Limitation of C3-CAM shift in the common ice plant under high irradiance.

In the halophytic plant Mesembryanthemum crystallinum salinity or drought can change the mode of photosynthesis from C(3) to crassulacean acid metabolism (CAM). These two stress factors are linked to oxidative stress, however, the induction of CAM by oxidative stress per se is not straightforward. Treatment with high light (HL) did not lead to the induction of CAM, as documented by a low night/day difference in malate level and a low expression of the CAM-related form of phosphoenolcarboxylase (Ppc1), despite causing some oxidative damage (elevated MDA level, malondialdehyde). In contrast to the action of high salinity (0.4M NaCl), HL treatment did not activate neither the cytosolic NADP-malic enzyme nor the chloroplastic form of NADP-dependent malate dehydrogenase (NADP-MDH). In plastids of HL-treated plants a huge amount of starch was accumulated. This was associated with a weak stimulation of hydrolytic and phosphorolytic starch-degrading enzymes, in contrast to their strong up-regulation under high salinity. It is concluded that HL alone is not able to activate starch degradation necessary for CAM performance. Moreover, in the absence of salinity in C(3)M. crystallinum plants an age-dependent increase in energy dissipation from PSII was documented under high irradiance, as illustrated by non-photochemical quenching (NPQ). Obtained data suggest that in this halophytic species several photoprotective strategies are strictly salinity-dependent.

Responses of chlorophyll fluorescence parameters of the facultative halophyte and C3-CAM intermediate species Mesembryanthemum crystallinum to salinity and high irradiance stress.

Mesembryanthemum crystallinum L. (Aizoaceae) is a facultative annual halophyte and a C(3)-photosynthesis/crassulacean acid metabolism intermediate species currently used as a model plant in stress physiology. Both salinity and high light irradiance stress are known to induce CAM in this species. The present study was performed to provide a diagnosis of alterations at the photosystem II level during salinity and irradiance stress. Plants were subjected for up to 13 days to either 0.4M NaCl salinity or high irradiance of 1000 micromol m(-2)s(-1), as well as to both stress factors combined (LLSA=low light plus salt; HLCO=high light of 1000 micromol m(-2)s(-1), no salt; HLSA=high light plus salt). A control of LLCO=low light of 200 micromol m(-2)s(-1), no salt was used. Parameters of chlorophyll a fluorescence of photosystem II (PSII) were measured with a pulse amplitude modulated fluorometer. HLCO and LLSA conditions induced a weak degree of CAM with day/night changes of malate levels (Deltamalate) of approximately 12mM in the course of the experiment, while HLSA induced stronger CAM of Deltamalate approximately 20 mM. Effective quantum yield of PSII, DeltaF/F'(m), was only slightly affected by LLSA, somewhat reduced during the course of the experiment by HLCO and clearly reduced by HLSA. Potential quantum efficiency of PSII, F(v)/F(m), at predawn times was not affected by any of the conditions, always remaining at 0.8, showing that there was no acute photoinhibition. During the course of the days HL alone (HLCO) also did not elicit photoinhibition; salt alone (LLSA) caused acute photoinhibition which was amplified by the combination of the two stresses (HLSA). Non-photochemical, NPQ, quenching remained low (<0.5) under LLCO, LLSA and HLCO and increased during the course of the experiment under HLSA to 1-2. Maximum apparent photosynthetic electron transport rates, ETR(max), declined during the daily courses and were reduced by LLSA and to a similar extent by HLSA. It is concluded that M. crystallinum expresses effective stress tolerance mechanisms but photosynthetic capacity is reduced by the synergistic effects of salinity and light irradiance stress combined.

Redox changes in the chloroplast and hydrogen peroxide are essential for regulation of C(3)-CAM transition and photooxidative stress responses in the facultative CAM plant Mesembryanthemum crystallinum L.

Mesembryanthemum crystallinum, a facultative halophyte and C(3)-Crassulacean acid metabolism (CAM) intermediate plant, has become a favoured plant for studying stress response mechanisms during C(3)-CAM shifts. One hour of exposure to excess light (EL) caused inhibition of photosynthetic electron transport in M. crystallinum leaves as indicated by chlorophyll a fluorescence measurements. This was accompanied by an increase in NADP-malic enzyme (ME), one of the key cytosolic enzymes involved in CAM, and by a general increase in superoxide dismutase (SOD) activity. In contrast, NAD-ME activity (the mitochondrial form of ME) was not affected by EL. Exposure to EL and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) treatment of a whole plant in low light induced hydrogen peroxide (H(2)O(2)) and C(3) to CAM transition. In contrast, treatment with 3-3,4-dichlorophenyl-1,1-dimethyl urea (DCMU) has blocked high light-induced H(2)O(2) accumulation and C(3)-CAM transition. Moreover, the abundance of transcripts encoding different SODs, ascorbate peroxidase and SOD activity was differently regulated by DCMU and DBMIB. Results of applying EL or high light, H(2)O(2) and photosynthetic electron transport inhibitors suggest that the redox events in the vicinity of PSII and/or PSI and photo-produced H(2)O(2) play a major role in the regulation of C(3)-CAM transition and photooxidative stress responses in M. crystallinum.

Integrating diel starch metabolism with the circadian and environmental regulation of Crassulacean acid metabolism in Mesembryanthemum crystallinum.

The diel (24-h) Crassulacean acid metabolism (CAM) cycle in Mesembryanthemum crystallinum (L.) requires rhythmic patterns of transitory starch degradation to produce carbon skeletons for phospho enolpyruvate (PEP) synthesis during the nocturnal Phase I, when PEP carboxylase (PEPc) mediates CO(2) fixation. Under a normal light-dark cycle, nocturnal malate accumulation and nocturnal CO(2) uptake were observed for CAM-induced, but not C(3), M. crystallinum. In both C(3) and CAM plants, transcripts encoding beta-amylase and starch phosphorylase accumulated during the afternoon and declined nocturnally. Under a continuous light regime, ribulose-1,5-bisphosphate carboxylase/oxygenase activity remained co-ordinated with the CAM phases, and circadian abundance patterns were observed for transcripts encoding starch degradative enzymes. Despite circadian PEPc kinase expression, the accumulation of vacuolar malate ceased under continuous light. Exposure to CO(2)-free air for 24 h inhibited starch accumulation over the photoperiod, but re-fixation of respiratory CO(2) resulted in the overnight accumulation of malate to levels comparable to those of control plants. Upon return to normal air, the depleted starch concentration led to stoichiometric decreases in Phase-I CO(2) uptake and malate accumulation. The up-regulation of PEPc kinase transcripts under these conditions was ineffective at sustaining Phase-I CO(2) uptake under starch-limited conditions. We conclude that starch turnover regulates and limits carbon flux through the diel CAM cycle.