Pre-concentration and measurement of low levels of gamma-ray emitting radioisotopes in coastal waters.

We describe extensive testing of a large-volume, high-speed water sampler for the concentration and measurement of radionuclides using high-resolution gamma ray spectrometry. The sampler processed hundreds to thousands of liters of natural waters with variable suspended sediment and salinity loads at flow rates of 10-201/min. Extraction of most radionuclides in the water column was accomplished through the combination of physical filtration down to 0.1 microm particle size and chemical separation of dissolved species on cellulose-based inorganic sorbent beds without recourse to complex, or hazardous chemistry. Performance and extraction efficiencies for suites of radioisotopes were determined in the laboratory and in the field with river and coastal ocean water samples. Extraction and recovery efficiencies are better than 90% for most fission and activation product radioisotopes. This methodology has broad application to the study of the distribution and fate of radioisotopes in coastal waterways.

Induction of delta-cadinene synthase and sesquiterpenoid phytoalexins in cotton by Verticillium dahliae.

Phytoalexin biosynthesis occurred earlier in the resistant cotton cultivar Seabrook Sea Island 12B2 (SBSI) (Gossypium barbadense) than in the susceptible cotton cultivar Rowden (G. hirsutum) after inoculation with a defoliating isolate of the pathogen Verticillium dahliae. This was demonstrated by significantly higher levels of phytoalexins in SBSI 12 h after inoculation. Furthermore, by 48 h after inoculation of SBSI, the phytoalexins hemigossypol and desoxyhemigossypol achieved levels (23.9 and 10.5 microgram/g of fresh tissue, respectively) sufficient to completely inhibit conidial germination. Rowden required 96 h to attain comparable levels. Similarly, the activity of delta-cadinene synthase, a key enzyme required for the biosynthesis of the terpenoid phytoalexins, increased more rapidly in the resistant cotton cultivar than in the susceptible one. The changes in phytoalexin concentrations and enzyme activity are consistent with the hypothesis that phytoalexins are an essential component in protecting the plant from infection by V. dahliae.

Regulation of HMG-CoA reductase activity in plants.

This brief review summarizes the current literature on the regulation of HMG-CoA reductase (HMGR) in plants. The mevalonate pathway, which starts with the synthesis of mevalonate by HMGR, has more branch pathways in plants than in most other organisms, leading to a tremendous variety of isoprenoid products. Evidence suggests that HMGR is an important control point for the synthesis of many of these plant isoprenoids, including some that are vital for primary metabolism and pest resistance. Plant HMGR activity responds in vivo to a variety of developmental and environmental signals, such as cell division, light, and infection. Plants regulate HMGR activity at the level of mRNA by differential induction of HMGR gene family members, and posttranslationally by enzyme modification. Calcium, calmodulin, and proteolytic degradation may also have a role in regulation of plant HMGR.

Isolation of adenylate cyclase mutants from Rhizobium meliloti deficient in nodulation.

Six Rhizobium meliloti mutants were isolated after Tn5-mediated mutagenesis as resistant to inhibition by a mixture of amino acids (serine, methionine, glycine and leucine). All were defective in adenylate cyclase activity and failed to form nodules in infected roots of Medicago sativa. Furthermore, like other nodulation mutants, they showed altered motility and increased secretion of exopolysaccharides; addition of cAMP to the growth medium abolished some of these phenotypic defects. The possibility that adenylate cyclase participates in the transduction of signals inducing nodulation is discussed.

Adenylate cyclase activity in cyanobacteria: activation by Ca(2+)-calmodulin and a calmodulin-like activity.

An adenylate cyclase activity was partially characterized in the cyanobacterium Anabaena sp. The enzyme activity is found in soluble cell fractions and shows an apparent molecular weight of about 183,400. This adenylate cyclase is activated by Ca2+ and bovine brain or spinach calmodulin and it is inhibited by EGTA and some phenothiazine derivatives. Furthermore, Anabaena sp. extracts contain a calmodulin-like activity which stimulates bovine brain cyclic AMP phosphodiesterase and the Anabaena adenylate cyclase. EGTA and phenothiazine derivatives block the cyanobacterial modulator effect.

Reconstitution of a light-stimulated adenylate cyclase from retina and Neurospora crassa preparations. Characterization of the heterologous systems using normal and degenerative retinas.

Adenylate cyclase catalytic subunits from Neurospora crassa membranes may interact with regulatory factors from membranes of bovine retinal rod outer segments (pretreated with N-ethylmaleimide), reconstituting a heterologous system which, in the presence of light, is catalytically active in assay mixtures containing MgATP. Maximal activation was observed at 550 nm. Transducin-depleted retinal membranes were not capable of reconstituting the heterologous light-stimulated adenylate cyclase system. Addition of a transducin preparation to depleted membranes restored the reconstitution capacity of these membranes. A similar heterologous adenylate cyclase system was reconstituted with Neurospora and mouse retinal whole membranes (pretreated with N-ethylmaleimide). Membranes from mice suffering photoreceptor degeneration (rd homozygotes) did not reconstitute an heterologous adenylate cyclase system.

Adenylate cyclase activity in a higher plant, alfalfa (Medicago sativa).

An adenylate cyclase activity in Medicago sativa L. (alfalfa) roots was partially characterized. The enzyme activity remains in the supernatant fluid after centrifugation at 105,000 g and shows in crude extracts an apparent Mr of about 84,000. The enzyme is active with Mg2+ and Ca2+ as bivalent cations, and is inhibited by EGTA and by chlorpromazine. Calmodulin from bovine brain or spinach leaves activates this adenylate cyclase.