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Phosphoenolpyruvate Carboxylase During Development of Crassulacean Acid Metabolism and During a Diurnal Cycle in Mesembryanthemum Crystallinum
Authors:GREENWAY  H; WINTER  K; LUTTGE  U
Abstract:Crassulacean acid metabolism (CAM) in Mesembryanthemum crystallinumwas induced by transfer of plants from 100 to 400 mM NaCl. Diurnalmalate fluctuations developed slowly; maximum rates of net malatesynthesis in the dark were reached only on the 10th day afterNaCl was increased to 400 mM. In contrast, phosphoenolpyruvatecarboxylase (PEPC) activity, assayed at optimum pH of 8–0,had nearly reached its maximum on the 5th day after plants weretransferred to 400 mM NaCl. Characteristics of PEPC changedduring the first 12 d of exposure of plants to 400 mM NaCl.There were increases in the ratio of PEPC activity at pH 7 0/PEPCactivity at pH 8.0, and decreases in the Km for PEP measuredat pH 7.0, and possibly in the degree of malate inhibition.All further measurements were made once CAM was well established.In vivo rates of malate synthesis were 14–18 times smallerthan PEPC activity at 2 mM PEP, both processes being measuredat 15 °C. It is suggested that the high PEPC levels favourrapid, preferential flow of carbon to malate, by maintainingvery low PEP levels in the cytoplasm. PEPC changed in characteristicsduring the diurnal cycle. During the first few minutes afterisolation, extracts made during the first hours of the day,when malate was consumed, showed very low PEPC activity at pH7.0 but high activity at pH 8.0. The activity of PEPC at pH7.0 rose gradually during storage of the extracts at 0 °C,usually reaching the activity at pH 8.0 after about 30–50min. In contrast, extracts obtained during the first hours ofthe night, when malate was synthesized, showed high PEPC activityat both pH 7.0 and 8–0 within 30–50 s after extraction.The results indicate that PEPC of M. crystallinum, performingdistinct CAM, may exist in two states. One state would favourrapid malate synthesis and transport to the vacuoles and wouldfunction during the night. The second state, with little activitybelow pH 7.5, would occur during the day, thus preventing complicationsof continued synthesis of malate while it is converted to carbohydrates.
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