30-year progress of membrane transport in plants

Hedrich, Rainer; Marten, Irene

doi:10.1007/s00425-006-0341-x

Cited by 18 publications

(18 citation statements)

References 198 publications

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“…In this study on CLC‐7, we propose the central lytic vacuole of Arabidopsis mesophyll protoplasts as a novel system for the functional characterization of intracellular transporters from animal cells. This large organelle constitutes a convenient system for detailed patch‐clamp studies (Hedrich, 1995; Hedrich & Marten, 2006). In addition to macroscopic current recordings from the whole vacuole, many reports investigated single channel behaviour and modulation in excised vacuolar membrane patches, both in the cytosolic‐side‐out (Scholz‐Starke et al 2006) and vacuolar‐side‐out configurations (Pottosin & Martinez‐Estevez, 2003).…”

Section: Discussionmentioning

confidence: 99%

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7

Costa

Gutla

Boccaccio

et al. 2012

The Journal of Physiology

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Key points• Ion transport proteins in intracellular membranes of eukaryotic cells play key roles in many physiological and pathological processes.• The function of many of these transporters is poorly understood, because their intracellular localization makes them difficult to study.• Here, we used the large organelle of plant cells, the central vacuole, as a novel system to study an intracellular transporter from animal cells.• Our data showed that the lysosomal chloride transporter CLC-7 from rat constitutes a functional transport protein in the central vacuole of the model plant Arabidopsis thaliana (thale cress).• This novel approach has the potential to elucidate the transport properties of further, poorly studied intracellular ion channels and transporters.Abstract Functional characterization of intracellular transporters is hampered by the inaccessibility of animal endomembranes to standard electrophysiological techniques. Here, we used Arabidopsis mesophyll protoplasts as a novel heterologous expression system for the lysosomal chloride-proton exchanger CLC-7 from rat. Following transient expression of a rCLC-7:EGFP construct in isolated protoplasts, the fusion protein efficiently targeted to the membrane of the large central vacuole, the lytic compartment of plant cells. Membrane currents recorded from EGFP-positive vacuoles were almost voltage independent and showed time-dependent activation at elevated positive membrane potentials as a hallmark. The shift in the reversal potential of the current induced by a decrease of cytosolic pH was compatible with a 2Cl − /1H + exchange stoichiometry. Mutating the so-called gating glutamate into alanine (E245A) uncoupled chloride fluxes from the movement of protons, transforming the transporter into a chloride channel-like protein. Importantly, CLC-7 transport activity in the vacuolar expression system was recorded in the absence of the auxiliary subunit Ostm1, differently to recent data obtained in Xenopus oocytes using a CLC-7 mutant with partial plasma membrane expression. We also show that plasma membrane-targeted CLC-7 E245A is non-functional in Xenopus oocytes when expressed without Ostm1. In summary, our data suggest the existence of an alternative CLC-7 operating mode, which is active when the protein is not in complex with Ostm1. The vacuolar expression system has the potential to become a valuable tool for functional studies on intracellular ion channels and transporters from animal cells.

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Section: Discussionmentioning

confidence: 99%

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7

Costa

Gutla

Boccaccio

et al. 2012

The Journal of Physiology

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“…27, and references therein), the molecular identification of anion permeable channels is still in progress. Recently, the guard cell slow anion channel (SLAC1), an aluminum-activated malate channel, and a nitrate-permeable transporter of the CLC family have been identified (28 -31).…”

Section: Discussionmentioning

confidence: 99%

Perception of the Arabidopsis Danger Signal Peptide 1 Involves the Pattern Recognition Receptor AtPEPR1 and Its Close Homologue AtPEPR2

Król

Mentzel

Chinchilla

et al. 2010

Journal of Biological Chemistry

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347

298

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Plasma membrane-borne pattern recognition receptors, which recognize microbe-associated molecular patterns and endogenous damage-associated molecular patterns, provide the first line of defense in innate immunity. In plants, leucine-rich repeat receptor kinases fulfill this role, as exemplified by FLS2 and EFR, the receptors for the microbe-associated molecular patterns flagellin and elongation factor Tu. Here we examined the perception of the damage-associated molecular pattern peptide 1 (AtPep1), an endogenous peptide of Arabidopsis identified earlier and shown to be perceived by the leucine-rich repeat protein kinase PEPR1. Using seedling growth inhibition, elicitation of an oxidative burst and induction of ethylene biosynthesis, we show that wild type plants and the pepr1 and pepr2 mutants, affected in PEPR1 and in its homologue PEPR2, are sensitive to AtPep1, but that the double mutant pepr1/pepr2 is completely insensitive. As a central body of our study, we provide electrophysiological evidence that at the level of the plasma membrane, AtPep1 triggers a receptor-dependent transient depolarization through activation of plasma membrane anion channels, and that this effect is absent in the double mutant pepr1/pepr2. The double mutant also fails to respond to AtPep2 and AtPep3, two distant homologues of AtPep1 on the basis of homology screening, implying that the PEPR1 and PEPR2 are responsible for their perception too. Our findings provide a basic framework to study the biological role of AtPep1-related danger signals and their cognate receptors.In plant immunity, a first line of defense is based on the perception of a group of conserved, pathogen-derived molecules, called microbe-associated molecular patterns (MAMPs) 4 by pattern recognition receptors, which cause the expression of defense genes as well as metabolic rearrangements, and ultimately activate basal resistance to potential pathogens (1, 2). In Arabidopsis, the best studied MAMPs are the bacterial flagellin (active epitope flg22) and elongation factor Tu (active EF-Tu epitopes elf13, elf18, and elf26), which are recognized by their cognate leucine rich repeat-receptor-like kinases FLS2 (flagellin-sensitive 2) and EFR (EF-Tu receptor), respectively (reviewed in Ref. 1). Perception of these MAMPs leads to a set of responses that can be used to monitor the recognition process, including the triggering of ion fluxes, the generation of reactive oxygen species (ROS), accumulation of ethylene, and finally up-regulation of defense-related genes; investment into increased resistance against bacterial pathogens negatively feeds back on plant growth (3). In addition to these MAMP/ pattern recognition receptor systems, another class of surveillance system recognizes plant-derived molecules previously known as "endogenous elicitors" and now as DAMPs (damageassociated molecular patterns): DAMPs are endogenous molecules that newly appear in the intercellular space in response to the damage caused by a pathogen attack, e.g. cell wall fragments or effectors deri...

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“…Several contracted families were functionally enriched in exocytosis and transmembrane transport functions, which are usually involved in the release of secondary metabolites, hormones, and numerous other compounds (Hedrich and Marten, 2006). Interestingly, different multi-antimicrobial extrusion protein families either expanded or contracted within the genome of C. argyrosperma.…”

Section: Molecular Plantmentioning

confidence: 99%

The Genome of Cucurbita argyrosperma (Silver-Seed Gourd) Reveals Faster Rates of Protein-Coding Gene and Long Noncoding RNA Turnover and Neofunctionalization within Cucurbita

et al. 2019

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Whole-genome duplications are an important source of evolutionary novelties that change the mode and tempo at which genetic elements evolve within a genome. The Cucurbita genus experienced a wholegenome duplication around 30 million years ago, although the evolutionary dynamics of the coding and noncoding genes in this genus have not yet been scrutinized. Here, we analyzed the genomes of four Cucurbita species, including a newly assembled genome of Cucurbita argyrosperma, and compared the gene contents of these species with those of five other members of the Cucurbitaceae family to assess the evolutionary dynamics of protein-coding and long intergenic noncoding RNA (lincRNA) genes after the genome duplication. We report that Cucurbita genomes have a higher protein-coding gene birth-death rate compared with the genomes of the other members of the Cucurbitaceae family. C. argyrosperma gene families associated with pollination and transmembrane transport had significantly faster evolutionary rates. lincRNA families showed high levels of gene turnover throughout the phylogeny, and 67.7% of the lincRNA families in Cucurbita showed evidence of birth from the neofunctionalization of previously existing protein-coding genes. Collectively, our results suggest that the whole-genome duplication in Cucurbita resulted in faster rates of gene family evolution through the neofunctionalization of duplicated genes.

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30-year progress of membrane transport in plants

Cited by 18 publications

References 198 publications

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7

Perception of the Arabidopsis Danger Signal Peptide 1 Involves the Pattern Recognition Receptor AtPEPR1 and Its Close Homologue AtPEPR2

The Genome of Cucurbita argyrosperma (Silver-Seed Gourd) Reveals Faster Rates of Protein-Coding Gene and Long Noncoding RNA Turnover and Neofunctionalization within Cucurbita

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30-year progress of membrane transport in plants

Cited by 18 publications

References 198 publications

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl−/H+ exchanger CLC‐7

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl−/H+ exchanger CLC‐7

Perception of the Arabidopsis Danger Signal Peptide 1 Involves the Pattern Recognition Receptor AtPEPR1 and Its Close Homologue AtPEPR2

The Genome of Cucurbita argyrosperma (Silver-Seed Gourd) Reveals Faster Rates of Protein-Coding Gene and Long Noncoding RNA Turnover and Neofunctionalization within Cucurbita

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The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7

The Arabidopsis central vacuole as an expression system for intracellular transporters: functional characterization of the Cl⁻/H⁺ exchanger CLC‐7