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81.
Harry J. Klee Maria B. Hayford Stephen G. Rogers 《Molecular & general genetics : MGG》1987,210(2):282-287
Summary Cloning of genes by transformation with genomic banks and rescue of a phenotype has been extensively used in bacterial systems. This approach has not been possible in plant systems because of the large genome sizes and poor transformation frequencies of most plant species. Recent advances in plant transformation permit the generation of large numbers of transformants in petunia. We have used this system to rescue a model gene encoding resistance to kanamycin by shotgun cloning. The gene encoding neomycin phosphotransferase (NPTII) was introduced into the genome of Arabidopsis thaliana by Agrobacterium tumefaciens-mediated transformation. A genomic bank of DNA from this tissue was constructed in a cosmid vector containing features which would allow its use in plant transformation. The unselected genomic bank was mobilized from Escherichia coli to A. tumefaciens and used to retransform petunia leaf discs. The rescued gene was identified by its ability to confer a kanamycin-resistant phenotype in petunia tissue. The presence of the NPTII gene was confirmed by nopaline assay and Southern blot analysis. This experiment demonstrates the feasibility of gene rescue, in certain circumstances, in plants. 相似文献
82.
The action of the herbicide glyphosate [N-(phosphonomethyl)-glycine] on phenolic metabolism and phenylalanine ammonia lyase (PAL; EC 4.3.1.5) activity was investigated in yellow nutsedge (Cyperus esculentus L.). Glyphosate caused significant increases in the amount of total soluble hydroxyphenolics in the three fractions studied (neutral, acid and residual). Qualitative and quantitative differences in relation to these fractions and the amount of applied glyphosate were observed. Most of the phenolic compounds which increased after glyphosate treatment were benzoic acids (gentisic. p -OH-benzoic, salicylic and vanillic). Gentisic acid showed the greatest increase in neutral and acid fractions, being twenty- and four-fold, respectively, of the amount found in the control. PAL activity was not affected by the lowest doses of glyphosate (10−4 and 10−3 M) , but a dramatic decrease in PAL activity was observed after 10−2 M treatment. These findings, together with the low levels of cinnamic acids measured in treated yellow nutsedge plants, suggest that PAL activity is only marginally involved in glyphosate action. Since the herbicidal action probably takes place at 5-enol-pyruvylshikimate-3-P synthase (EC 2.5.1.19), an alternative pathway to PAL in phenolic biosynthesis should be activated yielding benzoic acids. 相似文献
83.
The effects of Rhizobium strain and its interaction with plant cultivar were examined in glasshouse-grownPhaseolus vulgaris in two experiments where the physiological attributes defining the symbiotic efficiency were determined.
Strains of Rhizobium significantly affected nodulation, rates of N accumulation, partitioning of N within the mature shoot
and remobilizaton of the N stored in the vegetative organs to the seeds.
The most efficient symbiosis (strain CO5 with Negro Argel), in comparison with the least efficient symbiosis (strain 127 K-17
with Venezuela-350) showed higher rates of C2H2 reduction from flowering to mid pod fill stage, evolved less hydrogen from nodules and showed higher rates of N transport
as well as higher percentages of ureide-N in the xylem sap. At maturity, the best cultivar/strain association exceeded the
total N accumulated in the seed and the harvest index of the poorest symbiosis in 88% and 20%, respectively. The other symbiotic
combinations were intermediate in all characteristics.
Nitrogen accumulation in plant shoot showed highly significant correlation with acetylene reduction rates, nodule relative
efficiency, total N transport in the xylem sap and percentage of N transported as ureides. 相似文献
84.
Carlos Eduardo Falavigna da Rocha Maria Helena Gonzaga de Carvalho Bjornber 《Hydrobiologia》1987,153(2):97-107
Cyclopid copepods collected mainly in aquatic microcosms and semiterrestrial habitats in the Juréia Ecological Reserve are studied. Hesperocyclops herbsti and Bryocyclops campaneri are described as new species and their taxonomical relationships discussed. Females of Muscocyclops operculatus (Chappuis) are redescribed and the males described for the first time. An emended diagnosis for Muscocyclops is proposed. 相似文献
85.
A monoclonal antibody, 3F12, was generated which reacted specifically against infective or metacyclic stage Leishmania major promastigotes, but not with noninfective promastigotes obtained from log phase cultures. The antibody recognized a cell surface and released molecule that could be metabolically labeled with [14C]glucose, [3H]mannose, [3H]galactose, and [3H]palmitic acid, but not with [35S]methionine or [3H]leucine. The molecule was the major species surface-labeled by [3H]sodium borohydride after periodate treatment. The glycolipid appeared to be shed primarily as free carbohydrate because 70% of the released material partitioned in the aqueous fraction after phase separation in TX-114. The molecule could be distinguished from the L. major glycolipid which has already been extensively described because its migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was of higher relative m.w. However, a close relationship between the two molecules was indicated by the finding that another monoclonal antibody, WIC-79.3, recognized both forms of the glycolipid; one produced and released only by log phase promastigotes, and one produced and released only by metacyclic promastigotes. The loss of agglutination with peanut agglutinin which has been shown to accompany metacyclogenesis was found to be caused by the loss of expression of the log form of the glycolipid which in most cases appeared to be the result of the developmental modification of this molecule. A survey of a number of virulent and avirulent. L. major strains and clones reinforced an absolute association between the ability of these promastigotes to initiate infection in BALB/c mice and their expression and release of the 3F12-binding, developmentally regulated form of the glycolipid. Not only does this glycolipid serve as the first well defined molecular marker for infective stage metacyclic promastigotes, but its unique structure is very likely to contribute to the adaptive changes that allow these parasites to survive within the vertebrate host. 相似文献
86.
Pertussis Toxin Attenuates D2 Inhibition and Enhances D1 Stimulation of Adenylate Cyclase by Dopamine in Rat Striatum 总被引:2,自引:1,他引:1
The response of adenylate cyclase to GTP and to dopamine (DA) was investigated in synaptic plasma membranes isolated from rat striatum injected with pertussis toxin, which inactivates the inhibitory guanine nucleotide-binding regulatory protein (Ni) of adenylate cyclase. Pertussis toxin treatment reverted the inhibitory effects on the enzyme activity elicited by micromolar concentrations of GTP and reduced by 50% the DA inhibition of cyclase activity via D2 receptors. The toxin treatment enhanced the net stimulation of enzyme activity by DA in the presence of micromolar concentrations of GTP. However, the stimulatory effect of the selective D1 receptor agonist SKF 38393 was not significantly affected. The data indicate that Ni mediates D2 inhibition of striatal adenylate cyclase and participates in the modulation of D1 stimulation of the enzyme activity by DA. 相似文献
87.
88.
Yeast submitochondrial particles, in a Pi- and NADH-dependent reaction, produced low concentrations of free ATP in the absence of added ADP. This formation of free ATP, as measured by the luciferin-luciferase method, was strongly stimulated by oligomycin. For maximal stimulation, oligomycin was to be added not earlier than 5–10 min after the addition of NADH. Upon addition of antimycin or FCCP the system was completely inhibited. The amount of free ATP formed corresponded to one-third of the amount of bound ATP in submitochondrial particles. The stimulatory effect of oligomycin disappeared if the submitochondrial particles were spun down after oligomycin stimulation and then resuspended in the reaction medium, whereas submitochondrial particles with no oligomycin added initially were stimulated by oligomycin after the same procedure. A different picture emerged with addition of ADP. If the submitochondrial particles were preenergized with NADH in the presence of oligomycin before the addition of ADP the formation of free ATP upon subsequent addition of ADP was inhibited by oligomycin. In the presence of oligomycin, but lacking preenergization with NADH, a stimulation of free ATP formation was achieved with added ADP. A possible explanation for the stimulating effect of oligomycin on ATP formation in the absence of added ADP is that it enhances the release of bound ATP in an energy-requiring process. The release of only about one-third of the bound ATP could indicate that one of three nucleotide-binding subunits involved in the mechanism of ATP formation by ATP synthase is in a state suitable for such an energy-dependent release of ATP. 相似文献
89.
90.
M Roghani C Da Silva M Castagna 《Biochemical and biophysical research communications》1987,142(3):738-744
The major interaction site for tumor-promoting phorbol esters is the calcium-activated, phospholipid-dependent protein kinase (protein kinase C), a key-element in signal transduction. Binding of phorbol esters results in enzyme activation which mediates, at least in part, the action of these agents. We have investigated the effects of tumor promoter chloroform on protein kinase C activity. Like thrombin and 12-O-tetradecanoylphorbol-13-acetate (TPA), chloroform was able to activate protein kinase C in intact rabbit platelets. In addition, chloroform stimulated enzyme activity as well as TPA binding capacity in cell-free system. Scatchard analysis of the data has shown that chloroform increased the number of phorbol ester binding sites. Structurally related compounds, carbon tetrachloride and methylene chloride, activated the enzyme similarly. 相似文献