Samanea

Abstract: Leaflet movements in Samanea saman are driven by the shrinking and swelling of cells in opposing (extensor and flexor) regions of the motor organ (pulvinus). Changes in cell volume, in turn, depend upon large changes in motor cell content of K+, Cl− and other ions. We performed patch-clamp experiments on extensor and flexor protoplasts, to determine whether their plasma membranes contain channels capable of carrying the large K+ currents that flow during leaflet movement. Recordings in the “whole-cell” mode reveal depolarization-activated K+ currents in extensor and flexor cells that increase slowly (t½ = ca. 2 seconds) and remain active for minutes. Recordings from excised patches reveal a single channel conductance of ca. 20 picosiemens in both cell types. The magnitude of the K+ currents is adequate to account quantitatively for K+ loss, previously measured in vivo during cell shrinkage. The K+ channel blockers tetraethylammonium (5 millimolar) or quinine (1 millimolar) blocked channel opening and decreased light- and dark-promoted movements of excised leaflets. These results provide evidence for the role of potassium channels in leaflet movement.

Abstract: In a search for potassium channels involved in light- and clock-regulated leaf movements, we cloned four putative K channel genes from the leaf-moving organs, pulvini, of the legume Samanea saman. The S. saman SPOCK1 is homologous to KCO1, an Arabidopsis two-pore-domain K channel, the S. saman SPORK1 is similar to SKOR and GORK, Arabidopsis outward-rectifying Shaker-like K channels, and the S. saman SPICK1 and SPICK2 are homologous to AKT2, a weakly-inward-rectifying Shaker-like Arabidopsis K channel. All four S. saman sequences possess the universal K-channel-specific pore signature, TXXTXGYG, strongly suggesting a role in transmembrane K(+) transport. The four S. saman genes had different expression patterns within four leaf parts: "extensor" and "flexor" (the motor tissues), the leaf blades (mainly mesophyll), and the vascular bundle ("rachis"). Based on northern blot analysis, their transcript level was correlated with the rhythmic leaf movements: (a) all four genes were regulated diurnally (Spick2, Spork1, and Spock1 in extensor and flexor, Spick1 in extensor and rachis); (b) Spork1 and Spock1 rhythms were inverted upon the inversion of the day-night cycle; and (c) in extensor and/or flexor, the expression of Spork1, Spick1, and Spick2 was also under a circadian control. These findings parallel the circadian rhythm shown to govern the resting membrane K(+) permeability in extensor and flexor protoplasts and the susceptibility of this permeability to light stimulation (Kim et al., 1993). Thus, Samanea pulvinar motor cells are the first described system combining light and circadian regulation of K channels at the level of transcript and membrane transport.

Abstract: Leaflet movement in legumes depends on rhythmic, light-regulated ion fluxes in opposing regions of the leaf-moving organ. In flexor and extensor protoplasts from Samanea saman Merrill, opening and closing of K+ channels were rhythmic in constant darkness. When channels were open in flexor protoplasts they were closed in extensor protoplasts, and vice versa. The rhythms were shifted by a delay in the onset of constant darkness, a response typical of endogenous circadian rhythms. During the light period, the channels in flexor protoplasts were sensitive to red light that was followed by premature darkness; phytochrome was implicated as the photoreceptor.

Abstract: NYCTINASTIC plants such as Albizzia julibrissin, Mimosa pudica and Samanea saman have compound leaves with paired leaflets which are usually horizontal (open) during daylight and vertical (closed) at night, oscillating between these positions with a circadian rhythm during long dark periods1–3. The angle of a leaflet is determined by the relative turgour of motor cells on opposite sides of the pulvinus, an organ which subtends the leaflet. Extensor cells are turgid and flexor cells are flaccid when leaflets are open and vice versa when they are closed. This is regulated by movements of potassium2,4–8, which is high in turgid cells and low in flaccid cells. Inhibitors of oxidative phosphorylation and low temperatures impede opening and promote closure, suggesting that active transport predominates during opening, while diffusion is favoured during closure4,8–10.