J.T.M. Elzenga

Characterization of a Light-Controlled Anion Channel in the Plasma Membrane of Mesophyll Cells of Pea

In leaf mesophyll cells of pea (Pisum sativum) light induces a transient depolarization that is at least partly due to an increased plasma membrane conductance for anions. Several channel types were identified in the plasma membrane of protoplasts from mesophyll cells using the patch-clamp technique. One of these was an anion channel with a single-channel conductance of 32 picasiemens in symmetrical 100/100 KCl solutions. In asymmetrical solutions the reversal potential indicates a high selectivity for Cl- over K+ at high cytoplasmic Cl-. At negative membrane voltages the channel openings were interrupted by very short closures. In the open channel conductance several substrates were identified. At a cytoplasmic negative logarithm of Ca concentration higher than 6.3, no channel openings were observed. When the protoplast was illuminated in the cell-attached configuration, at least one channel type had a higher opening probability. This channel can tentatively be identified as the above-described anion channel based on conductance and the characteristic short closures at negative membrane potentials. This light activation of the 32-picasiemen anion channel is a strong indication that this channel conducts the light-induced depolarizing current. Because channel activity is strongly Ca2+-dependent, a role of cytoplasmic Ca2+ concentration changes in the light activation of the conductance is discussed.

Characterization of ion channels in the plasma membrane of epidermal cells of expanding pea (Pisum sativum arg) leaves

Ion channels in isolated patches of the plasma membrane of pea (Pisum sativum arg) epidermal cells were studied with the patch-clamp technique. One anion and one cation channel were dominantly present in most trials. The anion channel conducts nitrate, halides and malate, with a conductance in symmetrical 100 mm Cl− of 300 pS and can be blocked by SITS when applied to the cytoplasmic side of the membrane. The cation channel poorly discriminates between potassium, sodium and lithium, is not blocked by either TEA or Ba2+, and has a conductance of 35 pS in symmetrical 100 mm K+. The open probability of the cation channel increases with increase of the Ca2+ concentration on the cytoplasmic side of the membrane from 0.1 to 1 μm. The possible role of these two channels in the physiology of epidermal cells is discussed.

Kinetics of Ca2+- and ATP-dependent, voltage-controlled anion conductance in the plasma membrane of mesophyll cells of Pisum sativum

Abstract. Whole-cell patch-clamp techniques were used to measure anion currents through the plasma membrane of protoplasts of mesophyll cells of expanding pea (Pisum sativum L.) leaves. Voltage-induced changes of the currents could be modelled with single exponential activation and deactivation kinetics. The anion currents were activated at negative membrane potentials. The time constant of activation, τact, increased from 145 ms at −140 mV to 380 ms at −20 mV. A Boltzmann fit to the activation curve, n∞ (ΔGVm/ΔGmax), yielded a half-activation voltage of +27 mV. Opening and closing rate constants, α and β respectively, were calculated from the values of τ and n∞. The currents depended on the presence of cytoplasmic Ca2+ concentrations higher than 10−6 M. Including 3 mM MgATP in the intracellular solution resulted in a voltage-dependent inactivation of the anion current. The conductance-voltage relation resulting from the voltage-dependent activation and inactivation had a maximum at about −25 mV. The relations of the current in pea are discussed with respect to the anion currents in guard cells and suspension-cultured tobacco cells, and its possible role in growing leaf cells.

Competition for pollinators and intra-communal spectral dissimilarity of flowers

Competition for pollinators occurs when, in a community of flowering plants, several simultaneously flowering plant species depend on the same pollinator. Competition for pollinators increases interspecific pollen transfer rates, thereby reducing the number of viable offspring. In order to decrease interspecific pollen transfer, plant species can distinguish themselves from competitors by having a divergent phenotype. Floral colour is an important signalling cue to attract potential pollinators and thus a major aspect of the flower phenotype. In this study, we analysed the amount of spectral dissimilarity of flowers among pollinator-competing plants in a Dutch nature reserve. We expected pollinator-competing plants to exhibit more spectral dissimilarity than non-competing plants. Using flower visitation data of 2 years, we determined the amount of competition for pollinators by different plant species. Plant species that were visited by the same pollinator were considered specialist and competing for that pollinator, whereas plant species visited by a broad array of pollinators were considered non-competing generalists. We used principal components analysis to quantify floral reflectance, and found evidence for enhanced spectral dissimilarity among plant species within specialist pollinator guilds (i.e. groups of plant species competing for the same pollinator). This is the first study that examined intra-communal dissimilarity in floral reflectance with a focus on the pollination system.