Citrulline and DRIP-1 Protein (ArgE Homologue) in Drought Tolerance of Wild Watermelon

Drought-affected plants experience more than just desiccationof their organs due to water deficit. Plants transpire 1000times more molecules of water than of CO₂ fixed by photosynthesisin full sunlight. One effect of transpiration is to cool theleaves. Accordingly, drought brings about such multi-stressesas high temperatures, excess photoradiation and other factorsthat affect plant viability. Wild watermelon serves as a suitablemodel system to study drought responses of C₃ plants, sincethis plant survives drought by maintaining its water contentwithout any wilting of leaves or desiccation even under severedrought conditions. Under drought conditions in the presenceof strong light, wild watermelon accumulates high concentrationsof citrulline, glutamate and arginine in its leaves. The accumulationof citrulline and arginine may be related to the induction ofDRIP-1, a homologue of ArgE in Escherichia coli, where it functionsto incorporate the carbon skeleton of glutamate into the ureacycle. Immunogold electron microscopy reveals the enzyme tobe confined exclusively to the cytosol. DRIP-1 is also inducedby treating wild watermelon with 150 mM NaCl, but is not inducedfollowing treatment with 100 µM abscisic acid. The salttreatment causes the accumulation of -aminobutyrate, glutamineand alanine, in addition to a smaller amount of citrulline.Citrulline may function as a potent hydroxyl radical scavenger.     

Studies On Cyclic Nucleotide Metabolism In Tetrahymena Pyriformis: Partial Characterization of Cyclic Amp- and Cyclic Gmp-Dependent Phosphodiesterases*

SYNOPSIS. Cyclic nucleotide phosphodiesterase [EC 3.1.4.17] was examined in Tetrahymena pyriformis strain NT-1. Enzymic activity was associated with the soluble and the particulate fractions, whereas most of the cyclic GMP phosphodiesterase activity was localized in the soluble fraction: the activities were optimal at pH 8.0–9.0. Although very low activities were detected in the absence of divalent cations, they were significantly increased by the addition of either Mg²⁺ or Mn^2-. A kinetic analysis of the properties of the enzymes yielded 2 apparent K_III values ranging in concentration from 0.5 to 50 μM and from 0.1 to 62 μ M for cyclic AMP and GMP. respectively. A Ca²⁺-dependent activating factor for cyclic nucleotide phosphodiesterase was extracted from Tetrahymena cells, but this factor did not stimulate guanylate cyclase [EC 4.6.1.2] activity in this organism. On the other hand, Tetrahymena also contained a protein activator which stimulated guanylate cyclase in the presence of Ca²⁺, although this activator did not stimulate the phosphodiesterase. the results suggested that Tetrahymena might contain 2 types of Ca²⁺-dependent activators, one specific for phosphodiesterase and the other for guanylate cyclase.     

The long-term responses of the photosynthetic proton circuit to drought

Proton motive force (pmf) across thylakoid membranes is not only for harnessing solar energy for photosynthetic CO₂ fixation, but also for triggering feedback regulation of photosystem II antenna. The mechanisms for balancing these two roles of the proton circuit under the long-term environmental stress, such as prolonged drought, have been poorly understood. In this study, we report on the response of wild watermelon thylakoid 'proton circuit' to drought stress using both in vivo spectroscopy and molecular analyses of the representative photosynthetic components. Although drought stress led to enhanced proton flux via a ∼34% increase in cyclic electron flow around photosystem I (PS I), an observed ∼fivefold decrease in proton conductivity, g_H⁺, across thylakoid membranes suggested that decreased ATP synthase activity was the major factor for sustaining elevated q_E. Western blotting analyses revealed that ATP synthase content decreased significantly, suggesting that quantitative control of the complex plays a pivotal role in down-regulation of g_H⁺. The expression level of cytochrome b ₆ f complex – another key control point in photosynthesis – also declined, probably to prevent excess-reduction of PS I electron acceptors. We conclude that plant acclimation to long-term environmental stress involves global changes in the photosynthetic proton circuit, in which ATP synthase represents the key control point for regulating the relationship between electron transfer and pmf.