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61.
Alison M. Berry James R. Thayer Carol S. Enderlin A. Daniel Jones 《Archives of microbiology》1990,154(5):510-513
Nitrogen-starved cells of Frankia strain HFPArl3 incorporated [13N]-labeled ammonium into glutamine serine (glutamate, alanine, aspartate), after five-minute radioisotope exposures. High initial endogenous pools of glutamate were reduced, while total glutamine increased, during short term NH
inf4
sup+
incubation. Preincubation of cells in methionine sulfoximine (MSX) resulted in [13N]glutamine reduced by more than 80%, while [13N]glutamate and [13N]alanine levels increased. The results suggest that glutamine synthetase is the primary enzyme of ammonium assimilation, and that glutamate dehydrogenase and alanine dehydrogenase may also function in ammonium assimilation at low levels. Efflux of [13N]serine and lesser amounts of [13N]glutamine was detected from the Frankia cells. The identity of both Ser and Gln in the extracellular compartment was confirmed with gas chromatography/mass spectrometry. Serine efflux may be of significance in nitrogen transfer in Frankia.Abbreviations Pthr
phosphothreonine
- Aad
-amino-adipate
- MSX
methionine sulfoximine 相似文献
62.
Medicago truncatula,a model plant for studying the molecular genetics of theRhizobium-legume symbiosis 总被引:7,自引:0,他引:7
David G. Barker Sylvie Bianchi François Blondon Yvette Dattée Gérard Duc Sadi Essad Pascal Flament Philippe Gallusci Gérard Génier Pierre Guy Xavier Muel Jacques Tourneur Jean Dénarié Thierry Huguet 《Plant Molecular Biology Reporter》1990,8(1):40-49
Medicago truncatula has all the characteristics required for a concerted analysis of nitrogen-fixing symbiosis withRhizobium using the tools of molecular biology, cellular biology and genetics.M. truncatula is a diploid and autogamous plant has a relatively small genome, and preliminary molecular analysis suggests that allelic
heterozygosity is minimal compared with the cross-fertilising tetraploid alfalfa (Medicago sativa). TheM. truncatula cultivar Jemalong is nodulated by theRhizobium meliloti strain 2011, which has already served to define many of the bacterial genes involved in symbiosis with alfalfa. A genotype
of Jemalong has been identified which can be regenerated after transformation byAgrobacterium, thus allowing the analysis ofin-vitro-modified genes in an homologous transgenic system. Finally, by virtue of the diploid, self-fertilising and genetically homogeneous
character ofM. truncatula, it should be relatively straightforward to screen for recessive mutations in symbiotic genes, to carry out genetic analysis,
and to construct an RFLP map for this plant. 相似文献
63.
Kunio Yonemasu Takako Sasaki Yoshiko Dohi Charles M. Lapière Betty Nusgens 《生物化学与生物物理学报:疾病的分子基础》1990,1096(1):47-51
C1q, a collagen-like complement protein, was purified from the serum of a ddermatosparactic calf which lacks procollagen N-terminal proteinase (pN-proteinase). The specific hemolytic activity of the serum Clq from the dermatosparactic animal was identical to that of C1q from a normal calf. Gel-filtration of serum from dermatosparactic calf, on Sepharose 6B, showed the presence of C1q-antigenic material at only one position which was identical to the elution position of normal bovine C1q. No differdence, under dissociating conditions, could be seen in the size of the chains of C1q in specific immunoprecipitates isolated from the sera of dermatosparactic and normal animals, as judged by polyacrylamidegel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS). The C1q from the dermatosparactic animal showed the same N-terminal amino acid and typtic-digest peptide pattern on HPLC as C1q from the normal calf. These results strongly suggest that pN-proteinase is not involved in the extracellular processing of C1q. 相似文献
64.
In goodeid teleosts, prolonged embryonic development takes place within the ovarian cavity. Apposed maternal and embryonic epithelia interface via a nutritive liquid (embryotrophe) and facilitate aplacental matrotrophy. The role of the internal ovarian epithelium (IOE) in providing proteins for the embryotrophe has been studied using transmission electron-microscopic examinations of both the resting and the active ovarian lining, and isoelectric focusing of embryotrophe and maternal blood serum. The simple IOE is apparently composed of only one, filament-containing cell-type. In the non-gravid ovary these cells are cuboidal to columnar in shape, and are either compact and electron-dense or oedematous and light. During gestation, swelling of the ovarian connective tissue gives rise to dovetailing of the IOE with the subjacent capillary plexus. Part of the IOE overlying the capillaries becomes stretched, resulting in a thin endothelium-like demarcation. The nuclei and the bulk of the cytoplasm are usually recessed between the meshes of the protruding capillary network. The blood-embryotrophe pathway is thus reduced in places, to less than one m. The active form of the IOE contains a well-developed vacuolar apparatus composed of small vesicles, vacuoles, multivesicular bodies, and a few lysosomes. Elements of the RER are sparsely distributed throughout the cytoplasm. Endocytotic activity is observable at the apical and basolateral plasma membrane. Isoelectric focusing of both serum and embryotrophe produces numerous bands each between pI 4–8, which reveal many homologies. The intensity of corresponding bands varies considerably. It is concluded that the cells of the IOE provide a transport pathway for serum-derived macromolecular substances rather than produce proteinaceous secretions. 相似文献
65.
Azotobacter vinelandii was grown diazotrophically in sucrose-limited chemostat cultures at either 12, 48, 108, 144 or 192 M dissolved oxygen. Steady state protein levels and growth yield coefficients (Y) on sucrose increased with increasing dilution rate (D). Specific rate of sucrose consumption (q) increased in direct proportion to D. Maintenance coefficients (m) extrapolated from plots of q versus D, as well as from plots of 1/Y versus 1/D exhibited a nonlinear relationship to the dissolved oxygen concentration. Constant maximal theoretical growth yield coefficients (Y
G) of 77.7 g cells per mol of sucrose consumed were extrapolated irrespective of differences in ambient oxygen concentration. For comparison, glucose-, as well as acetate-limited cultures were grown at 108 M oxygen. Fairly identical m- and Y
G-values, when based on mol of substrate-carbon with glucose and sucrose grown cells, indicated that both substrates were used with the same efficiency. However, acetate-limited cultures showed significantly lower m- and, at comparable, D, higher Y-values than cultures limited by either sucrose or glucose. Substrate concentrations (K
s) required for half-maximal growth rates on sucrose were not constant, they increased when the ambient oxygen concentration was raised and, at a given oxygen concentration, when D was decreased. Since biomass levels varied in linear proportion to K
s these results are interpreted in terms of variable substrate uptake activity of the culture.Abbreviations
D
dilution rate
-
K
s
substrate concentration required for half maximal growth rate
-
m
maintenance coefficient
-
q
specific rate of substrate consumption
-
Y
growth yield coefficient
-
Y
G
maximum theoretical growth yield coefficient 相似文献
66.
Barbara J. Javor 《Archives of microbiology》1988,149(5):433-440
Seven strains of extremely halophilic bacteria (Halobacterium spp., Halococcus spp., and Haloarcula sp.) fixed CO2 under light and dark conditions. Light enhanced CO2 fixation in Halobacterium halobium but inhibited it in Halobacterium volcanii and Haloarcula strain GN-1. Propionate stimulated 14CO2 incorporation in some strains, but inhibited it in others. Semi-starvation in basal salts plus glycerol induced enhanced CO2 fixation rates. 14CO2 fixation in semi-starved cells was stimulated by NH
4
+
or pyruvate and inhibited by succinate and acetate in most strains. No possible reductant was found. In cell-free extracts of H. halobium, NH
4
+
but not propionate stimulated 14CO2 fixation. No RuBP carboxylase activity was detected. The main 14C-labeled -keto acid detected after a 2-min incubation with 14CO2 and pyruvate was pyruvate. Little or no -ketobutyrate was detected among the early products of propionate-stimulated CO2 fixation. Glycine was the major amino acid synthesized during a 2-min incubation with NH
4
+
, propionate, and 14CO2. Propionate-stimulated CO2 fixation was sensitive to trimethoprim and insensitive to avidin. A novel pathway for non-reductive CO2 fixation involving a glycine synthase reaction with CO2, NH
4
+
, and a methyl carbon derived from the -carbon cleavage of propionate is tentatively proposed.Abbreviations used BBS
buffered basal salts
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- MOPS
3-(N-morpholino)propanesulfonic acid
- DNPH
2,4-dinitrophenylhydrazine
- DNP
dinitrophenyl
- TLC
thin-layer chromatography
- FH4
tetrahydrofolate
This work was supported by National Science Foundation grant PCM-8116330 and Petroleum Research Fund grant PRF 13704-AC2 相似文献
67.
A role for haemoglobin in all plant roots? 总被引:4,自引:2,他引:2
Abstract. We have found haemoglobin in plant roots whereas previously it has been recorded only in nitrogen fixing nodules of plants. Haemoglobin occurs not only in the roots of those plants that are capable of nodulation but also in the roots of species that are not known to nodulate. We suggest that a haemoglobin gene may be a component of the genome of all plants. The gene structure and sequence in two unrelated families of plants suggests that the plant haemoglobins have had a single origin and that this origin relates to the haemoglobin gene of the animal kingdom. At present we cannot completely rule out the possibility of a horizontal transfer of the gene from the animal kingdom to a progenitor of the dicotyledonous angiosperms but we favour a single origin of the gene from a progenitor organism to both the plant and animal kingdoms. We speculate about the possible functions of haemoglobin in plant roots and put the case that it is unlikely to have a function in facilitating oxygen diffusion. We suggest that haemoglobin may act as a signal molecule indicating oxygen deficit and the consequent need to shift plant metabolism from an oxidative to a fermentative pathway of energy generation. 相似文献
68.
Micropropagation of the actinorhizal plant Hippophae rhamnoides L. (sea-buckthorn) was achieved on Murashige and Skoog (MS) medium supplemented with 1 M of benzylaminopurine (BA). A multiplication frequency of three to five shoots per explant was observed after 28 days. Rooting of these shoots was achieved in a medium containing 1/4 strength MS without growth regulators. The rooted plants were transferred to Turface R artificial substrate and inoculated with pure cultures of two Frankia strains. These plantlets subsequently developed nodules which fixed nitrogen. 相似文献
69.
Sources,fates, and impacts of nitrogen inputs to terrestrial ecosystems: review and synthesis 总被引:1,自引:0,他引:1
The relative importance of nitrogen inputs from atmospheric deposition and biological fixation is reviewed in a number of diverse, non-agricultural terrestrial ecosystems. Bulk precipitation inputs of N (l–l2 kg N ha–1 yr–1) are the same order of magnitude as, or frequently larger than, the usual range of inputs from nonsymbiotic fixation (< 1=" –=" 5=" kg=" n=">–1 yr–1), especially in areas influenced by industrial activity. Bulk precipitation measurements may underestimate total atmospheric deposition by 30–40% because they generally do not include all forms of wet and dry deposition. Symbiotic fixation generally ranges from 10–160 kg N ha–1 yr–1) in ecosystems where N-fixing species are present during early successional stages, and may exceed the range under unusual conditions.Rates of both symbiotic and nonsymbiotic fixation appear to be greater during early successional stages of forest development, where they have major impacts on nitrogen dynamics and ecosystem productivity. Fates and impacts of these nitrogen inputs are important considerations that are inadequately understood. These input processes are highly variable in space and time, and few sites have adequate comparative information on both nitrogen deposition and fixation.
相似文献
| - more intensive studies of total atmospheric deposition, especially of dry deposition, are needed over a wide range of ecosystems; |
| - additional studies of symbiotic fixation are needed that carefully quantify variation over space and time, examine more factors regulating fixation, and focus upon the availability of N and its effects upon productivity and other nutrient cycling processes; |
| - process-level studies of associative N-fixation should be conducted over a range of ecosystems to determine the universal importance of rhizosphere fixation; |
| - further examination of the role of free-living fixation in wood decomposition and soil organic matter genesis is needed, with attention upon spatial and temporal variation; and |
| - investigations of long-term biogeochemical impacts of these inputs must be integrated with process-level studies using modern modelling techniques. |
70.
J D Kowit R W Linck D M Kenney 《Biology of the cell / under the auspices of the European Cell Biology Organization》1988,64(3):283-291
Detergent extraction of human blood platelets pre-treated with Taxol to stabilize microtubules allows isolation of marginal band (MB) cytoskeletons. We studied MB cytoskeleton structure using dark-field light microscopy and negative stain electron microscopy (EM). Dark-field illumination clearly demonstrated the "hoop" shape of MB cytoskeletons in unfixed suspensions where the microtubule coils had a mean diameter of 2.87 microns (+/- 0.18 micron, SD). Microtubules were uncoiled by brief exposure to trypsin (2 ng/micrograms protein) or by NaCl (154-600 mM) but not by DNase I, which removed approximately 40% of total actin, but had no effect on dark-field images of microtubule coils. As microtubules uncoiled, a single fiber emerged from the hoop and gradually lengthened as the brightness of the hoop diminished; these fibers correspond to the single microtubules seen by EM. Polypeptides of coiled and uncoiled MB cytoskeletons were analyzed by SDS-PAGE. When microtubules became uncoiled, no changes in the major components (alpha- and beta-tubulin, IEF-51K, or actin) were found. However, a number (greater than 10) of minor polypeptides, each less than 5% of total cytoskeletal protein and with an Mr ranging from 80,000- greater than 260,000, were decreased in "uncoiled" MB cytoskeletons. These results implicate one or more of these minor polypeptides in maintenance of hoop integrity. Dark-field light microscopy thus provides an approach toward investigating the mechanism(s) involved in maintaining the microtubule coil of the platelet marginal band. 相似文献