (including mannitol) and did not affect the activity of sucrose phos-phate synthase. The results indicate that AKINβ1 is involved in the regulation of nitrogen metabolism and sugar signaling.

  1. Identification, cloning and characterization of sis7 and sis10 sugar-insensitive mutants of Arabidopsis

    Directory of Open Access Journals (Sweden)

    Biddle Kelly D

    2008-10-01

    Full Text Available Abstract Background The levels of soluble sugars, such as glucose and sucrose, help regulate many plant metabolic, physiological and developmental processes. Genetic screens are helping identify some of the loci involved in plant sugar response and reveal extensive cross-talk between sugar and phytohormone response pathways. Results A forward genetic screen was performed to identify mutants with increased resistance to the inhibitory effects of high levels of exogenous sugars on early Arabidopsis seedling development. The positional cloning and characterization of two of these sugar insensitive (sis mutants, both of which are also involved in abscisic acid (ABA biosynthesis or response, are reported. Plants carrying mutations in SIS7/NCED3/STO1 or SIS10/ABI3 are resistant to the inhibitory effects of high levels of exogenous Glc and Suc. Quantitative RT-PCR analyses indicate transcriptional upregulation of ABA biosynthesis genes by high concentrations of Glc in wild-type germinating seeds. Gene expression profiling revealed that a significant number of genes that are expressed at lower levels in germinating sis7-1/nced3-4/sto1-4 seeds than in wild-type seeds are implicated in auxin biosynthesis or transport, suggesting cross-talk between ABA and auxin response pathways. The degree of sugar insensitivity of different sis10/abi3 mutant seedlings shows a strong positive correlation with their level of ABA insensitivity during seed germination. Conclusion Mutations in the SIS7/NCED3/STO1 gene, which is primarily required for ABA biosynthesis under drought conditions, confer a sugar-insensitive phenotype, indicating that a constitutive role in ABA biosynthesis is not necessary to confer sugar insensitivity. Findings presented here clearly demonstrate that mutations in ABI3 can confer a sugar-insensitive phenotype and help explain previous, mixed reports on this topic by showing that ABA and sugar insensitivity exhibit a strong positive correlation in

  2. 77 FR 18827 - Draft Guidance for Industry on Bioequivalence Recommendations for Iron Sucrose Injection...

    Science.gov (United States)

    2012-03-28

    ... Recommendations for Iron Sucrose Injection; Availability AGENCY: Food and Drug Administration, HHS. ACTION: Notice... industry entitled ``Bioequivalence Recommendations for Iron Sucrose.'' The recommendations provide specific...) for iron sucrose injection. DATES: Although you can comment on any guidance at any time (see 21 CFR...

  3. Reinforcement Value and Substitutability of Sucrose and Wheel Running: Implications for Activity Anorexia

    Science.gov (United States)

    Belke, Terry W.; Duncan, Ian D.; Pierce, W. David

    2006-01-01

    Choice between sucrose and wheel-running reinforcement was assessed in two experiments. In the first experiment, ten male Wistar rats were exposed to concurrent VI 30 s VI 30 s schedules of wheel-running and sucrose reinforcement. Sucrose concentration varied across concentrations of 2.5, 7.5, and 12.5%. As concentration increased, more behavior…

  4. Elucidating the Role of Transport Processes in Leaf Glucosinolate Distribution

    DEFF Research Database (Denmark)

    Madsen, Svend Roesen; Olsen, Carl Erik; Nour-Eldin, Hussam Hassan;

    2014-01-01

    In Arabidopsis (Arabidopsis thaliana), a strategy to defend its leaves against herbivores is to accumulate glucosinolates along the midrib and at the margin. Although it is generally assumed that glucosinolates are synthesized along the vasculature in an Arabidopsis leaf, thereby suggesting...... that the margin accumulation is established through transport, little is known about these transport processes. Here, we show through leaf apoplastic fluid analysis and glucosinolate feeding experiments that two glucosinolate transporters, GTR1 and GTR2, essential for long-distance transport of glucosinolates...... in Arabidopsis, also play key roles in glucosinolate allocation within a mature leaf by effectively importing apoplastically localized glucosinolates into appropriate cells. Detection of glucosinolates in root xylem sap unambiguously shows that this transport route is involved in root-to-shoot glucosinolate...

  5. The H+-sucrose cotransporter NtSUT1 is essential for sugar export from tobacco leaves

    International Nuclear Information System (INIS)

    In many species translocation of sucrose from the mesophyll to the phloem is carrier mediated. A sucrose/H+-symporter cDNA, NtSUT1, was isolated from tobacco (Nicotiana tabacum) and shown to be highly expressed in mature leaves and at low levels in other tissues, including floral organs. To study the in vivo function of NtSUT1, tobacco plants were transformed with a SUT1 antisense construct under control of the cauliflower mosaic virus 355 promoter. Upon maturation, leaves of transformants expressing reduced amounts of SUT1 mRNA curled downward, and strongly affected plants developed chloroses and necroses that led to death. The leaves exhibited impaired ability to export recently fixed 14CO2 and were unable to export transient starch during extended periods of darkness. As a consequence, soluble carbohydrates accumulated and photosynthesis was reduced. Autoradiographs of leaves show a heterogenous pattern of CO2 fixation even after a 24-h chase. The 14C pattern does not change with time, suggesting that movement of photosynthate between mesophyll cells may also be impaired. The affected lines show a reduction in the development of the root system and delayed or impaired flowering. Taken together, the effects observed in a seed plant (tobacco) demonstrate the importance of SUT1 for sucrose loading into the phloem via an apoplastic route and possibly for intermesophyll transport as well

  6. Arabidopsis thaliana peroxidase N

    DEFF Research Database (Denmark)

    Mirza, Osman Asghar; Henriksen, A; Ostergaard, L;

    2000-01-01

    The structure of the neutral peroxidase from Arabidopsis thaliana (ATP N) has been determined to a resolution of 1.9 A and a free R value of 20.5%. ATP N has the expected characteristic fold of the class III peroxidases, with a C(alpha) r.m.s.d. of 0.82 A when compared with horseradish peroxidase C...... (HRP C). HRP C is 54% identical to ATP N in sequence. When the structures of four class III plant peroxidases are superimposed, the regions with structural differences are non-randomly distributed; all are located in one half of the molecule. The architecture of the haem pocket of ATP N is very similar...... to that of HRP C, in agreement with the low small-molecule substrate specificity of all class III peroxidases. The structure of ATP N suggests that the pH dependence of the substrate turnover will differ from that of HRP C owing to differences in polarity of the residues in the substrate-access channel. Since...

  7. Streptococcus mutans in a wild, sucrose-eating rat population.

    Science.gov (United States)

    Coykendall, A L; Specht, P A; Samol, H H

    1974-07-01

    Streptococcus mutans, an organism implicated in dental caries and not previously found outside of man and certain laboratory animals, was isolated from the mouths of wild rats which ate sugar cane. The strains isolated fermented mannitol and sorbitol, and failed to grow in 6.5% NaCl or at 45 C. They formed in vitro plaques on nichrome wires when grown in sucrose broth. They also stored intracellular polysaccharide which could be catabolized by washed, resting cells. Deoxyribonucleic acid-deoxyribonucleic acid reassociations revealed two genetic types. One type shared extensive deoxyribonucleic acid base sequences with S. mutans strains HS6 and OMZ61, two members of a genetic type found in man and laboratory hamsters. The other type seemed unrelated to any S. mutans genetic type previously encountered. It is concluded that the ecological triad of tooth-sucrose-S. mutans is not a phenomenon unique to man and experimental animals. PMID:4601769

  8. Sucrose solution freezing studied by magnetic resonance imaging.

    Science.gov (United States)

    Mahdjoub, Rachid; Chouvenc, Pierre; Seurin, Marie José; Andrieu, Julien; Briguet, André

    2006-03-20

    Ice formation of a 20% w/v sucrose solution was monitored during the freezing process by magnetic resonance imaging (MRI). An original experimental setup was designed with oil as a cooling fluid that allows accurate control of the temperature. The NMR signal intensity of particular sampled volumes was observed during the entire cooling period, from 0 to -50 degrees C, showing a peak characteristic to a transition before the loss of the signal. Moreover, spatial ice distribution of the frozen matrix was observed by high resolution MRI with an isotropic resolution of 78x78x78microm(3). MRI has proved to be a novel technique for determining the glass transition temperature of frozen sucrose solutions, in the concentration range where calorimetric measurements are not feasible. PMID:16430876

  9. A Sucrose Solution Application to the Study of Model Biological Membranes

    CERN Document Server

    Kiselev, M A; Kisselev, A M; Lombardo, D; Killany, M; Lesieur, S; Ollivon, M

    2001-01-01

    The small-angle X-ray and neutron scattering, time resolved X-ray small-angle and wide-angle diffraction coupled with differential scanning calorimetry have been applied to the investigation of unilamellar and multilamellar dimyristoylphosphatidylcholine (DMPC) vesicles in sucrose buffers with sucrose concentrations from 0 to 60%. Sucrose buffer decreased vesicle size and polydispersity and increased an X-ray contrast between phospholipid membrane and bulk solvent sufficiently. No influence of sucrose on the membrane thickness or mutual packing of hydrocarbon chains has been detected. The region of sucrose concentrations 30%-40% created the best experimental conditions for X-ray small-angle experiments with phospholipid vesicles.

  10. Sucrose polyesters from poultry fat as non-ionic emulsifiers

    Directory of Open Access Journals (Sweden)

    Megahed, Mohamed G.

    2000-12-01

    Full Text Available Poultry fats are rich in palmitic and oleic acids are produced as by-products from poultry industries. These fats can be utilized in the preparation of emulsifiers. Sucrose esters of poultry fat are prepared from low-cost poultry fat and sucrose by esterification. The yield of sucrose esters prepared in this work exceeds than 85%. The hydrophilic-lipophilic balance (HLB, surface tension, interfacial tension and stability index (SI were evaluated and compared with standard emulsifiers prepared from pure palmitate and oleate esters. Concerning the stability of emulsions, the values of interfacial tension and HLB were higher for the sucrose esters of poultry fat.Las grasas de aves de corral son ricas en ácidos palmítico y oleico y son producidas como subproductos de las industrias avícolas. Estas grasas pueden ser utilizadas en la preparación de emulsionantes. Los ésteres de sacarosa de grasas de aves de corral fueron preparadas a partir de grasas de bajo coste y sacarosa por esterificación. El rendimiento de los ésteres de sacarosa preparados en este trabajo superó el 85%. El balance hidrofílicolipofílico (HLB, tensión superficial, tensión interfacial e índice de estabilidad (SI fueron evaluados y comparados con emulsionantes estandar preparados de ésteres de palmitato y oleato puros. Los valores de tensión interfacial y el HLB fueron más altos en los poliésteres de sacarosa de las grasas de aves de corral.

  11. Central melanocortins regulate the motivation for sucrose reward.

    Directory of Open Access Journals (Sweden)

    Rahul Pandit

    Full Text Available The role of the melanocortin (MC system in feeding behavior is well established. Food intake is potently suppressed by central infusion of the MC 3/4 receptor agonist α-melanocyte stimulating hormone (α-MSH, whereas the MC 3/4 receptor inverse-agonist Agouti Related Peptide (AGRP has the opposite effect. MC receptors are widely expressed in both hypothalamic and extra-hypothalamic brain regions, including nuclei involved in food reward and motivation, such as the nucleus accumbens (NAc and the ventral tegmental area. This suggests that MCs modulate motivational aspects of food intake. To test this hypothesis, rats were injected intracerebroventricularly with α-MSH or AGRP and their motivation for sucrose was tested under a progressive ratio schedule of reinforcement. Food motivated behavior was dose-dependently decreased by α-MSH. Conversely, AGRP increased responding for sucrose, an effect that was blocked by pretreatment with the dopamine receptor antagonist α-flupenthixol. In contrast to progressive ratio responding, free intake of sucrose remained unaltered upon α-MSH or AGRP infusion. In addition, we investigated whether the effects of α-MSH and AGRP on food motivation were mediated by the NAc shell. In situ hybridization of MC3 and MC4 receptor expression confirmed that the MC4 receptor was expressed throughout the NAc, and injection of α-MSH and AGRP into the NAc shell caused a decrease and an increase in motivation for sucrose, respectively. These data show that the motivation for palatable food is modulated by MC4 receptors in the NAc shell, and demonstrate cross-talk between the MC and dopamine system in the modulation of food motivation.

  12. Orosensory self-stimulation by sucrose involves brain dopaminergic mechanisms.

    Science.gov (United States)

    Schneider, L H

    1989-01-01

    The most convincing body of evidence supporting a role for brain dopaminergic mechanisms in sweet taste reward has been obtained using the sham-feeding rat. In rats prepared with a chronic gastric fistula and tested with the cannula open, intake is a direct function of the palatability of the solution offered as well as of the state of food deprivation. Because essentially none of the ingested fluid passes on to the intestine, negative postingestive feedback is eliminated. Thus, the relative orosensory/hedonic potency of the food determines and sustains the rate of sham intake; long periods of food deprivation are not required. In this way, the sham feeding of sweet solutions may be considered a form of oral self-stimulation behavior and afford a preparation through which the neurochemical and neuranatomical substrates of sweet taste reward may be identified. The results obtained in the series of experiments summarized in this paper clearly indicate that central D-1 and D-2 receptor mechanisms are critical for the orosensory self-stimulation by sucrose in the rat. In conclusion, I suggest that such investigations of the roles of brain dopaminergic mechanisms in the sucrose sham-feeding rat preparation may further our understanding of normal and aberrant attractions to sweet fluids in humans (see Cabanac, Drewnowski, and Halmi, this volume), as an innate, positive affective response of human neonates to sucrose and the sustained positive hedonic ratings for glucose when tasted but not when consumed have demonstrated. PMID:2699194

  13. Sucrose-replacement by rebaudioside a in a model beverage.

    Science.gov (United States)

    Majchrzak, Dorota; Ipsen, Annika; Koenig, Juergen

    2015-09-01

    Rebaudioside A (RA), a component of Stevia rebaudiana, is a non-caloric sweetener of natural origin, suitable to meet consumers' demand for sweet taste, but undesirable flavors were reported at high concentrations. Aim of this study was to create a model beverage (ice-tea) in which sucrose was replaced increasingly by RA to identify optimal sensory profile for consumer acceptance. Samples with 20 % and 40 % sucrose replacement by RA, respectively, showed very similar sensory profiles but were significantly higher in some flavor attributes, such as artificial sweetness, licorice-like and metallic, as well as in sweet and bitter aftertaste (p < 0.05) compared to the reference ice-tea. In both hedonic tests, preference and acceptance samples with RA have been judged as comparable to the reference despite perception of some undesirable notes. In view of the results of our study it can be stated that a replacement of 20 % or 40 % sucrose by RA in an ice-tea is achievable. PMID:26345024

  14. Pioglitazone can ameliorate insulin resistance in low-dose streptozotocin and high sucrose-fat diet induced obese rats

    Institute of Scientific and Technical Information of China (English)

    Shi-ying DING; Zhu-fang SHEN; Yue-teng CHEN; Su-juan SUN; Quan Liu; Ming-zhi XIE

    2005-01-01

    Aim: To investigate the effect of the peroxisome proliferator-activator receptor (PPAR)-γ agonist, pioglitazone, on insulin resistance in low-dose streptozotocin and high sucrose-fat diet induced obese rats. Methods: Normal female Wistar rats were injected intraperitoneally with low-dose streptozotocin (STZ, 30 mg/kg) and fed with a high sucrose-fat diet for 8 weeks. Pioglitazone (20 mg/kg) was administered orally to the obese and insulin-resistant rats for 28 d. Intraperitoneal glucose tolerance tests, insulin tolerance tests and gluconeogenesis tests were carried out over the last 14 d. At the end of d 28 of the treatment, serums were collected for biochemical analysis. Glucose transporter 4 (GLUT4) and insulin receptor substrate-1 (IRS-1) protein expression in the liver and skeletal muscle were detected using Western blotting. Results: Significant insulin resistance and obesity were observed in low-dose STZ and high sucrose-fat diet induced obese rats. Pioglitazone (20 mg/kg) treatment significantly decreased serum insulin,triglyceride and free fatty acid levels, and elevated high density lipoprotein-cholesterol (HDL-C) levels. Pioglitazone also lowered the lipid contents in the liver and muscles of rats undergoing treatment. Gluconeogenesis was inhibited and insulin sensitivity was improved markedly. The IRS-1 protein contents in the liver and skeletal muscles and the GLUT4 contents in skeletal muscle were elevated significantly. Conclusion: The data suggest that treatment with pioglitazone improves insulin sensitivity in low-dose STZ and high sucrose-fat diet induced obese rats. The insulin sensitizing effect may be associated with ameliorating lipid metabolism, reducing hyperinsulinemia, inhibiting gluconeogenesis, and increasing IRS-1 and GLUT4 protein expression in insulin-sensitive tissues.

  15. On the role of a Lipid-Transfer Protein. Arabidopsis ltp3 mutant is compromised in germination and seedling growth.

    OpenAIRE

    Pagnussat, Luciana A; Oyarburo, Natalia; Cimmino, Carlos; Pinedo, Marcela L; de la Canal, Laura

    2015-01-01

    Plant Lipid-Transfer Proteins (LTPs) exhibit the ability to reversibly bind/transport lipids in vitro. LTPs have been involved in diverse physiological processes but conclusive evidence on their role has only been presented for a few members, none of them related to seed physiology. Arabidopsis seeds rely on storage oil breakdown to supply carbon skeletons and energy for seedling growth. Here, Arabidopsis ltp3 mutant was analyzed for its ability to germinate and for seedling establishment. Lt...

  16. Episodic sucrose intake during food restriction increases synaptic abundance of AMPA receptors in nucleus accumbens and augments intake of sucrose following restoration of ad libitum feeding.

    Science.gov (United States)

    Peng, X-X; Lister, A; Rabinowitsch, A; Kolaric, R; Cabeza de Vaca, S; Ziff, E B; Carr, K D

    2015-06-01

    Weight-loss dieting often leads to loss of control, rebound weight gain, and is a risk factor for binge pathology. Based on findings that food restriction (FR) upregulates sucrose-induced trafficking of glutamatergic AMPA receptors to the nucleus accumbens (NAc) postsynaptic density (PSD), this study was an initial test of the hypothesis that episodic "breakthrough" intake of forbidden food during dieting interacts with upregulated mechanisms of synaptic plasticity to increase reward-driven feeding. Ad libitum (AL) fed and FR subjects consumed a limited amount of 10% sucrose, or had access to water, every other day for 10 occasions. Beginning three weeks after return of FR rats to AL feeding, when 24-h chow intake and rate of body weight gain had normalized, subjects with a history of sucrose intake during FR consumed more sucrose during a four week intermittent access protocol than the two AL groups and the group that had access to water during FR. In an experiment that substituted noncontingent administration of d-amphetamine for sucrose, FR subjects displayed an enhanced locomotor response during active FR but a blunted response, relative to AL subjects, during recovery from FR. This result suggests that the enduring increase in sucrose consumption is unlikely to be explained by residual enhancing effects of FR on dopamine signaling. In a biochemical experiment which paralleled the sucrose behavioral experiment, rats with a history of sucrose intake during FR displayed increased abundance of pSer845-GluA1, GluA2, and GluA3 in the NAc PSD relative to rats with a history of FR without sucrose access and rats that had been AL throughout, whether they had a history of episodic sucrose intake or not. A history of FR, with or without a history of sucrose intake, was associated with increased abundance of GluA1. A terminal 15-min bout of sucrose intake produced a further increase in pSer845-GluA1 and GluA2 in subjects with a history of sucrose intake during FR

  17. Nitrosative stress and apoptosis in non-anemic healthy rats induced by intravenous iron sucrose similars versus iron sucrose originator.

    Science.gov (United States)

    Toblli, Jorge E; Cao, Gabriel; Angerosa, Margarita

    2015-04-01

    Iron can both induce and inhibit nitrosative stress. Intracellular iron levels play an important role in nitric oxide (NO(•)) signaling mechanisms. Depending on various factors, such as the cell's redox state and transition metal levels, NO(•) generation may lead to lipid peroxidation and DNA damage as well as both anti- and pro-apoptotic effects. Administration of intravenous iron sucrose originator (IS(ORIG)) has been shown not to cause significant tyrosine nitration or significantly increased caspase 3 levels in non-anemic rats. In this study, the potential of several marketed iron sucrose similars (ISSs) to induce tyrosine nitration and caspase 3 expression in non-anemic rats was assessed. Although the physico-chemical properties of most of the analyzed ISSs complied with the United States Pharmacopeia for iron sucrose injection, all ISSs resulted in higher levels of tyrosine nitration and increased the expression of caspase 3 versus IS(ORIG). Moreover, significant differences were detected in tissue iron distribution between IS(ORIG)- and ISS-treated animals. In general, ISORIG resulted in higher levels of ferritin deposits versus ISSs whereas ISSs showed higher Prussian blue-stainable iron(III) deposits than IS(ORIG). This result suggests that some iron from ISSs bypassed the tightly regulated pathway through resident macrophages of the liver, spleen and bone marrow thus, ending up in the cellular compartment that favors oxidative and or nitrosative stress as well as apoptosis. The results also confirm that polynuclear iron(III)-oxyhydroxide carbohydrates, such as iron sucrose, cannot be fully characterized by physico-chemical methods alone.

  18. Characterization of Sugar Insensitive (sis) Mutants of Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Gibson, Susan I.

    2009-06-08

    Despite the fact that soluble sugar levels have been postulated to play an important role in the control of a wide variety of plant metabolic and developmental pathways, the mechanisms by which plants respond to soluble sugar levels remain poorly understood. Plant responses to soluble sugar levels are also important in bioenergy production, as plant sugar responses are believed to help regulate both carbon fixation and carbon partitioning. For example, accumulation of soluble sugars, such as sucrose and glucose, in source tissues leads to feedback inhibition of photosynthesis, thereby decreasing rates of carbon fixation. Soluble sugar levels can also affect sink strengths, affecting the rates of accumulation of carbon-based compounds into both particular molecular forms (e.g. carbohydrates versus lipids versus proteins) and particular plant organs and tissues. Mutants of Arabidopsis that are defective in the ability to respond to soluble sugar levels were isolated and used as tools to identify some of the factors involved in plant sugar response. These sugar insensitive (sis) mutants were isolated by screening mutagenized seeds for those that were able to germinate and develop relatively normal shoot systems on media containing 0.3 M glucose or 0.3 M sucrose. At these sugar concentrations, wild-type Arabidopsis germinate and produce substantial root systems, but show little to no shoot development. Twenty-eight sis mutants were isolated during the course of four independent mutant screens. Based on a preliminary characterization of all of these mutants, sis3 and sis6 were chosen for further study. Both of these mutations appear to lie in previously uncharacterized loci. Unlike many other sugar-response mutants, sis3 mutants exhibit a wild-type or near wild-type response in all phytohormone-response assays conducted to date. The sis6-1 mutation is unusual in that it appears to be due to overexpression of a gene, rather than representing a loss of function mutation

  19. Exploiting Natural Variation in Arabidopsis

    NARCIS (Netherlands)

    Molenaar, J.A.; Keurentjes, J.J.B.

    2014-01-01

    Natural variation for many traits is present within the species Arabidopsis thaliana . This chapter describes the use of natural variation to elucidate genes underlying the regulation of quantitative traits. It deals with the development and use of mapping populations, the detection and handling of

  20. Exploiting natural variation in Arabidopsis

    NARCIS (Netherlands)

    J.A. Molenaar; J.J.B. Keurentjes

    2014-01-01

    Natural variation for many traits is present within the species Arabidopsis thaliana. This chapter describes the use of natural variation to elucidate genes underlying the regulation of quantitative traits. It deals with the development and use of mapping populations, the detection and handling of g

  1. High sucrose consumption promotes obesity whereas its low consumption induces oxidative stress in Drosophila melanogaster.

    Science.gov (United States)

    Rovenko, Bohdana M; Kubrak, Olga I; Gospodaryov, Dmytro V; Perkhulyn, Natalia V; Yurkevych, Ihor S; Sanz, Alberto; Lushchak, Oleh V; Lushchak, Volodymyr I

    2015-08-01

    The effects of sucrose in varied concentrations (0.25-20%) with constant amount of yeasts in larval diet on development and metabolic parameters of adult fruit fly Drosophila melanogaster were studied. Larvae consumed more food at low sucrose diet, overeating with yeast. On high sucrose diet, larvae ingested more carbohydrates, despite consuming less food and obtaining less protein derived from yeast. High sucrose diet slowed down pupation and increased pupa mortality, enhanced levels of lipids and glycogen, increased dry body mass, decreased water content, i.e. resulted in obese phenotype. Furthermore, it suppressed reactive oxygen species-induced oxidation of lipids and proteins as well as the activity of superoxide dismutase. The activity of catalase was gender-related. In males, at all sucrose concentrations used catalase activity was higher than at its concentration of 0.25%, whereas in females sucrose concentration virtually did not influence the activity. High sucrose diet increased content of protein thiols and the activity of glucose-6-phosphate dehydrogenase. The increase in sucrose concentration also enhanced uric acid level in females, but caused opposite effects in males. Development on high sucrose diets was accompanied by elevated steady-state insulin-like peptide 3 mRNA level. Finally, carbohydrate starvation at yeast overfeeding on low sucrose diets resulted in oxidative stress reflected by higher levels of oxidized lipids and proteins accompanied by increased superoxide dismutase activity. Potential mechanisms involved in regulation of redox processes by carbohydrates are discussed.

  2. High sucrose consumption promotes obesity whereas its low consumption induces oxidative stress in Drosophila melanogaster.

    Science.gov (United States)

    Rovenko, Bohdana M; Kubrak, Olga I; Gospodaryov, Dmytro V; Perkhulyn, Natalia V; Yurkevych, Ihor S; Sanz, Alberto; Lushchak, Oleh V; Lushchak, Volodymyr I

    2015-08-01

    The effects of sucrose in varied concentrations (0.25-20%) with constant amount of yeasts in larval diet on development and metabolic parameters of adult fruit fly Drosophila melanogaster were studied. Larvae consumed more food at low sucrose diet, overeating with yeast. On high sucrose diet, larvae ingested more carbohydrates, despite consuming less food and obtaining less protein derived from yeast. High sucrose diet slowed down pupation and increased pupa mortality, enhanced levels of lipids and glycogen, increased dry body mass, decreased water content, i.e. resulted in obese phenotype. Furthermore, it suppressed reactive oxygen species-induced oxidation of lipids and proteins as well as the activity of superoxide dismutase. The activity of catalase was gender-related. In males, at all sucrose concentrations used catalase activity was higher than at its concentration of 0.25%, whereas in females sucrose concentration virtually did not influence the activity. High sucrose diet increased content of protein thiols and the activity of glucose-6-phosphate dehydrogenase. The increase in sucrose concentration also enhanced uric acid level in females, but caused opposite effects in males. Development on high sucrose diets was accompanied by elevated steady-state insulin-like peptide 3 mRNA level. Finally, carbohydrate starvation at yeast overfeeding on low sucrose diets resulted in oxidative stress reflected by higher levels of oxidized lipids and proteins accompanied by increased superoxide dismutase activity. Potential mechanisms involved in regulation of redox processes by carbohydrates are discussed. PMID:26050918

  3. The F130S point mutation in the Arabidopsis high-affinity K+ transporter AtHAK5 increases K+ over Na+ and Cs+ selectivity and confers Na+ and Cs+ tolerance to yeast under heterologous expression

    Directory of Open Access Journals (Sweden)

    Fernando eAleman

    2014-09-01

    Full Text Available Potassium (K+ is an essential macronutrient required for plant growth, development and high yield production of crops. Members of group I of the KT/HAK/KUP family of transporters, such as HAK5, are key components for K+ acquisition by plant roots at low external K+ concentrations. Certain abiotic stress conditions such as salinity or Cs+-polluted soils may jeopardize plant K+ nutrition because HAK5-mediated K+ transport is inhibited by Na+ and Cs+. Here, by screening in yeast a randomly-mutated collection of AtHAK5 transporters, a new mutation in AtHAK5 sequence is identified that greatly increases Na+ tolerance. The single point mutation F130S, affecting an amino acid residue conserved in HAK5 transporters from several species, confers high salt tolerance, as well as Cs+ tolerance. This mutation increases more than 100-fold the affinity of AtHAK5 for K+ and reduces the Ki values for Na+ and Cs+, suggesting that the F130 residue may contribute to the structure of the pore region involved in K+ binding. In addition, this mutation increases the Vmax for K+. All this changes occur without increasing the amount of the AtHAK5 protein in yeast and support the idea that this residue is contributing to shape the selectivity filter of the AtHAK5 transporter.

  4. CvADH1, a member of short-chain alcohol dehydrogenase family, is inducible by gibberellin and sucrose in developing watermelon seeds.

    Science.gov (United States)

    Kim, Joonyul; Kang, Hong-Gyu; Jun, Sung-Hoon; Lee, Jinwon; Yim, Jieun; An, Gynheung

    2003-01-01

    To understand the molecular mechanisms that control seed formation, we selected a seed-preferential gene (CvADH1) from the ESTs of developing watermelon seeds. RNA blot analysis and in situ localization showed that CvADH1 was preferentially expressed in the nucellar tissue. The CvADH1 protein shared about 50% homology with short-chain alcohol dehydrogenase including ABA2 in Arabidopsis thaliana, stem secoisolariciresinol dehydrogenase in Forsythia intermedia, and 3beta-hydroxysterol dehydrogenase in Digitalis lanata. We investigated gene-expression levels in seeds from both normally pollinated fruits and those made parthenocarpic via N-(2-chloro-4-pyridyl)-N'-phenylurea treatment, the latter of which lack zygotic tissues. Whereas the transcripts of CvADH1 rapidly started to accumulate from about the pre-heart stage in normal seeds, they were not detectable in the parthenocarpic seeds. Treating the parthenogenic fruit with GA(3) strongly induced gene expression, up to the level accumulated in pollinated seeds. These results suggest that the CvADH1 gene is induced in maternal tissues by signals made in the zygotic tissues, and that gibberellin might be one of those signals. We also observed that CvADH1 expression was induced by sucrose in the parthenocarpic seeds. Therefore, we propose that the CvADH1 gene is inducible by gibberellin, and that sucrose plays an important role in the maternal tissues of watermelon during early seed development. PMID:12552151

  5. Protective mechanism of desiccation tolerance in Reaumuria soongorica: Leaf abscission and sucrose accumulation in the stem

    Institute of Scientific and Technical Information of China (English)

    LIU YuBing; ZHANG TengGuo; LI XingRong; WANG Gang

    2007-01-01

    Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub, is widely found in semi-arid areas in northwestern China and can survive severe desiccation of its vegetative organs. In order to study the protective mechanism of desiccation tolerance in R. Soongorica, diurnal patterns of net photosynthetic rate (Pn), water use efficiency (WUE) and chlorophyll fluorescence parameters of Photosystem II (PSII), and sugar content in the source leaf and stem were investigated in 6-year-old plants during progressive soil drought imposed by the cessation of watering. The results showed that R. Soongorica was characterized by very low leaf water potential, high WUE, photosynthesis and high accumulation of sucrose in the stem and leaf abscission under desiccation. The maximum Pn increased at first and then declined during drought, but intrinsic WUE increased remarkably in the morning with increasing drought stress. The maximal photochemical efficiency of PSII (Fv/Fm) and the quantum efficiency of noncyclic electric transport of PSII(Φpsii) decreased significantly under water stress and exhibited an obvious phenomenon of photoinhibition at noon. Drought stressed plants maintained a higher capacity of dissipation of the excitation energy (measured as NPQ) with the increasing intensity of stress. Conditions of progressive drought promoted sucrose and starch accumulation in the stems but not in the leaves. However, when leaf water potential was less than -21.3 Mpa, the plant leaves died and then abscised. But the stem photosynthesis remained and, afterward the plants entered the dormant state. Upon rewatering, the shoots reactivated and the plants developed new leaves. Therefore, R. Soongorica has the ability to reduce water loss through leaf abscission and maintain the vigor of the stem cells to survive desiccation.

  6. Protective mechanism of desiccation tolerance in Reaumuria soongorica: Leaf abscission and sucrose accumulation in the stem

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub, is widely found in semi-arid areas in northwestern China and can survive severe desiccation of its vegetative organs. In order to study the protective mechanism of desiccation tolerance in R. soongorica, diurnal patterns of net photosynthetic rate (Pn), water use efficiency (WUE) and chlorophyll fluorescence parameters of Photosystem II (PSII), and sugar content in the source leaf and stem were investigated in 6-year-old plants during progressive soil drought imposed by the cessation of watering. The results showed that R. soongorica was char-acterized by very low leaf water potential, high WUE, photosynthesis and high accumulation of sucrose in the stem and leaf abscission under desiccation. The maximum Pn increased at first and then de-clined during drought, but intrinsic WUE increased remarkably in the morning with increasing drought stress. The maximal photochemical efficiency of PSII (Fv/Fm) and the quantum efficiency of noncyclic electric transport of PSII(ΦPSII) decreased significantly under water stress and exhibited an obvious phenomenon of photoinhibition at noon. Drought stressed plants maintained a higher capacity of dis-sipation of the excitation energy (measured as NPQ) with the increasing intensity of stress. Conditions of progressive drought promoted sucrose and starch accumulation in the stems but not in the leaves. However, when leaf water potential was less than –21.3 MPa, the plant leaves died and then abscised. But the stem photosynthesis remained and, afterward the plants entered the dormant state. Upon re-watering, the shoots reactivated and the plants developed new leaves. Therefore, R. soongorica has the ability to reduce water loss through leaf abscission and maintain the vigor of the stem cells to survive desiccation.

  7. Effect of sucrose starvation on sycamore (Acer pseudoplatanus) cell carbohydrate and Pi status.

    Science.gov (United States)

    Rébeillé, F; Bligny, R; Martin, J B; Douce, R

    1985-03-15

    The mobilization of stored carbohydrates during sucrose starvation was studied with sycamore (Acer pseudoplatanus) cells. When sucrose was omitted from the nutrient medium, the intracellular sucrose pool decreased rapidly during the first hours of the experiment, whereas the starch content remained practically unchanged. After 10h of sucrose starvation, starch hydrolysis replaced sucrose breakdown. From this moment, the phosphate-ester pool and respiration rate decreased with time. Conversely, the intracellular Pi concentration increased. 31P n.m.r. of intact sycamore cells indicated that, under these conditions, most of the Pi accumulated in the vacuole. These results strongly suggest that starch breakdown, in contrast with sucrose hydrolysis, is not rapid enough to maintain a high cellular metabolism.

  8. Effect of sucrose on physical properties of spray-dried whole milk powder.

    Science.gov (United States)

    Ma, U V Lay; Ziegler, G R; Floros, J D

    2008-11-01

    Spray-dried whole milk powders were prepared from whole condensed milk with various sucrose concentrations (0%, 2.5%, 5%, 7.5%, and 10% w/w), and their glass transition temperature and some physical properties of importance in chocolate manufacture were evaluated. In milk powder samples, the glass transition temperature and free-fat content decreased in a nonlinear manner with sucrose addition. Moreover, increasing sucrose concentration reduced the formation of dents on the particle surface. Addition of sucrose in whole condensed milk increased linearly the apparent particle density and in a nonlinear manner the particle size of spray-dried milk powders. The particle size volume distribution of milk powders with the highest sucrose concentration differed from the log-normal distribution of the other samples due to the formation of large agglomerates. Neither vacuole volume, nor the amorphous state of milk powders was affected by sucrose addition.

  9. Comparative Proteomic Analysis of Arabidopsis Mature Pollen and Germinated Pollen

    Institute of Scientific and Technical Information of China (English)

    Junjie Zou; Lianfen Song; Wenzheng Zhang; Yi Wang; Songlin Ruan; Wei-Hua Wu

    2009-01-01

    Proteomic analysis was applied to generating the map of Arabidopsis mature pollen proteins and analyzing the differentially expressed proteins that are potentially involved in the regulation of Arabidopsis pollen germination. By applying 2-D electrophoresis and silver staining, we resolved 499 and 494 protein spots from protein samples extracted from pollen grains and pollen tubes, respectively. Using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, we identified 189 distinct proteins from 213 protein spots expressed in mature pollen or pollen tubes, and 75 new identified proteins that had not been reported before in research into the Arabidopsis pollen proteome. Comparative analysis revealed that 40 protein spots exhibit reproducible significant changes between mature pollen and pollen tubes. And 21 proteins from 17 downregulated and six upregulated protein spots were identified. Functional category analysis indicated that these differentially expressed proteins mainly involved in signaling, cellular structure, transport, defense/stress responses, transcription, metabolism, and energy production. The patterns of changes at protein level suggested the important roles for energy metabolism-related proteins in pollen tube growth, accompanied by the activation of the stress response pathway and modifications to the cell wall.

  10. Deer Frozen Semen Quality in Tris Sucrose and Tris Glucose Extender with Different Glycerol Concentrations

    OpenAIRE

    W. M. M. Nalley; R. Handarini; R.I Arifiantini; T.L. Yusuf; B. Purwantara; G. Semiadi

    2011-01-01

    In order to improve Timor deer (Cervus timorensis) frozen semen quality, the influence of sugar and glycerol concentration on semen characteristics of sperm was investigated. The semen was collected from five sexually mature Timor deer using an electroejaculator. The semen was evaluated and divided into six equal tubes and diluted with Tris sucrose glycerol 10% (TSG10); Tris sucrose glycerol 12% (TSG12); Tris sucrose glycerol 14% (TSG14); Tris glucose glycerol 10% (TGG10); Tris glucose glyce...

  11. [Succinic acid production from sucrose and sugarcane molasses by metabolically engineered Escherichia coli].

    Science.gov (United States)

    Li, Feng; Ma, Jiangfeng; Wu, Mingke; Ji, Yaliang; Chen, Wufang; Ren, Xinyi; Jiang, Min

    2015-04-01

    Sugarcane molasses containing large amounts of sucrose is an economical substrate for succinic acid production. However, Escherichia coli AFP111 cannot metabolize sucrose although it is a promising candidate for succinic acid production. To achieve sucrose utilizing ability, we cloned and expressed cscBKA genes encoding sucrose permease, fructokinase and invertase of non-PTS sucrose-utilization system from E. coli W in E. coli AFP111 to generate a recombinant strain AFP111/pMD19T-cscBKA. After 72 h of anaerobic fermentation of the recombinant in serum bottles, 20 g/L sucrose was consumed and 12 g/L succinic acid was produced. During dual-phase fermentation comprised of initial aerobic growth phase followed by anaerobic fermentation phase, the concentration of succinic acid from sucrose and sugarcane molasses was 34 g/L and 30 g/L, respectively, at 30 h of anaerobic phase in a 3 L fermentor. The results show that the introduction of non-PTS sucrose-utilization system has sucrose-metabolizing capability for cell growth and succinic acid production, and can use cheap sugarcane molasses to produce succinic acid.

  12. Photocatalytic properties of hierarchical ZnO flowers synthesized by a sucrose-assisted hydrothermal method

    Science.gov (United States)

    Lv, Wei; Wei, Bo; Xu, Lingling; Zhao, Yan; Gao, Hong; Liu, Jia

    2012-10-01

    In this work, hierarchical ZnO flowers were synthesized via a sucrose-assisted urea hydrothermal method. The thermogravimetric analysis/differential thermal analysis (TGA-DTA) and Fourier transform infrared spectra (FTIR) showed that sucrose acted as a complexing agent in the synthesis process and assisted combustion during annealing. Photocatalytic activity was evaluated using the degradation of organic dye methyl orange. The sucrose added ZnO flowers showed improved activity, which was mainly attributed to the better crystallinity as confirmed by X-ray photoelectron spectroscopy (XPS) analysis. The effect of sucrose amount on photocatalytic activity was also studied.

  13. Biological Sensor for Sucrose Availability: Relative Sensitivities of Various Reporter Genes

    OpenAIRE

    Miller, William G; Brandl, Maria T; Quiñones, Beatriz; Lindow, Steven E.

    2001-01-01

    A set of three sucrose-regulated transcriptional fusions was constructed. Fusions p61RYTIR, p61RYlac, and p61RYice contain the scrR sucrose repressor gene and the promoterless gfp, lacZ, and inaZ reporter genes, respectively, fused to the scrY promoter from Salmonella enterica serovar Typhimurium. Cells of Erwinia herbicola containing these fusions are induced only in media amended with sucrose, fructose, or sorbose. While a large variation in sucrose-dependent reporter gene activity was obse...

  14. HEXOKINASE 1 is required for glucose-induced repression of bZIP63, At5g22920 and BT2 in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Sabine eKunz

    2015-07-01

    Full Text Available Simple sugars, like glucose and sucrose, act as signals to modulate the expression of hundreds of genes in plants. Frequently, however, it remains unclear whether this regulation is induced by the sugars themselves or by their derivatives generated in the course of carbohydrate (CH metabolism. In the present study, we tested the relevance of different CH metabolism and allocation pathways affecting expression patterns of five selected sugar-responsive genes (bZIP63, At5g22920, BT2, MGD2 and TPS9 in Arabidopsis thaliana. In general, the expression followed diurnal changes in the overall sugar availability. However, under steady growth conditions, this response was hardly impaired in the mutants for CH metabolizing/ transporting proteins (adg1, sex1, sus1-4, sus5/6 and tpt2, including also hexokinase1 (HXK1 loss- and gain-of-function plants - gin2.1 and oe3.2, respectively. In addition, transgenic plants carrying pbZIP63::GUS showed no changes in reporter-gene-expression when grown on sugar under steady-state conditions. In contrast, short-term treatments of agar-grown seedlings with 1% glucose or sucrose induced pbZIP63::GUS repression, which became even more apparent in seedlings grown in liquid media. Subsequent analyses of liquid-grown gin2.1 and oe3.2 seedlings revealed that glucose-dependent regulation of the five selected genes was not affected in gin2.1, whereas it was enhanced in oe3.2 plants for bZIP63, At5g22920 and BT2. The sugar treatments had no effect on ATP/ADP ratio, suggesting that changes in gene expression were not linked to cellular energy status. Overall, the data suggest that HXK1 does not act as glucose sensor controlling bZIP63, At5g22920 and BT2 expression, but it is nevertheless required for production of a downstream metabolic signal regulating their expression.

  15. In vivo localization in Arabidopsis protoplasts and root tissue.

    Science.gov (United States)

    Lee, Myoung Hui; Lee, Yongjik; Hwang, Inhwan

    2013-01-01

    In eukaryotic cells, a large number of proteins are transported to their final destination after translation by a process called intracellular trafficking. Transient gene expression, either in plant protoplasts or in specific plant tissues, is a fast, flexible, and reproducible approach to study the cellular function of proteins, protein subcellular localizations, and protein-protein interactions. Here we describe the general method of protoplast isolation, polyethylene glycol-mediated protoplast transformation and immunostaining of protoplast or intact root tissues for studying the localization of protein in Arabidopsis.

  16. Strigolactone acts downstream of auxin to regulate bud outgrowth in pea and Arabidopsis.

    Science.gov (United States)

    Brewer, Philip B; Dun, Elizabeth A; Ferguson, Brett J; Rameau, Catherine; Beveridge, Christine A

    2009-05-01

    During the last century, two key hypotheses have been proposed to explain apical dominance in plants: auxin promotes the production of a second messenger that moves up into buds to repress their outgrowth, and auxin saturation in the stem inhibits auxin transport from buds, thereby inhibiting bud outgrowth. The recent discovery of strigolactone as the novel shoot-branching inhibitor allowed us to test its mode of action in relation to these hypotheses. We found that exogenously applied strigolactone inhibited bud outgrowth in pea (Pisum sativum) even when auxin was depleted after decapitation. We also found that strigolactone application reduced branching in Arabidopsis (Arabidopsis thaliana) auxin response mutants, suggesting that auxin may act through strigolactones to facilitate apical dominance. Moreover, strigolactone application to tiny buds of mutant or decapitated pea plants rapidly stopped outgrowth, in contrast to applying N-1-naphthylphthalamic acid (NPA), an auxin transport inhibitor, which significantly slowed growth only after several days. Whereas strigolactone or NPA applied to growing buds reduced bud length, only NPA blocked auxin transport in the bud. Wild-type and strigolactone biosynthesis mutant pea and Arabidopsis shoots were capable of instantly transporting additional amounts of auxin in excess of endogenous levels, contrary to predictions of auxin transport models. These data suggest that strigolactone does not act primarily by affecting auxin transport from buds. Rather, the primary repressor of bud outgrowth appears to be the auxin-dependent production of strigolactones. PMID:19321710

  17. Arabidopsis thaliana—Aphid Interaction

    OpenAIRE

    Louis, Joe; Singh, Vijay,; Shah, Jyoti

    2012-01-01

    Aphids are important pests of plants that use their stylets to tap into the sieve elements to consume phloem sap. Besides the removal of photosynthates, aphid infestation also alters source-sink patterns. Most aphids also vector viral diseases. In this chapter, we will summarize on recent significant findings in plant-aphid interaction, and how studies involving Arabidopsis thaliana and Myzus persicae (Sülzer), more commonly known as the green peach aphid (GPA), are beginning to provide impor...

  18. Stem cell organization in Arabidopsis

    OpenAIRE

    Wendrich, J.R.

    2016-01-01

    Growth of plant tissues and organs depends on continuous production of new cells, by niches of stem cells. Stem cells typically divide to give rise to one differentiating daughter and one non-differentiating daughter. This constant process of self-renewal ensures that the niches of stem cells or meristems stay active throughout plant-life. Specification of stem cells occurs very early during development of the emrbyo and they are maintained during later stages. The Arabidopsis embryo is a hig...

  19. Arabidopsis CDS blastp result: AK240730 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240730 J043030K09 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 2e-11 ...

  20. Arabidopsis CDS blastp result: AK288052 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288052 J075151I09 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 6e-14 ...

  1. Arabidopsis CDS blastp result: AK240911 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240911 J065037E05 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 4e-22 ...

  2. Arabidopsis CDS blastp result: AK241119 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241119 J065094C22 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 2e-13 ...

  3. Arabidopsis CDS blastp result: AK243149 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243149 J100032I21 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 7e-12 ...

  4. Arabidopsis CDS blastp result: AK241581 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241581 J065181K09 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 4e-15 ...

  5. Arabidopsis CDS blastp result: AK287479 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287479 J043023O14 At2g32440.1 68415.m03963 ent-kaurenoic acid hydroxylase, putati...ve / cytochrome P450, putative identical to ent-kaurenoic acid hydroxylase / cytochrome P450 CYP88A (GI:1302...1856) [Arabidopsis thaliana]; similar to ent-kaurenoic acid hydroxylase [Arabidopsis thaliana] GI:13021853 1e-17 ...

  6. Using "Arabidopsis" Genetic Sequences to Teach Bioinformatics

    Science.gov (United States)

    Zhang, Xiaorong

    2009-01-01

    This article describes a new approach to teaching bioinformatics using "Arabidopsis" genetic sequences. Several open-ended and inquiry-based laboratory exercises have been designed to help students grasp key concepts and gain practical skills in bioinformatics, using "Arabidopsis" leucine-rich repeat receptor-like kinase (LRR RLK) genetic…

  7. Effect of Sucrose Concentration on Sucrose-Dependent Adhesion and Glucosyltransferase Expression of S. mutans in Children with Severe Early-Childhood Caries (S-ECC

    Directory of Open Access Journals (Sweden)

    Wei Zhao

    2014-09-01

    Full Text Available Sucrose, extracellular polysaccharide, and glucosyltransferases (GTFs are key factors in sucrose-dependent adhesion and play important roles in the process of severe early-childhood caries (S-ECC. However, whether sucrose concentration regulates gtf expression, extracellular polysaccharide synthesis, and sucrose-dependent adhesion is related to the different genotypes of S. mutans isolated from ECC in children and still needs to be investigated. In this study, 52 strains of S. mutans were isolated from children with S-ECC and caries-free (CF children. Water-insoluble glucan (WIG synthesis was detected by the anthrone method, adhesion capacity by the turbidimetric method, and expression of gtf by RT-PCR in an in vitro model containing 1%–20% sucrose. The genotypes of S. mutans were analyzed by AP-PCR. The results showed that WIG synthesis, adhesion capacity, and gtf expression increased significantly when the sucrose concentration was from 1% to 10%. WIG synthesis and gtfB as well as gtfC expression of the 1% and 5% groups were significantly lower than those of the 10% and 20% groups (p < 0.05. There were no significant differences between the 10% and 20% groups. The fingerprints of S. mutans detected from individuals in the S-ECC group exhibited a significant difference in diversity compared with those from CF individuals (p < 0.05. Further, the expression of gtfB and gtfC in the S-ECC group was significantly different among the 1- to 5-genotype groups (p < 0.05. It can be concluded that sucrose-dependent adhesion might be related to the diversity of genotypes of S. mutans, and the 10% sucrose level can be seen as a “turning point” and essential factor for the prevention of S-ECC.

  8. An International Bioinformatics Infrastructure to Underpin the Arabidopsis Community

    Science.gov (United States)

    The future bioinformatics needs of the Arabidopsis community as well as those of other scientific communities that depend on Arabidopsis resources were discussed at a pair of recent meetings held by the Multinational Arabidopsis Steering Committee (MASC) and the North American Arabidopsis Steering C...

  9. Identification of sucrose synthase as an actin-binding protein

    Science.gov (United States)

    Winter, H.; Huber, J. L.; Huber, S. C.; Davies, E. (Principal Investigator)

    1998-01-01

    Several lines of evidence indicate that sucrose synthase (SuSy) binds both G- and F-actin: (i) presence of SuSy in the Triton X-100-insoluble fraction of microsomal membranes (i.e. crude cytoskeleton fraction); (ii) co-immunoprecipitation of actin with anti-SuSy monoclonal antibodies; (iii) association of SuSy with in situ phalloidin-stabilized F-actin filaments; and (iv) direct binding to F-actin, polymerized in vitro. Aldolase, well known to interact with F-actin, interfered with binding of SuSy, suggesting that a common or overlapping binding site may be involved. We postulate that some of the soluble SuSy in the cytosol may be associated with the actin cytoskeleton in vivo.

  10. Sugaring the pill: ethics and uncertainties in the use of sucrose for newborn infants.

    Science.gov (United States)

    Wilkinson, Dominic J C; Savulescu, Julian; Slater, Rebeccah

    2012-07-01

    Sucrose is widely used for the management of procedural pain in newborn infants, including capillary blood sampling, venepuncture, and vascular cannulation. Multiple randomized controlled trials have demonstrated that sweet-tasting solutions reduce behavioral responses to acute painful stimuli. It has been claimed that sucrose should be a standard of care in neonatal units and that further placebo-controlled trials of sucrose are unnecessary and unethical. However, recently published data cast doubt on the analgesic properties of sucrose. We review this new evidence and analyze the philosophical and ethical questions that it raises, including the "problem of other minds." Sugar may be better understood not as an analgesic, removing or relieving pain, but as a compensating pleasure. There is a need for further research on the mechanism of sucrose's effect on pain behavior and on the long-term effects of sucrose treatment. Such trials will require comparison with placebo or with other interventions. Given uncertainty about the benefit of sucrose, it may be wise to use alternative analgesics or nonpharmacological interventions where these are available and appropriate. Sucrose may not be the answer to procedural pain in newborns.

  11. 40 CFR 180.1222 - Sucrose octanoate esters; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Sucrose octanoate esters; exemption... FOOD Exemptions From Tolerances § 180.1222 Sucrose octanoate esters; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for residues of...

  12. Coordination of sucrose uptake and respiration in the yeast Debaryomyces yamadae

    NARCIS (Netherlands)

    Kaliterna, J.; Weusthuis, R.A.; Castrillo, J.I.; Dijken, van J.P.; Pronk, J.T.

    1995-01-01

    Screening in batch cultures identified Debaryomyces yamadae as a yeast that exhibits the Kluyver effect for sucrose: this disaccharide can be respired but, even under oxygen-limited conditions, alcoholic fermentation of sucrose does not occur. Ethanol, glycerol and arabitol were the main fermentatio

  13. Rats' preferences for high fructose corn syrup vs. sucrose and sugar mixtures.

    Science.gov (United States)

    Ackroff, Karen; Sclafani, Anthony

    2011-03-28

    High fructose corn syrup (HFCS) has replaced sucrose in many food products, which has prompted research comparing these two sweeteners in rodents. The present study examined the relative palatability of HFCS and sucrose for rats, offering 11% carbohydrate solutions to match the content of common beverages for human consumption. The animals initially preferred HFCS to sucrose but after separate experience with each solution they switched to sucrose preference. Approximating the composition of HFCS with a mixture of fructose and glucose (55:45) yielded a solution that was less attractive than sucrose or HFCS. However, HFCS contains a small amount of glucose polymers, which are very attractive to rats. A 55:42:3 mixture of fructose, glucose and glucose polymers (Polycose) was equally preferred to HFCS and was treated similarly to HFCS in comparisons vs. sucrose. Post-oral effects of sucrose, which is 50% fructose and 50% glucose, may be responsible for the shift in preference with experience. This shift, and the relatively small magnitude of differences in preference for HFCS and sucrose, suggest that palatability factors probably do not contribute to any possible difference in weight gain responses to these sweeteners.

  14. Comparison of Oral Iron and IV Iron Sucrose for Treatment of Anemia in Postpartum Indian Women

    Directory of Open Access Journals (Sweden)

    Aggarwal Rohina S, Mishra Vineet V, Panchal Navin A, Patel Nital H, Deshchougule Vrushali V, Jasani Anil F

    2012-01-01

    Full Text Available Objectives: To determine the efficacy of intravenous iron sucrose and oral iron in anaemic postpartum women, presenting at Institute of Kidney Disease and Research Centre. Method: Descriptive case series. The study was carried out in the department of Obstetrics and Gynaecology at Institute of Kidney Disease and Research Centre over a period of six months from 02-02-2010 to 02-08-2010. 50 cases with proven iron deficiency with Hb ?7gm% were included in the study. Total iron deficit was calculated using a standard formula. Target haemoglobin was 12 gm %. Iron sucrose was administered by intravenous infusion. Haemoglobin was repeated 1, 2, 3, and 4 weeks after the last dose of intravenous iron sucrose. Results: On inclusion, the 2 groups were comparable in terms of both anthropometric and biologic data Distribution of cases by economic status showed, 21 patients (42.0% belonged to lower class, 20 patients (40% belonged to middle class and 09 patients (18% were of upper class.Target hemoglobin levels were achieved in 4 weeks time in 20 (80% patients in iron sucrose group as compared to 10 (40% of patients in oral iron group .There was significant improvement in the various hematological parameters in iv sucrose group as compared to patients in oral iron group. There were no significant allergic reactions in IV sucrose group. Conclusion: This study has shown a significant improvement in the iron sucrose group. Iron sucrose is safe and well tolerated.

  15. Pursuing the Pavlovian Contributions to Induction in Rats Responding for 1% Sucrose Reinforcement

    Science.gov (United States)

    Weatherly, Jeffrey N.; Huls, Amber; Kulland, Ashley

    2007-01-01

    The present study investigated whether Pavlovian conditioning contributes, in the form of the response operandum serving as a conditioned stimulus, to the increase in the rate of response for 1% liquid-sucrose reinforcement when food-pellet reinforcement is upcoming. Rats were exposed to conditions in which sign tracking for 1% sucrose was…

  16. Sucrose regulation of ADP-glucose pyrophosphorylase subunit genes transcript levels in leaves and fruits

    Science.gov (United States)

    Li, Xiangyang; Xing, Jinpeng; Gianfagna, Thomas J.; Janes, Harry W.

    2002-01-01

    ADP-glucose pyrophosphorylase (AGPase, EC2.7.7.27) is a key regulatory enzyme in starch biosynthesis. The enzyme is a heterotetramer with two S and two B subunits. In tomato, there are three multiple forms of the S subunit gene. Agp S1, S2 and B are highly expressed in fruit from 10 to 25 days after anthesis. Agp S3 is only weakly expressed in fruit. Sucrose significantly elevates expression of Agp S1, S2 and B in both leaves and fruits. Agp S1 exhibits the highest degree of regulation by sucrose. In fact, sucrose may be required for Agp S1 expression. For excised leaves incubated in water, no transcripts for Agp S1 could be detected in the absence of sucrose, whereas it took up to 16 h in water before transcripts were no longer detectable for Agp S2 and B. Neither Agp S3 nor the tubulin gene is affected by sucrose, demonstrating that this response is specifically regulated by a carbohydrate metabolic signal, and is not due to a general increase in metabolism caused by sucrose treatment. Truncated versions of the promoter for Agp S1 indicate that a specific region 1.3-3.0 kb upstream from the transcription site is responsible for sucrose sensitivity. This region of the S1 promoter contains several cis-acting elements present in the promoters of other genes that are also regulated by sucrose. c2002 Elsevier Science Ireland Ltd. All rights reserved.

  17. Biosynthesis of fructo-oligosaccharides by Sporotrichum thermophile during submerged batch cultivation in high sucrose media

    DEFF Research Database (Denmark)

    Katapodis, P.; Kalogeris, E.; Kekos, D.;

    2004-01-01

    Biosynthesis of fructo-oligosaccharides (FOS) was observed during growth of the thermophilic fungus Sporotrichum thermophile on media containing high sucrose concentrations. Submerged batch cultivation with the optimum initial sucrose concentration of 250 g/l allowed the production of 12.5 g FOS...

  18. Oxytocin Differentially Affects Sucrose Taking and Seeking in Male and Female Rats

    Science.gov (United States)

    Zhou, Luyi; Ghee, Shannon M.; See, Ronald E.; Reichel, Carmela M.

    2015-01-01

    Oxytocin has a modulatory role in natural and drug reward processes. While the role of oxytocin in pair bonding and reproduction has been extensively studied, sex differences in conditioned and unconditioned behavioral responses to oxytocin treatment have not been fully characterized. Here, we determined whether male and female rats would show similar dose response curves in response to acute oxytocin on measures of locomotor activity, sucrose seeking, and sucrose intake. Male and freely cycling female rats received vehicle or oxytocin (0.1, 0.3, 1, 3 mg/kg, IP) injections before behavioral tests designed to assess general motor activity, as well as sucrose self-administration and seeking. Lower doses of oxytocin decreased motor activity in a novel environment in females relative to males. Likewise, lower doses of oxytocin in females decreased responding for sucrose during maintenance of sucrose self-administration and reinstatement to sucrose-conditioned cues. However, sucrose seeking in response to a sucrose prime was only decreased by the highest oxytocin dose in both sexes. In general, oxytocin had similar effects in both sexes. However, females were more sensitive to lower doses of oxytocin than males. These findings are consistent with the notion that oxytocin regulates many of the same behaviors in males and females, but that the effects are typically more profound in females. Therapeutic use of oxytocin should include sex as a factor in determining dose regimens. PMID:25647756

  19. Feedback inhibition of starch degradation in Arabidopsis leaves mediated by trehalose 6-phosphate.

    Science.gov (United States)

    Martins, Marina Camara Mattos; Hejazi, Mahdi; Fettke, Joerg; Steup, Martin; Feil, Regina; Krause, Ursula; Arrivault, Stéphanie; Vosloh, Daniel; Figueroa, Carlos María; Ivakov, Alexander; Yadav, Umesh Prasad; Piques, Maria; Metzner, Daniela; Stitt, Mark; Lunn, John Edward

    2013-11-01

    Many plants accumulate substantial starch reserves in their leaves during the day and remobilize them at night to provide carbon and energy for maintenance and growth. In this paper, we explore the role of a sugar-signaling metabolite, trehalose-6-phosphate (Tre6P), in regulating the accumulation and turnover of transitory starch in Arabidopsis (Arabidopsis thaliana) leaves. Ethanol-induced overexpression of trehalose-phosphate synthase during the day increased Tre6P levels up to 11-fold. There was a transient increase in the rate of starch accumulation in the middle of the day, but this was not linked to reductive activation of ADP-glucose pyrophosphorylase. A 2- to 3-fold increase in Tre6P during the night led to significant inhibition of starch degradation. Maltose and maltotriose did not accumulate, suggesting that Tre6P affects an early step in the pathway of starch degradation in the chloroplasts. Starch granules isolated from induced plants had a higher orthophosphate content than granules from noninduced control plants, consistent either with disruption of the phosphorylation-dephosphorylation cycle that is essential for efficient starch breakdown or with inhibition of starch hydrolysis by β-amylase. Nonaqueous fractionation of leaves showed that Tre6P is predominantly located in the cytosol, with estimated in vivo Tre6P concentrations of 4 to 7 µm in the cytosol, 0.2 to 0.5 µm in the chloroplasts, and 0.05 µm in the vacuole. It is proposed that Tre6P is a component in a signaling pathway that mediates the feedback regulation of starch breakdown by sucrose, potentially linking starch turnover to demand for sucrose by growing sink organs at night.

  20. Pronounced phenotypic changes in transgenic tobacco plants overexpressing sucrose synthase may reveal a novel sugar signaling pathway

    Directory of Open Access Journals (Sweden)

    Quynh Anh eNguyen

    2016-01-01

    Full Text Available Soluble sugars not only serve as nutrients, but also act as signals for plant growth and development, but how sugar signals are perceived and translated into physiological responses in plants remains unclear. We manipulated sugar levels in transgenic plants by overexpressing sucrose synthase (SuSy, which is a key enzyme believed to have reversible sucrose synthesis and sucrose degradation functions. The ectopically expressed SuSy protein exhibited sucrose-degrading activity, which may change the flux of sucrose demand from photosynthetic to non-photosynthetic cells, and trigger an unknown sucrose signaling pathway that lead to increased sucrose content in the transgenic plants. An experiment on the transition from heterotrophic to autotrophic growth demonstrated the existence of a novel sucrose signaling pathway, which stimulated photosynthesis, and enhanced photosynthetic synthesis of sucrose, which was the direct cause or the sucrose increase. In addition, a light/dark time treatment experiment, using different day length ranges for photosynthesis/respiration showed the carbohydrate pattern within a 24-hour day and consolidated the role of sucrose signaling pathway as a way to maintain sucrose demand, and indicated the relationships between increased sucrose and upregulation of genes controlling development of the shoot apical meristem (SAM. As a result, transgenic plants featured a higher biomass and a shorter time required to switch to reproduction compared to those of control plants, indicating altered phylotaxis and more rapid advancement of developmental stages in the transgenic plants.

  1. Tape-Arabidopsis Sandwich - a simpler Arabidopsis protoplast isolation method

    Directory of Open Access Journals (Sweden)

    Lee Shu-Hong

    2009-11-01

    Full Text Available Abstract Background Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP, is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast isolation method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments. Results In this report, we present a new procedure that we call the Tape-Arabidopsis Sandwich. This is a simple and fast mesophyll protoplast isolation method. Two kinds of tape (Time tape adhered to the upper epidermis and 3 M Magic tape to the lower epidermis are used to make a "Tape-Arabidopsis Sandwich". The Time tape supports the top side of the leaf during manipulation, while tearing off the 3 M Magic tape allows easy removal of the lower epidermal layer and exposes mesophyll cells to cell wall digesting enzymes when the leaf is later incubated in an enzyme solution. The protoplasts released into solution are collected and washed for further use. For TEAMP, plasmids carrying a gene expression cassette for a fluorescent protein can be successfully delivered into protoplasts isolated from mature leaves grown under optimal conditions. Alternatively, these protoplasts may be used for bimolecular fluorescence complementation (BiFC to investigate protein-protein interactions in vivo, or for Western blot analysis. A significant advantage of this protocol over the current method is that it allows the generation of protoplasts in less than 1 hr

  2. A sucrose-rich diet induces mutations in the rat colon

    DEFF Research Database (Denmark)

    Dragsted, Lars O.; Daneshvar, Bahram; Vogel, Ulla;

    2002-01-01

    A sucrose-rich diet has repeatedly been observed to have cocarcinogenic actions in the colon and liver of rats and to increase the number of aberrant crypt foci in rat colon. To investigate whether sucrose-rich diets might directly increase the genotoxic response in the rat colon or liver, we have...... added sucrose to the diet of Big Blue rats, a strain of Fischer rats carrying 40 copies of the lambda-phage on chromosome 4. Dietary sucrose was provided to the rats for 3 weeks at four dose levels including the background level in the purified diet [3.4% (control), 6.9%, 13.8%, or 34.5%] without...... of a sucrose-rich diet. No significant increase in mutations was observed in the liver. To seek an explanation for this finding, a variety of parameters were examined representing different mechanisms, including increased oxidative stress, changes in oxidative defense, effects on DNA repair, or changes...

  3. Sucrose Solutions as Prospective Medium to Study the Vesicle Structure SAXS and SANS study

    CERN Document Server

    Kiselev, M A; Kisselev, A M; Lombardo, D; Killany, M; Lesieur, S

    2001-01-01

    The possibility to use sucrose solutions as medium for X-ray and neutron small-angle scattering experiments has been explored for dimyristoylphosphatidylcholine (DMPC) vesicles and mixed DMPC/C_(12)E_(8) aggregates. The influence of sucrose concentration on phospholipid vesicles size and polydispersity has been investigated by complimentary X-ray and neutron scattering. Sucrose solutions decreased vesicle size and polydispersity and increased a contrast between phospholipid membrane and bulk solvent sufficiently for X-rays. 40% sucrose in H2O increased X-ray contrast by up to 10 times compared to pure H2O. The range of sucrose concentration 30%-40% created the best experimental conditions for the X-ray small-angle experiment with phospholipid vesicles.

  4. Intestinal sugar transport

    Institute of Scientific and Technical Information of China (English)

    Laurie A Drozdowski; Alan BR Thomson

    2006-01-01

    Carbohydrates are an important component of the diet.The carbohydrates that we ingest range from simple monosaccharides (glucose, fructose and galactose) to disaccharides (lactose, sucrose) to complex polysaccharides. Most carbohydrates are digested by salivary and pancreatic amylases, and are further broken down into monosaccharides by enzymes in the brush border membrane (BBM) of enterocytes. For example, lactase-phloridzin hydrolase and sucraseisomaltase are two disaccharidases involved in the hydrolysis of nutritionally important disaccharides. Once monosaccharides are presented to the BBM, mature enterocytes expressing nutrient transporters transport the sugars into the enterocytes. This paper reviews the early studies that contributed to the development of a working model of intestinal sugar transport, and details the recent advances made in understanding the process by which sugars are absorbed in the intestine.

  5. Identification of actively filling sucrose sinks. [Solanum tuberosum; Phaseolus lunatus; Manihot esculenta; Liquidambar styraciflua L. ; Carya illinoinensis

    Energy Technology Data Exchange (ETDEWEB)

    Sung, Shijean S.; Xu, Dianpeng; Black C.C. (Univ. of Georgia, Athens (USA))

    1989-04-01

    Certain actively filling plant sucrose sinks such as a seed, a tuber, or a root can be identified by measuring the uridine diphosphate and pyrophosphate-dependent metabolism of sucrose. Sucrolysis in both active and quiescent sucrose sinks was tested and sucrose synthase was found to be the predominant sucrose breakdown activity. Sucrolysis via invertases was low and secondary in both types of sinks. Sucrose synthase activity dropped markedly, greater than fivefold, in quiescent sinks. The test are consistent with the hypothesis that the sucrose filling activity, i.e. the sink strength, of these plant sinks can be measured by testing the uridine diphosphate and pyrophosphate-dependent breakdown of sucrose. Measuring the initial reactions of sucrolysis shows much promise for use in agriculture crop and tree improvement research as a biochemical test for sink strength.

  6. Jasmonate Signal Pathway in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Xiao-Yi Shan; Zhi-Long Wang; Daoxin Xie

    2007-01-01

    Jasmonates (JAs), which include jasmonic acid and its cyclopentane derivatives are synthesized from the octadecanoid pathway and widely distributed throughout the plant kingdom. JAs modulate the expression of numerous genes and mediate responses to stress, wounding, insect attack, pathogen infection, and UV damage. They also affect a variety of processes in many plant developmental processes. The JA signal pathway involves two important events: the biosynthesis of JA and the transduction of JA signal. Several important Arabidopsis mutants in jasmonate signal pathway were described in this review.

  7. Detecting autophagy in Arabidopsis roots by membrane-permeable cysteine protease inhibitor E-64d and endocytosis tracer FM4–64

    OpenAIRE

    Oh-ye, Yuumi; Inoue, Yuko; Moriyasu, Yuji

    2011-01-01

    Autophagy is the process by which cells degrade their own components in lysosomes or vacuoles. Autophagy in tobacco BY-2 cells cultured in sucrose-free medium takes place in formed, autolysosomes in the presence of a cysteine protease inhibitor. The autolysosomes in BY-2 cells are located in the endocytotic pathway and thus can be stained with fluorescent endocytosis marker FM4–64. In the present study, in order to detect autophagy in the root cells of Arabidopsis, we incubated root tips from...

  8. Exploring the neutral invertase-oxidative stress defence connection in Arabidopsis thaliana.

    Science.gov (United States)

    Xiang, Li; Le Roy, Katrien; Bolouri-Moghaddam, Mohammad-Reza; Vanhaecke, Mieke; Lammens, Willem; Rolland, Filip; Van den Ende, Wim

    2011-07-01

    Over the past decades, considerable advances have been made in understanding the crucial role and the regulation of sucrose metabolism in plants. Among the various sucrose-catabolizing enzymes, alkaline/neutral invertases (A/N-Invs) have long remained poorly studied. However, recent findings have demonstrated the presence of A/N-Invs in various organelles in addition to the cytosol, and their importance for plant development and stress tolerance. A cytosolic (At-A/N-InvG, At1g35580) and a mitochondrial (At-A/N-InvA, At1g56560) member of the A/N-Invs have been analysed in more detail in Arabidopsis and it was found that At-A/N-InvA knockout plants show an even more severe growth phenotype than At-A/N-InvG knockout plants. The absence of either A/N-Inv was associated with higher oxidative stress defence gene expression, while transient overexpression of At-A/N-InvA and At-A/N-InvG in leaf mesophyll protoplasts down-regulated the oxidative stress-responsive ascorbate peroxidase 2 (APX2) promoter. Moreover, up-regulation of the APX2 promoter by hydrogen peroxide or abscisic acid could be blocked by adding metabolizable sugars or ascorbate. A hypothetical model is proposed in which both mitochondrial and cytosolic A/N-Invs can generate glucose as a substrate for mitochondria-associated hexokinase, contributing to mitochondrial reactive oxygen species homeostasis.

  9. Elucidating the Roles of Transport Processes in Glucosinolate Distribution

    DEFF Research Database (Denmark)

    Madsen, Svend Roesen

    Glucosinolates are plant defense compounds characteristic of the economically important plant family of Brassicaceae, which comprises crops as oilseed rape, cabbage, broccoli and the model plant Arabidopsis thaliana (Arabidopsis). Recently, two Arabidopsis glucosinolate transporters, GTR1 and GTR2......, were identified. In this thesis, we show that GTR1 and GTR2 are not required for obtaining the strategic margin distribution of glucosinolates in a mature Arabidopsis leaf, and we suggest a model for leaf allocation of glucosinolates. As glucosinolates are defense compounds, we asked if eliminating GTR......1 and GTR2 would influence the ability of the plant to defend itself against attackers. Infecting WT and gtr1gtr2 dKO leaves with the necrotic fungus Botrytis cinerea showed an increased susceptibility of the transporter mutant compared to WT. In a second biotic interaction, we infested WT and gtr1...

  10. Use of the Biphasic 13C-Sucrose/Glucose Breath Test to Assess Sucrose Maldigestion in Adults with Functional Bowel Disorders

    Science.gov (United States)

    Balesh, Albert M.; Shelby, Harold T.

    2016-01-01

    Sucrase insufficiency has been observed in children with of functional bowel disorders (FBD) and symptoms of dietary carbohydrate intolerance may be indistinguishable from those of FBD. A two-phase 13C-sucrose/13C-glucose breath test (13C-S/GBT) was used to assess sucrase activity because disaccharidase assays are seldom performed in adults. When 13C-sucrose is hydrolyzed to liberate monosaccharides, oxidation to 13CO2 is a proportional indicator of sucrase activity. Subsequently, 13C-glucose oxidation rate was determined after a secondary substrate ingestion (superdose) to adjust for individual habitus effects (Phase II). 13CO2 enrichment recovery ratio from 13C-sucrose and secondary 13C-glucose loads reflect the individualized sucrase activity [Coefficient of Glucose Oxidation for Sucrose (CGO-S)]. To determine if sucrase insufficiency could be a factor in FBD, 13C-S/GBT was validated using subjects with known sucrase gene mutation status by comparing 13CO2-breath enrichment with plasma 13C-glucose enrichment. 13C-S/GBT was used to assess sucrose digestion in FBD patients and asymptomatic controls. 13CO2-breath enrichment correlated with the appearance of 13C-sucrose-derived glucose in plasma (r2 = 0.80). Mean, control group CGO-S-enrichment outcomes were 1.01 at 60′, 0.92 at 75′, and 0.96 at mean 60′–75′ with normal CGO-S defined as >0.85 (95% C.I.). In contrast, FBD patients demonstrated lower CGO-S values of 0.77 at 60′, 0.77 at 75′, and 0.76 at mean 60′–75′ (Chi Square: 6.55; p < 0.01), which points to sucrose maldigestion as a cause of FBD. PMID:27579322

  11. Expression analysis of genes associated with sucrose accumulation in sugarcane (Saccharum spp. hybrids) varieties differing in content and time of peak sucrose storage.

    Science.gov (United States)

    Chandra, A; Verma, P K; Islam, M N; Grisham, M P; Jain, R; Sharma, A; Roopendra, K; Singh, K; Singh, P; Verma, I; Solomon, S

    2015-05-01

    Sucrose synthesis/accumulation in sugarcane is a complex process involving many genes and regulatory sequences that control biochemical events in source-sink tissues. Among these, sucrose synthase (SuSy), sucrose phosphate synthase (SPS), soluble acid (SAI) and cell wall (CWI) invertases are important. Expression of these enzymes was compared in an early (CoJ64) and late (BO91) maturing sugarcane variety using end-point and qRT-PCR. Quantitative RT-PCR at four crop stages revealed high CWI expression in upper internodes of CoJ64, which declined significantly in both top and bottom internodes with maturity. In BO91, CWI expression was high in top and bottom internodes and declined significantly only in top internodes as the crop matured. Overall, CWI expression was higher in CoJ64 than in BO91. During crop growth, there was no significant change in SPS expression in bottom internodes in CoJ64, whereas in BO91 it decreased significantly. Apart from a significant decrease in expression of SuSy in mature bottom internodes of BO91, there was no significant change. Similar SAI expression was observed with both end-point and RT-PCR, except for significantly increased expression in top internodes of CoJ64 with maturity. SAI, being a major sucrose hydrolysing enzyme, was also monitored with end-point PCR expression in internode tissues of CoJ64 and BO91, with higher expression of SAI in BO91 at early crop stages. Enzyme inhibitors, e.g. manganese chloride (Mn(++) ), significantly suppressed expression of SAI in both early- and late-maturing varieties. Present findings enhance understanding of critical sucrose metabolic gene expression in sugarcane varieties differing in content and time of peak sucrose storage. Thus, through employing these genes, improvement of sugarcane sucrose content is possible.

  12. Use of the Biphasic (13)C-Sucrose/Glucose Breath Test to Assess Sucrose Maldigestion in Adults with Functional Bowel Disorders.

    Science.gov (United States)

    Opekun, Antone R; Balesh, Albert M; Shelby, Harold T

    2016-01-01

    Sucrase insufficiency has been observed in children with of functional bowel disorders (FBD) and symptoms of dietary carbohydrate intolerance may be indistinguishable from those of FBD. A two-phase (13)C-sucrose/(13)C-glucose breath test ((13)C-S/GBT) was used to assess sucrase activity because disaccharidase assays are seldom performed in adults. When (13)C-sucrose is hydrolyzed to liberate monosaccharides, oxidation to (13)CO2 is a proportional indicator of sucrase activity. Subsequently, (13)C-glucose oxidation rate was determined after a secondary substrate ingestion (superdose) to adjust for individual habitus effects (Phase II). (13)CO2 enrichment recovery ratio from (13)C-sucrose and secondary (13)C-glucose loads reflect the individualized sucrase activity [Coefficient of Glucose Oxidation for Sucrose (CGO-S)]. To determine if sucrase insufficiency could be a factor in FBD, (13)C-S/GBT was validated using subjects with known sucrase gene mutation status by comparing (13)CO2-breath enrichment with plasma (13)C-glucose enrichment. (13)C-S/GBT was used to assess sucrose digestion in FBD patients and asymptomatic controls. (13)CO2-breath enrichment correlated with the appearance of (13)C-sucrose-derived glucose in plasma (r (2) = 0.80). Mean, control group CGO-S-enrichment outcomes were 1.01 at 60', 0.92 at 75', and 0.96 at mean 60'-75' with normal CGO-S defined as >0.85 (95% C.I.). In contrast, FBD patients demonstrated lower CGO-S values of 0.77 at 60', 0.77 at 75', and 0.76 at mean 60'-75' (Chi Square: 6.55; p < 0.01), which points to sucrose maldigestion as a cause of FBD. PMID:27579322

  13. Proteomics Analysis Reveals Post-Translational Mechanisms for Cold-Induced Metabolic Changes in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Tian Li; Alma L.Burlingame; Zhi-Ping Deng; Zhi Yong Wang; Shou-Ling Xu; Juan A.Oses-Prieto; Sunita Putil; Peng Xu; Rui-Ju Wang; Kathy H.Li; David A.Malty; Liz-He An

    2011-01-01

    Cold-induced changes of gene expression and metabolism are critical for plants to survive freezing. Largely by changing gene expression, exposure to a period of non-freezing low temperatures increases plant tolerance to freezing-a phenomenon known as cold acclimation. Cold also induces rapid metabolic changes, which provide instant protection before temperature drops below freezing point. The molecular mechanisms for such rapid metabolic responses to cold remain largely unknown. Here, we use two-dimensional difference gel electrophoresis (2-D DIGE) analysis of sub-cellular fractions of Arabidopsis thaliana proteome coupled with spot identification by tandem mass spectrometry to identify early cold-responsive proteins in Arabidopsis. These proteins include four enzymes involved in starch degradation, three HSP100 proteins, several proteins in the tricarboxylic acid cycle, and sucrose metabolism. Upon cold treatment, the Disproportionating Enzyme 2 (DPE2), a cytosolic transglucosidase metabolizing maltose to glucose, increased rapidly in the centrifugation pellet fraction and decreased in the soluble fraction. Consistent with cold-induced inactivation of DPE2 enzymatic activity, the dpe2 mutant showed increased freezing tolerance without affecting the C-repeat binding transcription factor (CBF) transcriptional pathway. These results support a model that cold-induced inactivation of DPE2 leads to rapid accumulation of maltose, which is a cold-induced compatible solute that protects cells from freezing damage. This study provides evidence for a key role of rapid post-translational regulation of carbohydrate metabolic enzymes in plant protection against sudden temperature drop.

  14. Ectopic expression of a tobacco vacuolar invertase inhibitor in guard cells confers drought tolerance in Arabidopsis.

    Science.gov (United States)

    Chen, Su-Fen; Liang, Ke; Yin, Dong-Mei; Ni, Di-An; Zhang, Zhi-Guo; Ruan, Yong-Ling

    2016-12-01

    There are several hypotheses that explain stomatal behavior. These include the concept of osmoregulation mediated by potassium and its counterions malate and chlorine and the more recent starch-sugar hypothesis. We have previously reported that the activity of the sucrose cleavage enzyme, vacuolar invertase (VIN), is significantly higher in guard cells than in other leaf epidermal cells and its activity is correlated with stomatal aperture. Here, we examined whether VIN indeed controls stomatal movement under normal and drought conditions by transforming Arabidopsis with a tobacco vacuolar invertase inhibitor homolog (Nt-inhh) under the control of an abscisic acid-sensitive and guard cell-specific promoter (AtRab18). The data obtained showed that guard cells of transgenic Arabidopsis plants had lower VIN activity, stomatal aperture and conductance than that of wild-type plants. Moreover, the transgenic plants also displayed higher drought tolerance than wild-type plants. The data indicate that VIN is a promising target for manipulating stomatal function to increase drought tolerance. PMID:26899912

  15. Polyploidy in the Arabidopsis genus.

    Science.gov (United States)

    Bomblies, Kirsten; Madlung, Andreas

    2014-06-01

    Whole genome duplication (WGD), which gives rise to polyploids, is a unique type of mutation that duplicates all the genetic material in a genome. WGD provides an evolutionary opportunity by generating abundant genetic "raw material," and has been implicated in diversification, speciation, adaptive radiation, and invasiveness, and has also played an important role in crop breeding. However, WGD at least initially challenges basic biological functions by increasing cell size, altering relationships between cell volume and DNA content, and doubling the number of homologous chromosome copies that must be sorted during cell division. Newly polyploid lineages often have extensive changes in gene regulation, genome structure, and may suffer meiotic or mitotic chromosome mis-segregation. The abundance of species that persist in nature as polyploids shows that these problems are surmountable and/or that advantages of WGD might outweigh drawbacks. The molecularly especially tractable Arabidopsis genus has several ancient polyploidy events in its history and contains several independent more recent polyploids. This genus can thus provide important insights into molecular aspects of polyploid formation, establishment, and genome evolution. The ability to integrate ecological and evolutionary questions with molecular and genetic understanding makes comparative analyses in this genus particularly attractive and holds promise for advancing our general understanding of polyploid biology. Here, we highlight some of the findings from Arabidopsis that have given us insights into the origin and evolution of polyploids. PMID:24788061

  16. The Measurement of Sucrose Content of Sugar Cane Using Ultrasonic Waves

    Directory of Open Access Journals (Sweden)

    Amoranto Trisnobudi, Tjia Liong Hoei, Enung Rosihan Nugraha

    2001-04-01

    Full Text Available The measurement of sucrose content of sugar cane is usually carried out by using polarimeter and Brix Wager scale. These two apparatus are operated manually so that the accuracy of the measurement results is depended on the operator skill. To overcome this problem we have developed an alternative method that can measure the sucrose content more quickly and accurately than the conventional methods. This new method was carried out by using ultrasonic waves whose velocity depends on the sucrose content. Firstly, the electronic apparatus used was calibrated with 37 samples of sugar cane with various sucrose content from 4.46 % to 7.29 %. The result of this calibration was an empirical equation between the ultrasonic wave velocity V and the sucrose content R, i.e. R = 2.65 V2 - 11,95 V + 17,65 where R in % and V in km/s. Then this equation was stored as database in a computer program that will be used to calculate the sucrose content. Finally, this sucrose content measurement system was tested by using 30 samples of sugar cane. The maximum error of the measurement result was 6.4 %.

  17. Differential motivational profiles following adolescent sucrose access in male and female rats.

    Science.gov (United States)

    Reichelt, Amy C; Abbott, Kirsten N; Westbrook, R Fred; Morris, Margaret J

    2016-04-01

    Adolescents are the highest consumers of sugar sweetened drinks. Excessive consumption of such drinks is a likely contributor to the development of obesity and may be associated with enduring changes in the systems involved in reward and motivation. We examined the impact of daily sucrose consumption in young male and female rats (N=12 per group) across the adolescent period on the motivation to perform instrumental responses to gain food rewards as adults. Rats were or were not exposed to a sucrose solution for 2 h each day for 28 days across adolescence [postnatal days (P) 28-56]. They were then trained as adults (P70 onward) to lever press for a palatable 15% cherry flavored sucrose reward and tested on a progressive ratio (PR) schedule to assess motivation to respond for reinforcement. Female rats exposed to sucrose had higher breakpoints on the PR schedule than controls, whereas male rats exposed to sucrose had lower breakpoints than controls. These results show that consumption of sucrose during adolescence produced sex-specific behavioral changes in responding for sucrose as adults. PMID:26826605

  18. Enamel and dentine demineralization by a combination of starch and sucrose in a biofilm - caries model.

    Science.gov (United States)

    Botelho, Juliana Nunes; Villegas-Salinas, Mario; Troncoso-Gajardo, Pía; Giacaman, Rodrigo Andrés; Cury, Jaime Aparecido

    2016-05-20

    Sucrose is the most cariogenic dietary carbohydrate and starch is considered non-cariogenic for enamel and moderately cariogenic for dentine. However, the cariogenicity of the combination of starch and sucrose remains unclear. The aim of this study was to evaluate the effect of this combination on Streptococcus mutans biofilm composition and enamel and dentine demineralization. Biofilms of S. mutans UA159 were grown on saliva-coated enamel and dentine slabs in culture medium containing 10% saliva. They were exposed (8 times/day) to one of the following treatments: 0.9% NaCl (negative control), 1% starch, 10% sucrose, or 1% starch and 10% sucrose (starch + sucrose). To simulate the effect of human salivary amylase on the starch metabolization, the biofilms were pretreated with saliva before each treatment and saliva was also added to the culture medium. Acidogenicity of the biofilm was estimated by evaluating (2 times/day) the culture medium pH. After 4 (dentine) or 5 (enamel) days of growth, biofilms (n = 9) were individually collected, and the biomass, viable microorganism count, and polysaccharide content were quantified. Dentine and enamel demineralization was assessed by determining the percentage of surface hardness loss. Biofilms exposed to starch + sucrose were more acidogenic and caused higher demineralization (p < 0.0001) on either enamel or dentine than those exposed to each carbohydrate alone. The findings suggest that starch increases the cariogenic potential of sucrose. PMID:27223133

  19. CHEMOENZYMATIC SYNTHESIS OF BIODEGRADABLE POLY(1′-O-VINYLADIPOYL-SUCROSE)

    Institute of Scientific and Technical Information of China (English)

    Des-hui Lu; Qi Wu; Xian-fu Lin

    2002-01-01

    A novel polymer containing the sucrose group was synthesized by radical polymerization from an enzymaticallyprepared monomer, 1′-O-vinyladipoyl-sucrose (VAS). Transesterification reaction of sucrose with divinyl adipate inanhydrous pyridine catalyzed by an alkaline protease from Bacillus subtilis at 60℃ for 7 days gave VAS (yield 55%) withoutany blocking/deblocking steps. The vinyl sucrose ester could be polymerized with potassium persulfate and H2O2 as initiatorto give poly(1′-O-vinyladipoyl-sucrose) with Mn = 33,000 and Mw = 53,200, Mw/Mn = 1.61. The polymer was biodegradable.After 6 days in aqueous buffer (pH 7), this alkaline protease could degrade poly(1′-O-vinyladipoyl-sucrose) to Mn of ca.1080, Mw/Mn = 3.30 (37℃), and Mn of ca. 5200, Mw/Mn = 2.44 (4℃). The polymer containing the sucrose branch would be afunctional material in various application fields.

  20. Combined compared to dissociated oral and intestinal sucrose stimuli induce different brain hedonic processes

    Directory of Open Access Journals (Sweden)

    Caroline eClouard

    2014-08-01

    Full Text Available The characterization of brain networks contributing to the processing of oral and/or intestinal sugar signals in a relevant animal model might help to understand the neural mechanisms related to the control of food intake in humans and suggest potential causes for impaired eating behaviors. This study aimed at comparing the brain responses triggered by oral and/or intestinal sucrose sensing in pigs. Seven animals underwent brain single photon emission computed tomography (99mTc-HMPAO further to oral stimulation with neutral or sucrose artificial saliva paired with saline or sucrose infusion in the duodenum, the proximal part of the intestine. Oral and/or duodenal sucrose sensing induced differential cerebral blood flow (CBF changes in brain regions known to be involved in memory, reward processes and hedonic (i.e. pleasure evaluation of sensory stimuli, including the dorsal striatum, prefrontal cortex, cingulate cortex, insular cortex, hippocampus and parahippocampal cortex. Sucrose duodenal infusion only and combined sucrose stimulation induced similar activity patterns in the putamen, ventral anterior cingulate cortex and hippocampus. Some brain deactivations in the prefrontal and insular cortices were only detected in the presence of oral sucrose stimulation. Finally, activation of the right insular cortex was only induced by combined oral and duodenal sucrose stimulation, while specific activity patterns were detected in the hippocampus and parahippocampal cortex with oral sucrose dissociated from caloric load. This study sheds new light on the brain hedonic responses to sugar and has potential implications to unravel the neuropsychological mechanisms underlying food pleasure and motivation.

  1. Reconstruction and analysis of nutrient-induced phosphorylation networks in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Guangyou eDuan

    2013-12-01

    Full Text Available Elucidating the dynamics of molecular processes in living organisms in response to external perturbations is a central goal in modern systems biology. We investigated the dynamics of protein phosphorylation events in Arabidopsis thaliana exposed to changing nutrient conditions. Phosphopeptide expression levels were detected at five consecutive time points over a time interval of 30 minutes after nutrient resupply following prior starvation. The three tested inorganic, ionic nutrients NH4+, NO3-, PO43- elicited similar phosphosignaling responses that were distinguishable from those invoked by the sugars mannitol, sucrose. When embedded in the protein-protein interaction network of Arabidopsis thaliana, phosphoproteins were found to exhibit a higher degree compared to average proteins. Based on the time-series data, we reconstructed a network of regulatory interactions mediated by phosphorylation. The performance of different network inference methods was evaluated by the observed likelihood of physical interactions within and across different subcellular compartments and based on gene ontology semantic similarity. The dynamic phosphorylation network was then reconstructed using a Pearson correlation method with added directionality based on partial variance differences. The topology of the inferred integrated network corresponds to an information dissemination architecture, in which the phosphorylation signal is passed on to an increasing number of phosphoproteins stratified into an initiation, processing, and effector layer. Specific phosphorylation peptide motifs associated with the distinct layers were identified indicating the action of layer-specific kinases. Despite the limited temporal resolution, combined with information on subcellular location, the available time-series data proved useful for reconstructing the dynamics of the molecular signaling cascade in response to nutrient stress conditions in the plant Arabidopsis thaliana.

  2. An Arabidopsis mutant impaired in coenzyme A biosynthesis is sugar dependent for seedling establishment.

    Science.gov (United States)

    Rubio, Silvia; Larson, Tony R; Gonzalez-Guzman, Miguel; Alejandro, Santiago; Graham, Ian A; Serrano, Ramón; Rodriguez, Pedro L

    2006-03-01

    Once the plant coenzyme A (CoA) biosynthetic pathway has been elucidated by comparative genomics, it is feasible to analyze the physiological relevance of CoA biosynthesis in plant life. To this end, we have identified and characterized Arabidopsis (Arabidopsis thaliana) T-DNA knockout mutants of two CoA biosynthetic genes, HAL3A and HAL3B. The HAL3A gene encodes a 4'-phosphopantothenoyl-cysteine decarboxilase that generates 4'-phosphopantetheine. A second gene, HAL3B, whose gene product is 86% identical to that of HAL3A, is present in the Arabidopsis genome. HAL3A appears to have a predominant role over HAL3B according to their respective mRNA expression levels. The hal3a-1, hal3a-2, and hal3b mutants were viable and showed a similar growth rate as that in wild-type plants; in contrast, a hal3a-1 hal3b double mutant was embryo lethal. Unexpectedly, seedlings that were null for HAL3A and heterozygous for HAL3B (aaBb genotype) displayed a sucrose (Suc)-dependent phenotype for seedling establishment, which is in common with mutants defective in beta-oxidation. This phenotype was genetically complemented in aaBB siblings of the progeny and chemically complemented by pantethine. In contrast, seedling establishment of Aabb plants was not Suc dependent, proving a predominant role of HAL3A over HAL3B at this stage. Total fatty acid and acyl-CoA measurements of 5-d-old aaBb seedlings in medium lacking Suc revealed stalled storage lipid catabolism and impaired CoA biosynthesis; in particular, acetyl-CoA levels were reduced by approximately 80%. Taken together, these results provide in vivo evidence for the function of HAL3A and HAL3B, and they point out the critical role of CoA biosynthesis during early postgerminative growth. PMID:16415216

  3. MADS on the move : a study on MADS domain protein function and movement during floral development in Arabidopsis thaliana

    NARCIS (Netherlands)

    Urbanus, S.L.

    2010-01-01

    In this thesis we investigated the behaviour of fluorescently-tagged MADS domain proteins during floral development in the model plant Arabidopsis thaliana, and explored the importance of intercellular transport via plasmodesmata for MADS domain transcription factor functioning. The MADS domain tran

  4. EVIDENCE OF DIFFERENTIAL PH REGULATION OF THE ARABIDOPSIS VACUOLAR CA2+/H+ ANTIPORTERS CAX1 AND CAX2

    Science.gov (United States)

    The Arabidopsis Ca(2+)/H(+) antiporters cation exchanger (CAX) 1 and 2 utilise an electrochemical gradient to transport Ca(2+) into the vacuole to help mediate Ca(2+) homeostasis. Previous whole plant studies indicate that activity of Ca(2+)/H(+) antiporters is regulated by pH. However, the pH regul...

  5. Sucrose/bovine serum albumin mediated biomimetic crystallization of calcium carbonate

    Indian Academy of Sciences (India)

    Cheng-Li Yao; Wang-Hua Xu; Ai-Min Ding; Jin-Mao Zhu

    2009-01-01

    To understand the role of the sucrose/bovine serum albumin system in the biomineralization process, we have tested the influence of different concentration of the sucrose/bovine serum albumin (BSA) on calcium carbonate (CaCO3) precipitation. The CaCO3 crystals were characterized by scanning electron microscope (SEM), Fourier transform infrared spectrograph (FT-IR) and powder X-ray diffractometry (XRD). The possible formation mechanism of CaCO3 in the sucrose/bovine serum albumin system was discussed.

  6. Sucrose, glucose and fructose have similar genotoxicity in the rat colon and affect the metabolism

    DEFF Research Database (Denmark)

    Hansen, Max; Baunsgaard, D.; Autrup, H.;

    2008-01-01

    We have shown previously that a high sucrose intake increases the background level of somatic mutations and the level of bulky DNA adducts in the colon epithelium of rats. The mechanism may involve either glucose or fructose formed by hydrolysis of sucrose. Male Big Blue (R) rats were fed 30......% sucrose, glucose, fructose or potato starch as part of the diet. Mutation rates and bulky DNA adduct levels were determined in colon and liver. The concentration of short-chain fatty acids and pH were deter-mined in caecum, C-peptide was determined in plasma, biomarkers for oxidative damage...

  7. Synthesis of a sucrose dimer with enone tether; a study on its functionalization

    Directory of Open Access Journals (Sweden)

    Zbigniew Pakulski

    2014-05-01

    Full Text Available The reaction of appropriately functionalized sucrose phosphonate with sucrose aldehyde afforded a dimer composed of two sucrose units connected via their C6-positions (‘the glucose ends’. The carbonyl group in this product (enone was stereoselectively reduced with zinc borohydride and the double bond (after protection of the allylic alcohol formed after reduction was oxidized with osmium tetroxide to a diol. Absolute configurations of the allylic alcohol as well as the diol were determined by circular dichroism (CD spectroscopy using the in situ dimolybdenum methodology.

  8. COMPARATIVE STUDY BETWEEN INTRAVENOUS IRON SUCROSE AND ORAL IRON FOR TREATMENT OF POST PARTUM ANEMIA

    Directory of Open Access Journals (Sweden)

    Samta

    2014-03-01

    Full Text Available AIMS & OBJECTIVES: The aim of study was to compare the efficacy, safety and compliance of intravenous iron sucrose complex with oral Iron therapy in treatment of postpartum anemia. MATERIAL AND METHODS: 60 postpartum women who had delivered within 24 -48 hours and having hemoglobin 0.05. Intravenous iron sucrose did not result in any serious adverse reactions. CONCLUSION: Intravenous iron sucrose is more effective, rapid and safe in increasing hemoglobin level in women with postpartum anemia in comparison with oral iron therapy. It also replenishes iron stores more rapidly without any serious adverse effects.

  9. Biological sensor for sucrose availability: relative sensitivities of various reporter genes.

    Science.gov (United States)

    Miller, W G; Brandl, M T; Quiñones, B; Lindow, S E

    2001-03-01

    A set of three sucrose-regulated transcriptional fusions was constructed. Fusions p61RYTIR, p61RYlac, and p61RYice contain the scrR sucrose repressor gene and the promoterless gfp, lacZ, and inaZ reporter genes, respectively, fused to the scrY promoter from Salmonella enterica serovar Typhimurium. Cells of Erwinia herbicola containing these fusions are induced only in media amended with sucrose, fructose, or sorbose. While a large variation in sucrose-dependent reporter gene activity was observed in cells harboring all gene fusions, fusions to the inaZ reporter gene yielded a much wider range of activity and were responsive to lower levels of sucrose than either lacZ or gfp. The lacZ reporter gene was found to be more efficient than gfp, requiring approximately 300-fold fewer cells for a detectable response over all concentrations of sucrose. Similarly, inaZ was found to be more efficient than lacZ, requiring 30-fold fewer cells at 1.45 microM sucrose and 6,100-fold fewer cells at 29 mM sucrose for a quantifiable response. The fluorescence of individual cells containing p61RYTIR was quantified following epifluorescence microscopy in order to relate the fluorescence exhibited by populations of cells in batch cultures with that of individual cells in such cultures. While the mean fluorescence intensity of a population of individual cells increased with increasing concentrations of sucrose, a wide range of fluorescence intensity was seen among individual cells. For most cultures the distribution of fluorescence intensity among individual cells was log-normally distributed, but cells grown in intermediate concentrations of sucrose exhibited two distinct populations of cells, one having relatively low fluorescence and another with much higher fluorescence. When cells were inoculated onto bean leaves, whole-cell ice nucleation and gfp-based biological sensors for sucrose each indicated that the average concentration of sucrose on moist leaf surfaces was about 20 micro

  10. Effects of Vacuum Impregnation with Sucrose Solution on Mango Tissue.

    Science.gov (United States)

    Lin, Xian; Luo, Cailian; Chen, Yulong

    2016-06-01

    The influences of vacuum impregnation (VI) on the tissue of mango cubes during atmospheric immersion in sucrose solution were investigated. Results showed that VI effectively facilitated water loss (WL) and sugar gain (SG) during the 300min immersion process, with increases of 20.59% and 31.26%, respectively. A pectin solubilization/degradation phenomenon was observed in the immersion process. The intercellular space and cross section area in the VI-treated mango tissue increased immediately after being released to atmospheric pressure. And it was noted that after experiencing shrinkage-relaxation period twice in the 300 min immersion process, the size of VI-treated mango cells recovered to the original level of fresh ones. Major variations in WL, protopectin content, water soluble pectin content, firmness and microstructure of mango cubes appeared within the first 60 min. In addition, the firmness of mango cubes was positively correlated with the protopectin content (P < 0.01), but negatively correlated with WL and the water soluble pectin content (P < 0.01), indicating that WL and degradation of protopectin contributed greatly to the loss of firmness. PMID:27100561

  11. Analysis of Protein Oligomeric Species by Sucrose Gradients.

    Science.gov (United States)

    Tenreiro, Sandra; Macedo, Diana; Marijanovic, Zrinka; Outeiro, Tiago Fleming

    2016-01-01

    Protein misfolding, aggregation, and accumulation are a common hallmark in various neurodegenerative diseases. Invariably, the process of protein aggregation is associated with both a loss of the normal biological function of the protein and a gain of toxic function that ultimately leads to cell death. The precise origin of protein cytotoxicity is presently unclear but the predominant theory posits that smaller oligomeric species are more toxic than larger aggregated forms. While there is still no consensus on this subject, this is a central question that needs to be addressed in order to enable the design of novel and more effective therapeutic strategies. Accordingly, the development and utilization of approaches that allow the biochemical characterization of the formed oligomeric species in a given cellular or animal model will enable the correlation with cytotoxicity and other parameters of interest.Here, we provide a detailed description of a low-cost protocol for the analysis of protein oligomeric species from both yeast and mammalian cell lines models, based on their separation according to sedimentation velocity using high-speed centrifugation in sucrose gradients. This approach is an adaptation of existing protocols that enabled us to overcome existing technical issues and obtain reliable results that are instrumental for the characterization of the types of protein aggregates formed by different proteins of interest in the context of neurodegenerative disorders. PMID:27613047

  12. Rationalization of Sucrose Solution Using During the Fruit Osmotic Dehydration

    Directory of Open Access Journals (Sweden)

    Mirko Babić

    2009-12-01

    Full Text Available The model of sustainable energy production of dried fruit conducted by using combined technology – the model that has been developed at the Faculty of Agriculture in Novi Sad – includes osmotic dehydration of fruit in sucrose solution. During the process of dehydration the moisture content of the solution is increased due to mass transfer of moisture from fruit. This article examines different models of recycling and concentrating of the solution. Thus, the model for concentrating of the solution has been chosen according to this analysis, and it has been applied within its own technology. Evaporators of the low temperature solution have been used and they are based on the solar energy source. Two types of devices have been made on the basis of the heating process of evaporating. One type is filled with the stainless steel shavings, while the other type is based on the fillings by plates. The paper presents the evaluation model of the benefits of this concentrating manner as well as the evaluation criterion of the evaporators’ fillings types. The energy support used here was an original solar air heater of semi-concentrated type.

  13. The three-dimensional structure of TrmB, a transcriptional regulator of dual function in the hyperthermophilic archaeon Pyrococcus furiosus in complex with sucrose.

    Science.gov (United States)

    Krug, Michael; Lee, Sung-Jae; Boos, Winfried; Diederichs, Kay; Welte, Wolfram

    2013-06-01

    TrmB is a repressor that binds maltose, maltotriose, and sucrose, as well as other α-glucosides. It recognizes two different operator sequences controlling the TM (Trehalose/Maltose) and the MD (Maltodextrin) operon encoding the respective ABC transporters and sugar-degrading enzymes. Binding of maltose to TrmB abrogates repression of the TM operon but maintains the repression of the MD operon. On the other hand, binding of sucrose abrogates repression of the MD operon but maintains repression of the TM operon. The three-dimensional structure of TrmB in complex with sucrose was solved and refined to a resolution of 3.0 Å. The structure shows the N-terminal DNA binding domain containing a winged-helix-turn-helix (wHTH) domain followed by an amphipathic helix with a coiled-coil motif. The latter promotes dimerization and places the symmetry mates of the putative recognition helix in the wHTH motif about 30 Å apart suggesting a canonical binding to two successive major grooves of duplex palindromic DNA. This suggests that the structure resembles the conformation of TrmB recognizing the pseudopalindromic TM promoter but not the conformation recognizing the nonpalindromic MD promoter.

  14. Arabidopsis CDS blastp result: AK061395 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061395 006-305-E02 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multip...lication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-125 ...

  15. Arabidopsis CDS blastp result: AK104882 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104882 001-044-H04 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multip...lication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-119 ...

  16. Arabidopsis CDS blastp result: AK066854 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066854 J013075C10 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multipl...ication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-119 ...

  17. Arabidopsis CDS blastp result: AK101318 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101318 J033034D12 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multipl...ication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-125 ...

  18. Arabidopsis CDS blastp result: AK069960 [KOME

    Lifescience Database Archive (English)

    Full Text Available thyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltrans...T1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 5e-60 ...

  19. Arabidopsis CDS blastp result: AK064768 [KOME

    Lifescience Database Archive (English)

    Full Text Available thyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltrans...T1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 1e-112 ...

  20. Arabidopsis CDS blastp result: AK061551 [KOME

    Lifescience Database Archive (English)

    Full Text Available ethyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltran...MT1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 2e-67 ...

  1. Arabidopsis CDS blastp result: AK104764 [KOME

    Lifescience Database Archive (English)

    Full Text Available ethyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltran...MT1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 2e-67 ...

  2. Arabidopsis CDS blastp result: AK098998 [KOME

    Lifescience Database Archive (English)

    Full Text Available thyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltrans...T1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 8e-57 ...

  3. Arabidopsis CDS blastp result: AK061859 [KOME

    Lifescience Database Archive (English)

    Full Text Available ethyltransferase 1 / caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1) identical to O-methyltran...MT1) (Flavonol 3- O-methyltransferase 1) (Caffeic acid/5-hydroxyferulic acid O- methyltransferase) {Arabidopsis thaliana} 1e-100 ...

  4. Arabidopsis CDS blastp result: AK102695 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102695 J033103F21 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  5. Arabidopsis CDS blastp result: AK102134 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102134 J033085F12 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  6. Arabidopsis CDS blastp result: AK066835 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066835 J013087I16 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 1e-171 ...

  7. Arabidopsis CDS blastp result: AK065259 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065259 J013002J18 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  8. Arabidopsis CDS blastp result: AK100523 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100523 J023100P04 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  9. Arabidopsis CDS blastp result: AK242550 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242550 J080319D10 At2g35630.1 68415.m04369 microtubule organization 1 protein (MO...R1) identical to microtubule organization 1 protein GI:14317953 from [Arabidopsis thaliana] 5e-44 ...

  10. Arabidopsis CDS blastp result: AK241043 [KOME

    Lifescience Database Archive (English)

    Full Text Available upted by a stop codon, creating non-consensus donor and acceptor splice sites. 2e-41 ... ...tical to SP|P92997 Germin-like protein subfamily 1 member 13 precursor {Arabidopsis thaliana}; exon 2 interr

  11. Arabidopsis CDS blastp result: AK243135 [KOME

    Lifescience Database Archive (English)

    Full Text Available upted by a stop codon, creating non-consensus donor and acceptor splice sites. 7e-43 ... ...tical to SP|P92997 Germin-like protein subfamily 1 member 13 precursor {Arabidopsis thaliana}; exon 2 interr

  12. The fifth international conference on Arabidopsis research

    Energy Technology Data Exchange (ETDEWEB)

    Hangarter, R.; Scholl, R.; Davis, K.; Feldmann, K.

    1993-12-31

    This volume contains abstracts of oral and poster presentations made in conjunction with the Fifth International Conference on Arabidopsis Research held August 19--22, 1993 at the Ohio State University, Columbus, Ohio.

  13. Arabidopsis CDS blastp result: AK101526 [KOME

    Lifescience Database Archive (English)

    Full Text Available ucosaminyltransferase, putative similar to N-acetylglucosaminyltransferase I from Arabidopsis thaliana [gi:5139335]; contains AT-AC non-consensus splice sites at intron 13 1e-179 ...

  14. Arabidopsis CDS blastp result: AK119708 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK119708 002-157-E08 At1g28330.1 dormancy-associated protein, putative (DRM1) identical to dormancy...-associated protein [Arabidopsis thaliana] GI:2995990; similar to dormancy-associated protei

  15. Arabidopsis CDS blastp result: AK060981 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060981 006-202-H08 At1g28330.1 dormancy-associated protein, putative (DRM1) identical to dormancy...-associated protein [Arabidopsis thaliana] GI:2995990; similar to dormancy-associated protei

  16. Arabidopsis CDS blastp result: AK111576 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK111576 J013075J23 At1g01510.1 C-terminal binding protein (ANGUSTIFOLIA) nearly id...entical to C-terminal binding protein ANGUSTIFOLIA [Arabidopsis thaliana] GI:15408535; contains Pfam profile

  17. Arabidopsis CDS blastp result: AK120838 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120838 J023022B11 At1g01510.1 C-terminal binding protein (ANGUSTIFOLIA) nearly id...entical to C-terminal binding protein ANGUSTIFOLIA [Arabidopsis thaliana] GI:15408535; contains Pfam profile

  18. Arabidopsis CDS blastp result: AK111921 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK111921 001-013-A10 At1g01510.1 C-terminal binding protein (ANGUSTIFOLIA) nearly i...dentical to C-terminal binding protein ANGUSTIFOLIA [Arabidopsis thaliana] GI:15408535; contains Pfam profil

  19. Characterization of xanthophyll pigments, photosynthetic performance, photon energy dissipation, reactive oxygen species generation and carbon isotope discrimination during artemisinin-induced stress in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    M Iftikhar Hussain

    Full Text Available Artemisinin, a potent antimalarial drug, is phytotoxic to many crops and weeds. The effects of artemisinin on stress markers, including fluorescence parameters, photosystem II photochemistry, photon energy dissipation, lipid peroxidation, reactive oxygen species generation and carbon isotope discrimination in Arabidopsis thaliana were studied. Arabidopsis ecotype Columbia (Col-0 seedlings were grown in perlite and watered with 50% Hoagland nutrient solution. Adult plants of Arabidopsis were treated with artemisinin at 0, 40, 80, 160 μM for one week. Artemisinin, in the range 40-160 μM, decreased the fresh biomass, chl a, b and leaf mineral contents. Photosynthetic efficiency, yield and electron transport rate in Arabidopsis were also reduced following exposure to 80 and 160 μM artemisinin. The ΦNPQ and NPQ were less than control. Artemisinin treatment caused an increase in root oxidizability and lipid peroxidation (MDA contents of Arabidopsis. Calcium and nitrogen contents decreased after 80 and 160 μM artemisinin treatment compared to control. δ13C values were less negative following treatment with artemisinin as compared to the control. Artemisinin also decreased leaf protein contents in Arabidopsis. Taken together, these data suggest that artemisinin inhibits many physiological and biochemical processes in Arabidopsis.

  20. Terpene Specialized Metabolism in Arabidopsis thaliana

    OpenAIRE

    Tholl, Dorothea; Lee, Sungbeom

    2011-01-01

    Terpenes constitute the largest class of plant secondary (or specialized) metabolites, which are compounds of ecological function in plant defense or the attraction of beneficial organisms. Using biochemical and genetic approaches, nearly all Arabidopsis thaliana (Arabidopsis) enzymes of the core biosynthetic pathways producing the 5-carbon building blocks of terpenes have been characterized and closer insight has been gained into the transcriptional and posttranscriptional/translational mech...

  1. Nitric oxide induces the alternative oxidase pathway in Arabidopsis seedlings deprived of inorganic phosphate

    OpenAIRE

    Royo, Beatriz; Moran, Jose F.; Ratcliffe, R. George; Gupta, Kapuganti J.

    2015-01-01

    Phosphate starvation compromises electron flow through the cytochrome pathway of the mitochondrial electron transport chain, and plants commonly respond to phosphate deprivation by increasing flow through the alternative oxidase (AOX). To test whether this response is linked to the increase in nitric oxide (NO) production that also increases under phosphate starvation, Arabidopsis thaliana seedlings were grown for 15 d on media containing either 0 or 1mM inorganic phosphate. The effects of th...

  2. Molecular Biology, Biochemistry and Cellular Physiology of Cysteine Metabolism in Arabidopsis thaliana

    OpenAIRE

    Hell, Rüdiger; Wirtz, Markus

    2011-01-01

    Cysteine is one of the most versatile molecules in biology, taking over such different functions as catalysis, structure, regulation and electron transport during evolution. Research on Arabidopsis has contributed decisively to the understanding of cysteine synthesis and its role in the assimilatory pathways of S, N and C in plants. The multimeric cysteine synthase complex is present in the cytosol, plastids and mitochondria and forms the centre of a unique metabolic sensing and signaling sys...

  3. Blocking the Metabolism of Starch Breakdown Products in Arabidopsis Leaves Triggers Chloroplast Degradation

    OpenAIRE

    Stettler, Michaela; Eicke, Simona; Mettler, Tabea; Messerli, Gaëlle; Hörtensteiner, Stefan; Zeeman, Samuel C.

    2009-01-01

    In most plants, a large fraction of photo-assimilated carbon is stored in the chloroplasts during the day as starch and remobilized during the subsequent night to support metabolism. Mutations blocking either starch synthesis or starch breakdown in Arabidopsis thaliana reduce plant growth. Maltose is the major product of starch breakdown exported from the chloroplast at night. The maltose excess 1 mutant (mex1), which lacks the chloroplast envelope maltose transporter, accumulates high levels...

  4. Sucrose in Cyanobacteria: From a Salt-Response Molecule to Play a Key Role in Nitrogen Fixation

    Directory of Open Access Journals (Sweden)

    María A. Kolman

    2015-01-01

    Full Text Available In the biosphere, sucrose is mainly synthesized in oxygenic photosynthetic organisms, such as cyanobacteria, green algae and land plants, as part of the carbon dioxide assimilation pathway. Even though its central position in the functional biology of plants is well documented, much less is known about the role of sucrose in cyanobacteria. In those prokaryotes, sucrose accumulation has been associated with salt acclimation, and considered as a compatible solute in low-salt tolerant strains. In the last years, functional characterizations of sucrose metabolizing enzymes, metabolic control analysis, cellular localization of gene expressions, and reverse genetic experiments have revealed that sucrose metabolism is crucial in the diazotrophic growth of heterocystic strains, and besides, that it can be connected to glycogen synthesis. This article briefly summarizes the current state of knowledge of sucrose physiological functions in modern cyanobacteria and how they might have evolved taking into account the phylogenetic analyses of sucrose enzymes.

  5. Synthesis of oligo- and polysaccharides by Lactobacillus reuteri 121 reuteransucrase at high concentrations of sucrose.

    Science.gov (United States)

    Meng, Xiangfeng; Dobruchowska, Justyna M; Gerwig, Gerrit J; Kamerling, Johannis P; Dijkhuizen, Lubbert

    2015-09-23

    GTFA, a glucansucrase enzyme of the probiotic bacterium Lactobacillus reuteri 121, is capable of synthesizing an α-glucan polysaccharide with (1 → 4) and (1 → 6) linkages from sucrose. With respect to its biosynthesis, the present study has shown that the ratio of oligosaccharide versus polysaccharide synthesized was directly proportional to the concentration of sucrose. It appears that the size distribution of products is kinetically controlled, but the linkage distribution in the polysaccharide material is not changed. At high sucrose concentrations the sucrose isomers leucrose and trehalulose were synthesized, using the accumulated fructose as acceptor, together with 4'- and 6'-α-D-glucosyl-leucrose and 6'-α-D-glucosyl-trehalulose. The finding of an additional branched hexasaccharide demonstrates that the enzyme is able to introduce branch-points already in relatively short oligosaccharides.

  6. Large deformation properties of short doughs: Effect of sucrose in relation to mixing time

    NARCIS (Netherlands)

    Baltsavias, A.; Jurgens, A.; Vliet, T. van

    1999-01-01

    Large deformation rheological properties of short doughs of various composition prepared under various mixing times were determined in uniaxial compression. Sucrose-syrup doughs exhibited prominent yielding and flow behaviour. Their apparent biaxial extensional viscosity decreased with increasing su

  7. Microplate assay for rapid determination of sucrose, glucose, fructose and raffinose

    Science.gov (United States)

    Current methods for the quantification of carbohydrates in sugarbeet roots have limitations. Polarimetry and refractometry measure only sucrose content and are inaccurate with deteriorated roots. High performance liquid chromatography (HPLC) and gas chromatography (GC) quantify all simple carbohy...

  8. What´s cheapest, intravenous iron sucrose- or intravenous iron carboxymaltose treatment in IBD patients?

    DEFF Research Database (Denmark)

    Bager, Palle; Dahlerup, Jens Frederik

      What´s cheapest, intravenous iron sucrose- or intravenous iron carboxymaltose treatment in IBD patients? It dependent on the economic evaluation perspective!   Aim: To evaluate the health care cost for intravenous iron sucrose (Venofer®, Vifor) and intravenous iron carboxymaltose (Ferinject......®, Vifor) treatment to IBD patients in an outpatient setting.   Background: Intravenous iron sucrose can be given as a maximum of 200 mg Fe++ per infusion vs. intravenous iron carboxymaltose that can be given as a maximum of 1000 mg Fe++ in a single infusion leading to fewer infusions and visits. The drug......-cost per mg iron is for iron carboxymaltose approximately double the cost of iron sucrose.   Patients and Methods: Data related to 111 IBD-patients treated with intravenous iron at Aarhus University Hospital from August 2005 until October 2009 was used for the economic evaluation. Analysis included...

  9. Starch and sucrose synthesis in Phaseolus vulgaris as affected by light, CO2, and abscisic acid

    International Nuclear Information System (INIS)

    Phaseolus vulgaris L. leaves were subjected to various light, CO2, and O2 levels and abscisic acid, then given a 10 minute pulse of 14CO2 followed by a 5 minute chase with unlabeled CO2. After the chase period, very little label remained in the ionic fractions except at low CO2 partial pressure. Most label was found in the neutral, alcohol soluble fraction or in the insoluble fraction digestable by amyloglucosidase. Sucrose formation was linearly related to assimilation rate. Starch formation increased linearly with assimilation rate, but did not occur if the assimilation rate was below 4 micromoles per square meter per second. Neither abscisic acid, nor high CO2 in combination with low O2 caused significant perturbations of the sucrose/starch formation ratio. These studies indicate that the pathways for starch and sucrose synthesis both are controlled by the rate of net CO2 assimilation, with sucrose the preferred product at very low assimilation rates

  10. Effects of Replacing Sucrose with Various Sugar Alcohols on Quality Properties of Semi-dried Jerky.

    Science.gov (United States)

    Jang, Sung-Jin; Kim, Hyun-Wook; Hwang, Ko-Eun; Song, Dong-Heon; Kim, Yong-Jae; Ham, Youn-Kyung; Lim, Yun-Bin; Jeong, Tae-Jun; Kim, Si-Young; Kim, Cheon-Jei

    2015-01-01

    The objective of this study was to investigate the effects of replacing sucrose with sugar alcohols (sorbitol, glycerol and xylitol) on the quality properties of semi-dried jerky. Total 7 treatments of jerkies were prepared as follows: control with sucrose, and treatments with 2.5 and 5.0% of sucrose replaced by each sugar alcohol, respectively. Drying yield, pH, water activity, moisture content, shear force, myofibrillar fragmentation index (MFI), 2-thiobarbituric acid reactive substance (TBARS) value, sugar content, and sensory evaluation were evaluated. Xylitol slightly decreased the pH when compared to the other sugar alcohols (p>0.05). The water activity of the semi-dried jerky was significantly reduced by treatment with glycerol and xylitol (psorbitol and glycerol treatments, but that replacing sucrose with 5.0% xylitol demonstrated the lowest shear force (psorbitol and glycerol were lower than the control and xylitol treatment (pmanufacturing meat products.

  11. Diverse Exopolysaccharide Producing Bacteria Isolated from Milled Sugarcane: Implications for Cane Spoilage and Sucrose Yield.

    Directory of Open Access Journals (Sweden)

    Stanton Hector

    Full Text Available Bacterial deterioration of sugarcane during harvesting and processing is correlated with significant loss of sucrose yield and the accumulation of bacterial polysaccharides. Dextran, a homoglucan produced by Leuconostoc mesenteroides, has been cited as the primary polysaccharide associated with sugarcane deterioration. A culture-based approach was used to isolate extracellular polysaccharide (EPS producing bacterial strains from milled sugarcane stalks. Ribosomal RNA sequencing analysis grouped 25 isolates into 4 genera. This study identified 2 bacterial genera not previously associated with EPS production or sucrose degradation. All isolates produced polysaccharide when grown in the presence of sucrose. Monosaccharide analysis of purified polymers by Gas Chromatography revealed 17 EPSs consisting solely of glucose (homoglucans, while the remainder contained traces of mannose or fructose. Dextranase treatment of polysaccharides yielded full digestion profiles for only 11 extracts. Incomplete hydrolysis profiles of the remaining polysaccharides suggest the release of longer oligosaccharides which may interfere with sucrose crystal formation.

  12. ESR investigation of L-α-alanine and sucrose radicals produced by heavy-ion irradiation

    International Nuclear Information System (INIS)

    We investigated sucrose and L-α-alanine radicals produced by heavy ion irradiation with various LETs (linear energy transfer). The impact of the heavy ions on the samples produced stable free radicals, which were analyzed by ESR (electron spin resonance). The obtained spectral patterns were the same as those for carbon (C), neon (Ne), and silicon (Si) ion irradiations. The absorbed dose dependences for the irradiated sucrose and alanine samples were examined. The ESR response has a linear relation with the absorbed dose for both compounds. In addition, the total spin concentration obtained by heavy-ion irradiation correlated logarithmically with the LET. Let Qualitative ESR analyses showed that the production of sucrose and alanine radical depended on both different particle irradiation and the LET under the same dose. Thus, the present ESR results imply that sucrose together with L-α-alanine can be used to investigate radical production processes induced by ionizing particles. (author)

  13. The interaction of temperature and sucrose concentration on foraging preferences in bumblebees.

    Science.gov (United States)

    Whitney, Heather M; Dyer, Adrian; Chittka, Lars; Rands, Sean A; Glover, Beverley J

    2008-09-01

    Several authors have found that flowers that are warmer than their surrounding environment have an advantage in attracting pollinators. Bumblebees will forage preferentially on warmer flowers, even if equal nutritional reward is available in cooler flowers. This raises the question of whether warmth and sucrose concentration are processed independently by bees, or whether sweetness detectors respond to higher sugar concentration as well as higher temperature. We find that bumblebees can use lower temperature as a cue to higher sucrose reward, showing that bees appear to process the two parameters strictly independently. Moreover, we demonstrate that sucrose concentration takes precedence over warmth, so that when there is a difference in sucrose concentration, bees will typically choose the sweeter feeder, even if the less sweet feeder is several degrees warmer.

  14. Heat capacity and transition behavior of sucrose by standard, fast scanning and temperature-modulated calorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Magoń, A. [Department of Chemistry, University of Technology, 35-959 Rzeszów (Poland); Wurm, A.; Schick, C. [Department of Physics, University of Rostock, 18057 Rostock (Germany); Pangloli, Ph.; Zivanovic, S. [Department of Food Science and Technology, University of Tennessee, Knoxville, TN 37996 (United States); Skotnicki, M. [Department of Pharmaceutical Technology, Poznan University of Medical Sciences, 60-780 Poznań (Poland); Pyda, M., E-mail: mpyda@utk.edu [Department of Chemistry, University of Technology, 35-959 Rzeszów (Poland)

    2014-08-10

    Highlights: • Experimental, apparent heat capacity of sucrose was investigated by advanced thermal analysis. • Vibrational heat capacity of solid state was linked with a low temperature experimental heat capacity of sucrose. • Equilibrium melting parameters of sucrose were determined. • Decomposition, superheating of crystalline sucrose during melting process were presented. • TGA, DSC, TMDSC, and FSC are useful tools for characterization of sucrose. - Abstract: The heat capacity (C{sub p}) of crystalline and amorphous sucrose was determined using standard and quasi-isothermal temperature modulated differential scanning calorimetry. The results were combined with the published data determined by adiabatic calorimetry, and the C{sub p} values are now reported for the wide 5–600 K range. The experimental C{sub p} of solid sucrose at 5–300 K was used to calculate the vibrational, solid C{sub p} based on the vibrational molecular motions. The calculated solid and liquid C{sub p} together with the transition parameters for equilibrium conditions were used as references for detailed quantitative thermal analysis of crystalline and amorphous sucrose. Melting temperature (T{sub m}) of the crystalline sucrose was identified in a broad 442–465 K range with a heat of fusion of 40–46 J/mol determined at heating rates 0.5–20 K/min, respectively. The equilibrium T{sub m} and heat of fusion of crystalline sucrose were estimated at zero heating rate as T{sup o}{sub m} = 424.4 K and ΔH{sup o}{sub f} = 32 kJ/mol, respectively. The glass transition temperature (T{sub g}) of amorphous sucrose was at 331 K with a change in C{sub p} of 267 J/(mol K) as it was estimated from reversing heat capacity by quasi-isothermal TMDSC on cooling. At heating rates less than 30 K/min, thermal decomposition occurred during melting, while at extreme rate of 1000 K/s, degradation was not observed. Data obtained by fast scanning calorimetry (FSC) at 1000 K/s, showed that T{sub m} was

  15. Heat capacity and transition behavior of sucrose by standard, fast scanning and temperature-modulated calorimetry

    International Nuclear Information System (INIS)

    Highlights: • Experimental, apparent heat capacity of sucrose was investigated by advanced thermal analysis. • Vibrational heat capacity of solid state was linked with a low temperature experimental heat capacity of sucrose. • Equilibrium melting parameters of sucrose were determined. • Decomposition, superheating of crystalline sucrose during melting process were presented. • TGA, DSC, TMDSC, and FSC are useful tools for characterization of sucrose. - Abstract: The heat capacity (Cp) of crystalline and amorphous sucrose was determined using standard and quasi-isothermal temperature modulated differential scanning calorimetry. The results were combined with the published data determined by adiabatic calorimetry, and the Cp values are now reported for the wide 5–600 K range. The experimental Cp of solid sucrose at 5–300 K was used to calculate the vibrational, solid Cp based on the vibrational molecular motions. The calculated solid and liquid Cp together with the transition parameters for equilibrium conditions were used as references for detailed quantitative thermal analysis of crystalline and amorphous sucrose. Melting temperature (Tm) of the crystalline sucrose was identified in a broad 442–465 K range with a heat of fusion of 40–46 J/mol determined at heating rates 0.5–20 K/min, respectively. The equilibrium Tm and heat of fusion of crystalline sucrose were estimated at zero heating rate as Tom = 424.4 K and ΔHof = 32 kJ/mol, respectively. The glass transition temperature (Tg) of amorphous sucrose was at 331 K with a change in Cp of 267 J/(mol K) as it was estimated from reversing heat capacity by quasi-isothermal TMDSC on cooling. At heating rates less than 30 K/min, thermal decomposition occurred during melting, while at extreme rate of 1000 K/s, degradation was not observed. Data obtained by fast scanning calorimetry (FSC) at 1000 K/s, showed that Tm was 483 K and Tg was 364 K. Superheating effects were observed during the melting with

  16. Role of metabolite transporters in source-sink carbon allocation.

    Science.gov (United States)

    Ludewig, Frank; Flügge, Ulf-Ingo

    2013-01-01

    Plants assimilate carbon dioxide during photosynthesis in chloroplasts. Assimilated carbon is subsequently allocated throughout the plant. Generally, two types of organs can be distinguished, mature green source leaves as net photoassimilate exporters, and net importers, the sinks, e.g., roots, flowers, small leaves, and storage organs like tubers. Within these organs, different tissue types developed according to their respective function, and cells of either tissue type are highly compartmentalized. Photoassimilates are allocated to distinct compartments of these tissues in all organs, requiring a set of metabolite transporters mediating this intercompartmental transfer. The general route of photoassimilates can be briefly described as follows. Upon fixation of carbon dioxide in chloroplasts of mesophyll cells, triose phosphates either enter the cytosol for mainly sucrose formation or remain in the stroma to form transiently stored starch which is degraded during the night and enters the cytosol as maltose or glucose to be further metabolized to sucrose. In both cases, sucrose enters the phloem for long distance transport or is transiently stored in the vacuole, or can be degraded to hexoses which also can be stored in the vacuole. In the majority of plant species, sucrose is actively loaded into the phloem via the apoplast. Following long distance transport, it is released into sink organs, where it enters cells as source of carbon and energy. In storage organs, sucrose can be stored, or carbon derived from sucrose can be stored as starch in plastids, or as oil in oil bodies, or - in combination with nitrogen - as protein in protein storage vacuoles and protein bodies. Here, we focus on transport proteins known for either of these steps, and discuss the implications for yield increase in plants upon genetic engineering of respective transporters. PMID:23847636

  17. Role of metabolite transporters in source-sink carbon allocation

    Directory of Open Access Journals (Sweden)

    Frank eLudewig

    2013-07-01

    Full Text Available Plants assimilate carbon dioxide during photosynthesis in chloroplasts. Assimilated carbon is subsequently allocated throughout the plant. Generally, two types of organs can be distinguished, mature green source leaves as net photoassimilate exporters, and net importers, the sinks, e.g. roots, flowers, small leaves and storage organs like tubers. Within these organs, different tissue types developed according to their respective function, and cells of either tissue type are highly compartmentalized. Photoassimilates are allocated to distinct compartments of these tissues in all organs, requiring a set of metabolite transporters mediating this intercompartmental transfer.The general route of photoassimilates can be briefly described as follows. Upon fixation of carbon dioxide in chloroplasts of mesophyll cells, triose phosphates either enter the cytosol for mainly sucrose formation or remain in the stroma to form transiently stored starch which is degraded during the night and enters the cytosol as maltose or glucose to be further metabolized to sucrose. In both cases, sucrose enters the phloem for long distance transport or is transiently stored in the vacuole, or can be degraded to hexoses which also can be stored in the vacuole.In the majority of plant species, sucrose is actively loaded into the phloem via the apoplast. Following long distance transport, it is released into sink organs, where it enters cells as source of carbon and energy. In storage organs, sucrose can be stored, or carbon derived from sucrose can be stored as starch in plastids, or as oil in oil bodies, or - in combination with nitrogen - as protein in protein storage vacuoles and protein bodies.Here, we focus on transport proteins known for either of these steps, and discuss the implications for yield increase in plants upon genetic engineering of respective transporters.

  18. The effects of fruiting positions on cellulose synthesis and sucrose metabolism during cotton (Gossypium hirsutum L. fiber development.

    Directory of Open Access Journals (Sweden)

    Yina Ma

    Full Text Available Cotton (Gossypium hirsutum L. boll positions on a fruiting branch vary in their contribution to yield and fiber quality. Fiber properties are dependent on deposition of cellulose in the fiber cell wall, but information about the enzymatic differences in sucrose metabolism between these fruiting positions is lacking. Therefore, two cotton cultivars with different sensitivities to low temperature were tested in 2010 and 2011 to quantify the effect of fruit positions (FPs on fiber quality in relation to sucrose content, enzymatic activities and sucrose metabolism. The indices including sucrose content, sucrose transformation rate, cellulose content, and the activities of the key enzymes, sucrose phosphate synthase (SPS, acid invertase (AI and sucrose synthase (SuSy which inhibit cellulose synthesis and eventually affect fiber quality traits in cotton fiber, were determined. Results showed that as compared with those of FP1, cellulose content, sucrose content, and sucrose transformation rate of FP3 were all decreased, and the variations of cellulose content and sucrose transformation rate caused by FPs in Sumian 15 were larger than those in Kemian 1. Under FP effect, activities of SPS and AI in sucrose regulation were decreased, while SuSy activity in sucrose degradation was increased. The changes in activities of SuSy and SPS in response to FP effect displayed different and large change ranges between the two cultivars. These results indicate that restrained cellulose synthesis and sucrose metabolism in distal FPs are mainly attributed to the changes in the activities of these enzymes. The difference in fiber quality, cellulose synthesis and sucrose metabolism in response to FPs in fiber cells for the two cotton cultivars was mainly determined by the activities of both SuSy and SPS.

  19. Sucrose radical-production cross section regarding heavy-ion irradiation

    OpenAIRE

    Nakagawa, Kouichi; Ikota, Nobuo; Anzai, Kazunori

    2008-01-01

    We investigated the sucrose radical-production cross section induced by heavy-ion irradiation. L-alanine was also used in order to compare radical yield and cross section. The stable free radicals after irradiation were analyzed by EPR (electron paramagnetic resonance). The radical yield obtained by the irradiated samples had a logarithmic correlation with the LET (linear energy transfer). Quantitative EPR analyses showed that radical productions for sucrose and L-alanine vary both by differe...

  20. Dosakaya Juice Assuages Development of Sucrose Induced Impaired Glucose Tolerance and Imbalance in Antioxidant Defense

    OpenAIRE

    Kumar, Dommati Anand; Pisupati S.R Sweeya; Shukla, Srishti; Anusha, Sanga Venkata; Akshara, Dasari; Madhusudana, Kuncha; Tiwari, Ashok Kumar

    2015-01-01

    Objective: The objective was to explore the effect of Dosakaya (DK) (Cucumis melo var. chito) juice on sucrose induced dysglycemia and disturbances in antioxidant defense in rats. Materials and Methods: Rats were preconditioned with DK juice before administration of sucrose beverage continuously for 1-month. Blood glucose tolerance test and glutathione (GSH) homeostasis pathways in kidney were analyzed in different group of animals at the end of the study. Results: DK juice diffused (P < 0.00...

  1. Role of plaque in the clearance of salivary sucrose and its influence on salivary ph

    OpenAIRE

    Kumar, A; R Hedge; U Dixit

    2011-01-01

    Background: The prevalence of dental caries in children, in India, is higher than many of the industrialized countries. The sugar most commonly associated with dental caries is sucrose, as the microorganisms in the dental plaque have the ability to convert this dietary constituent into various organic acids. Aims and Objectives: This study was conducted to study the effect of the presence of plaque on the salivary clearance of sucrose and to study the effect of the presence of plaque on saliv...

  2. Optimal Control for Industrial Sucrose Crystallization with Action Dependent Heuristic Dynamic Programming

    OpenAIRE

    Xiaofeng Lin; Heng Zhang; Li Wei; Huixia Liu

    2009-01-01

    This paper applies a neural-network-based approximate dynamic programming (ADP) method, namely, the action dependent heuristic dynamic programming (ADHDP), to an industrial sucrose crystallization optimal control problem. The industrial sucrose crystallization is a nonlinear and slow time-varying process. It is quite difficult to establish a precise mechanism model of the crystallization, because of complex internal mechanism and interacting variables. We developed a neural network model of t...

  3. Experimental Study of Sorbitol Production by Zymomonas mobilis in High Sucrose Concentration

    OpenAIRE

    Rui Sérgio dos Santos Ferreira da Silva; João Batista Buzato; Maria Antonia Pedrine Colabone Celligoi; Marcos Roberto de Oliveira

    2004-01-01

    The sorbitol presents several industrial applications and its conventional production is of high cost and low yield. Sorbitol production by Zymomonas mobilis production has attracted attention as both production cost and environmental impact are low. The sorbitol plays an osmo-protective rule so that its production is promoted by high sugar concentrations. This work has evaluated the effect of high sucrose concentration in the sorbitol production. The raise of sucrose concentration from 100 t...

  4. Electrical stimulation of the lateral habenula produces an inhibitory effect on sucrose self-administration

    OpenAIRE

    Friedman, Alexander; Lax, Elad; Dikshtein, Yahav; Abraham, Lital; Flaumenhaft, Yakov; Sudai, Einav; Ben-Tzion, Moshe; Yadid, Gal

    2010-01-01

    The lateral habenula (LHb) plays a role in prediction of negative reinforcement, punishment and aversive responses. In the current study, we examined the role that the LHb plays in the regulation of negative reward responses and aversion. First, we tested the effect of intervention in LHb activity on sucrose reinforcing behavior. An electrode was implanted into the LHb and rats were trained to self-administer sucrose (20%; 16 days) until at least three days of stable performance were achieved...

  5. Comparison and validation of two analytical methods for measurement of urinary sucrose and fructose excretion

    OpenAIRE

    Song, Xiaoling; Navarro, Sandi L.; Diep, Pho; Thomas, Wendy K.; Razmpoosh, Elena C.; Schwarz, Yvonne; Wang, Ching-Yun; Kratz, Mario; Neuhouser, Marian L; Lampe, Johanna W.

    2013-01-01

    Urinary sugars excretion has been proposed as a potential biomarker for intake of sugars. In this study we compared two analytical methods [gas chromatography (GC) and enzymatic reactions – UV absorption] for quantifying urinary fructose and sucrose using 24-hour urine samples from a randomized cross-over controlled feeding study. All samples were successfully quantified by the GC method; however 21% and 1.9% of samples were below the detection limit of the enzymatic method for sucrose and fr...

  6. Sucrose exposure in early life alters adult motivation and weight gain.

    Directory of Open Access Journals (Sweden)

    Cristianne R M Frazier

    Full Text Available The cause of the current increase in obesity in westernized nations is poorly understood but is frequently attributed to a 'thrifty genotype,' an evolutionary predisposition to store calories in times of plenty to protect against future scarcity. In modern, industrialized environments that provide a ready, uninterrupted supply of energy-rich foods at low cost, this genetic predisposition is hypothesized to lead to obesity. Children are also exposed to this 'obesogenic' environment; however, whether such early dietary experience has developmental effects and contributes to adult vulnerability to obesity is unknown. Using mice, we tested the hypothesis that dietary experience during childhood and adolescence affects adult obesity risk. We gave mice unlimited or no access to sucrose for a short period post-weaning and measured sucrose-seeking, food consumption, and weight gain in adulthood. Unlimited access to sucrose early in life reduced sucrose-seeking when work was required to obtain it. When high-sugar/high-fat dietary options were made freely-available, however, the sucrose-exposed mice gained more weight than mice without early sucrose exposure. These results suggest that early, unlimited exposure to sucrose reduces motivation to acquire sucrose but promotes weight gain in adulthood when the cost of acquiring palatable, energy dense foods is low. This study demonstrates that early post-weaning experience can modify the expression of a 'thrifty genotype' and alter an adult animal's response to its environment, a finding consistent with evidence of pre- and peri-natal programming of adult obesity risk by maternal nutritional status. Our findings suggest the window for developmental effects of diet may extend into childhood, an observation with potentially important implications for both research and public policy in addressing the rising incidence of obesity.

  7. Interaction of Metabolic Stress with Chronic Mild Stress in Altering Brain Cytokines and Sucrose Preference

    OpenAIRE

    Remus, Jennifer L.; Stewart, Luke T.; Camp, Robert M.; Novak, Colleen M.; Johnson, John D.

    2015-01-01

    There is growing evidence that metabolic stressors increase an organism’s risk of depression. Chronic mild stress is a popular animal model of depression and several serendipitous findings have suggested that food deprivation prior to sucrose testing in this model is necessary to observe anhedonic behaviors. Here, we directly tested this hypothesis by exposing animals to chronic mild stress and used an overnight two bottle sucrose test (food ad libitum) on day 5 and 10, then food and water de...

  8. Tethering Complexes in the Arabidopsis Endomembrane System.

    Science.gov (United States)

    Vukašinović, Nemanja; Žárský, Viktor

    2016-01-01

    Targeting of endomembrane transport containers is of the utmost importance for proper land plant growth and development. Given the immobility of plant cells, localized membrane vesicle secretion and recycling are amongst the main processes guiding proper cell, tissue and whole plant morphogenesis. Cell wall biogenesis and modification are dependent on vectorial membrane traffic, not only during normal development, but also in stress responses and in plant defense against pathogens and/or symbiosis. It is surprising how little we know about these processes in plants, from small GTPase regulation to the tethering complexes that act as their effectors. Tethering factors are single proteins or protein complexes mediating first contact between the target membrane and arriving membrane vesicles. In this review we focus on the tethering complexes of the best-studied plant model-Arabidopsis thaliana. Genome-based predictions indicate the presence of all major tethering complexes in plants that are known from a hypothetical last eukaryotic common ancestor (LECA). The evolutionary multiplication of paralogs of plant tethering complex subunits has produced the massively expanded EXO70 family, indicating a subfunctionalization of the terminal exocytosis machinery in land plants. Interpretation of loss of function (LOF) mutant phenotypes has to consider that related, yet clearly functionally-specific complexes often share some common core subunits. It is therefore impossible to conclude with clarity which version of the complex is responsible for the phenotypic deviations observed. Experimental interest in the analysis of plant tethering complexes is growing and we hope to contribute with this review by attracting even more attention to this fascinating field of plant cell biology. PMID:27243010

  9. Tethering complexes in the Arabidopsis endomembrane system

    Directory of Open Access Journals (Sweden)

    Nemanja eVukasinovic

    2016-05-01

    Full Text Available AbstractTargeting of endomembrane transport containers is of the utmost importance for proper land plant growth and development. Given the immobility of plant cells, localized membrane vesicle secretion and recycling are amongst the main processes guiding proper cell, tissue and whole plant morphogenesis. Cell wall biogenesis and modification are dependent on vectorial membrane traffic, not only during normal development, but also in stress responses and in plant defence against pathogens and/or symbiosis. It is surprising how little we know about these processes in plants, from small GTPase regulation to the tethering complexes that act as their effectors. Tethering factors are single proteins or protein complexes mediating first contact between the target membrane and arriving membrane vesicles. In this review we focus on the tethering complexes of the best-studied plant model – Arabidopsis thaliana. Genome-based predictions indicate the presence of all major tethering complexes in plants that are known from a hypothetical last eukaryotic common ancestor (LECA. The evolutionary multiplication of paralogs of plant tethering complex subunits has produced the massively expanded EXO70 family, indicating a subfunctionalization of the terminal exocytosis machinery in land plants. Interpretation of loss of function (LOF mutant phenotypes has to consider that related, yet clearly functionally-specific complexes often share some common core subunits. It is therefore impossible to conclude with clarity which version of the complex is responsible for the phenotypic deviations observed. Experimental interest in the analysis of plant tethering complexes is growing and we hope to contribute with this review by attracting even more attention to this fascinating field of plant cell biology.

  10. Amyloplast movement and gravityperception in Arabidopsis endoderm

    Science.gov (United States)

    Tasaka, M.; Saito, T.; Morita, M. T.

    Gravitropism of higher plant is a growth response regulating the orientation of organs elongation, which includes four sequential steps, the perception of gravistimulus, transduction of the physical stimulus to chemical signal, transmission of the signal, and differential cell elongation depending on the signal. To elucidate the molecular mechanism of these steps, we have isolated a number of Arabidopsis mutants with abnormal shoot gravitropic response. zig (zigzag)/sgr4(shoot gravitropism 4) shows little gravitropism in their shoots. Besides, their inflorescence stems elongate in a zigzag-fashion to bend at each node. ZIG encodes a SNARE, AtVTI11. sgr3 with reduced gravitropic response in inflorescence stems had a missense mutation in other SNARE, AtVAM3. These two SNAREs make a complex in the shoot endoderm cells that are gravity-sensing cells, suggesting that the vesicle transport from trans-Golgi network (TGN) to prevacuolar compartment (PVC) and/or vacuole is involved in gravitropism. Abnormal vesicular/vacuolar structures were observed in several tissues of both mutants. Moreover, SGR2 encodes phospholipase A1-like protein that resides in the vacuolar membrane. Endodermis-specific expression of these genes could complement gravitropism in each mutant. In addition, amyloplasts thought to be statoliths localized abnormally in their endoderm cells. These results strongly suggest that formation and function of vacuole in the endoderm cells are important for amyloplasts sedimentation, which is involved in the early process of shoot gravitropism. To reveal this, we constructed vertical stage microscope system to visualize the behavior of amyloplasts and vacuolar membrane in living endodermal cells. We hope to discuss the mechanism of gravity perception after showing their movements.

  11. Mutations in a new Arabidopsis cyclophilin disrupt its interaction with protein phosphatase 2A

    Science.gov (United States)

    Jackson, K.; Soll, D.; Evans, M. L. (Principal Investigator)

    1999-01-01

    The heterotrimeric protein phosphatase 2A (PP2A) is a component of multiple signaling pathways in eukaryotes. Disruption of PP2A activity in Arabidopsis is known to alter auxin transport and growth response pathways. We demonstrated that the regulatory subunit A of an Arabidopsis PP2A interacts with a novel cyclophilin, ROC7. The gene for this cyclophilin encodes a protein that contains a unique 30-amino acid extension at the N-terminus, which distinguishes the gene product from all previously identified Arabidopsis cyclophilins. Altered forms of ROC7 cyclophilin with mutations in the conserved DENFKL domain did not bind to PP2A. Unlike protein phosphatase 2B, PP2A activity in Arabidopsis extracts was not affected by the presence of the cyclophilin-binding molecule cyclosporin. The ROC7 transcript was expressed to high levels in all tissues tested. Expression of an ROC7 antisense transcript gave rise to increased root growth. These results indicate that cyclophilin may have a role in regulating PP2A activity, by a mechanism that differs from that employed for cyclophilin regulation of PP2B.

  12. Bimodal Porous Scaffolds by Sequential Electrospinning of Poly(glycolic acid with Sucrose Particles

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    B. Wulkersdorfer

    2010-01-01

    Full Text Available Electrospinning is a method to produce fine, biopolymer mesh with a three-dimensional architecture that mimics native extra-cellular matrix. Due to the small fiber diameter created in this process, conventional electrospun scaffolds have pore sizes smaller than the diameter of most cells. These scaffolds have limited application in tissue engineering due to poor cell penetration. We developed a hybrid electrospinning/particulate leaching technique to create scaffolds with increased porosity and improved cellular ingrowth. Poly(glycolic acid (PGA and a sucrose-ethanol suspension were electrospun in equal, alternating sequences at intervals of one, two, and ten minutes each. The scaffolds revealed fiber mesh with micropores of 10 m and uniformly distributed sucrose particles. Particulate leaching of sucrose from the one- or two-minute scaffolds revealed honeycomb structures with interconnected macropores between 50 and 250 m. Sucrose leaching from the ten-minute scaffolds resulted in laminated structures with isolated macropores between 200 and 350 m. Macropore size was directly proportional to the duration of the sucrose spinning interval. After 24 hours of cell culture, conventionally spun scaffolds demonstrated no cellular penetration. Conversely, the PGA/sucrose scaffolds demonstrated deep cellular penetration. This hybrid technique represents a novel method of generating electrospun scaffolds with interconnected pores suitable for cellular ingrowth.

  13. Chronic pramipexole treatment increases tolerance for sucrose in normal and ventral tegmental lesioned rats.

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    David eDARDOU

    2015-01-01

    Full Text Available The loss of dopamine neurons observed in Parkinson’s disease (PD elicits severe motor control deficits which are reduced by the use of dopamine agonists. However, recent works have indicated that D3-preferential agonists such as pramipexole can induce impulse control disorders such as food craving or compulsive eating. In the present study, we performed an intermittent daily feeding experiment to assess the effect of chronic treatment by pramipexole and VTA bilateral lesion on tolerance for sucrose solution. The impact of such chronic treatment on spontaneous locomotion and spatial memory was also examined. Changes in sucrose tolerance could indicate the potential development of a change in food compulsion or addiction related to the action of pramipexole. Neither the bilateral lesion of the VTA nor chronic treatment with pramipexole altered the spontaneous locomotion or spatial memory in rats. Rats without pramipexole treatment quickly developed a stable intake of sucrose solution in the 12h access phase. On the contrary, when under daily pramipexole treatment, rats developed a stronger and ongoing escalation of their sucrose solution intakes. In addition, we noted that the change in sucrose consumption was sustained by an increase of the expression of the dopamine 3 receptor in the core and the shell regions of the nucleus accumbens. The present results may suggest that long term stimulation of the dopamine 3 receptor in animals induces a strong increase in sucrose consumption, indicating an effect of this receptor on certain pathological aspects of food eating.

  14. Chronic pramipexole treatment increases tolerance for sucrose in normal and ventral tegmental lesioned rats.

    Science.gov (United States)

    Dardou, David; Chassain, Carine; Durif, Franck

    2014-01-01

    The loss of dopamine neurons observed in Parkinson's disease (PD) elicits severe motor control deficits which are reduced by the use of dopamine agonists. However, recent works have indicated that D3-preferential agonists such as pramipexole can induce impulse control disorders (ICDs) such as food craving or compulsive eating. In the present study, we performed an intermittent daily feeding experiment to assess the effect of chronic treatment by pramipexole and VTA bilateral lesion on tolerance for sucrose solution. The impact of such chronic treatment on spontaneous locomotion and spatial memory was also examined. Changes in sucrose tolerance could indicate the potential development of a change in food compulsion or addiction related to the action of pramipexole. Neither the bilateral lesion of the VTA nor chronic treatment with pramipexole altered the spontaneous locomotion or spatial memory in rats. Rats without pramipexole treatment quickly developed a stable intake of sucrose solution in the 12 h access phase. On the contrary, when under daily pramipexole treatment, rats developed a stronger and ongoing escalation of their sucrose solution intakes. In addition, we noted that the change in sucrose consumption was sustained by an increase of the expression of the Dopamine D3 receptor in the core and the shell regions of the nucleus accumbens. The present results may suggest that long-term stimulation of the Dopamine D3 receptor in animals induces a strong increase in sucrose consumption, indicating an effect of this receptor on certain pathological aspects of food eating. PMID:25610366

  15. A study on alfalfa-dodder relationships by 14C-sucrose incorporation in their tissues

    International Nuclear Information System (INIS)

    Plant-parasite relationships between lesser-dodder (Cuscuta epithymum) and alfalfa (Medicago sativa) are investigated. 14C-sucrose was incorporated via the cut stems of host and parasite into their tissues and the content of the free sugars, keto acids and free amino acids are determined. 14C-sucrose was introduced also in lesser-dodder previously taken from the host-plant and immersed directly into radioactive sucrose. It is shown that the parasite intensively absorbs sucrose from the host-plant tissues using it as a source of energy and as a substrat for different synthetic processes. In the case when the parasite was previously taken away from the host-plant 14C-sucrose taken up directly from the solution was used for the synthesis of various compounds (carbohydrates, amino- and keto acids). The suggestion was made that the respective enzyme systems are present in dodder tissues. The 14C-glucose and 14C-fructose content is an evidence for high invertase hydrolytic activity. The presence of 14C-keto acids shows that the 14C-sucrose incorporated in lesser-dodder tissues without the mediation of the host-plant was used as a respiratory substrate by the degradation following Krebs cycle. (author)

  16. Degradation of hydroxycinnamic acid mixtures in aqueous sucrose solutions by the Fenton process.

    Science.gov (United States)

    Nguyen, Danny M T; Zhang, Zhanying; Doherty, William O S

    2015-02-11

    The degradation efficiencies and behaviors of caffeic acid (CaA), p-coumaric acid (pCoA), and ferulic acid (FeA) in aqueous sucrose solutions containing the mixture of these hydroxycinnamic acids (HCAs) were studied by the Fenton oxidation process. Central composite design and multiresponse surface methodology were used to evaluate and optimize the interactive effects of process parameters. Four quadratic polynomial models were developed for the degradation of each individual acid in the mixture and the total HCAs degraded. Sucrose was the most influential parameter that significantly affected the total amount of HCA degraded. Under the conditions studied there was a sucrose in all reactions. The optimal values of the process parameters for a 200 mg/L HCA mixture in water (pH 4.73, 25.15 °C) and sucrose solution (13 mass %, pH 5.39, 35.98 °C) were 77% and 57%, respectively. Regression analysis showed goodness of fit between the experimental results and the predicted values. The degradation behavior of CaA differed from those of pCoA and FeA, where further CaA degradation is observed at increasing sucrose and decreasing solution pH. The differences (established using UV/vis and ATR-FTIR spectroscopy) were because, unlike the other acids, CaA formed a complex with Fe(III) or with Fe(III) hydrogen-bonded to sucrose and coprecipitated with lepidocrocite, an iron oxyhydroxide.

  17. Production of L-ornithine from sucrose and molasses by recombinant Corynebacterium glutamicum.

    Science.gov (United States)

    Zhang, Yuan-Yuan; Bu, Yi-Fan; Liu, Jian-Zhong

    2015-09-01

    Sucrose and molasses are attractive raw materials for industrial fermentation. Although Corynebacterium glutamicum shows sucrose-utilizing activity, sucrose or molasses is only a fraction of carbon source used in the fermentation medium in most works. An engineered C. glutamicum strain was constructed for producing L-ornithine with sucrose or molasses as a sole carbon source by transferring Mannheimia succiniciproducens β-fructofuranosidase gene (sacC). The engineered strain, C. glutamicum ΔAPE6937R42 (pEC-sacC), produced 22.0 g/L of L-ornithine with sucrose as the sole carbon source, which is on par with that obtained by the parent strain C. glutamicum ΔAPE6937R42 with glucose as the sole carbon. The resulting strain C. glutamicum ΔAPE6937R42 (pEC-sacC) produced 27.0 g/L of L-ornithine with molasses as the sole carbon source, which is higher than that obtained by the parent strain C. glutamicum ΔAPE6937R42 with glucose as the sole carbon. This strategy can be applied for developing sucrose- or molasses-utilizing industrial strains.

  18. PIF genes mediate the effect of sucrose on seedling growth dynamics.

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    Jodi L Stewart

    Full Text Available As photoautotrophs, plants can use both the form and amount of fixed carbon as a measure of the light environment. In this study, we used a variety of approaches to elucidate the role of exogenous sucrose in modifying seedling growth dynamics. In addition to its known effects on germination, high-resolution temporal analysis revealed that sucrose could extend the number of days plants exhibited rapid hypocotyl elongation, leading to dramatic increases in ultimate seedling height. In addition, sucrose changed the timing of daily growth maxima, demonstrating that diel growth dynamics are more plastic than previously suspected. Sucrose-dependent growth promotion required function of multiple phytochrome-interacting factors (PIFs, and overexpression of PIF5 led to growth dynamics similar to plants exposed to sucrose. Consistent with this result, sucrose was found to increase levels of PIF5 protein. PIFs have well-established roles as integrators of response to light levels, time of day and phytohormone signaling. Our findings strongly suggest that carbon availability can modify the known photomorphogenetic signaling network.

  19. Inulin and erythritol as sucrose replacers in short-dough cookies: sensory, fracture, and acoustic properties.

    Science.gov (United States)

    Laguna, Laura; Primo-Martín, Cristina; Salvador, Ana; Sanz, Teresa

    2013-05-01

    The effect of sucrose replacement by erythritol and inulin was studied in short-dough cookies using instrumental and sensory analysis. Two levels of replacement were used (25% and 50% of total sucrose content). Descriptive sensory analysis showed that the sucrose replacement affects visual and texture cookies characteristics, being the differences perceived by mouth greater than by hand. In general, sucrose substitutes produced a less crispy cookie and lower consumer acceptability, with the exception of 25% sucrose replacement by inulin. Matrix aeration attributes such as open and crumbly obtained by trained panel were important properties, and correlated positively with consumer acceptance and negatively with maximum force at break (hardness). Inulin cookies sensory properties were more similar to the control than the erythritol cookies. Also, consumer overall acceptance decreased significantly with sucrose replacement by erythritol. The analysis of texture and sound revealed that inulin cookies were softer whereas erythritol cookies were harder in comparison with control cookies; despite this difference, inulin cookies had similar sound characteristics to erythritol cookies.

  20. [Properties of sucrose phosphorylase from recombinant Escherichia coli and enzymatic synthesis of alpha-arbutin].

    Science.gov (United States)

    Wan, Yuejia; Ma, Jiangfeng; Xu, Rong; He, Aiyong; Jiang, Min; Chen, Kequan; Jiang, Yin

    2012-12-01

    Sucrose phosphorylase (EC 2.4.1.7, Sucrose phosphorylase, SPase) can be produced by recombinant strain Escherichia coli Rosetta(DE3)/Pet-SPase. Crude enzyme was obtained from the cells by the high pressure disruption and centrifugation. Sucrose phosphorylase was purified by Ni-NTA affinity column chromatography and desalted by ultrafiltration. The specific enzyme activity was 1.1-fold higher than that of the crude enzyme, and recovery rate was 82.7%. The purified recombinant SPase had a band of 59 kDa on SDS-PAGE. Thermostability of the enzyme was shown at temperatures up to 37 degrees C, and pH stability between pH 6.0 and 6.7. The optimum temperature and pH were 37 degrees C and 6.7, respectively. The K(m) of SPase for sucrose was 7.3 mmol/L, and Vmax was 0.2 micromol/(min x mg). Besides, alpha-arbutin was synthesized from sucrose and hydroquinone by transglucosylation with recombinant SPase. The optimal conditions for synthesis of alpha-arbutin were 200 U/mL of recombinant SPase, 20% of sucrose, and 1.6% hydroquinone at pH 6-6.5 and 25 degrees C for 21 h. Under these conditions, alpha-arbutin was obtained with a 78.3% molar yield with respect to hydroquinone, and the concentration of alpha-arbutin was about 31 g/L.

  1. Modulation of receptors and adenylate cyclase activity during sucrose feeding, food deprivation, and cold exposure

    International Nuclear Information System (INIS)

    Thermogenesis in brown adipose tissue (BAT) serves as a regulator of body temperature and weight maintenance. Thermogenesis can be stimulated by catecholamine activation of adenylate cyclase through the β-adrenergic receptor. To investigate the effects of sucrose feeding, food deprivation, and cold exposure on the β-adrenergic pathway, adenylate cyclase activity and β-adrenergic receptors were assessed in rat BAT after 2 wk of sucrose feeding, 2 days of food deprivation, or 2 days of cold exposure. β-Adrenergic receptors were identified in BAT using [125I]iodocyanopindolol. Binding sites had the characteristics of mixed β1- and β2-type adrenergic receptors at a ratio of 60/40. After sucrose feeding or cold exposure, there was the expected increase in BAT mitochondrial mass as measured by total cytochrome-c oxidase activity but a decrease in β-adrenergic receptor density due to a loss of the β1-adrenergic subtype. This BAT β-adrenergic receptor downregulation was tissue specific, since myocardial β-adrenergic receptors were unchanged with either sucrose feeding or cold exposure. Forskolin-stimulated adenylate cyclase activity increased in BAT after sucrose feeding or cold exposure but not after food deprivation. These data suggest that in BAT, sucrose feeding or cold exposure result in downregulation of β-adrenergic receptors and that isoproterenol-stimulated adenylate cyclase activity was limited by receptor availability

  2. Coplanar waveguides loaded with a split ring resonator-based microwave sensor for aqueous sucrose solutions

    International Nuclear Information System (INIS)

    In this study, a coplanar waveguide (CPW) loaded with a split ring resonator (SRR) based microwave sensor was developed for the detection of aqueous sucrose solutions. The fabrication and testing enabled the identification of the sucrose concentration. The CPW loaded with a SRR structure design was produced using electromagnetic models to improve its sensitivity by increasing the magnitude of transmission coefficient (S 21). The resonance behavior, based on S 21 characteristics of the microwave sensor, was analyzed in the range from 2–3 GHz with air and deionized water containing different sucrose concentrations in the range from 0–1 g ml−1. The experimental results showed that the proposed system has great potential to determine the sucrose concentration. It was shown that the proposed sensor has a high dynamic range and linearity for sucrose concentration sensing. The feature characteristic based on the CPW loaded with SRR sensing was excellent as defined by a T-circuit model as an inductor, capacitor, and resistor. It also provides an opportunity for the development of a low-cost sucrose meter system as an electronic tongue. (paper)

  3. Advances in Arabidopsis research in China from 2006 to 2007

    Institute of Scientific and Technical Information of China (English)

    LIANG Yan; ZUO JianRu; YANG WeiCai

    2007-01-01

    @@ Arabidopsis thaliana, a model plant species, has a number of advantages over other plant species as an experimental organism due to many of its genetic and genomic features. The Chinese Arabidopsis community has made significant contributions to plant biology research in recent years[1,2]. In 2006, studies of plant biology in China received more attention than ever before, especially those pertaining to Arabidopsis research. Here we briefly summarize recent advances in Arabidopsis research in China.

  4. The ACR11 encodes a novel type of chloroplastic ACT domain repeat protein that is coordinately expressed with GLN2 in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Hsu Chih-Ping

    2011-08-01

    Full Text Available Abstract Background The ACT domain, named after bacterial aspartate kinase, chorismate mutase and TyrA (prephenate dehydrogenase, is a regulatory domain that serves as an amino acid-binding site in feedback-regulated amino acid metabolic enzymes. We have previously identified a novel type of ACT domain-containing protein family, the ACT domain repeat (ACR protein family, in Arabidopsis. Members of the ACR family, ACR1 to ACR8, contain four copies of the ACT domain that extend throughout the entire polypeptide. Here, we describe the identification of four novel ACT domain-containing proteins, namely ACR9 to ACR12, in Arabidopsis. The ACR9 and ACR10 proteins contain three copies of the ACT domain, whereas the ACR11 and ACR12 proteins have a putative transit peptide followed by two copies of the ACT domain. The functions of these plant ACR proteins are largely unknown. Results The ACR11 and ACR12 proteins are predicted to target to chloroplasts. We used protoplast transient expression assay to demonstrate that the Arabidopsis ACR11- and ACR12-green fluorescent fusion proteins are localized to the chloroplast. Analysis of an ACR11 promoter-β-glucuronidase (GUS fusion in transgenic Arabidopsis revealed that the GUS activity was mainly detected in mature leaves and sepals. Interestingly, coexpression analysis revealed that the GLN2, which encodes a chloroplastic glutamine synthetase, has the highest mutual rank in the coexpressed gene network connected to ACR11. We used RNA gel blot analysis to confirm that the expression pattern of ACR11 is similar to that of GLN2 in various organs from 6-week-old Arabidopsis. Moreover, the expression of ACR11 and GLN2 is highly co-regulated by sucrose and light/dark treatments in 2-week-old Arabidopsis seedlings. Conclusions This study reports the identification of four novel ACT domain repeat proteins, ACR9 to ACR12, in Arabidopsis. The ACR11 and ACR12 proteins are localized to the chloroplast, and the expression

  5. Positive impact of sucrose supplementation during slow freezing of cat ovarian tissues on cellular viability, follicle morphology, and DNA integrity.

    Science.gov (United States)

    Tanpradit, Nae; Comizzoli, Pierre; Srisuwatanasagul, Sayamon; Chatdarong, Kaywalee

    2015-06-01

    The objectives of the study were to (1) examine and optimize the impact of sucrose during slow freezing and (2) compare the results of two freezing methods (slow freezing and vitrification) on cellular viability (germinal and stromal cells), follicle morphology, DNA integrity, and gap junction protein expression (connexin 43 [Cx 43]). Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm(2)) than the group without sucrose (1.9 ± 0.2; P sucrose-treated groups (0.1 M, 47.4% and 0.3 M, 43.5%) than the group without sucrose (0 M, 33.8%; P sucrose groups (0.1 M, 1.2% and 0.3 M, 1.9%) than the group without sucrose (7.7%; P sucrose concentrations. In terms of the freezing methods used, vitrified ovarian tissues had fewer viable follicles (3.2 ± 0.6) than the slow-freezing method (4.6 ± 0.6; P sucrose supplementation and slow-freezing method on the follicular viability, follicular histologic appearances of follicles, and apoptosis of the follicles and stromal cells in cat ovarian tissues.

  6. The influence of temperature, daylength and sucrose concentration on the growth and development of Alstroemeria 'Zebra' in vitro

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    Eleonora Gabryszewska

    2013-12-01

    Full Text Available The influence of temperature, daylength and sucrose concentration on the growth and development of Alstroemeria 'Zebra' in vitro was investigated. Only slightly more lateral rhizomes were formed at 25°C in comparison with 17°C on the medium with BA. Presence of BA in the medium strongly increased number of upright growing shoots and more shoots were formed in temperature 25°C than in temperature 17°C. Rhizome multiplication and formation of upright growing shoots were not significantly influenced by daylength. Sucrose was required for the formation of lateral rhizomes, upright growing shoots and roots. The highest number Of lateral rhizomes was observed on the medium containing 60 and 80 gl-1 sucrose and BA. Presence of BA in the medium with different sucrose concentrations markedly influenced the formation of upright growing shoots; the highest number of shoots was found on the explants cultured on the media with 20 and 30 gl-1 sucrose. Low and high concentrations of sucrose inhibited upright growing shoots formation and their elongation. Rhizogenesis was not observed on rhizomes growing on the medium without sucrose and with 10 gl-1 sucrose, either in the presence or absence of NAA„ The number of roots per plantlet on the medium with NAA increased with increasing sucrose levels. The highest sucrose concentrations slightly inhibited roots formation.

  7. Differences in bingeing behavior and cocaine reward following intermittent access to sucrose, glucose or fructose solutions.

    Science.gov (United States)

    Rorabaugh, J M; Stratford, J M; Zahniser, N R

    2015-08-20

    Daily intermittent access to sugar solutions results in intense bouts of sugar intake (i.e. bingeing) in rats. Bingeing on sucrose, a disaccharide of glucose and fructose, has been associated with a "primed" mesolimbic dopamine (DA) pathway. Recent studies suggest glucose and fructose engage brain reward and energy-sensing mechanisms in opposing ways and may drive sucrose intake through unique neuronal circuits. Here, we examined in male Sprague-Dawley rats whether or not (1) intermittent access to isocaloric solutions of sucrose, glucose or fructose results in distinctive sugar-bingeing profiles and (2) previous sugar bingeing alters cocaine locomotor activation and/or reward, as determined by conditioned place preference (CPP). To encourage bingeing, rats were given 24-h access to water and 12-h-intermittent access to chow plus an intermittent bottle that contained water (control) or 8% solutions of sucrose, glucose or fructose for 9days, followed by ad libitum chow diet and a 10-day cocaine (15mg/kg; i.p.) CPP paradigm. By day 4 of the sugar-bingeing diet, sugar bingeing in the fructose group surpassed the glucose group, with the sucrose group being intermediate. All three sugar groups had similar chow and water intake throughout the diet. In contrast, controls exhibited chow bingeing by day 5 without altering water intake. Similar magnitudes of cocaine CPP were observed in rats with a history of sucrose, fructose or chow (control) bingeing. Notably, the glucose-bingeing rats did not demonstrate a significant cocaine CPP despite showing similar cocaine-induced locomotor activity as the other diet groups. Overall, these results show that fructose and glucose, the monosaccharide components of sucrose, produce divergent degrees of bingeing and cocaine reward.

  8. Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose.

    Science.gov (United States)

    Park, Si Jae; Jang, Young-Ah; Noh, Won; Oh, Young Hoon; Lee, Hyuk; David, Yokimiko; Baylon, Mary Grace; Shin, Jihoon; Yang, Jung Eun; Choi, So Young; Lee, Seung Hwan; Lee, Sang Yup

    2015-03-01

    A sucrose utilization pathway was established in Ralstonia eutropha NCIMB11599 and R. eutropha 437-540 by introducing the Mannheimia succiniciproducens MBEL55E sacC gene that encodes β-fructofuranosidase. These engineered strains were examined for the production of poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)], respectively, from sucrose as a carbon source. It was found that β-fructofuranosidase excreted into the culture medium could hydrolyze sucrose to glucose and fructose, which were efficiently used as carbon sources by recombinant R. eutropha strains. When R. eutropha NCIMB11599 expressing the sacC gene was cultured in nitrogen-free chemically defined medium containing 20 g/L of sucrose, a high P(3HB) content of 73.2 wt% could be obtained. In addition, R. eutropha 437-540 expressing the Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene accumulated P(3HB-co-21.5 mol% LA) to a polymer content of 19.5 wt% from sucrose by the expression of the sacC gene and the Escherichia coli ldhA gene. The molecular weights of P(3HB) and P(3HB-co-21.5 mol%LA) synthesized in R. eutropha using sucrose as a carbon source were 3.52 × 10(5) (Mn ) and 2.19 × 10(4) (Mn ), respectively. The engineered R. eutropha strains reported here will be useful for the production of polyhydroxyalkanoates (PHAs) from sucrose, one of the most abundant and relatively inexpensive carbon sources.

  9. Sucrose-induced analgesia during early life modulates adulthood learning and memory formation.

    Science.gov (United States)

    Nuseir, Khawla Q; Alzoubi, Karem H; Alabwaini, Jehad; Khabour, Omar F; Kassab, Manal I

    2015-06-01

    This study is aimed at examining the long-term effects of chronic pain during early life (postnatal day 0 to 8weeks), and intervention using sucrose, on cognitive functions during adulthood in rats. Pain was induced in rat pups via needle pricks of the paws. Sucrose solution or paracetamol was administered for analgesia before the paw prick. Control groups include tactile stimulation to account for handling and touching the paws, and sucrose alone was used. All treatments were started on day one of birth and continued for 8weeks. At the end of the treatments, behavioral studies were conducted to test the spatial learning and memory using radial arm water maze (RAWM), as well as pain threshold via foot-withdrawal response to a hot plate apparatus. Additionally, the hippocampus was dissected, and blood was collected. Levels of neurotrophins (BDNF, IGF-1 and NT-3) and endorphins were assessed using ELISA. The results show that chronic noxious stimulation resulted in comparable foot-withdrawal latency between noxious and tactile groups. On the other hand, pretreatment with sucrose or paracetamol increased pain threshold significantly both in naive rats and noxiously stimulated rats (Psucrose treatment prevented such impairment (PSucrose significantly increased serum levels of endorphin and enkephalin. Chronic pain decreased levels of BDNF in the hippocampus and this decrease was prevented by sucrose and paracetamol treatments. Hippocampal levels of NT-3 and IGF-1 were not affected by any treatment. In conclusion, chronic pain induction during early life induced short memory impairment, and pretreatment with sucrose prevented this impairment via mechanisms that seem to involve BDNF. As evident in the results, sucrose, whether alone or in the presence of pre-noxious stimulation, increases pain threshold in such circumstances; most likely via a mechanism that involves an increase in endogenous opioids.

  10. Generation Means Analysis of Seed Sucrose Content in Cowpea (Vigna unguiculata L. Walp.

    Directory of Open Access Journals (Sweden)

    J.B. Noubissie Tchiagam

    2011-11-01

    Full Text Available The sucrose concentration of seeds is an important component of the taste in cowpea (Vigna unguiculata L. Walp.. The objective of this research was to estimate the heritability, to investigate the mode of genetic control and to evaluate the potential for genetic improvement of sucrose content in cowpea using generation’s means analysis. F1 and F2 generations as well as backcross populations (BCP1 and BCP2 were produced in three hybrid combinations by crosses of four selected cultivars. The sucrose level was measured via a High Performance Liquid Chromatography (HPLC system. The range of variation in sucrose content among tested varieties was from 1.88 mg/g (Lori to 5.32 mg/g (C93W-24-125. Broad-sense heritability (h2 was of a moderate 58.50% to high 68.12% value and no transgressive segregation was observed. Narrow-sense heritability (39.47 to 47.83% and heterosis (-20.00 to - 5.94% values and gene effects analysis suggested that the sucrose content is controlled by additive and non-additive genes. At 10% level of selection, an increase of 19.60 to 33.64% was predicted after one cycle. Epistasis was significant in most of crosses and at least five genes affected the sucrose accumulation in cowpea seeds. These results suggested that breeding for increased sucrose content in the seeds of cowpea can be quite successful through recurrent selection in later generations.

  11. Efficacy and safety of intravenous iron sucrose in treating adults with iron deficiency anemia

    Directory of Open Access Journals (Sweden)

    Rodolfo Delfini Cançado

    2011-12-01

    Full Text Available BACKGROUND: Iron deficiency is the most common disorder in the world, affecting approximately 25% of the world`s population and the most common cause of anemia. OBJECTIVE: To evaluate the efficacy and safety of intravenous iron sucrose (IS in the treatment of adults with iron deficiency anemia METHODS: Eighty-six adult patients with iron deficiency anemia, who had intolerance or showed no effect with oral iron therapy, received a weekly dose of 200 mg of intravenous iron sucrose until the hemoglobin level was corrected or until receiving the total dose of intravenous iron calculated for each patient RESULTS: The mean hemoglobin and serum ferritin levels were 8.54 g/dL and 7.63 ng/mL (pre-treatment and 12.1 g/dL and 99.0 ng/mL (post-treatment (p-value < 0.0001, respectively. The average increases in hemoglobin levels were 3.29 g/dL for women and 4.58 g/dL for men; 94% of male and 84% of female patients responded (hemoglobin increased by at least 2 g/dL to intravenous iron therapy. Correction of anemia was obtained in 47 of 69 (68.1% female patients and in 12 of 17 male (70.6% patients. A total of 515 intravenous infusions of iron sucrose were administered and iron sucrose was generally well tolerated with no moderate or serious adverse drug reactions recorded by the investigators. CONCLUSIONS: Our data confirm that the use of intravenous iron sucrose is a safe and effective option in the treatment of adult patients with iron deficiency anemia who lack satisfactory response to oral iron therapy. Intravenous iron sucrose is well tolerated and with a clinically manageable safety profile when using appropriate dosing and monitoring. The availability of intravenous iron sucrose would potentially improve compliance and thereby reduce morbidities from iron deficiency.

  12. Efficacy and safety of intravenous iron sucrose in treating adults with iron deficiency anemia

    Science.gov (United States)

    Cançado, Rodolfo Delfini; de Figueiredo, Pedro Otavio Novis; Olivato, Maria Cristina Albe; Chiattone, Carlos Sérgio

    2011-01-01

    Background Iron deficiency is the most common disorder in the world, affecting approximately 25% of the world`s population and the most common cause of anemia. Objective To evaluate the efficacy and safety of intravenous iron sucrose (IS) in the treatment of adults with iron deficiency anemia Methods Eighty-six adult patients with iron deficiency anemia, who had intolerance or showed no effect with oral iron therapy, received a weekly dose of 200 mg of intravenous iron sucrose until the hemoglobin level was corrected or until receiving the total dose of intravenous iron calculated for each patient Results The mean hemoglobin and serum ferritin levels were 8.54 g/dL and 7.63 ng/mL (pre-treatment) and 12.1 g/dL and 99.0 ng/mL (post-treatment) (p-value < 0.0001), respectively. The average increases in hemoglobin levels were 3.29 g/dL for women and 4.58 g/dL for men; 94% of male and 84% of female patients responded (hemoglobin increased by at least 2 g/dL) to intravenous iron therapy. Correction of anemia was obtained in 47 of 69 (68.1%) female patients and in 12 of 17 male (70.6%) patients. A total of 515 intravenous infusions of iron sucrose were administered and iron sucrose was generally well tolerated with no moderate or serious adverse drug reactions recorded by the investigators. Conclusions Our data confirm that the use of intravenous iron sucrose is a safe and effective option in the treatment of adult patients with iron deficiency anemia who lack satisfactory response to oral iron therapy. Intravenous iron sucrose is well tolerated and with a clinically manageable safety profile when using appropriate dosing and monitoring. The availability of intravenous iron sucrose would potentially improve compliance and thereby reduce morbidities from iron deficiency. PMID:23049360

  13. Bioavailability of nanoparticulate hematite to Arabidopsis thaliana

    International Nuclear Information System (INIS)

    The environmental effects and bioavailability of nanoparticulate iron (Fe) to plants are currently unknown. Here, plant bioavailability of synthesized hematite Fe nanoparticles was evaluated using Arabidopsis thaliana (A. thaliana) as a model. Over 56-days of growing wild-type A. thaliana, the nanoparticle-Fe and no-Fe treatments had lower plant biomass, lower chlorophyll concentrations, and lower internal Fe concentrations than the Fe-treatment. Results for the no-Fe and nanoparticle-Fe treatments were consistently similar throughout the experiment. These results suggest that nanoparticles (mean diameter 40.9 nm, range 22.3–67.0 nm) were not taken up and therefore not bioavailable to A. thaliana. Over 14-days growing wild-type and transgenic (Type I/II proton pump overexpression) A. thaliana, the Type I plant grew more than the wild-type in the nanoparticle-Fe treatment, suggesting Type I plants cope better with Fe limitation; however, the nanoparticle-Fe and no-Fe treatments had similar growth for all plant types. -- Highlights: ► Iron nanoparticles were synthesized and assessed for bioavailability to Arabidopsis. ► Arabidopsis grew better in the presence of EDTA-bound iron than nanoparticulate iron. ► Arabidopsis grew the same in the presence of nanoparticulate iron compared to no iron. -- Synthesized iron nanoparticles were not bioavailable to Arabidopsis thaliana in agar nutrient media

  14. Mining the active proteome of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Renier A. L. Van Der Hoorn

    2011-11-01

    Full Text Available Assigning functions to the >30.000 proteins encoded by the Arabidopsis genome is a challenging task of the Arabidopsis Functional Genomics Network. Although genome-wide technologies like proteomics and transcriptomics have generated a wealth of information that significantly accelerated gene annotation, protein activities are poorly predicted by transcript or protein levels as protein activities are post-translationally regulated. To directly display protein activities in Arabidopsis proteomes, we developed and applied Activity-based Protein Profiling (ABPP. ABPP is based on the use of small molecule probes that react with the catalytic residues of distinct protein classes in an activity-dependent manner. Labeled proteins are separated and detected from proteins gels and purified and identified by mass spectrometry. Using probes of six different chemotypes we have displayed of activities of 76 Arabidopsis proteins. These proteins represent over ten different protein classes that contain over 250 Arabidopsis proteins, including cysteine- serine- and metallo-proteases, lipases, acyltransferases, and the proteasome. We have developed methods for identification of in vivo labeled proteins using click-chemistry and for in vivo imaging with fluorescent probes. In vivo labeling has revealed novel protein activities and unexpected subcellular activities of the proteasome. Labeling of extracts displayed several differential activities e.g. of the proteasome during immune response and methylesterases during infection. These studies illustrate the power of ABPP to display the functional proteome and testify to a successful interdisciplinary collaboration involving chemical biology, organic chemistry and proteomics.

  15. Safety and efficacy of aspartame-based liquid versus sucrose-based liquids used for dilution in oral sodium phosphate solutions for colonoscopy preparations.

    Science.gov (United States)

    Chamberlain, Sherman M; Balart, J Carter; Sideridis, Kostas; Salek, Jefrey; Sridhar, Subbaramiah; Thompson, William O

    2007-11-01

    The aim of this study was to investigate whether an oral sodium phosphate solution (OSPS) mixed with aspartame-based clear liquids as the diluent would yield improved colon cleansing results compared to an OSPS mixed with sucrose-based liquids as the diluent. Fifty-one patients undergoing colonoscopy were prospectively randomized into two groups to receive different OSPS colonoscopy preparations, with sucrose-based or aspartame-based liquids used as diluents. The primary end point was the quality of the colonoscopy preparation and secondary end points were serum electrolytes before and after preparations. No significant difference in colonoscopy preparation quality was seen between the two OSPS diluent groups (Mantel-Haenzel chi (2) = 0.795, P = 0.484). There were no significant differences in mean electrolyte shifts of sodium, potassium, blood urea nitrogen (BUN), creatinine (Cr), or BUN/Cr ratios between the two groups. There was a statistically significant increase in serum phosphorous in the aspartame-based group compared to the sucrose-based diluent group (P = 0.021). In conclusion, there was no clinically detectable difference in colonoscopy preparation quality between the two OSPS diluent groups. This study suggests that passive fluid transport by aquaporins may well be the major mediator of fluid shifts in the study subjects. This result suggests the potential importance of aquaporins and minimizes the importance of sodium glucose cotransporter SGLT1 in fluid and electrolyte transport in the human gastrointestinal tract. Aspartame or its constituent amino acids may enhance phosphate absorption across the human small intestine.

  16. Safety and efficacy of aspartame-based liquid versus sucrose-based liquids used for dilution in oral sodium phosphate solutions for colonoscopy preparations.

    Science.gov (United States)

    Chamberlain, Sherman M; Balart, J Carter; Sideridis, Kostas; Salek, Jefrey; Sridhar, Subbaramiah; Thompson, William O

    2007-11-01

    The aim of this study was to investigate whether an oral sodium phosphate solution (OSPS) mixed with aspartame-based clear liquids as the diluent would yield improved colon cleansing results compared to an OSPS mixed with sucrose-based liquids as the diluent. Fifty-one patients undergoing colonoscopy were prospectively randomized into two groups to receive different OSPS colonoscopy preparations, with sucrose-based or aspartame-based liquids used as diluents. The primary end point was the quality of the colonoscopy preparation and secondary end points were serum electrolytes before and after preparations. No significant difference in colonoscopy preparation quality was seen between the two OSPS diluent groups (Mantel-Haenzel chi (2) = 0.795, P = 0.484). There were no significant differences in mean electrolyte shifts of sodium, potassium, blood urea nitrogen (BUN), creatinine (Cr), or BUN/Cr ratios between the two groups. There was a statistically significant increase in serum phosphorous in the aspartame-based group compared to the sucrose-based diluent group (P = 0.021). In conclusion, there was no clinically detectable difference in colonoscopy preparation quality between the two OSPS diluent groups. This study suggests that passive fluid transport by aquaporins may well be the major mediator of fluid shifts in the study subjects. This result suggests the potential importance of aquaporins and minimizes the importance of sodium glucose cotransporter SGLT1 in fluid and electrolyte transport in the human gastrointestinal tract. Aspartame or its constituent amino acids may enhance phosphate absorption across the human small intestine. PMID:17406813

  17. Invertase and sucrose synthase activities in coffee plants sprayed with sucrose solution Atividade de invertases e sacarose sintase em plantas de cafeeiro pulverizadas com solução de sacarose

    OpenAIRE

    José Carlos da Silva; José Donizeti Alves; Amauri Alves de Alvarenga; Marcelo Murad Magalhães; Dárlan Einstein do Livramento; Daniela Deitos Fries

    2003-01-01

    One management practice of which the efficiency has not yet been scientifically tested is spraying coffee plants with diluted sucrose solutions as a source of carbon for the plant. This paper evaluates the effect of foliar spraying with sugar on the endogenous level of carbohydrates and on the activities of invertase and sucrose synthase in coffee (Coffea arabica L.) seedlings with reduced (low) and high (normal) levels of carbon reserve. The concentrations used were 0.5 and 1.0% sucrose, and...

  18. Effects of ruminal doses of sucrose, lactose, and corn starch on ruminal fermentation and expression of genes in ruminal epithelial cells.

    Science.gov (United States)

    Oba, M; Mewis, J L; Zhining, Z

    2015-01-01

    The objective was to evaluate effects of a ruminal dose of sucrose, lactose, and corn starch on ruminal fermentation and expression of genes in ruminal epithelial cells. Six ruminally cannulated nonlactating nonpregnant Holstein cows (body weight=725±69.6kg) were assigned to treatments in a 3×3 Latin square design with 7-d periods; 1d for data and sample collection followed by a 6-d washout period. Cows were fed a diet containing whole-crop barley silage and dry ground corn, and dietary neutral detergent fiber and crude protein contents were 41.8 and 13.2% [dry matter (DM) basis], respectively. Treatment was a pulse-dose of sucrose, lactose, and corn starch (3.0, 3.0, and 2.85kg of DM, respectively; providing similar amounts of hexose across the treatments) through the ruminal cannulas. All treatments were given with alfalfa silage (1.75kg DM) to prevent acute rumen acidosis. Rumen pH was continuously monitored, and rumen fluid was sampled at 0, 30, 60, 90, 120, 150, and 180min after the dose. In addition, ruminal papillae were sampled from the ventral sac at 180min after the dose. Ruminal dosing with sucrose and lactose, compared with corn starch, increased ruminal total volatile fatty acid concentration and molar proportion of butyrate from 60 to 180min after the dose, and expression of genes for sodium hydrogen exchanger isoforms 1 and 2, and ATPase isoform 1 in ruminal epithelial cells. Ruminal dosing with sucrose, compared with lactose and corn starch, decreased rumen pH from 120 to 180min after the dose and molar proportion of acetate in ruminal fluid from 60 to 150min after the dose, and increased molar proportion of propionate in ruminal fluid from 60 to 150min, and expression of genes involved in butyrate metabolism (3-hydroxy-3-methylglutaryl-coenzyme A synthase isoform 1) and anion exchange across ruminal apical cell membrane (putative anion transporter isoform 1). These results suggest that replacing dietary starch with sugars may affect ruminal

  19. The arabidopsis cyclic nucleotide interactome

    KAUST Repository

    Donaldson, Lara

    2016-05-11

    Background Cyclic nucleotides have been shown to play important signaling roles in many physiological processes in plants including photosynthesis and defence. Despite this, little is known about cyclic nucleotide-dependent signaling mechanisms in plants since the downstream target proteins remain unknown. This is largely due to the fact that bioinformatics searches fail to identify plant homologs of protein kinases and phosphodiesterases that are the main targets of cyclic nucleotides in animals. Methods An affinity purification technique was used to identify cyclic nucleotide binding proteins in Arabidopsis thaliana. The identified proteins were subjected to a computational analysis that included a sequence, transcriptional co-expression and functional annotation analysis in order to assess their potential role in plant cyclic nucleotide signaling. Results A total of twelve cyclic nucleotide binding proteins were identified experimentally including key enzymes in the Calvin cycle and photorespiration pathway. Importantly, eight of the twelve proteins were shown to contain putative cyclic nucleotide binding domains. Moreover, the identified proteins are post-translationally modified by nitric oxide, transcriptionally co-expressed and annotated to function in hydrogen peroxide signaling and the defence response. The activity of one of these proteins, GLYGOLATE OXIDASE 1, a photorespiratory enzyme that produces hydrogen peroxide in response to Pseudomonas, was shown to be repressed by a combination of cGMP and nitric oxide treatment. Conclusions We propose that the identified proteins function together as points of cross-talk between cyclic nucleotide, nitric oxide and reactive oxygen species signaling during the defence response.

  20. Elucidating the role of transport processes in leaf glucosinolate distribution.

    Science.gov (United States)

    Madsen, Svend Roesen; Olsen, Carl Erik; Nour-Eldin, Hussam Hassan; Halkier, Barbara Ann

    2014-11-01

    In Arabidopsis (Arabidopsis thaliana), a strategy to defend its leaves against herbivores is to accumulate glucosinolates along the midrib and at the margin. Although it is generally assumed that glucosinolates are synthesized along the vasculature in an Arabidopsis leaf, thereby suggesting that the margin accumulation is established through transport, little is known about these transport processes. Here, we show through leaf apoplastic fluid analysis and glucosinolate feeding experiments that two glucosinolate transporters, GTR1 and GTR2, essential for long-distance transport of glucosinolates in Arabidopsis, also play key roles in glucosinolate allocation within a mature leaf by effectively importing apoplastically localized glucosinolates into appropriate cells. Detection of glucosinolates in root xylem sap unambiguously shows that this transport route is involved in root-to-shoot glucosinolate allocation. Detailed leaf dissections show that in the absence of GTR1 and GTR2 transport activity, glucosinolates accumulate predominantly in leaf margins and leaf tips. Furthermore, we show that glucosinolates accumulate in the leaf abaxial epidermis in a GTR-independent manner. Based on our results, we propose a model for how glucosinolates accumulate in the leaf margin and epidermis, which includes symplasmic movement through plasmodesmata, coupled with the activity of putative vacuolar glucosinolate importers in these peripheral cell layers. PMID:25209984