Heat reduces nitric oxide production required for auxin-mediated gene expression and fate determination in tree tobacco guard cell protoplasts.

Tree tobacco (Nicotiana glauca) is an equatorial perennial with a high basal thermotolerance. Cultured tree tobacco guard cell protoplasts (GCPs) are useful for studying the effects of heat stress on fate-determining hormonal signaling. At lower temperatures (32°C or less), exogenous auxin (1-naphthalene acetic acid) and cytokinin (6-benzylaminopurine) cause GCPs to expand 20- to 30-fold, regenerate cell walls, dedifferentiate, reenter the cell cycle, and divide.

Heat suppresses activation of an auxin-responsive promoter in cultured guard cell protoplasts of tree tobacco.

Cultured guard cell protoplasts (GCP) of tree tobacco (Nicotiana glauca) comprise a novel system for investigating the cell signaling mechanisms that lead to acquired thermotolerance and thermoinhibition. At 32 degrees C in a medium containing an auxin (1-naphthaleneacetic acid [NAA]) and a cytokinin (6-benzylaminopurine), GCP expand, regenerate cell walls, dedifferentiate, and divide. At 38 degrees C, GCP acquire thermotolerance within 24 h, but their expansion is limited and they neither regenerate walls nor reenter the cell cycle.

Guard cell protoplasts: isolation, culture, and regeneration of plants.

Guard cell protoplasts have been used extensively in short-term experiments designed to elucidate the signal transduction mechanisms that regulate stomatal movements. The utility of uard cell protoplasts for other types of longer-term signal transduction experiments is just now being realized. Because highly purified, primary isolates of guard cell protoplasts are synchronous initially, they are uniform in their responses to changes in culture conditions.

Are diurnal patterns of stomatal movement the result of alternating metabolism of endogenous guard cell ABA and accumulation of ABA delivered to the apoplast around guard cells by transpiration?

Abscisic acid (ABA) prevents opening of closed stomata and causes open stomata to close. A dual-source model is proposed linking ABA to diurnal stomatal movements. Darkness would favour guard cell biosynthesis of endogenous ABA and disfavour ABA catabolism.

Thermotolerant guard cell protoplasts of tree tobacco do not require exogenous hormones to survive in culture and are blocked from reentering the cell cycle at the G1-to-S transition.

When guard cell protoplasts (GCPs) of tree tobacco [Nicotiana glauca (Graham)] are cultured at 32 degrees C with an auxin (1-napthaleneacetic acid) and a cytokinin (6-benzylaminopurine), they reenter the cell cycle, dedifferentiate, and divide. GCPs cultured similarly but at 38 degrees C and with 0.1 micro M +/- -cis,trans-abscisic acid (ABA) remain differentiated.

Red light activates a chloroplast-dependent ion uptake mechanism for stomatal opening under reduced CO concentrations in Vicia spp.

•  Under red light in ambient CO guard cells of faba bean (Vicia faba) fix CO and accumulate sucrose, causing stomata to open. We examined whether at [CO ] low enough to limit guard cell photosynthesis stomata would open when illuminated with red (R) or far-red (FR) light. •  After illumination with R or FR in buffered KCl solutions, net stomatal opening was c.