Sucrose transport across the vacuolar membrane of Ananas comosus

2002 ◽  
Vol 29 (6) ◽  
pp. 717 ◽  
Author(s):  
Shelley R. McRae ◽  
John T. Christopher ◽  
J. Andrew C. Smith ◽  
Joseph A. M. Holtum
Keyword(s):  
Atpase Activity ◽  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
External Medium ◽  
Soluble Sugars ◽  
Sugar Transport ◽  
Phase Iii ◽  
Ananas Comosus ◽  
Sucrose Uptake ◽  
Sucrose Transport

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. In Ananas comosus L. (Merr.) (pineapple), a widely cultivated bromeliad that exhibits crassulacean acid metabolism (CAM), much of the carbohydrate synthesized during the daytime appears to accumulate as soluble sugars in the vacuole. To investigate the mechanism of sugar transport into the vacuole, microsomal extracts were prepared from deacidifying leaves harvested during Phase III of the CAM cycle. The vesicle preparations exhibited features expected for a fraction highly enriched in vacuolar membranes (tonoplast), i.e. the ATPase activity of 16 ±�2�nkat mg-1 protein was inhibited 96% by 50 mm KNO3, an inhibitor of vacuolar ATPases, and was only 7% inhibited by 100μm NaN3 plus 100μm Na3VO4, inhibitors of mitochondrial and plasma membrane ATPases, respectively. Further, the microsomal ATPase activity showed a pH optimum between 7.0 and 8.0, typical of a vacuolar ATPase. When presented with Mg-ATP, vesicles established H+ gradients that could be maintained for at least 25 min. The vesicles were able to take up [14C]sucrose from an external medium. Sucrose uptake exhibited saturable kinetics with an apparent Km of 50 m sucrose and apparent Vmax of 171 ± 5 pkat mg-1 protein. Sucrose uptake was not dependent upon, nor stimulated by, Mg-ATP, suggesting that the mechanism of sucrose transport into the vacuole in A. comosus does not involve H+-coupled cotransport. However, the initial rates of sucrose uptake from the external medium were stimulated when vesicles were preloaded with sucrose. This trans-stimulation is consistent with characteristics expected for a sucrose uniporter capable of operating in an exchange mode. It is proposed that the accumulation of glucose and fructose in leaf vacuoles of Ananas during the light period involves at least two steps - transport of sucrose into the vacuole by a mechanism exhibiting characteristics of a sucrose uniporter, followed by cleavage of sucrose by a vacuolar acid invertase to form glucose and fructose.

Regulation of Rubisco activity in crassulacean acid metabolism plants: better late than never

2002 ◽  
Vol 29 (6) ◽  
pp. 689 ◽  
Author(s):  
Kate Maxwell ◽  
Howard Griffiths ◽  
Brent Helliker ◽  
Andrew Roberts ◽  
Richard P. Haslam ◽  
...  
Keyword(s):  
Electron Transport ◽  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
Rubisco Activase ◽  
Light Period ◽  
Phase Iii ◽  
Co2 Uptake ◽  
Rubisco Activity ◽  
Free Atmosphere ◽  
Phase Iv

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. The diurnal regulation of Rubisco was compared for a range of crassulacean acid metabolism (CAM) species in the context of high carboxylation and electron transport capacities, which may be an order of magnitude greater than rates of net CO2 uptake. Early in the light period, Rubisco activity and electron transport were limited when phosphoenolpyruvate carboxylase (PEPC) may have been operating, and maximal extractable activities and activation state for Rubisco were achieved at the end of Phase III, prior to the direct atmospheric uptake of CO2 during Phase IV. The delayed activation was associated with levels of Rubisco activase protein, which reached a maximum at midday, and may account for this pattern of Rubisco activation. This regulation may be modified by environmental conditions - processes that tend to restrict PEPC activity, such as drought stress or incubation of leaves overnight in an oxygen-free atmosphere, release Rubisco from inhibition early in the light period. The quantum yield of light use also tracks Rubisco carboxylation, being particularly low at dawn when PEPC is active. The plasticity in expression of the CAM cycle is therefore matched by the regulation of key carboxylases, with extractable Rubisco activity maximal when drawdown of atmospheric CO2 to cells in succulent CAM tissues is most likely to limit photon utilization shortly after midday, during Phase IV.

Carbohydrate partitioning in the leaves of Bromeliaceae performing C3 photosynthesis or Crassulacean acid metabolism

1998 ◽  
Vol 25 (3) ◽  
pp. 371 ◽  
Author(s):  
John T. Christopher ◽  
Joseph A. M. Holtum
Keyword(s):  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
Soluble Sugars ◽  
Carbohydrate Accumulation ◽  
Carbohydrate Partitioning ◽  
C3 Species ◽  
C3 Photosynthesis ◽  
Cam Species

Carbohydrate accumulation was measured in the leaves of 11 speciesrepresenting the three subfamilies of Bromeliaceae. In the Tillandsioideae the C3 species Vriesea carinata Wawra accumulated starch and sucrose while the Crassulacean acid metabolism (CAM)species Tillandsia tricolor Schlechtendal & Chamissoaccumulated mainly starch. In the Pitcairnioideae the C3species Pitcairnia paniculata Ruiz & Pavon and two CAM species Dyckia sp. andFosterella schidosperma Barker accumulated sucrose butnot starch. Of six CAM species in the Bromelioideae, threeCryptanthus zonatus (Visiani) Beer,Neoregalia spectabilis Moore andPortea petropolitana Wawra accumulated starch but notsoluble sugars while three (Ananus comosus Linnaeus,Orthophytum vagans M.B. Foster andNidularium bilbergioides Schultes filius) accumulatedstarch as well as soluble sugars. Carbohydrate accumulation patterns weresimilar for species within each subfamily in that the Pitcairnioideae speciesdid not accumulate starch but accumulated sucrose while species from theTillandsioideae and Bromelioideae all accumulated starch (some alsoaccumulated soluble sugars). Carbohydrate accumulation patterns were notsimilar for C3 species versus CAM species from thedifferent subfamilies. These data suggest that variations in carbohydratebiochemistry resulting from different evolutionary histories have a greaterinfluence on carbohydrate accumulation patterns in CAM bromeliads than theconstraints of the CAM pathway itself.

Metabolic control of photosynthetic electron transport in crassulacean acid metabolism-induced Mesembryanthemum crystallinum

2002 ◽  
Vol 29 (6) ◽  
pp. 697 ◽  
Author(s):  
Mark Aurel Schöttler ◽  
Helmut Kirchhoff ◽  
Engelbert Weis ◽  
Katharina Siebke
Keyword(s):  
Electron Transport ◽  
Chlorophyll A ◽  
Chlorophyll A Fluorescence ◽  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
Electron Flux ◽  
Photosynthetic Electron Transport ◽  
Mesembryanthemum Crystallinum ◽  
Phase Iii ◽  
Linear Electron

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. We investigated photosynthetic electron transport in leaves of the facultative crassulacean acid metabolism (CAM) plant Mesembryanthemum crystallinum L. After CAM induction, electron transport exhibited variable redox kinetics during the diurnal CAM cycle. In CAM Phase IV, most of PSI (P700) and chlorophyll a fluorescence relaxed with a halftime of 20 ms after a saturating light pulse. This time-constant may reflect the overall linear electron flux from PSII to PSI in saturating light. Comparable relaxation kinetics were also determined for C3 plants. At the end of CAM Phase I and during Phase II, slow components (> 50% of signal amplitude) appeared in both P700 reduction and fluorescence relaxation. They displayed halftimes > 250 ms and > 1 s, suggesting a strong restriction of the linear electron flux from H2O to NADP. The appearance of the slow redox components was accompanied by a decrease in the Fv/Fm ratio of chlorophyll a fluorescence, suggesting a reversible detachment of light-harvesting complex (LHC) II from PSII. The slow redox fractions and the depression of Fv/Fm disappeared again in parallel to malate decarboxylation during CAM Phase III. We discuss a reversible downregulation of linear electron flux during CAM Phases II and III, due to a reversible deprivation of cytochrome-b6f complexes (cyt-bfs) and PSI from the linear system. In parallel, a redistribution of some LHCIIs could also occur. This could be an adaptive response to a reduced metabolic demand for NADPH due to a limited carbon flux through the Calvin cycle, resulting from low Rubisco activation. Furthermore, the cyt-bfs and PSIs deprived of linear electron transport could support cyclic electron flux to cover an increased ATP demand during gluconeogenesis in CAM Phase III.

scholarly journals Effect of Elevated Carbon Dioxide on the Growth and Physiological Responses of Pineapple, A Species with Crassulacean Acid Metabolism

1997 ◽  
Vol 122 (2) ◽  
pp. 233-237 ◽  
Author(s):  
Jun Zhu ◽  
Duane P. Bartholomew ◽  
Guillermo Goldstein
Keyword(s):  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
Titratable Acidity ◽  
Early Morning ◽  
Dry Mass ◽  
Ananas Comosus ◽  
Carbon Isotopic ◽  
Diurnal Difference ◽  
Dark Fixation ◽  
Average Plant

Despite the potential impact of rising global CO2 levels, only a limited number of studies have been conducted on the effects of ambient and elevated CO2 on plants having Crassulacean acid metabolism (CAM). To our knowledge, there are no studies for pineapple [Ananas comosus (L.) Merr.], the most commercially important CAM plant. Pineapple plants were grown at CO2 levels of ≈330 (ambient) and ≈730 (elevated) μmol·mol-1 in open-top chambers for 4 months. The mean air temperature in the chambers was ≈39 °C day/24 °C night. Average plant dry mass at harvest was 180 g per plant at elevated CO2 and 146 g per plant at ambient CO2. More biomass was partitioned to stem and root but less to leaf for plants grown at elevated CO2; leaf thickness was 11% greater at elevated than at ambient CO2. The diurnal difference in leaf titratable acidity (H+) at elevated CO2 reached 347 mmol·m-2, which was up to 42% greater than levels in plants grown in ambient CO2. Carbon isotopic discrimination (Δ) of plants was 3.75% at ambient CO2 and 3.17% at elevated CO2, indicating that CO2 uptake via the CAM pathway was enhanced more by elevated CO2 than uptake via the C3 pathway. The nonphotochemical quenching coefficient (qN) of leaves was ≈45% lower in the early morning for plants grown at elevated than at ambient CO2, while afternoon values were comparable. The qN data suggested that the fixation of external CO2 was enhanced by elevated CO2 in the morning but not in the afternoon when leaf temperature was ≥40 °C. We found no effect of CO2 levels on leaf N or chlorophyll content. Pineapple dry matter gain was enhanced by elevated CO2, mainly due to increased CO2 dark fixation in environments with day temperatures high enough to suppress C3 photosynthesis.

Carbohydrate partitioning in crassulacean acid metabolism plants: reconciling potential conflicts of interest

2002 ◽  
Vol 29 (6) ◽  
pp. 707 ◽  
Author(s):  
Anne M. Borland ◽  
Antony N. Dodd
Keyword(s):  
Crassulacean Acid Metabolism ◽  
Acid Metabolism ◽  
Conflicts Of Interest ◽  
Critical Mass ◽  
Soluble Sugars ◽  
Carbon Budgets ◽  
Cam Plants ◽  
Carbohydrate Partitioning ◽  
Isotopic Signatures ◽  
High Level

This paper originates from a presentation at the IIIrd International Congress on Crassulacean Acid Metabolism, Cape Tribulation, Queensland, Australia, August 2001. The construction of diel leaf carbon budgets, together with analyses of the δ13C composition of biochemical fractions, was used to examine how crassulacean acid metabolism (CAM) plants adjust carbohydrate partitioning in response to shifting sink demands. For Mesembryanthemum crystallinum L., net carbon budgets indicated clear shifts in assimilate partitioning and in the relative proportions of day : night export as CAM was induced. Different patterns of carbohydrate partitioning in primary and axillary leaves of this species may reflect the different sink priorities of determinate and indeterminate growth. In primary leaves, the high level of diel starch turnover may be a strategy for ensuring production of a critical mass of juvenile tissue that poises the plant for CAM induction. In axillary leaves, the high day-night flux through soluble sugars may ensure ready availability of assimilates for export to reproductive sinks. Carbon isotope ratios were measured for various organic fractions isolated from leaves and fruits of two species of Clusia that differ in CAM expression. Similar and C3-like isotopic signatures were obtained for the structural material isolated from fruits of Clusia minor L. and Clusia rosea Jacq. The data suggest that the partitioning of C4- and C3-derived assimilates into discrete storage and transport pools of soluble sugars will ensure reproductive output, regardless of the level of CAM that is induced in these species.

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