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1.
Growing-season populations of rhizobia associated with annual host-plant roots and nearby soil were examined in a field soil showing a nodulation problem in the second year after establishment. Rhizobium lupini reached higher populations at a faster rate than R. trifolii. A sharp drop in the population of R. trifolii associated with subterranean clover roots early in the growing season was followed by a recovery to high numbers. No such phenomenon occurred with R. lupini. The numbers of rhizobia under patches of non-nodulated plants in second-year stands were very low, usually <5/g soil, whereas the numbers under healthy plants in problem stands were similar to those under established stands. Differences in the colonization of both root and soil by R. trifolii in the first year were reflected in the second-year nodulation.  相似文献   

2.
Soil cores from under senesced legume swards were partitioned into (1) soils, (2) legume roots, (3) nodules and (4) debris. The viable population of Rhizobium trifolii under subterranean clover was found to be predominantly in the nodule fraction; but with R. lupini under Serradella the distribution was more even. In commercially sown stands the rhizobia often failed to migrate from the original sown rows into the soil between the rows. The relative importance of the various fractions as contributors to the “pool” of rhizobia available for nodulation of subsequent generations of host-plants is discussed.  相似文献   

3.
Microorganisms (348 fungi, 388 actinomycetes and 319 bacteria) were isolated from a nodulation problem soil, a non-problem virgin soil, a cultivated problem soil and the rhizosphere of clover plants grown in the problem soil. Rhizobium trifolii TA 1 which failed to establish in problem soils was inhibited on laboratory media by a greater number of these soil microorganisms than the better soil colonizing R. trifolii (WU95 and WU290) and R. lupini (WU425). R. lupini was not inhibited or stimulated on agar by many soil or rhizosphere isolates. R. meliloti showed greater stimulation than either R. trifolii or R. lupini and was inhibited by relatively few soil microorganisms so that its poor soil survival was thought to be due to chemical or physical soil conditions rather than to biotic factors. The greatest incidence of rhizobial inhibitors, mainly associated with TA 1, was found among the isolates from the clover rhizosphere. There was a reduction in the relative numbers of rhizobial inhibitors isolated from the cultivated soil compared with the virgin problem soils, a result possibly due to the changed soil environment changing with cultivation, altered vegetation and the addition of superphosphate. Inhibitors of rhizobia were more frequent amongst the bacteria than fungi or actinomycetes. Strong stimulation was more commonly shown by fungi than by actinomycetes or bacteria. The interaction on agar between rhizobia and the soil microflora is related to soil colonization and persistence.  相似文献   

4.
Soil samples taken from 28 sites following varying periods of cropping in a crop-pasture rotation contained very low populations of Rhizobium trifolii. Populations were less than 103g?1R. trifolii of soil for 89% of the sites and were significantly correlated with soil pH. Application of lime resulted in a build-up of R. trifolii in the absence of the host legume, subterranean clover, but when inoculated clover seed was sown the populations built up to satisfactory levels after the first season's growth, regardless of soil pH.The number of nodules per plant was increased by the application of lime, but the plants growing in unlimed soil had fewer, larger nodules. The increase in nodulation with lime on these low-calcium acid soils persisted to the third growing season.  相似文献   

5.
Marked strains of Rhizobium trifolii, distinguishable from other strains antigenically and by streptomycin resistance, were introduced by seed inoculation of subterranean clover (Trifolium subterraneum L.) into a field environment having a natural population of R. trifolii. Isolates from nodules obtained periodically during the following 41 months were classified using both methods of identification in parallel. This procedure made it possible to determine the reliability of each method independently.There was a gradual disappearance of the inoculum strains which occurred more rapidly in plots of cv. Woogenellup than in plots seeded with cv. Mount Barker. At five harvests, there was 95% (or greater) correspondence between inoculum survival using either method of identification. There was evidence that a small proportion of the progeny of the inocula sustained independent loss of antigenic character and/or streptomycin resistance in the field or, alternatively, that strains occurring naturally acquired these characteristics. A few nodules contained more than one strain of rhizobia. These exceptions occurred at low frequency and did not interfere substantially with identification results. It is concluded that gel immune diffusion serology and the use of streptomycin-resistant mutants are both reliable methods for identifying strains of rhizobia reisolated from field environments.  相似文献   

6.
Abstract

Birdsfoot trefoil (Lotus tenius), red clover (Trifolium pratense) and white clover (Trifolium repens) were evaluated as potential forage legumes on andic soils. A greenhouse study was used to evaluate the influence of soil pH on: (1) the establishment and growth of these forage legumes, (2) N and P quantity and concentration, and (3) Rhizobium lupini and Rhizobium trifolii survival.

Aluminum sulfate and finely‐ground calcium carbonate were used to adjust soil pH. Soil pH's were 4.8, 5.1, 5.6, 6.2, 6.4, 6.9, 7.0 and 7.4. Birdsfoot trefoil, red clover and white clover were grown in 15 cm pots in the greenhouse and harvested five times at approximately 20‐day intervals. Yield was measured and plant material was analyzed for total N and P. Populations of R. lupini and R. trifolii were monitored using the MPN technique.

Manipulation of pH in the Mission soil was found to have a significant effect on the growth of the three forage legumes, tissue N and P concentration and uptake, and on the survival of R. lupini and R. trifolii in the soil. In general, tested parameters increased with increasing soil pH. Greatest forage yield occurred in the 6.9–7.0 pH range. The andic nature of the Mission soil requires a pH adjustment above pH 6.2 for acceptable establishment and yields of the three forage legumes studied. This is in sharp contrast to non‐andic northern Idaho soils where forage legume yields are usually not adversely affectea above pH 5.5. All three legumes appeared to have good potential ana should be evaluated under field conditions.  相似文献   

7.
Strains of Rhizobium trifolii incorporated into commercial peat inoculants were compared for their effect on the establishment and growth of oversown white clover (Trifolium repens) on soils devoid of infective rhizobia.There were marked differences in numbers of seedlings establishing and clover dry matter production per hectare with the various strains. However, when adjusted to a constant number of established seedlings, dry matter production from all strains, apart from one strain at one site, were similar indicating that the strains did not appear to influence the growth of individual clover plants.The marked differences in establishment of clover inoculated with the various strains could not be accounted for by differences in the number of rhizobia in the peat inoculant.Selecting strains of rhizobia for ability to increase establishment is considered important where clover is oversown onto soils devoid of rhizobia.  相似文献   

8.
An examination of 85 strains of bacteria from five species of rhizobia (Rhizobium sp., R. japonicum, R. lupini, R. meliloti and R. trifolii), using a new semi-quantitative assay procedure, disclosed wide diversity among the strains in their requirement for, and response to, vitamins, carbon sources, and nitrogen sources. Approximately half of the strains in the first four species grew as well without vitamins as they did when supplied with a vitamin mixture or with yeast extract, but the other strains showed considerable variation in their requirements. Some strains were inhibited by yeast extract, or showed best growth in basic media supplemented with only one vitamin. The strains within the species differed widely in their utilization of gluconate, mannitol and arabinose as C-sources; there was less diversity in their use of glutamine, histidine, NH4+-N and NO3?-N as N-sources. The significance of these observations in the culture of rhizobia in the laboratory, in their ecological adaptation to particular environments, and in their ability to form an effective symbiosis with particular host legumes, is discussed.  相似文献   

9.
The aim of this study was to identifiy the endophytic bacteria recovered from the foliage, tap roots and nodules of red clover plants (Trifolium pratense L.); and to assess the effects of the nodule bacteria, alone and in combination with Rhizobium spp., on the growth and development of red clover seedlings. Thirty-one bacteria species from 14 different genera were recovered from within the foliage, roots and nodules of red clover plants cv. AC Charlie. Genera diversity and species number were greatest in foliage tissues. Pantoea agglomerans (59.6%) was the most frequent species recovered in foliage tissues, Agrobacterium rhizogenes A in the tap root (49.2%) and Rhizobium leguminosarum BV phaseoli and R. loti B in the nodules (27.2% each). Recovery of Rhizobium species was not restricted to the nodules, and species of this genus were systemic throughout the plant. Clover root nodules were host to 12 bacteria species other than rhizobia, of which 8 were specific to this tissue. Using non-selective media, R. leguminosarum BV trifolii constituted only 8.8% of all the root nodule bacteria recovered. In root bacterization experiments, species of nodule bacteria promoted growth of red clover more often when applied in combination with R. leguminosarum BV trifolii than when applied singly. However, Bacillus megaterium, Bordetella avium and Curtobacterium luteum consistently promoted growth either individually or in combination with R. leguminosarum BV trifolii. Nodulation was promoted when R. leguminosarum BV trifolii was coinoculated with Bacillus insolitus, B. brevis or A. rhizogenes A. Single isolate applications of Rhizobium species to roots always led to the depression of clover growth, but mixtures of R. leguminosarum BV trifolii and R. leguminosarum BV phaseoli resulted in growth promotion. The latter is considered further evidence of the beneficial allelopathic side effect of strain competition for the same ecological niche. Received: 27 June 1996  相似文献   

10.
I ndole acetic acid (IAA) production from tryptophan by cell suspensions of Rhizobium trifolii, R. leguminosarum, R. phaseoli and R. lupini was studied in the presence or in the absence of 2-ketoglutaric acid. In R. lupini, production of IAA was strongly enhanced by the ketoacid, but in fast growing rhizobia it was less enhanced or unaffected. On the other hand, glutamic acid inhibits IAA production by R. meliloti, but stimulates IAA production in both R. leguminosarum and R. phaseoli. A hypothesis is proposed to explain these results.  相似文献   

11.
Evidence is presented of strain differences within Rhizobium trifolii in ability to colonize soil surrounding common pasture species, both legume and non-legume, in problem sandy soils in Western Australia. R. lupini and R. trifolii are shown to spread better than R. meliloti in these soils.  相似文献   

12.
Results from an innovative approach to improve remediation in the rhizosphere by encouraging healthy plant growth and thus enhancing microbial activity are reported. Mixed grass-legume systems, together with microbial inoculants, were used to remediate a polycyclic aromatic hydrocarbon (chrysene) spiked agricultural soil. Inoculants were symbiotic rhizobia, which may play an important role in rhizoremediation by increasing plant and root growth. An inoculum of an isolate of Rhizobium leguminosarum bv. trifolii, selected for PAH tolerance, was produced using a peat carrier. The inoculum and white clover (Trifolium repens L.), were planted into soils with ryegrass (Lolium perenne L.). The soils spiked with chrysene (500 mg kg−1) then aged for 4 weeks. Shoot- and root-biomass of plants, and the amount of root nodulation, were determined. Rhizobial populations, soil pH and soil nitrogen were also monitored throughout the trial. In addition, the ability of the inoculated rhizobial strain to utilise chrysene as a sole carbon source was assessed. Direct uptake and/or degradation of chrysene by the clover and ryegrass did not occur to a significant degree. Enhanced losses of chrysene were seen in planted, non-sterile soils that contained a rhizobial inoculum. No direct degradation of chrysene by R. leguminosarum bv. trifolii was observed and no enhanced losses of PAHs were detected in sterile soils after inoculation with rhizobia. Results suggest that the enhanced dissipation of chrysene, observed in the non-sterile planted inoculated pots, was not a result of degradation of chrysene by R. leguminosarum bv. trifolii. The symbiotic association with R. leguminosarum bv. trifolii improved plant vigour and growth in inoculated planted treatments. This may have stimulated the rhizospheric microflora to degrade chrysene.  相似文献   

13.
Five strains of R. trifolii were evaluated, at two inoculation levels in the presence or absence of lime pelleting, for their ability to compete and persist in a tussock grassland soil containing a naturalized population of rhizobia. Effects on the growth of the oversown white clover (Trifolium repens) were also investigated.Strains showed marked differences in their ability to form nodules on the host in competition with the naturalized population of rhizobia and also differed in their persistence over 15 months. The most competitive and persistent strain was PDDCC 2163 followed by 2153, 2666 and 2668 and the least competitive was 4144. Lime pelleting or increasing the rate of inoculation increased the competitive ability of strains. Strains that were highly competitive increased clover dry matter and N uptake.  相似文献   

14.
Glyphosale was added to samples of a sandy loam at rates of 0, 2, 5 and 10μg g?1soil. After 120 days, soil was transferred to pots which were planted with subterranean clover. Plants were inoculated with Rhizohium trifolii and N2 fixation (C2H2-reduction) was recorded after 9, 13, 15 and 19 weeks of growth. Nodule numbers and root weights were determined after the final C2H2-reduction assays had been performed. Decreased C2H2-reduction, nodule numbers and root weights associated with plants growing in glyphosate-treated soil indicated that this herbicide was not inactivated during the 120-days before planting.  相似文献   

15.
Liming an acid soil increased the yield and N content of subterranean clover in both field and glasshouse experiments. Application of Mo increased the N concentration of field-grown subterranean clover which corresponded with observed colour and growth differences, but did not change C2H2 reduction activity. Herbage Mo was not increased by liming, suggesting an absolute deficiency of Mo in these acid soils. In the glasshouse liming increased nodulation which increased the amount of N2 fixed but the lime had no direct effect on nitrogenase activity as measured by C2H2 reduction. In the field both inoculation and lime application increased soil populations of R. trifolii, but clover yield was greater with liming alone than with inoculation alone, indicating the sensitivity of the host plant to soil acidity.  相似文献   

16.
Leakage of materials reacting with the fluorescent probe 1-anilino-8-naphthalcne sulphonate (ANS) from root-nodule bacteria subjected to desiccation and rehydration suggests that death of these bacteria is associated with changes in membrane permeability. The slow-growing rhizobia and the fast-growing Rhizobium meliloti differ from other fast-growing bacteria of the R. leguminosarum group in that they exhibit little fluorescence in the presence of ANS. This probably reflects differences between these groups of bacteria in the polar/non-polar nature of the outer cell envelopes. Following desiccation and rehydration, all groups of root-nodule bacteria fluoresced strongly in the presence of ANS.Repair of desiccation-induced damage to the cell envelopes of R. japonicum QA372. R. trifolii SU297/32B and R. meliloti CC131 was not observed using a range of screening agents. However, repair of significant numbers of lysozyme-sensitive bacteria was observed with R. leguminosarum TA101 within an hour of rehydration.  相似文献   

17.
The role of cobalt in the processes leading to nodule initiation in Lupinus angustifolius L. cv. Uniharvest was studied. Survival and multiplication of Rhizobium lupini were examined in Lancelin soil and in the rhizosphere of L. angustifolius grown in it, with or without inoculation with antibiotic-resistant derivatives of R. lupini strain WU425. A clearing and staining technique was then used to record the incidence of nodule initiation in L. angustifolius seedlings.Within the constraints of experimental techniques, soil and rhizosphere numbers of R. lupini were unaffected by the cobalt status of the soil. Cobalt limitation was, however, inhibitory to nodule initiation in L. angustifolius, with reductions of up to 13 and 30% in nodule initiation for inoculated and uninoculated seedlings respectively.The severity of cobalt deficiency in L. angustifolius seems to be due solely to the plant's inability to provide cobalt to the rhizobia within the plant and not to any external limitation on their growth in soil or rhizosphere.  相似文献   

18.
The ability of 4 strains of Rhizobium trifolii to compete with naturalized strains in nodulating Trifolium subterruneum cv. Mt Barker and cv. Woogenellup was assessed at 5 sites in New South Wales. The populations of naturalized rhizobia at these sites ranged from 4 × 106 rhizobia/g to one where no rhizobia were detected. The introduced strains were inoculated singly or as mixed strain inocula onto seed of the host at 2 × 106 rhizobia/seed. There were marked differences in competitive ability between the strains but these differences were modified by the host cultivar and the site.At the R. trifolii-free site the inoculum strain formed 100% of the nodules in the 1st yr; by the second year serologically unrelated strains had invaded the plots and these formed almost all of the nodules in the 3rd yr. At the site where competition was greatest (4 × 106 naturalized rhizobia/g), there were no differences in the competitive abilities of the strains in the first year but at all other sites WU95 was superior whether used as a single strain or in a mixed strain inoculum. In these sites also the proportion of nodules formed by the inoculum strains declined markedly by the 2nd yr.  相似文献   

19.
The average number of survivors of fast-growing medic rhizobia (3 strains), fast-growing Rhizobium leguminosarum types (6 strains) and slow-growing species (9 strains) following desiccation of sandy soil inoculated with 106 bacteria·g?1 soil was 727, 795 and 15,682 bacteria·g?1 soil, respectively. Survival in desiccated sandy soil was not influenced by the degree of extracellular polysaccharide production in strains of R. trifolii, nor was it influenced by growth of R. meliloti and slow-growing species in media of low water activity before desiccation in sandy soil.A progressive increase in numbers of fast-growing bacteria surviving desiccation was observed in sandy soil amended with increasing concentrations of powdered montmorillonite, but not with mont-morillonite added as a suspension to the soil. The clay had either a detrimental effect or no effect on the survival of the slow-growing rhizobia. Maltose, sucrose and polyvinylpyrrolidonc provided a greater degree of protection to both fast- and slow-growing rhizobia than was obtained with montmorillonite. The effect of polyethylene glycol 6000 was similar to the effect of montmorillonite, as the polymer only protected the fast-growing rhizobia and not the slow-growing species.  相似文献   

20.
A major constraint to the renovation of forage legume‐based pastures on acidic soils of the Appalachian hill‐lands is thought to be the absence of effective rhizobia. A growth chamber experiment was done with aluminum (Al) toxic, low pH (≥ 4.2) soils from four series (Berks, Lily, Tate, and Westmoreland) that were planted with alfalfa (Medicago sativa L.), red clover (Trifolium pratense L.), white clover (Trifolium repens L.), or birdsfoot trefoil (Lotus corniculatus L.). These soils, without lime addition, were previously shown not to contain effective, naturalized populations of rhizobia for these plant species. However, a non‐toxic, pH 6.8, Watauga soil was shown to have such rhizobia but only for alfalfa. In the present study, these five soils were reexamined after liming to pH ≥ 5.5 for effective, naturalized populations of rhizobia and the efficacy of soil inoculation with commercially available rhizobia. In addition to effective, naturalized R. meliloti for alfalfa in the Watauga soil, similar populations of R. trifolii for red clover, and R. lotus for birdsfoot trefoil, were now found. Such rhizobia were also found for alfalfa in the Lily soil and for red clover in the Lily and Tate soil. Thus, liming allowed the expression of effectiveness of natural rhizobia that otherwise would not have been detected in soil pot experiments without lime. Inoculation of the toxic soils after lime addition with commercial rhizobia was effective in about half of the soil‐plant combinations that did not contain populations of effective, naturalized rhizobia. Asymbiotic shoot growth of all the plant species was significantly (P ≤ 0.05) correlated with soil pH over a range of 5.5–6.6. These results indicate that, in the absence of effective, naturalized populations of rhizobia, improvement of rhizobial inocula could increase forage production by ～34% for some species on some of the toxic soils, even after the pH of the soils is increased to ≥ 5.5.  相似文献   

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