Manfred H. Weisenseel

Ca2+ Regulates Reactive Oxygen Species Production and pH during Mechanosensing in Arabidopsis Roots

Mechanical stimulation of plants triggers a cytoplasmic Ca2+ increase that is thought to link the touch stimulus to appropriate growth responses. We found that in roots of Arabidopsis thaliana, external and endogenously generated mechanical forces consistently trigger rapid and transient increases in cytosolic Ca2+ and that the signatures of these Ca2+ transients are stimulus specific. Mechanical stimulation likewise elicited an apoplastic alkalinization and cytoplasmic acidification as well as apoplastic reactive oxygen species (ROS) production. These responses showed the same kinetics as mechanically induced Ca2+ transients and could be elicited in the absence of a mechanical stimulus by artificially increasing Ca2+ concentrations. Both pH changes and ROS production were inhibited by pretreatment with a Ca2+ channel blocker, which also inhibited mechanically induced elevations in cytosolic Ca2+. In trichoblasts of the Arabidopsis root hair defective2 mutant, which lacks a functional NADPH oxidase RBOH C, touch stimulation still triggered pH changes but not the local increase in ROS production seen in wild-type plants. Thus, mechanical stimulation likely elicits Ca2+-dependent activation of RBOH C, resulting in ROS production to the cell wall. This ROS production appears to be coordinated with intra- and extracellular pH changes through the same mechanically induced cytosolic Ca2+ transient.

Bioelectricity, gravity and plants

Abstract. This brief review summarizes gravity-induced changes in bioelectric parameters and evaluates their contribution to our understanding of the sensing of gravity, and the transduction and transmission of the gravity stimulus in plants. During the last few decades, information has accumulated demonstrating gravity-induced changes in surface potentials, membrane voltages, endogenous electric currents and ion fluxes. These changes point to the plasma membrane as the site of perception and transduction of the gravity signal. To date, it is reasonable to assume that gravity affects the state of ion channels (in particular, Ca2+ channels) and the activity of ion pumps (in particular, the electrogenic H+-ATPase) in the plasma membrane leading to intracellular and apoplasmic changes in ion activities and in membrane voltages. The flow of H+ and Ca2+ currents is probably the means by which information about gravity is amplified and transmitted from sensing to responding cells. No data are available so far about the effect of microgravity on bioelectric parameters. However, it would be interesting to learn if plants become hypersensitive to gravity during a prolonged stay in microgravity. If so, such plants might fire action potentials after return to earth, because more Ca2+ channels than usual may be activated by 1?g in microgravity-adapted plants.

Wound-Induced Changes of Membrane Voltage, Endogenous Currents, and Ion Fluxes in Primary Roots of Maize

The effects of mechanical wounding on membrane voltage, endogenous ion currents, and ion fluxes were investigated in primary roots of maize (Zea mays) using intracellular microelectrodes, a vibrating probe, and ion-selective electrodes. After a wedge-shaped wound was cut into the proximal elongation zone of the roots, a large inward current of approximately 60 [mu]A cm-2 was measured, together with a change in the current pattern along the root. The changes of the endogenous ion current were accompanied by depolarization of the membrane voltage of cortex cells up to 5 mm from the wound. Neither inhibitors of ion channels nor low temperature affected the large, wound-induced inward current. The fluxes of H+, K+, Ca2+, and Cl- contributed only about 7 [mu]A cm-2 to the wound-induced ion current. This suggests the occurrence of a large mass flow of negatively charged molecules, such as proteins, sulfated polysaccharides, and galacturonic acids, from the wound. Natural wounding of the root cortex by developing lateral roots caused an outwardly directed current, which was clearly different in magnitude and direction from the current induced by mechanical injury.

Electrotropism of Maize (Zea mays L.) Roots (Facts and Artifacts).

Intact and decapped primary roots of maize (Zea mays L.) were exposed to DC electric fields of 0.5 to 8.0 V/cm in low-salinity media to resolve conflicting results about the direction of electrotropism. In DC fields of 0.5 V/cm or 1.0 V/cm, intact roots always curved toward the cathode. In a field of 8.0 V/cm, intact roots curved toward the anode and stopped growth. Decapped roots also curved toward the anode both in weak and strong fields. The results indicate that growth toward the cathode is the true response of healthy roots.

MEMBRANE POTENTIAL AND ENDOGENOUS ION CURRENT OF PHYCOMYCES SPORANGIOPHORES

Glass microelectrodes were inserted into the growing zone of sporangiophores of Phycomyces blakesleeanus that had been submersed in artificial pond water. The membrane potential (inside negative) increased with increasing pH of the bathing solution from an average of −98 mV at pH 5 up to −131 mV at pH 7. Removal of Ca2+ from the medium hyperpolarized the membrane potential in the wild type, but caused a significant depolarization in the blue-light-insensitive madC mutant. KCN, diethylstilbestrol, and N,N′-dicyclohexylcarbodiimide depolarized the membrane potential in both the wild type and the madC mutant, while fusicoccin had no effect. Endogenous ion current of up to 2 μA cm−2 was measured in the growing zone of sporangiophores with an extracellular vibrating electrode. The current density and current pattern varied with the pH of the medium. At pH 5 most sporangiophores had weak inward current along the growing zone, whereas at pH 7 most sporangiophores had strong outward current. The response of the membrane potential to specific inhibitors and the presence of an endogenous ion current indicate an electrogenic H+-ATPase in the plasma membrane. The results show a negative correlation between growth rate of sporangiophores growing in buffered aqueous medium and magnitude of membrane potential, as well as density of outward current. They also indicate an important role of protons in controlling the growth of Phycomyces sporangiophores.

Weak AC-electric fields promote root growth and ER abundance of root cap cells

Abstract AC-electric fields of low strength and low frequency were applied to roots of garden cress ( Lepidium sativum L.) growing in aqueous medium. These fields strongly affected the growth, shape and statocyte structure of the roots. A maximum growth-promoting effect was observed with a field frequency of 10 Hz and field strengths of 10 −3 to 10 −1 V cm −1 . Growth rates of roots were similar to controls, i.e., roots growing without AC fields, at field strengths of 10 −4 and 1 V cm −1 . Ultrastructural analyses of root cap cells showed that an AC-electric field of 10 Hz caused a significant increase in abundance of endoplasmic reticulum (ER) in the statocytes. Both phenomena, growth promotion and ER volume increase, are most likely effected by stimulation of the plasma membrane H + -ATPase of the root cells.