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  • 1
    Electronic Resource
    Electronic Resource
    Palo Alto, Calif. : Annual Reviews
    Annual Review of Plant Physiology and Plant Molecular Biology 49 (1998), S. 727-760 
    ISSN: 1040-2519
    Source: Annual Reviews Electronic Back Volume Collection 1932-2001ff
    Topics: Biology
    Notes: Abstract While the concept of H+-coupling has dominated studies of energy-dependent organic solute transport in plants for over two decades, recent studies have demonstrated the existence of a group of organic solute transporters, belonging to the ATP-binding cassette (ABC) superfamily, that are directly energized by MgATP rather than by a transmembrane H+-electrochemical potential difference. Originally identified in microbial and animal cells, the ABC superfamily is one of the largest and most widespread protein families known. Competent in the transport of a broad range of substances including sugars, peptides, alkaloids, inorganic anions, and lipids, all ABC transporters are constituted of one or two copies each of an integral membrane sector and cytosolically oriented ATP-binding domain. To date, two major subclasses, the multidrug resistance-associated proteins (MRPs) and multidrug resistance proteins (MDRs) (so named because of the phenotypes conferred by their animal prototypes), have been identified molecularly in plants. However, only the MRPs have been defined functionally. This review therefore focuses on the functional capabilities, energetics, organization, and regulation of the plant MRPs. Otherwise known as GS-X pumps, or glutathione-conjugate or multispecific organic anion Mg2+-ATPases, the MRPs are considered to participate in the transport of exogenous and endogenous amphipathic anions and glutathionated compounds from the cytosol into the vacuole. Encoded by a multigene family and possessing a unique domain organization, the types of processes that likely converge and depend on plant MRPs include herbicide detoxification, cell pigmentation, the alleviation of oxidative damage, and the storage of antimicrobial compounds. Additional functional capabilities might include channel regulation or activity, and/or the transport of heavy metal chelates. The identification of the MRPs, in particular, and the demonstration of a central role for ABC transporters, in general, in plant function not only provide fresh insights into the molecular basis of energy-dependent solute transport but also offer the prospect for manipulating and investigating many fundamental processes that have hitherto evaded analysis at the transport level.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Intracellular compartmentation of 1-aminocyclopropane-1-carboxylic acid (ACC) and N-malonyl-1-aminocyclopropane-1-carboxylic acid (MACC) in wheat (Triticum aestivum L. cv. Kanzler) and barley (Hordeum vulgare L. cv. Gerbel) leaves was studied using different methods: first, the isolation of intact vacuoles from protoplasts and, second, a non-aqueous fractionation procedure. The two methods gave similar results. ACC concentrations were similar in the extravacuolar space and in the vacuole, whereas MACC was accumulated in the vacuolar space. Transport studies revealed that no specific carrier for ACC exists at the tonoplast. MACC transfer across the tonoplast was enhanced by 120% in the presence of ATP. MACC competitively inhibited malate transport into the vacuole indicating that the same transfer system catalyzes the transfer of the two dicarboxylates.It is concluded that malonylation of ACC is not a prerequisite for the transport of ACC through the tonoplast.
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  • 3
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] We have studied the utility of the yeast protein YCF1, which detoxifies cadmium by transporting it into vacuoles, for the remediation of lead and cadmium contamination. We found that the yeast YCF1-deletion mutant DTY167 was hypersensitive to Pb(II) as compared with wild-type yeast. DTY167 cells ...
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 289 (1981), S. 292-294 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Vacuoles were isolated from protoplasts as described in Fig. 1 legend and purity was confirmed by phase contrast microscopy (Fig. 1 c). Contamination with intact protoplasts never exceeded 5%. Purity was further tested using marker enzymes. The specific activity of a-mannosidase11,12 was much ...
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  • 5
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The glutathione S-conjugates of TV-ethylmaleimide (NEM-GS) and of metolachlor (metolachlor-GS), a glutathione conjugate of a chloroacetanilide herbicide, were barely taken up by isolated barley mesophyll vacuoles in the absence of Mg-ATP. In the presence of Mg-ATP, however, uptake was ...
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  • 6
    ISSN: 1432-2048
    Keywords: Hydrolases ; Nicotiana (hydrolase, protoplast) ; Protoplast ; Vacuolation ; Vacuole (regeneration)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts of tobacco (Nicotiana tabacum L. cv. Xanthi) were evacuolated by centrifugation in a density gradient. Evacuolation resulted in the quantitative loss of vacuolar hydrolytic activities. The evacuolated miniprotoplasts were cultivated under different conditions, and the regeneration of the central vacuole was investigated by light and electron microscopy as well as by the determination of activities of vacuolar marker enzymes. Vacuoles and hydrolytic activities, as well as cell wall material reappeared faster when the cells were cultivated at low osmotic strength. A newly synthesized tonoplast polypeptide could be detected using a polyspecific serum raised against tonoplast proteins of barley (Hordeum vulgare L.). Both vacuolar proton pumps, the ATPase as well as the pyrophosphatase appear to be newly synthesized during the regeneration of the vacuole.
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  • 7
    ISSN: 1432-2048
    Keywords: Nicotiana ; Protoplast ; Pyrophosphatase ; Vacuolar ATPase ; Vacuole (regeneration)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The re-formation of vacuoles in miniprotoplasts (evacuolated mesophyll protoplasts) of tobacco was investigated under different conditions. When a constant osmolarity was maintained, increasing the concentration of NaCl in the medium enhanced the regeneration of vacuoles compared to the control (0.5 M mannitol used as osmoticum). An enhanced growth rate of miniprotoplasts could also be observed under low-osmolarity conditions, by substitution of NaCl for KCl or NaNO3, or with different effectors (glycinebetaine and methyljasmonate). Using the polymerase chain reaction, one cDNA fragment of the B-subunit of the vacuolar ATPase and two fragments of the tonoplast-bound pyrophosphatase (PPase) of tobacco were cloned. Southern blot analyses indicates that for both proteins more than one gene is present in tobacco. During the regeneration of vacuoles the transcript level of the PPase increased earlier than that of the B-subunit of the vacuolar ATPase under all conditions tested (0.5 M mannitol, 0.3 M mannitol, and 0.25 M NaCl, respectively). Under salt-stress conditions (0.25 M NaCl used as osmoticum), the expression level of both proton pumps is enhanced compared to the control. This increase is not specifically due to salt stress but generally to an increased growth rate of the vacuole, since under low-osmolarity conditions the expression of the vacuolar pumps is enhanced, too.
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  • 8
    ISSN: 1432-2048
    Keywords: Key words: Acid phosphatase ; Lupinus ; Organic acids ; Phosphorus (deficiency ; uptake) ; Root exudates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Release of large amounts of citric acid from specialized root clusters (proteoid roots) of phosphorus (P)-deficient white lupin (Lupinus albus L.) is an efficient strategy for chemical mobilization of sparingly available P sources in the rhizosphere. The present study demonstrates that increased accumulation and exudation of citric acid and a concomitant release of protons were predominantly restricted to mature root clusters in the later stages of P deficiency. Inhibition of citrate exudation by exogenous application of anion-channel blockers such as ethacrynic- and anthracene-9-carboxylic acids may indicate involvement of an anion channel. Phosphorus-deficiency-induced accumulation and subsequent exudation of citric acid seem to be a consequence of both increased biosynthesis and reduced metabolization of citric acid in the proteoid root tissue, indicated by increased in-vitro activity and enzyme protein levels of phosphoenolpyruvate carboxylase (EC 4.1.1.31), and reduced activity of aconitase (EC 4.2.1.3) and root respiration. Similar to citric acid, acid phosphatase, which is secreted by roots and involved in the mobilization of the organic soil P fraction, was released predominantly from proteoid roots of P-deficient plants. Also 33Pi uptake per unit root fresh-weight was increased by approximately 50% in juvenile and mature proteoid root clusters compared to apical segments of non-proteoid roots. Kinetic studies revealed a K m of 30.7 μM for Pi uptake of non-proteoid root apices in P-sufficient plants, versus K m values of 8.5–8.6 μM for non-proteoid and juvenile proteoid roots under P-deficient conditions, suggesting the induction of a high-affinity Pi-uptake system. Obviously, P-deficiency-induced adaptations of white lupin, involved in P acquisition and mobilization of sparingly available P sources, are predominantly confined to proteoid roots, and moreover to distinct stages during proteoid root development.
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  • 9
    ISSN: 1432-2048
    Keywords: Key words: Catharanthus ; Chromate ; 4 ; 4-Diisothio cyanostilbene-2 ; 2′-Disulfonic acid ; Phosphate transport ; Tonoplast ; Vacuole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract.  Transport of inorganic orthophosphate (Pi) across the tonoplast membrane was studied using intact vacuoles isolated from suspension-cultured cells of Catharanthus roseus. Orthophosphate uptake was strongly stimulated in the presence of Mg-ATP and Mg-pyrophosphate and inhibited by bafilomycin and concanamycin which are potent inhibitors of the vacuolar H+-ATPase. These results indicated that the build-up of an electrochemical gradient by the H+ pumps was essential for the uptake of Pi. Potassium thiocyanate, which dissipates the membrane potential across the tonoplast, strongly inhibited the Mg-ATP-stimulated uptake of Pi, while only a weak inhibition was observed in the presence of NH4Cl, which dissipates the pH gradient. These results indicate that, as observed for other anions like malate or chloride, the electrical component is the driving force of Pi uptake, whereas the ΔpH plays only a minor role. Possible competitive inhibitors of Pi, MoO2− 4, VO3− 4 and CrO2− 4 were tested. Among them, CrO2− 4 strongly inhibited Pi uptake into the vacuoles. Various inhibitors of anion transport were also tested. Only 4,4-diisothiocyanostilbene-2,2′-disulfonic acid strongly inhibited Pi uptake into the vacuoles. The function of the vacuolar Pi transporters for cytoplasmic Pi homeostasis is discussed.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 1 (1982), S. 244-246 
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In primary leaves of barley allowed to senesce under natural conditions, carotenoids, like chlorophylls, disappear gradually. Commercial horse-radish peroxidase catalizes the oxidation of lutein to unknown colorless products. This reaction depends on the presence of 2,4 dichlorophenol. It is independent of peroxide but is nullified in the presence of catalase. Preparations of thylakoids from barley chloroplasts show an activity with features comparable to those of horse-radish peroxidase.
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