Lonnie J. Guralnick

Photosynthetic and anatomical characteristics in the C4–crassulacean acid metabolism-cycling plant Portulaca grandiflora

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. Portulaca grandiflora (Lind.) is a succulent species with C4 photosynthesis and crassulacean acid metabolism (CAM) cycling in leaves, and CAM-idling type photosynthesis in stems. We investigated the level and localization of carbon fixation enzymes and photosynthetic activity of leaves and stems of P. grandiflora under well-watered and drought conditions. As CAM activity increased during water stress, the leaf water-storage tissue collapsed, presumably transferring water to the bundle sheath and mesophyll cells, and so maintaining the C4 photosynthetic pathway. Tissue prints indicated an increase in phosphoenolpyruvate carboxylase (PEPC) in the water-storage tissue of leaves and the cortex of stems. Immunoblot analyses after 10 d of water stress showed that leaves had a slight decrease in the proteins of the C4-CAM pathway, while at the same time a new isoform of NADP-malic enzyme (NADP-ME) appeared. In contrast, the stem showed increases in proteins of the CAM pathway when water stressed. Under water stress, diurnal fluctuation in acidity in leaves was not accompanied by a net gain or loss of CO2 at night, and there was sustained, but decreased, fixation of CO2 during the day, characteristic of CAM cycling. High gross rates of O2 evolution were maintained during the day under water stress, suggesting induction of alternative electron sinks. With induced diurnal fluctuations in acidity in stems, there was no net carbon gain during the day or night. These results demonstrate, for the first time, that the stem of P. grandiflora is an inducible CAM-idling tissue. Our results also indicate that the C4 and CAM pathways operate independently of one another in P. grandiflora.

Evolutionary physiology: the extent of C4 and CAM photosynthesis in the genera Anacampseros and Grahamia of the Portulacaceae

The Portulacaceae is one of the few terrestrial plant families known to have both C(4) and Crassulacean acid metabolism (CAM) species. There may be multiple origins of the evolution of CAM within the Portulacaceae but the only clear evidence of C(4) photosynthesis is found in members of the genus Portulaca. In the Portulaca, CAM succulent tissue is overlaid with the C(4) tissue in a unique fashion where both pathways are operating simultaneously. Earlier reports have shown that the clade containing the genera Anacampseros and Grahamia may also contain C(4) photosynthetic species similar to the Portulaca, which would indicate multiple origins of C(4) photosynthesis within the family. The aim of the present study was to ascertain the true photosynthetic nature of these genera. An initial survey of the carbon isotope composition of the Anacampseros ranged from -12.6 per thousand to -24.0 per thousand, indicating very little CAM activity in some species, with other values close to the C(4) range. Anacampseros (=Grahamia) australiana which had been previously identified as a C(4) species had a carbon isotope composition value of -24.0 per thousand, which is more indicative of a C(3) species with a slight contribution of CAM activity. Other Anacampseros species with C(4)-like values have been shown to be CAM plants. The initial isotope analysis of the Grahamia species gave values in the range of -27.1 per thousand to -23.6 per thousand, placing the Grahamia species well towards the C(3) photosynthetic range. Further physiological studies indicated increased night-time CO(2) uptake with imposition of water stress, associated with a large diurnal acid fluctuation and a marked increased phosphoenolpyruvate carboxylase activity. This showed that the Grahamia species are actually facultative CAM plants despite their C(3)-like carbon isotope values. The results indicate that the Grahamia and Anacampseros species do not utilize the C(4) photosynthetic pathway. This is the first to identify that the Grahamia species are facultative CAM plants where CAM can be induced by water stress. This work supports earlier physiological work that indicates that this clade containing Anacampseros and Grahamia species comprises predominantly facultative CAM plants. This report suggests there may be only one clade which contains C(4) photosynthetic members with CAM-like characteristics.

Seasonal response to drought and rewatering in Portulacaria afra (L.) Jacq.

SummaryGas exchange characteristics of droughted and rewatered Portulacaria afra were studied during the seasonal shift from CAM to C3 photosynthesis. 14CO2 uptake, stomatal conductance, and total titratable acidity were determined for both irrigated and 2, 4, and 7.5 month waterstressed plants from summer 1984 to summer 1985. Irrigated P. afra plants were utilizing the CAM pathway throughout the summer and shifted to C3 during the winter and spring. Beginning in September, P. afra plants shifted from CAM to CAM-idling after 2 months of water-stress. When water-stress was initiated later in the fall, exogenous CO2 uptake was still measurable after 4 months of drought. After 7.5 months of stress, exogenous CO2 uptake was absent. The shift from CAM to CAM-idling or C3 in the fall and winter was related to when water stress was initiated and not to the duration of the stress. Gas exchange resumed within 24 h of rewatering regardless of the duration of the drought. In the winter and spring, rewatering resulted in a full resumption of daytime CO2 uptake. Whereas during the summer, rewatering quickly resulted in early morning CO2 uptake, but nocturnal CO2 uptake through the CAM pathway was observed after 7 days. Gas exchange measurements, rewatering characteristics, and transpirational water loss support the hypothesis that the C3 pathway was favored during the winter and spring. The CAM pathway was functional during the summer when potential for water loss was greater. Our investigations indicate that P. afra has a flexible photosynthetic system that can withstand long-term drought and has a rapid response to rewatering.

Photosynthetic Properties of the Three Major Tissue Layers of the CAM Plant, Peperomia Camptotricha

C4 plants are generally characterized by a spatial separation of the C4 and C4 modes of carbon fixation into bundle sheath cells and mesophyll cells, respectively. In contrast Cam plants typically exhibit a temporal separation of the C3 and C4 modes with initial carbon fixation by the C, pathway at night and decarboxylation of the C4 products and refixation or the released CO2 via the reductive pentose phosphate pathway during the day.