Phosphoenolpyruvate Carboxylase from the Crassulacean PlantBryophyllum fedtschenkoiHametet Perrier: ACTIVITY CHANGES AND KINETIC BEHAVIOUR IN CRUDE EXTRACTS

The total activity (capacity) of phosphoenolpyruvate carboxylasein crude extracts of leaves of Bryophyllum fedtschenkoi, measuredon a fresh weight basis, varies considerably with leaf positionand with photoperiodic treatment. No diurnal variation in capacitywas detected under long – day or short – day conditions.The enzyme in freshly prepared crude extracts is about 50 timesmore sensitive to inhibition by malate at pH 7 than that inaged extracts, or in the fully purified state. Desensitizationin extracts proceeds rapidly unless protective measures aretaken, and appears to be irreversible. A pH-dependence studyshows that the effect of desensitization on the kinetic parametersof malate inhibition is identical to that of increasing thepH by 2.0 units over a wide range, but the maximum velocityat pH 7.8 is virtually unaffected. The significance of the resultsfor both the experimental determination of cyclic changes inenzyme capacity, and the theories concerning the mechanism ofa circadian rhythm of phosphoenolpyruvate carboxylase activityin vivo are discussed.     

Overexpression of Arabidopsis acyl‐CoA‐binding protein ACBP2 enhances drought tolerance

Arabidopsis thaliana acyl‐CoA‐binding protein 2 (ACBP2) is a stress‐responsive protein that is also important in embryogenesis. Here, we assign a role for ACBP2 in abscisic acid (ABA) signalling during seed germination, seedling development and the drought response. ACBP2 was induced by ABA and drought, and transgenic Arabidopsis overexpressing ACBP2 (ACBP2‐OXs) showed increased sensitivity to ABA treatment during germination and seedling development. ACBP2‐OXs also displayed improved drought tolerance and ABA‐mediated reactive oxygen species (ROS) production in guard cells, thereby promoting stomatal closure, reducing water loss and enhancing drought tolerance. In contrast, acbp2 mutant plants showed decreased sensitivity to ABA in root development and were more sensitive to drought stress. RNA analyses revealed that ACBP2 overexpression up‐regulated the expression of Respiratory Burst Oxidase Homolog D (AtrbohD) and AtrbohF, two NAD(P)H oxidases essential for ABA‐mediated ROS production, whereas the expression of Hypersensitive to ABA1 (HAB1), an important negative regulator in ABA signalling, was down‐regulated. In addition, transgenic plants expressing ACBP2pro:GUS showed beta‐glucuronidase (GUS) staining in guard cells, confirming a role for ACBP2 at the stomata. These observations support a positive role for ACBP2 in promoting ABA signalling in germination, seedling development and the drought response.