Evidence that acid solutions induce plant cell elongation by acidifying the cytosol and stimulating the proton pump

AbstractAcetic acid (3 mM, pH 4.5) stimulated elongation growth of maizecoleoptiles at a much higher rate than citric acid at the same pH and concentration. The effect of these solutions on cytosolic pH and membrane potential of maize rhizodermis cells was measured with microelectrodes. Citric acid caused a decrease in cytosolic pH and a slow membrane hyperpolarization. Acetic acid induced a larger and more rapid cytosolic acidification and membrane hyperpolarization. Hence, the degree of growth stimulation by the acids was positively correlated with the extent of their cytosolic acidification and stimulation of the proton pump. We suggest the acids induce growth by acidifying the cytosol and stimulating the proton pump rather than via direct acidification of the cell wall.Acid-induced growthCell wall acidificationCytosolic acidificationIndoleacetic acidMembrane potentialZea may

Signal Transduction in Sinapis alba Root Hairs: Auxins as External Messengers

In developing root hairs of Sinapis alba the effects of externally applied indole-3-acetic acid (IAA) and other auxins have been investigated with respect to membrane potential, membrane conductance, cytosolic Ca2 + and pH. Following a delay of roughly 30s, 10- 12 to lO- IO M IAA slowly hyperpolarize, 10- 7 M IAA rapidly depolarize the root hairs, while 10- 9 M has hardly an effect. We show that these voltage responses are not the result of a change in membrane conductance or permeability, but are presumably caused by a change in H+ ATPase activity. The other tested auxins and analogues yielded comparable effects, but with much lower effectivity (IAA > 1-NAA > 2,4-D ~ 2-NAA > 2,3-D). Cytosolic Ca2+ and pH were decreased during depolarization by 0.2 and 0.4 units, respectively. No such changes were observed during hyperpolarization or about 1 h after the first encounter of the root hairs with IAA. We propose that IAA is a natural external signal for roots while competing with neighboring organisms for nutrients and salts, and suggest a signal chain with the plasma membrane H+ ATPase as a target protein. The delay in response to IAA, the time dependency, and the extremely low effective IAA concentrations point to the existence of a IAA receptor. Since the IAA-induced shifts in cytosolic pH and Ca2+ occur simultaneously with the depolarization, the question whether these ions are cellular messengers and part of an IAA-triggered signal chain is critically discussed

The role of the plasma-membrane Ca2+-ATPase in ca2+ homeostasis in Sinapis alba root hairs

The regulation of cytosolic Ca2+ has been investigated in growing root-hair cells of Sinapis alba L. with special emphasis on the role of the plasmamembrane Ca2+-ATPase. For this purpose, erythrosin B was used to inhibit the Ca2+-ATPase, and the Ca2+ ionophore A23187 was applied to manipulate cytosolic free [Ca2+] which was then measured with Ca2+-selective microelectrodes. (i) At 0.01 M, A23187 had no effect on the membrane potential but enhanced the Ca2+ permeability of the plasma membrane. Higher concentrations of this ionophore strongly depolarized the cells, also in the presence of cyanide. (ii) Unexpectedly, A23187 first caused a decrease in cytosolic Ca2+ by 0.2 to 0.3 pCa units and a cytosolic acidification by about 0.5 pH units, (iii) The depletion of cytosolic free Ca2+ spontaneously reversed and became an increase, a process which strongly depended on the external Ca2+ concentration, (iv) Upon removal of A23187, the cytosolic free [Ca2+] returned to its steady-state level, a process which was inhibited by erythrosin B. We suggest that the first reaction to the intruding Ca2+ is an activation of Ca2+ transporters (e.g. ATPases at the endoplasmic reticulum and the plasma membrane) which rapidly remove Ca2+ from the cytosol. The two observations that after the addition of A23187, (i) Ca2+ gradients as steep as-600 mV could be maintained and (ii) the cytosolic pH rapidly and immediately decreased without recovery indicate that the Ca2+-exporting plasma-membrane ATPase is physiologically connected to the electrochemical pH gradient, and probably works as an nH+/Ca2+-ATPase. Based on the finding that the Ca2+-ATPase inhibitor erythrosin B had no effect on cytosolic Ca2+, but caused a strong Ca2+ increase after the addion of A23187 we conclude that these cells, at least in the short term, have enough metabolic energy to balance the loss in transport activity caused by inhibition of the primary Ca2+-pump. We further conclude that this ATPase is a major Ca2+ regulator in stress situations where the cytosolic Ca2+ has been shifted from its steady-state level, as may be the case during processes of signal transduction

A blood atlas of COVID-19 defines hallmarks of disease severity and specificity.

Treatment of severe COVID-19 is currently limited by clinical heterogeneity and incomplete description of specific immune biomarkers. We present here a comprehensive multi-omic blood atlas for patients with varying COVID-19 severity in an integrated comparison with influenza and sepsis patients versus healthy volunteers. We identify immune signatures and correlates of host response. Hallmarks of disease severity involved cells, their inflammatory mediators and networks, including progenitor cells and specific myeloid and lymphocyte subsets, features of the immune repertoire, acute phase response, metabolism, and coagulation. Persisting immune activation involving AP-1/p38MAPK was a specific feature of COVID-19. The plasma proteome enabled sub-phenotyping into patient clusters, predictive of severity and outcome. Systems-based integrative analyses including tensor and matrix decomposition of all modalities revealed feature groupings linked with severity and specificity compared to influenza and sepsis. Our approach and blood atlas will support future drug development, clinical trial design, and personalized medicine approaches for COVID-19

Amine Transport in Riccia fluitans: Cytoplasmic and Vacuolar pH Recorded by a pH-Sensitive Microelectrode

The cytoplasmic and vacuolar pH and changes thereof in the presence of ammonia (NH(4)Cl) and methylamine (CH(3)NH(3)Cl) have been measured in rhizoid cells of Riccia fluitans by means of a pH-sensitive microelectrode. On addition of 1 micromolar NH(4)Cl, the cytoplasmic pH of 7.2 to 7.4 drops by 0.1 to 0.2 pH units, but shifts to pH 7.8 in the presence of 50 micromolar NH(4)Cl or 500 micromolar CH(3)NH(3)Cl. The pH of the vacuole increases drastically from 4.5 to 5.7 with these latter concentrations. Since a NH(4)(+)/CH(3)NH(3)(+) uniporter has been demonstrated in the plasmalemma of R. fluitans previously (Felle 1983 Biochim Biophys Acta 602:181-195), the concentration-dependent shifts of cytoplasmic pH are interpreted as results of two processes: first, acidification through deprotonation of the actively transported NH(4)(+); and second, alkalinization through protonation of NH(3) which is taken up to a significant extent from high external concentrations. Furthermore, it is concluded that the determination of intracellular pH by means of methylamine distribution is not a reliable method for eucaryotic systems