Cold-induced cytosolic free calcium ion concentration changes in wheat.

Relatively little is known about changes in the cytosolic free calcium ion concentration ([Ca(2+)](c)) in monocotyledonous plants. Therefore, we produced transgenic winter wheat lines stably expressing the calcium-sensitive photoprotein aequorin constitutively in the cytosol. [Ca(2+)](c) was detected in vivo by luminometry, and [Ca(2+)](c) elevations were imaged at video rate. Experiments with the transgenic seedlings focused on potential changes in [Ca(2+)](c) during cold exposure. Temperature-induced changes in [Ca(2+)](c) were found to be more dependent on the change in temperature (dT dt(-1)) than on the absolute value of temperature. [Ca(2+)](c) increased only at cooling rates higher than 8 degrees Cmin(-1), indicating that an overall cellular [Ca(2+)](c) increase is of minor relevance as a signal for cold acclimation in wheat under ecological conditions. The results are discussed with regard to the so-called 'calcium signature hypothesis'.

How does auxin enhance cell elongation? Roles of auxin-binding proteins and potassium channels in growth control.

Elongation growth and a several other phenomena in plant development are controlled by the plant hormone auxin. A number of recent discoveries shed light on one of the classical problems of plant physiology: the perception of the auxin signal. Two types of auxin receptors are currently known: the AFB/TIR family of F box proteins and ABP1. ABP1 appears to control membrane transport processes (H+ secretion, osmotic adjustment) while the TIR/AFBs have a role in auxin-induced gene expression. Models are proposed to explain how membrane transport (e.g., K+ and H+ fluxes) can act as a cross-linker for the control of more complex auxin responses such as the classical stimulation of cell elongation.

The auxin signal for protoplast swelling is perceived by extracellular ABP1.

Protoplasts of corn coleoptiles and Arabidopsis hypocotyls respond to the plant hormone auxin with a rapid change in volume. We checked the effect of antibodies directed against epitopes of auxin-binding protein 1 from Arabidopsis thaliana (AtERabp1) and Zea mays (ZmERabp1), respectively. Antibodies raised against the C-terminus of AtERabp1 inhibited the response to auxin, while antibodies raised against a part of box a, the putative auxin-binding domain, induced a swelling response similar to that caused by auxin treatment. Synthetic C-terminal oligopeptides of ZmERabp1 also caused a swelling response. These effects occurred regardless of whether the experiments were carried out with homologous (anti-AtERabp1 antibodies on Arabidopsis protoplasts or anti-ZmERabp1 antibodies in maize protoplasts) or heterologous immunological tools. The results indicate that the auxin signal for protoplast swelling is perceived by extracellular ABP1.

Fusicoccin- and IAA-induced elongation growth share the same pattern of K+ dependence.

The dependence of growth induced by the fungal toxin fusicoccin (FC) on the K+ content of the incubation medium was investigated in abraded maize coleoptiles. If the divalent ion Ca2+ was included in the bathing medium, no FC-induced growth occurred in the absence of K+, whereas a strong response was detected in presence of K+. The optimal K+ concentration was in the range of 1-10 mM. With the exception of Rb+, none of the other alkali ions (Na+, Li+, Cs+) could replace for K+ in sustaining FC-induced growth. The potassium channel blocker tetraethylammonium (TEA) reversibly inhibited FC-induced growth. As shown earlier for auxin-induced growth, no strict potassium dependence of FC-triggered elongation was observed in Ca2+ -free media. However, TEA abolished this apparently K+ independent FC-induced growth. It is concluded that FC-induced growth, like auxin-induced growth, requires K+ uptake through K+ channels.

Auxin-induced K+ channel expression represents an essential step in coleoptile growth and gravitropism.

Auxin-induced growth of coleoptiles depends on the presence of potassium and is suppressed by K+ channel blockers. To evaluate the role of K+ channels in auxin-mediated growth, we isolated and functionally expressed ZMK1 and ZMK2 (Zea mays K+ channel 1 and 2), two potassium channels from maize coleoptiles. In growth experiments, the time course of auxin-induced expression of ZMK1 coincided with the kinetics of coleoptile elongation. Upon gravistimulation of maize seedlings, ZMK1 expression followed the gravitropic-induced auxin redistribution. K+ channel expression increased even before a bending of the coleoptile was observed. The transcript level of ZMK2, expressed in vascular tissue, was not affected by auxin. In patch-clamp studies on coleoptile protoplasts, auxin increased K+ channel density while leaving channel properties unaffected. Thus, we conclude that coleoptile growth depends on the transcriptional up-regulation of ZMK1, an inwardly rectifying K+ channel expressed in the nonvascular tissue of this organ.

Oxidoreductases in plant plasma membranes.

Electron transporting oxidoreductases at biological membranes mediate several physiological processes. While such activities are well known and widely accepted as physiologically significant for other biological membranes, oxidoreductase activities found at the plasma membrane of plants are still being neglected. The ubiquity of the oxidoreductases in the plasma membrane suggests that the activity observed is of major importance in fact up to now no plant without redox activity at the plasmalemma is known. Involvement in proton pumping, membrane energization, ion channel regulation, iron reduction, nutrient uptake, signal transduction, and growth regulation has been proposed. However, positive proof for one of the numerous theories about the physiological function of the system is still missing. Evidence for an involvement in signalling and regulation of growth and transport activities at the plasma membrane is strong, but the high activity of the system displayed in some experiments also suggests function in defense against pathogens.

Reexamination of the Acid growth theory of auxin action.

Some crucial arguments against the acid growth theory of auxin action (U Kutschera, P Schopfer [1985] Planta 163: 483-493) have been reinvestigated by simultaneous measurements of proton fluxes and growth of maize (Zea mays L.) coleoptiles. Special care was taken to obtain a mild, effective, and reproducible abrasion of the cuticle. Proton secretion rates were determined in a computer-controlled pH-stat. In some experiments, equilibrium pH was measured. Growth rates were determined simultaneously in the same vessel using a transducer-type auxanometer. It was found that (a) the timing of auxin and fusicoccin-induced (FC) proton secretion and growth matches well, (b) the equilibrum external pHs in the presence of IAA and FC are lower than previously recorded and below the so-called ;threshold-pH,' (c) neutral or alkaline unbuffered solutions partially inhibit FC and IAA-induced growth in a similar manner, (d) the action of pH, FC, and IAA on growth are not additive. It is concluded that the acid-growth-theory correctly describes incidents taking place in the early phases of auxin-induced growth.

Hexachloroiridate IV as an Electron Acceptor for a Plasmalemma Redox System in Maize Roots.

Hexachloroiridate IV, a new artificial electron acceptor for the constitutive plant plasma membrane redox system has been investigated. It appeared not to permeate through biological membranes. Due to its higher redox potential, it is a more powerful electron acceptor than hexacyanoferrate III (ferricyanide) and even micromolar concentrations are rapidly reduced. Hexachloroiridate IV increased H(+) efflux over a concentration range of 0.05 to 0.1 millimolar. Lower concentrations slightly inhibited proton extrusion. Calcium stimulated both proton and electron transfer rates. Like hexacyanoferrate III-reduction, irridate reduction was inhibited by auxin.