Root phospholipids in Azospirillum-inoculated wheat seedlings exposed to water stress.

Azospirillum-plant association is accompanied by biochemical changes in roots which, in turn, promote plant-growth and tolerance to water stress. To shed light on the possible factors underlying these effects, roots from Azospirillum brasilense Sp245-inoculated Triticum aestivum seedlings growing in darkness under osmotic stress were analyzed for phospholipid (PL) composition, fatty acid (FA) distribution profiles and degree of unsaturation of the major PL classes. Azospirillum inoculation diminished ion leakage and increased 2,3,5-tripheniltetrazolium reducing ability in roots of well irrigated and water-stressed wheat seedlings. Total root PL content remained unaltered in all treatments. Six PL classes were detected, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) comprising over 80% of the total. While water stress increased PC content and diminished that of PE, none of these changes were observed either under Azospirillum inoculation alone or when both treatments were combined. The major FAs found in both PC and PE were 16:0, 18:0, 18:1, 18:2, and 18:3. Higher PC and lower PE unsaturation than in well irrigated controls were observed in roots from Azospirillum-inoculated, water-stressed seedlings. Azospirillum inoculation could contribute to protect wheat seedlings from water stress through changes in the FA distribution profiles of PC and PE major root phospholipids.

Determination of chlorimuron and metsulfuron residues in two soils of Argentina using a rapid seed-bioassay.

The presence of chlorimuron ethyl and metsulfuron methyl in two soils was determined by a modified petri dish bioassay. Pregerminated seeds of maize and sunflower were placed in petri dishes containing 85 to 100 g of treated soil. Radicle root lengths were measured after 24 h. Chlorimuron had no effect on maize on the Balcarce soil, however 0.007 microg g(-1) decreased sunflower root length. Chlorimuron decreased maize and sunflower root length regardless application dose on the San Cayetano soil. Metsulfuron decreased maize root length at 0.04 microg g(-1) and sunflower at 0.021 microg g(-1) on the Balcarce soil. On the San Cayetano soil metsulfuron at 0.001 microg g(-1) decreased maize and sunflower root length. The phytotoxicity of chlorimuron and metsulfuron changed according to soil type and dose. Maize and sunflower were 1.3-1.5 and 1.3-1.8 times respectively more sensitive to chlorimuron on the San Cayetano soil than on the Balcarce soil. In the case of metsulfuron, maize was similarly sensitive on both soils but sunflower was 1.7-2.0 times more sensitive on the San Cayetano soil than on the Balcarce soil. Phytotoxicity increased as organic matter (OM) content decreased and/or when the soil pH and concentration increased.

Immunological analysis of the heme proteins present in aerobically grown Escherichia coli.

Immunological methods were used to obtain information about Escherichia coli heme proteins. There is a membrane-bound catalase which consists of a single subunit (as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis) which is also present in the soluble fraction. Antibodies raised against purified, soluble cytochrome b562 showed that this cytochrome is not related to any of the membrane-bound cytochromes, including the b562 component of the cytochrome o complex. Cytochrome b556 is immunologically unrelated to the cytochrome b556 NR associated with the nitrate reductase system. Cytochrome b556 and cytochrome o are not present in a constant ratio in the membrane.

Succinate dehydrogenase in Rhodopseudomonas sphaeroides: subunit composition and immunocross-reactivity with other related bacteria.

Antibodies were raised against the succinate dehydrogenase (SDH) present in the chromatophores of phototrophically grown Rhodopseudomonas sphaeroides. Crossed immunoelectrophoresis experiments indicated that the SDH present in the cytoplasmic membranes of heterotrophically grown R. sphaeroides is probably the same enzyme observed in the chromatophores. The enzyme was extracted by Triton X-100 in a form which consisted of only two subunits (molecular weight, 68,000 and 30,000) and was not associated with a cytochrome b. The antibodies directed against SDH from R. sphaeroides showed no immunocross-reactivity with SDH from phylogenetically related bacterial species, including Rhodopseudomonas capsulata, Paracoccus denitrificans, Rhodopseudomonas palustris, Rhodospirillum rubrum, and Rhodospirillum fulvum.

Characterization of monoclonal antibodies directed against pyruvate oxidase from Escherichia coli: modulation of antibody-induced inhibition by enzyme conformation.

Monoclonal antibodies have been prepared against pyruvate oxidase, a flavoprotein dehydrogenase isolated from Escherichia coli. Six monoclonals were obtained, but only one was found to bind to the native form of the enzyme. This monoclonal, 1I1, was a potent inhibitor. Although this antibody inhibited the unactivated and lipid-activated forms of the enzyme, it had much less of an inhibitory effect on the protease-activated form of the enzyme, although the antibody still bound to this form. Hence, the coupling between antibody binding and the conformation at the active site can itself be modulated by the conformation of the protein.

Immunological characterization of an Escherichia coli strain which is lacking cytochrome d.

The isolated membranes from an Escherichia coli mutant strain which lacks spectroscopically detectable levels of cytochromes d, a1, and b558 also have abnormally low levels of N,N,N',N'-tetramethyl-p-phenylenediamine oxidase activity. In this paper, it is shown that the material previously identified as the N,N,N',N'-tetramethyl-p-phenylenediamine oxidase is, in fact, the two-subunit cytochrome d complex. Antisera directed against the native cytochrome d complex as well as against each of two subunits apparent on sodium dodecyl sulfate-polyacrylamide gels were used to show that the mutant strain lacks both subunits of the cytochrome d complex. Introduction of F-prime F152 into the mutant strain restored the two subunits along with the spectroscopic and enzymatic activity associated with the cytochrome d complex.

Modified, large-scale purification of the cytochrome o complex (bo-type oxidase) of Escherichia coli yields a two heme/one copper terminal oxidase with high specific activity.

The cytochrome o complex is a bo-type ubiquinol oxidase in the aerobic respiratory chain of Escherichia coli. This complex has a close structural and functional relationship with the eukaryotic and prokaryotic aa3-type cytochrome c oxidases. The specific activity, subunit composition, and metal content of the purified cytochrome o complex are not consistent for different preparative protocols reported in the literature. This paper presents a relatively simple preparation of the enzyme starting with a strain of Escherichia coli which overproduces the oxidase. The pure enzyme contains four subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Partial amino acid sequence data confirm the identities of subunit I, II, and III from the SDS-PAGE analysis as the cyoB, cyoA, and cyoC gene products, respectively. A slight modification of the purification protocol yields an oxidase preparation that contains a possible fifth subunit which may be the cyoE gene product. The pure four-subunit enzyme contains 2 equivs of iron but only 1 equiv of copper. There is no electron paramagnetic resonance detectable copper in the purified enzyme. Hence, the equivalent of CuA of the aa3-type cytochrome c oxidases is absent in this quinol oxidase. There is also no zinc in the purified quinol oxidase. Finally, monoclonal antibodies are reported that interact with subunit II. One of these monoclonals inhibits the quinol oxidase activity of the detergent-solubilized, purified oxidase. Hence, although subunit II does not contain CuA and does not interact with cytochrome c, it still must have an important function in the bo-type ubiquinol oxidase.