Isolation of Temperature-Sensitive Membrane Mutants of Escherichia coli K-12

Mutants with impaired biosynthesis of unsaturated fatty acids or altered metabolism of the phospholipids were isolated at a rather high frequency from a set of temperature-sensitive lysis mutants. It is suggested that preselection for the lysis phenotype makes it possible to isolate several kinds of mutants affected in the integrity of the cytoplasmic membrane.     

Physiological and Biochemical Mechanisms Involved in the Response to Abscisic Acid in Maize Coleoptiles

The role of abscisic acid (ABA) in controlling growth and developmenthas been studied in maize (Zea mays L.) coleoptile segments.Application of ABA reduces the elongation rate by about 50%and affects ion fluxes. In particular, proton extrusion is decreasedwhile potassium efflux is greatly enhanced. Apparently, ABAdoes not: seem to influence calcium influx from the apoplastinto the cytosol, but more likely it influences its efflux.Alteration of cytosolic calcium concentration may also be obtainedby increasing its release from internal stores. This possibilitymight be sustained by the increased hydrolysis of phosphatidylinositolupon ABA application. Change in the balance of ion fluxes shouldresult from regulation of transport mechanisms at the membranelevel and should produce changes in the transmembrane electricalpotential. The H⁺- ATPase and the ATP-dependent calcium transportactivities are both influenced by the treatment with ABA, –55%and –40%, respectively. Under these conditions [Ca²⁺]_cytand pH_cyt can be modified and, as a consequence of their regulation,they may play an important role in mediating the physiologicaland biochemical effects of ABA, acting as second intracellularmessengers. ¹Research supported by National Research Council of Italy, SpecialProject RAISA, Sub-Project N. 2, Paper n. 2782.     

Calcium-dependent binding of EDTA-extractable proteins to calf lens fiber membrane structures

Calf lens fiber membranes and fractions enriched in junction-like structures have been isolated in the absence and presence of EDTA. Their biochemical features have been studied. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting experiments have provided evidence that a distinct group of EDTA-extractable proteins, being one of the main protein components of calf lens fiber membranes and very likely also of junction-like structures, is bound to these membranes via calcium ions. In addition to these proteins, four polypeptides with apparent molecular weights between 14 000 and 17 000 are characteristic for detergent-insoluble lens fiber structures prepared in calcium-rich medium. The absence of EDTA-extractable proteins in the urea-soluble calcium-containing fraction implies that they are not components of the cytoskeleton and that the calcium-dependent binding of these proteins to the membrane is urea-resistant. The use of EDTA throughout the whole membrane isolation procedure results in their complete removal from the membranes which already starts during buffer washing. This indicates that EDTA-extractable proteins exclusively consist of extrinsic membrane proteins which probably are not involved in cytoskeleton binding.     

Allylic palladium(II) triazenido complexes: synthesis and spectroscopic studies

Reaction of [Pd(1-3-η-allyl)Cl]₂ with lithium triazenide (triazenide = p-XC₆H₄NN-NC₆H₄X-p; X = Cl, H, CH₃) affords dimeric complexes of the type [Pd(1-3-η-allyl)(triazenide)]₂. In the solid state the triazenido ligands are bridging two palladium atoms with their terminal nitrogen atoms, as shown by a preliminary X-ray determination of the complex with X = CH₃. The allyl groups are stereochemically equivalent. ¹H NMR spectra demonstrate the presence of two conformers in solution. The major component has the same configuration found in the solid. The other conformer has stereochemically non equivalent allyl groups. The concentration ratio of the two conformers is independent of the temperature, suggesting the absence of intramolecular processes and of palladium- triazenido bond breaking. This point is discussed also by comparing the (1-3-η-allyl)(triazenide)palladium (II) dimers with the closely related(1-3-η-allyl)(acetate)palladium(II) complexes.     

Thermographic visualization of cell death in tobacco and Arabidopsis

Pending cell death was visualized by thermographic imaging in bacterio‐opsin transgenic tobacco plants. Cell death in these plants was characterized by a complex lesion phenotype. Isolated cell death lesions were preceded by a colocalized thermal effect, as previously observed at sites infected by tobacco mosaic virus (TMV) ( Chaerle et al. 1999 Nature Biotechnology 17, 813–816). However, in most cases, a coherent front of higher temperature, trailed by cell death, initiated at the leaf base and expanded over the leaf lamina. In contrast to the homogenous thermal front, cell death was first visible close to the veins, and subsequently appeared as discrete spots on the interveinal tissue, as cell death spread along the veins. Regions with visible cell death had a lower temperature because of water evaporation from damaged cells. In analogy with previous observations on the localized tobacco–TMV interaction ( Chaerle et al. 1999 ), the kinetics of thermographic and continuous gas exchange measurements indicated that stomatal closure preceded tissue collapse. Localized spontaneous cell death could also be presymptomatically visualized in the Arabidopsis lsd2 mutant.