Regulation of malic-acid metabolism in Crassulacean-acid-metabolism plants in the dark and light: In-vivo evidence from (13)C-labeling patterns after (13)CO 2 fixation.
The labeling patterns in malic acid from dark (13)CO2 fixation in seven species of succulent plants with Crassulacean acid metabolism were analysed by gas chromatography-mass spectrometry and (13)C-nuclear magnetic resonance spectrometry. Only singly labeled malic-acid molecules were detected and on the average, after 12-14 h dark (13)CO2 fixation the ratio of [4-(13)C] to [1-(13)C] label was 2:1. However the 4-C carboxyl contained from 72 to 50% of the label depending on species and temperature. The (13)C enrichment of malate and fumarate was similar. These data confirm those of W. Cockburn and A. McAuley (1975, Plant Physiol. 55, 87-89) and indicate fumarase randomization is responsible for movement of label to 1-C malic acid following carboxylation of phosphoenolpyruvate. The extent of randomization may depend on time and on the balance of malic-acid fluxes between mitochondria and vacuoles. The ratio of labeling in 4-C to 1-C of malic acid which accumulated following (13)CO2 fixation in the dark did not change during deacidification in the light and no doubly-labeled molecules of malic acid were detected. These results indicate that further fumarase randomization does not occur in the light, and futile cycling of decarboxylation products of [(13)C] malic acid ((13)CO2 or [1-(13)C]pyruvate) through phosphoenolpyruvate carboxylase does not occur, presumably because malic acid inhibits this enzyme in the light in vivo. Short-term exposure to (13)CO2 in the light after deacidification leads to the synthesis of singly and multiply labeled malic acid in these species, as observed by E.W. Ritz et al. (1986, Planta 167, 284-291). In the shortest times, only singly-labeled [4-(13)C]malate was detected but this may be a consequence of the higher intensity and better detection statistics of this ion cluster during mass spectrometry. We conclude that both phosphoenolpyruvate carboxylase (EC 4.1.1.32) and ribulose-1,5-biphosphate carboxylase (EC 4.1.1.39) are active at this time.
Plant Environmental Biology Group, Research School of Biological Sciences, Australian National University, P.O. Box 475, 2601, Canberra, A.C.T., Australia.
C B
CB
Osmond
J A
JA
Holtum
M H
MH
O'Leary
C
C
Roeske
O C
OC
Wong
R E
RE
Summons
P N
PN
Avadhani