共查询到20条相似文献,搜索用时 31 毫秒
1.
Detection of genetic variation in Bradyrhizobium japonicum USDA 110 variants using DNA fingerprints generated with GC rich arbitrary PCR primers 总被引:2,自引:0,他引:2
Bradyrhizobium japonicum USDA 110 has been shown to contain several genetically similar naturally occurring colony morphology variants. These variants differ in symbiotic nitrogen fixation ability and in the utilization of various carbon substrates. They have been shown to share extensive DNA homology and appear to be derived from a common ancestor. Despite these similarities certain B. japonicum USDA 110 variants have been shown to be devoid of symbiotic nitrogen fixation. One of these variants (L2-110), however, was recently shown to possess significant levels of explanta nitrogen fixation and to synthesize functional dinitrogenase enzyme within bacteroids. In an effort to identify genetic markers which could explain differences in symbiotic nitrogen fixation between B. japonicum variants, DNA fingerprints were generated by PCR using arbitrary primers. Two of these primers with GC rich sequences were able to differentiate between B. japonicum USDA 110 variants I-110, L2-110, and MN-110. Unique markers have now been identified which could be examined further to determine if they explain the differences in symbiotic nitrogen fixation between USDA 110 variants. 相似文献
2.
Mathis JN Barbour WM Miller TB Israel DW Elkan GH 《Applied and environmental microbiology》1986,52(1):81-85
We have isolated a colonial derivative of Bradyrhizobium japonicum USDA 110 (designated MN-110) that is both mannitol utilizing and N(2) fixing. Derivative MN-110 showed growth on mannitol and glucose similar to that of non-N(2)-fixing, mannitol-utilizing L2-110. Derivative MN-110 showed high constitutive and induced d-mannitol dehydrogenase activity (similar to L2-110) relative to N(2)-fixing, non-mannitol-utilizing I-110. Hybridization to EcoRI and HindIII total DNA digests with cloned USDA 110 nif DK and nif H genes revealed similar patterns for non-N(2)-fixing mannitol-utilizing derivative L1-110 and derivative MN-110. Symbiotic tests with soybean cultivars Ransom and Lee indicate MN-110 to be a superior N(2)-fixing derivative compared with derivative I-110 and the parent strain USDA 110. However, these differences were not revealed when comparing 28-day-old soybean-B. japonicum associations but were apparent in 49-day-old associations. It was apparent from this work that mannitol utilization was not necessarily correlated to symbiotic effectiveness in B. japonicum and that gene rearrangements were not responsible for differences in N(2) fixation between L1-110 or L2-110 and MN-110. 相似文献
3.
Analysis of the Symbiotic Performance of Bradyrhizobium japonicum USDA 110 and Its Derivative I-110 and Discovery of a New Mannitol-Utilizing, Nitrogen-Fixing USDA 110 Derivative 总被引:2,自引:1,他引:1
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
James N. Mathis Daniel W. Israel W. Mark Barbour Brion D. W. Jarvis Gerald H. Elkan 《Applied microbiology》1986,52(1):75-80
Previously, Bradyrhizobium japonicum USDA 110 was shown to contain colony morphology variants which differed in nitrogen-fixing ability. Mannitol-utilizing derivatives L1-110 and L2-110 have been shown to be devoid of symbiotic nitrogen fixation ability, and non-mannitol-utilizing derivatives I-110 and S-110 have been shown to be efficient at nitrogen fixation. The objectives of this study were to determine the effect of media carbon sources on the symbiotic N2-fixing ability of strain USDA 110 and to compare the effectiveness of strain USDA 110 and derivative I-110. Based on acetylene reduction activity and the nitrogen content of 41-day-old soybean plants, neither derivative I-110 nor cultures of USDA 110 grown in media favoring non-mannitol-using derivatives had symbiotic nitrogen fixation that was statistically superior to that of cultures of USDA 110 grown in media favoring mannitol-using derivatives. In another experiment 200 individual nodules formed by strain USDA 110 grown in yeast extract gluconate were screened for colony morphology of occupying variant(s) and acetylene reduction activity. Nodules occupied by mannitol-using derivatives (large colony type on 0.1% yeast extract-0.05% K2HPO4-0.08% MgSO4 · 7H2O-0.02% NaCl-0.001% FeCl3 · 6H2O [pH 6.7] with 1% mannitol [YEM] plates) had a mean acetylene reduction activity equal to that of nodules occupied by non-mannitol-using derivatives (small colony type on YEM plates). A total of 20 large colonial derivatives and 10 small colonial derivatives (I-110-like) were isolated and purified by repeated culture in YEM and YEG (same as YEM except 1% gluconate instead of 1% mannitol) media, respectively, followed by dilution in solutions containing 0.05% Tween 40. After 25 days of growth, soybean plants inoculated with the large colony isolates had mean whole-plant acetylene reduction activity, whole-plant dry weight, and whole-plant nitrogen contents equal to or better than those of plants inoculated with either the small colony isolates (I-110-like) or the I-110 (non-mannitol-using) derivative. Hence, the existence of a mannitol-utilizing derivative that fixes nitrogen in a culture of strain USDA 110 obtained from the U.S. Department of Agriculture, Beltsville, Md., was established. This new USDA 110 derivative was designated as MN-110 because it was a mannitol-utilizing nitrogen-fixing USDA 110 derivative. This derivative was morphologically indistinguishable from the non-nitrogen-fixing derivative L2-110 found in cultures obtained earlier from the U.S. Department of Agriculture, Beltsville. DNA-DNA homology and restriction enzyme analyses indicated that MN-110 is genetically related to other USDA 110 derivatives that have been characterized previously. 相似文献
4.
The objective of this investigation was to examine the effect of an elevated atmospheric CO2 partial pressure ( p CO2 ) on the N-sink strength and performance of symbiotic N2 fixation in Trifolium repens L. cv. Milkanova. After initial growth under ambient p CO2 in a nitrogen-free nutrient solution, T. repens in the exponential growth stage was exposed to ambient and elevated p CO2 (35 and 60 Pa) and two levels of mineral N (N-free and 7·5 mol m–3 N) for 36 d in single pots filled with silica sand in growth chambers. Elevated p CO2 evoked a significant increase in biomass production from day 12 after the start of CO2 enrichment. For plants supplied with 7·5 mol m–3 N, the relative contribution of symbiotically fixed N (%Nsym ) as opposed to N assimilated from mineral sources (15 N-isotope-dilution method), dropped to 40%. However, in the presence of this high level of mineral N, %Nsym was unaffected by atmospheric p CO2 over the entire experimental period. In plants fully dependent on N2 fixation, the increase in N yield reflects a stimulation of symbiotic N2 fixation that was the result of the formation of more nodules rather than of higher specific N2 fixation. These results are discussed with regard to physiological processes governing symbiotic N2 fixation and to the response of symbiotic N2 fixation to elevated p CO2 in field-grown T. repens . 相似文献
5.
L. D. Kuykendall, M. E. Barnett and J. N. Mathis. 1997. RSα is a repeated DNA sequence found within the nitrogen-fixation gene cluster of Bradyrhizobium japonicum , a symbiotic nitrogen-fixing bacterium that nodulates soybean. Bradyrhizobium japonicum strain 110 spc 4 contains 12 repeats, each located on a separate Xho I DNA restriction fragment between 1.2 and 14 kb in length. Although Fix+ and Fix− derivatives of B. japonicum USDA 110 were first reported more than two decades ago, genotypic differentiation, on the basis of RSα hybridization pattern, was reported only recently. Bradyrhizobium japonicum strain USDA 110 had only single copies of the RSα-hybridizing bands, but a particular Fix− derivative, MSDJGl, carried doublets of two distinct Xho I fragments that carry RSα3 and RSα4. In this study, RSα hybridization patterns were analysed further in both Fix+ and Fix− derivatives of strain 110 to test for duplication of these particular genomic regions. It was concluded that the duplication, or not, of genetic regions carrying RSα3 and RSα4 in strain USDA 110 derivatives is unrelated to symbiotic nitrogen-fixation ability. Like Fix− MSDJGl, Fix+ strain 110 derivatives I-110 and MN-110 had duplications of the Xho I DNA restriction fragments carrying RSα3 and RSα4, but Fix− strain 110 derivative L2–110 lacked these duplications. Thus, it is now clear that Fix− derivatives MSDJG1 and L2–110 arose via distinct genetic mechanisms. Interestingly, Fix+ derivatives of strain 110 from the laboratories of Elkan and Hennecke differed in RSα hybridization profile. 相似文献
6.
Milford S. Brown Supaporn Thamsurakul Gabor J. Bethlenfalvay 《Physiologia plantarum》1988,74(1):159-163
Soybean [ Glycine max (L.) Merr. cv. Hobbit] plants nodulated by Bradyhizobium japonicum strain USDA 110 were grown in pot cultures in severely P- and N-deficient soil and either colonized by the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae (Nicol. & Gerd.) Gerd. and Trappe or fertilized with a high (HP) or low (LP) level of KH2 PO4 (0.6 or 0.3 m M , respectively), After 7 weeks of growth, nodule and chloroplast activities (C2 H2 reduction and CO2 exchange rate) were determined. Photosynthetic P-use efficiency of CO2 fixation was significantly higher in VAM than in HP plants, while that of nitrogenase activity was lower. The LP plants were intermediate in both respects. The ratio of nodule to chloroplast activity [mol C2 H2 reduced (mol CO2 fixed)−1 ] was highest in HP and lowest in VAM plants. Root colonization by the VAM fungus significantly increased nodule number and dry weight and reduced nodule specific mass and activity in comparison to HP plants. In spite of lower nodule activity, VAM plants were significantly larger and had higher N concentrations than the HP plants. The results suggest nonnutritional. VAM-elicited and host-mediated effects on the symbiotic functions of the legume association. 相似文献
7.
Carbon and nitrogen partitioning in young nodulated pea (wild type and nitrate reductase-deficient mutant) plants exposed to NH4 NO3 总被引:1,自引:0,他引:1
David C. Taylor Barry J. Shelp Louise M. Nelson Bernard Grodzinski 《Physiologia plantarum》1988,74(4):593-601
Carbon and nitrogen partitioning was examined in a wild-type and a nitrate reductase-deficient mutant (A317) of Pisum sativum L. (ev. Juneau), effectively inoculated with two strains of Rhizobium leguminosarum (128C23 and 128C54) and grown hydroponically in medium without nitrogen for 21 days, followed by a further 7 days in medium without and with 5 mM NH4 NO3 . In wild-type symbioses the application of NH4 NO3 significantly reduced nodule growth, nitrogenase (EC 1.7.99.2) activity, nodule carbohydrates (soluble sugars and starch) and allocation of [14 C]-labelled (NO3 − , NH4 + , amino acids) in roots. In nodules, there was a decline in amino acids together with an increase in inorganic nitrogen concentration. In contrast, symbioses involving A317 exhibited no change in nitrogenase activity or nodule carbohydrates, and the concentrations of all nitrogenous solutes measured (including asparagine) in roots and nodules were enhanced. Photosynthate allocation to the nodule was reduced in the 128C23 symbiosis. Nitrite accumulation was not detected in any case. These data cannot be wholly explained by either the carbohydrate deprivation hypothesis or the nitrite hypothesis for the inhibition of symbiotic nitrogen fixation by combined nitrogen. Our result with A317 also provided evidence against the hypothesis that NO3 − and NH4 + or its assimilation products exert a direct effect on nitrogenase activity. It is concluded that more than one legume host and Rhizobium strain must be studied before generalizations about Rhizobium /legume interactions are made. 相似文献
8.
In Myrica gale L. plants the assimilation of ammonia released by symbiotic Frankia was observed by 15 N2 labelling and subsequent analysis of the isotopic enrichment of nodule amino acids over time by single ion monitoring gas chromatography-mass spectrometry. In detached nodules of Myrica , glutamine was the first amino acid labelled at 30 s and subsequently the amino acids glutamate, aspartate, alanine and γ-amino butyric acid (GABA) became labelled. This pattern of labelling is consistent with the incorporation of ammonium via glutamine synthetase [GS; EC 6.3.1.2]. No evidence for the ammonium assimilation via glutamate dehydrogenase [GDH; EC 1.4.1.2] was observed as glutamate became labelled only after glutamine. Using attached nodules and pulse-chase labelling, we observed synthesis of glutamine, glutamate, aspartate, alanine, GABA and asparagine, and followed the transport of fixed nitrogen in the xylem largely as glutamine and asparagine. Estimation of the cost of nitrogen fixation and asparagine synthesis in Myrica nodules suggests a minimum of one sucrose required per asparagine produced. Rapid translocation of recently fixed nitrogen was observed in Myrica gale nodules as 80% of the nitrogen fixed during a 1-h period was translocated out of the nodules within 9 h. The large pool of asparagine that is present in nodules may buffer the transport of nitrogen and thus act to regulate nitrogen fixation via a feedback mechanism. 相似文献
9.
Medicago ciliaris (L.) All., a salt-tolerant legume, was not nodulated by Rhizobium meliloti (2011), a strain commonly used for field inoculation of alfalfas. A strain of Rhizobium meliloti (ABS7) was isolated from saline Algerian soils. It is generally more salt-resistant than strain 2011, exhibits a higher rate of growth and induces the formation of nodules on M. ciliaris . C2 H2 reduction activity of M. ciliaris nodules was inhibited by 50% in the presence of 200 m M NaCl in the culture medium. whereas 100 m M NaCl was sufficient to inhibit the activity of nodules of M. sativa (L. cv. Europe). C2 H2 reduction by bacteroids, isolated from nodules of the two species of alfalfa, was directly inhibited by the presence of NaCl in the incubation medium. In both cases, glucose could support bacteroid nitrogen fixation, but only in a narrow range of O2 tensions. Bacteriods from M. ciliaris were more tolerant to salt than M. sativa ones. The salt resistance of bacteroids from nodules of plants watered with NaCl solutions was not improved in either species. Salt directly added to the incubation mixture of bacteroids or to the culture medium of plants inhibited O2 uptake of bacteroids isolated from nodules of both M. ciliaris and M. sativa . The depressive effect of NaCl on bacteroid C2 H2 reduction could be directly related to the drop in bacteroid respiration. The nitrogen fixation capacity of the M. ciliaris-Rhizobium meliloti (ABS7) symbiosis under saline conditions leads us to recommend the introduction of this association in salt-troubled areas. 相似文献
10.
Symbiotic N2 fixation in a high Alpine grassland: effects of four growing seasons of elevated CO2 总被引:1,自引:0,他引:1
J. A. Arnone Iii 《Functional ecology》1999,13(3):383-387
1. Increasing carbon dioxide concentration (E: 680 μl CO2 litre–1 vs ambient, A: 355 μl CO2 litre–1 ) around late-successional Alpine sedge communities of the Swiss Central Alps (2450 m) for four growing seasons (1992–1995) had no detectable effect on symbiotic N2 fixation in Trifolium alpinum —the sole N2 -fixing plant species in these communities (74 ± 30 mg N m–2 year–1 , A and E plots pooled).
2. This result is based on data collected in the fourth growing season showing that elevated CO2 had no effect on Trifolium above-ground biomass (4·4 ± 1·7 g m–2 , A and E plots pooled, n = 24) or N content per unit land area (124 ± 51 mg N m–2 , A and E pooled), or on the percentage of N Trifolium derived from the atmosphere through symbiotic N2 fixation (%Ndfa: 61·0 ± 4·1 across A and E plots) estimated using the 15 N dilution method.
3. Thus, it appears that N inputs to this ecosystem via symbiotic N2 fixation will not be dramatically affected in the foreseeable future even as atmospheric CO2 continues to rise. 相似文献
2. This result is based on data collected in the fourth growing season showing that elevated CO
3. Thus, it appears that N inputs to this ecosystem via symbiotic N
11.
Entry of the divalent cations Ni2+ , Co2+ and Zn2+ into cells of maize ( Zea mays L. cv. Dekalb XL 85) root tissue is accompanied by an acidification of the incubation medium, a decrease in both the pH of the cell sap and the level of malate in the cells, and by an inhibition of dark fixation of CO2 . K+ , on the contrary, induces only a very low acidification of the incubation medium, does not change either the pH of the cell sap or the malate level in the cells, and induces an increase in CO2 dark fixation. Different mechanisms are postulated for the stimulation of proton extrusion by divalent cations and K+ . 相似文献
12.
Expression of Bradyrhizobium japonicum wild-type strain USDA110 nirK , norC and nosZ denitrification genes in soybean root nodules was studied by in situ histochemical detection of β -galactosidase activity. Similarly, PnirK - lacZ , PnorC - lacZ , and PnosZ - lacZ fusions were also expressed in bacteroids isolated from root nodules. Levels of β -galactosidase activity were similar in both bacteroids and nodule sections from plants that were solely N2 -dependent or grown in the presence of 4 m M KNO3 . These findings suggest that oxygen, and not nitrate, is the main factor controlling expression of denitrification genes in soybean nodules. In plants not amended with nitrate, B. japonicum mutant strains GRK308, GRC131, and GRZ25, that were altered in the structural nirK , norC and nosZ genes, respectively, showed a wild-type phenotype with regard to nodule number and nodule dry weight as well as plant dry weight and nitrogen content. In the presence of 4 m M KNO3 , plants inoculated with either GRK308 or GRC131 showed less nodules, and lower plant dry weight and nitrogen content, relative to those of strains USDA110 and GRZ25. Taken together, the present results revealed that although not essential for nitrogen fixation, mutation of either the structural nirK or norC genes encoding respiratory nitrite reductase and nitric oxide reductase, respectively, confers B. japonicum reduced ability for nodulation in soybean plants grown with nitrate. Furthermore, because nodules formed by each the parental and mutant strains exhibited nitrogenase activity, it is possible that denitrification enzymes play a role in nodule formation rather than in nodule function. 相似文献
13.
Werner Selbitschka Uwe Dresing Margit Hagen Stefan Niemann Alfred Pühler 《FEMS microbiology ecology》1995,16(3):223-232
Abstract Using a newly developed integration vector, the Escherichia coli gusA gene conferring GUS-activity or the firefly ( Photinus pyralis ) luc gene mediating bioluminescence were integrated into a non-essential site of the chromosome of Rhizobium meliloti 2011. The integration of the constitutively expressed marker genes into the chosen site per se did not affect the strains' ability to perform homologous recombination, its growth characteristics or its symbiotic nitrogen fixation. Comparative microcosm analyses between the bioluminescent, recombination-proficient (RecA+ ) R. meliloti strain L33 whose construction is reported in this paper, and its previously described recombination-deficient (RecA− ) isogenic counterpart L1 indicate that RecA− strains of Rhizobium are safe hosts for deliberate release experiments. 相似文献
14.
We report a novel use of the 15 N dilution technique to detail the translocation of amino compounds in the legume Sesbania rostrata . The conventional 15 N dilution technique follows the dilution of 15 N within a labelled plant, as 14 N2 is fixed by symbiotic bacteria. In our experiments, stem-nodulated Sesbania rostrata were enriched by feeding with 15 N ammonium nitrate for 2 weeks, followed by a 1 week period where the only N available to the plants was via nitrogen fixation of atmospheric N2 . We measured the composition, concentration and 15 N enrichment of amino compounds in various plant tissues, both above and below the stem nodules, using GC-MS and isotopic abundance mass spectrometry techniques. Approximately 28% of the total N in the stem nodules was derived from internal plant sources. The ureides allantoic acid and allantoin were not abundant in xylem, leaf or nodule tissues. The amides asparagine and glutamine were the major export products from stem nodules although a wide range of other amino compounds are also synthesized. Amino acids within the nodules had a low level of enrichment, demonstrating that a small fraction (≈ 11%) was derived from outside the nodules, and significant cycling of N (28% of xylem N) through the root system was revealed by measurements of 15 N distribution and amino acid concentrations. 相似文献
15.
Benjamin Gourion Sandra Sulser Julia Frunzke Anne Francez-Charlot Philipp Stiefel Gabriella Pessi Julia A. Vorholt Hans-Martin Fischer 《Molecular microbiology》2009,73(2):291-305
PhyR is an unusual type of response regulator consisting of a receiver domain and an extracytoplasmic function (ECF) sigma factor-like domain. It was recently described as a master regulator of general stress response in Methylobacterium extorquens . Orthologues of this regulator are present in essentially all free-living Alphaproteobacteria. In most of them, phyR is genetically closely linked to a gene encoding an ECF σ factor. Here, we investigate the role of these two regulators in the soybean symbiont Bradyrhizobium japonicum USDA110. Using deletion mutants and phenotypic assays, we showed that PhyR and the ECF σ factor σEcfG are involved in heat shock and desiccation resistance upon carbon starvation. Both mutants had symbiotic defects on the plant hosts Glycine max (soybean) and Vigna radiata (mungbean). They induced fewer nodules than the wild type and these nodules were smaller, less pigmented, and their specific nitrogenase activity was drastically reduced 2 or 3 weeks after inoculation. Four weeks after infection, soybean nodule development caught up to a large extent whereas most mungbean nodules remained defective even 5 weeks after infection. Remarkably, both mutants triggered aberrant nodules on the different host plants with ectopically emerging roots. Microarray analysis revealed that PhyR and σEcfG control congruent regulons suggesting both regulators are part of the same signalling cascade. This finding was further substantiated by in vitro protein–protein interaction studies which are in line with a partner-switching mechanism controlling gene regulation triggered by phosphorylation of PhyR. The large number of genes of unknown function present in the PhyR/σEcfG regulon and the conspicuous symbiotic phenotype suggest that these regulators are involved in the Bradyrhizobium –legume interaction via yet undisclosed mechanisms. 相似文献
16.
Nodulation and nitrogenase activity in nitrogen-fixing woody plants stimulated by CO2 enrichment of the atmosphere 总被引:5,自引:0,他引:5
Richard J. Norby 《Physiologia plantarum》1987,71(1):77-82
The responses of three species of nitrogen-fixing trees to CO2 enrichment of the atmosphere were investigated under nutrient-poor conditions. Seedlings of the legume, Robinia pseudoacacia L. and the actinorhizal species, Alnus glutinosa (L.) Gaertn. and Elaeagnus angustifolia L. were grown in an infertile forest soil in controlled-environment chambers with atmospheric CO2 concentrations of 350 μl −1 (ambient) or 700 μl −1 . In R. pseudoacacia and A. glutinosa , total nitrogenase (N2 reduction) activity per plant, assayed by the acetylene reduction method, was significantly higher in elevated CO2 , because the plants were larger and had more nodule mass than did plants in ambient CO2 . The specific nitrogenase activity of the nodules, however, was not consistently or significantly affected by CO2 enrichment. Substantial increases in plant growth occurred with CO2 enrichment despite probable nitrogen and phosphorus deficiencies. These results support the premises that nutrient limitations will not preclude growth responses of woody plants to elevated CO2 and that stimulation of symbiotic activity by CO2 enrichment of the atmosphere could increase nutrient availability in infertile habitats. 相似文献
17.
18.
Measurements of the short-term response of nodulated roots of soybean ( Glycine max L. Merr, cv. Harosoy: Bradyrhizobium japonicum USDA 16) to rapid changes in surrounding pO2 indicate that their ability to reversibly adjust gaseous diffusive resistance is retained whether plants are cultured in rhizospheres of very low (2.8%) or very high (61.2%) pO2 . Thus the capacity for reversible short-term diffusion adjustment is additional to structural changes in the fixed diffusional barriers of nodules which allow their continued fixation of N2 in unfavourably high or low external pO2 . Anatomical evidence, involving quantitative measurement of intercellular spaces in the cortical tissues using electron microscopy of thin sections, indicates that the major fixed diffusional barrier is a boundary layer of cells in the inner cortex which may be as small as one cell thick in nodules from 2.8% O2 to 5 or 6 cells thick, and almost completely devoid of intercellular spaces, in those from 61.2% O2 . The data are interpreted to indicate that the variable diffusion harrier is distinct from the boundary layer and is most likely to be a property of cells and/or intercellular spaces inside the boundary layer of the nodule cortex. 相似文献
19.
A perturbation of the histidine biosynthetic pathway in legume microsymbionts can abolish their symbiotic competence. Twenty-one histidine-requiring (His− ) mutants were isolated from berseem clover-nodulating, symbiotically-competent (Nod+ , Fix+ ) Rhizobium leguminosarum bv. ' trifolii ' strain RTH 48 Smr by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) mutagenesis followed by enrichment. These mutants were analysed for their biochemical defect and the corresponding effect, if any, on their symbiotic abilities. Cross-feeding, supplementation and enzymatic studies identified three types of mutants. Group 1 mutants, His-2 and His-12, grew with histidine supplementation but not with the addition of either L -histidinol or L -histidinol phosphate to the medium ; they lacked histidinol dehydrogenase (EC 1.1.1.23) activity and consequently formed only ineffective, or 'non-fixing' nodules. Group 2 mutant, His-17, grew when supplemented with either L -histidinol or L -histidine, had low histidinol phosphate phosphatase (EC 3.1.3.15) activity (37% of wild-type), and consequently failed to nodulate berseem clover. Group 3, the remaining 18 mutants, grew when supplemented with L -histidinol phosphate, L -histidinol or histidine, and did not nodulate. Typically, reversion rates were between 10−7 and 10−8 . Defects in early steps of the pathway abolished nodulating ability, whereas lesions in the last step did not. The last step, however, was required for symbiotic nitrogen fixation. It is hypothesized that histidine may be supplied by the host in sufficient quantity for nodulation by histidinol dehydrogenase mutants to occur, whereas the amount provided in the nodule may be insufficient to support bacteroid development and nitrogen fixation. 相似文献
20.
The possibility to induce nitrate reductase (NR; EC 1.6.6.2) in needles of Scots pine ( Pinus sylvestris L.) seedlings was studied. The NR activity was measured by an in vivo assay. Although increased NR activities were found in the roots after application of NO3 − , no such increase could be detected in the needles. Detached seedlings placed in NO3 − solution showed increasing NR activities with increasing NO3 − concentrations. Exposure of seedlings to NOx (70–80 ppb NO2 and 8–12ppb NO) resulted in an increase of the NR activity from 10–20 nmol NO2 − (g fresh weight)−1 h−1 to about 400 nmol NO2 − (g fresh weight)−1 h−1 . This level was reached after 2–4 days of exposure, thereafter the NR activity decreased to about 200 nmol NO2 − (g fresh weight)−1 h−1 . Analyses of free amino acids showed low concentrations of arginine and glutamine in NOx -fumigated seedlings compared to corresponding controls. 相似文献