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1.
Summary Bacteria isolated from the root zones of field-grown soybean plants [Glycine max (L.) Merr.] were examined in a series of glasshouse experiments for an ability to affect nodulation competition among three strains of Bradyrhizobium japonicum (USDA 31, USDA 110, and USDA 123). Inocula applied at planting contained competing strains of B. japonicum with or without one of eleven isolates of rhizosphere bacteria. Tap-root nodules were harvested 28 days after planting, and nodule occupancies were determined for the bradyrhizobia strains originally applied. Under conditions of low iron availability, five isolates (four Pseudomonas spp. plus one Serratia sp.) caused significant changes in nodule occupancy relative to the corresponding control which was not inoculated with rhizosphere bacteria. During subsequent glasshouse experiments designed to verify and further characterize these effects, three fluorescent Pseudomonas spp. consistently altered nodulation competition among certain combinations of bradyrhizobia strains when the rooting medium did not contain added iron. This alteration typically reflected enhanced nodulation by USDA 110. Two of these isolates produced similar, although less pronounced, effects when ferric hydroxide was added to the rooting medium. The results suggest that certain rhizosphere bacteria, particularly fluorescent Pseudomonas spp., can affect nodulation competition among strains of R. japonicum. An additional implication is that iron availability may be an important factor modifying interactions involving the soybean plant, B. japonicum, and associated microorganisms in the host rhizosphere.Paper No. 10648 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7601, USA  相似文献   

2.
N2 fixation, photosynthesis of whole plants and yield increases in soybeans inoculated with mixed cultures of Bradyrhizobium japonicum 110 and Pseudomonas fluorescens 20 or P. fluorescens 21 as well as Glomus mosseae were found in pot experiments in gray forest soil carried out in a growth chamber. The effects of pseudomonads and vesicular-arbuscular (VA) mycorrhizal fungus on these parameters were found to be the same. Dual inoculation of soybeans with mixed cultures of microorganisms stimulated nodulation, nitrogenase activity of nodules and enhanced the amount of biological nitrogen in plants as determined by the 15N dilution method in comparison to soybeans inoculated with nodule bacteria alone. An increased leaf area in dually infected soybeans was estimated to be the major factor increasing photosynthesis. P. fluorescens and G. mosseae stimulated plant growth, photosynthesis and nodulation probably due to the production of plant growth-promoting substances. Increasing phosphorus fertilizer rates within the range of 5–40 mg P 100 g-1 1:1 (v/v) soil: sand in a greenhouse experiment led to a subsequent improvement in nodulation, and an enhancement of N2 fixation and yield in soybeans dually inoculated with B. japonicum 110 and P. fluorescens 21. These indexes were considerably higher in P-treated plants inoculated with mixed bacterial culture than in plants inoculated with nodule bacteria alone.  相似文献   

3.
Summary Six fast-growing soybean rhizobia (Rhizobium fredii) and thirteen slow-growing soybean rhizobia (Bradyrhizobium japonicum) were examined for resistance to 10 antibiotics. Axenic studies were carried out to determine the competitiveness of dual-strain inocula consisting of fast- and slow-growing rhizobia isolated from subtropical-tropical soils for nodule occupancy on a hybrid of Asian and US soybean cultivars. Nodule occupancy was determined by intrinsic resistance to erythromycin and neomycin. The results showed wide variability in resistance to 10 antibiotics for fast- and slow-growing rhizobia. The intrinsic antibiotic resistance of fast- and slow-growing rhizobia was extremely high against nalidixic acid (400 g ml–1) and penicillin (200 g ml–1). The competitive ability of inoculant strains for nodule occupancy varied for different combination sets and with the plant growing media. Our results show that fast-growing rhizobia nodulate a hybrid of Asian and US soybean cultivars. Fast-growing soybean rhizobia did not completely exclude nodulation by the slow-growing strains, which formed 0–79% nodules, depending on the strain used in the inoculum.  相似文献   

4.
Soybean (Glycine max L. Merr.) cultivars Akisirome and Peking were inoculated with Bradyrhizobium japonicum Is-21 and Rhizobium fredii USDA 194, respectively, and were grown in cylindrical pots containing sterilized vermiculite which were aerated with CO2-free air or ambient air to study the effects of CO2 deficiency in the rhizosphere on plant growth, nodulation, and nitrogen fixation. The repressive effects of CO2-free air treatment were more conspicuous in Peking than Akisirome, and nodule number, nodule mass, amount of biologically fixed N and plant growth of Peking were reduced remarkably by the CO2-free air treatment.

Acetylene reduction activity (ARA) of Peking inoculated with USDA 194 and Akisirome inoculated Is-21 was assayed in the absence and presence of CO2, ARA of Akisirome was not affected by the absence of CO2, while that of Peking decreased drastically.

Based on these results, it was concluded that nitrogen fixation by Akisirome inoculated with B. japonicum Is-21 was not appreciably influenced by CO2 deficiency in the rhizosphere, while that of Peking inoculated with R. fredii USDA 194 was severely repressed, and the decrease was estimated to be due to both the reduction of the nodule mass and specific nitrogen-fixing activity.  相似文献   

5.
Summary A field experiment was condutced in a clay loam soil to study the performance of three Bradyrhizobium japonicum strains; USDA 110, USDA 138 and TAL 379, in relation to their N2-fixing potential and competitiveness on two soybean cultivars (Clark and Calland). Inoculation of soybean cultivars with these strains, either singly or in combination, induced significant increases in plant dry weight, N2 fixation and seed yields. However, no significant differences were found between the rhizobial strains and/or their mixtures in N2 fixation and increased seed yield for both cultivars. The two soybean cultivars gave similar responses to inoculation. No significant differences in seed yield were observed between Clark and Calland cultivars. The interaction between inoculant strain and soybean cultivar was not significant. The competition between strains for nodulation was assessed. Strain USDA 110 was the most competitive, followed by USDA 138. Strain TAL 379 was always less competitive on both cultivars. The incidence of double-strain occupancy of nodules varied from 8% to 40%.  相似文献   

6.
Summary Experiments to assess the ability of free-living cells of six strains of soybean rhizobia (Bradyrhizobium japonicum USDA 76, 94, 110, 122, 123, and 135) to denitrify nitrate in five soils showed that although some strains ofB. japonicum have the capacity to rapidly denitrify nitrate in soils under anaerobic conditions, it is unlikely that the numbers of soybean rhizobia commonly found under field conditions are sufficient to significantly influence either the extent or the products of denitrification in soil. It is our general conclusion that the advantages, if any, that the ability to denitrify conveys to rhizobia or to the rhizobia-legume symbiosis are not offset by increased losses of plant-available N when denitrifying strains of rhizobia are present as free-living cells in soil.  相似文献   

7.
Soybean plant is characterized by a systemic autoregulatory control system of nodulation (autoregulation) by initial infection with rhizobia, and plants commonly display a systemic acquired resistance (SAR) to pathogenic microbe infection related to salicylic acid (SA) signal transmission. We investigated the effect of exogenous SA supply on soybean nodulation to determine whether SA affects the autoregulation of nodulation. Seedlings of the hypernodulating mutants NOD1-3, NOD2-4 and their parent cv. Williams were treated or not treated (control) with a 100 μmS-SA solution at 5 d before the inoculation of Bradyrhizobium japonicum strain USDA110. The nodule dry weight and the number of nodules of the wild type soybean Williams exhibiting autoregulation drastically decreased by the addition of 100 μm SA. The decrease in the nodule number was not caused by the reduction of the rhizobium number in the medium. Salicylic acid inhibited only early nodule formation and did not affect the growth of formed nodules. The inhibitory effect of SA on the nodulation of NOD1-3 and NOD2-4 was significantly less pronounced than that in Williams. These results indicate that SA is directly involved in signal transmission in the autoregulation, and that SA or the SAR induced by SA stimulates the autoregulation of nodulation in soybean.  相似文献   

8.
Summary Mixed infections of Bradyrhizobium japonicum strains in early and late nodules of four soybean cultivars were studied in a field soil. Nodule occupants were identified by immunofluorescence using serogroup specific antibodies prepared against B. japonicum strains USDA 110, USDA 123, and USDA 138. Double infection was determined directly by combined examination of the same microscopic field by fluorescence and phase contrast microscopy. Double strain occupancy was observed consistently, and its occurrence did not differ substantially in pouch, soil pot, and field experiments, ranging in incidence from 12% to 32%. No significant differences in the incidence or nature of double infection could be attributed to cultivar, seed inoculation, or plant maturity. Strains reactive to strain USDA 123-fluorescent antibody were dominant in both singly and doubly infected nodules irrespective of cultivar, plant age, or seed inoculation with strain USDA 110.Paper no. 15092 in the Scientific Journal Series of the Minnesota Agricultural Experiment Station, St. Paul  相似文献   

9.
Volcanic ash soil, which is widely distributed in Japan, contains a large amount of well-structured soil aggregates. By using these aggregates as carrier materials, we prepared (brady)rhizobial inoculants for red kidney bean (Phaseolus vulgaris) and soybean (Glycine max). Autoclaved soil aggregates were inoculated with Rhizobium tropici CIATS99R or Bradyrhizobium japonicum USDA110R, incubated for 15 or 21 d at 30°C, slowly air-dried at 20°C to prepare the aggregate-based inoculants, and stored at various temperatures. The populations of CIATS99R and USDA110R in the aggregate-based inoculants were maintained during several months of storage at 20°C. When the aggregate-based inoculants were mixed with soil, CIATS99R and USDA110R cells showed a remarkably improved survival in soils compared with those mixed with soil without carrier material. The effect of the aggregate-based inoculants on the growth of red kidney bean and soybean was examined in pot experiments. By placing a small amount of the inoculant just beneath the seeds at the time of sowing, plant growth was significantly enhanced compared with the use of traditional peat-based inoculant. In addition, nodule formation on the upper part of soybean roots and nodule occupancy by the inoculated strain were remarkably enhanced by the aggregate-based inoculant. It is suggested that soil aggregates might be suitable carrier materials for preparing cheap and effective (brady)rhizobial inoculants.  相似文献   

10.
We examined the competitiveness of five effective Rhizobium leguminosarum biovar phaseoli strains in the nodulation of kidney beans (Phaseolus vulgaris L.), either alone or in pairwise combination, against the indigenous strains. The results showed that the introduced Rhizobium sp. strains (B2, B17, B36, T2, or CIAT 652) occupying 64–79% of the total nodules (as single inocula) were more competitive in nodulation than the native rhizobia. However, the competitiveness of the individual Rhizobium sp. strain either increased or decreased when used in a pairwise combination of double-strain inocula. For example, strain B17, although quite competitive against the indigenous population (68% nodule occupancy), became poorly competitive in the presence of strain B2 (reduced from 68 to 2.5%). A similar reduction in nodule occupancy by strain B17 was observed in the presence of B36 or CIAT 652, indicating that two competitive strains may not always be compatible. These results suggest that it is important to co-select competitive as well as compatible rhizobia for multistrain inoculant formulation.  相似文献   

11.
 Most soils sown with field beans (Phaseolus vulgaris L.) contain indigenous rhizobia which might interfere with the establishment of inoculated strains. As a consequence, the benefits of bean inoculation are usually questioned, and the use of N fertilizer is gradually becoming a common practice. The present study had the objective of evaluating the effectiveness of inoculation and N fertilization in field soil with (site 1) and without (site 2) a previous bean-cropping history. At site 1, which had a rhizobial population of 7×102 cells g–1 soil, inoculation had no effect on nodulation or yield, whereas at site 2 (<10 cells g–1 soil) inoculation increased nodulation, nodule occupancy by the inoculated strain and grain yield. N fertilizer decreased nodulation at both sites, but increased grain yield at site 1 but not at site 2, indicating that the response to inoculation and N fertilization depends on the cropping history. When bean was cultivated for the first time, indigenous populations of rhizobia were low and high yields were accomplished solely with seed inoculation, with no further response to N fertilizer. In contrast, previous cultivation of bean increases soil rhizobia, preventing nodule formation by inoculated strains, and N fertilizer may be necessary for maximum yields. A significant interaction effect between N fertilizer and inoculation was detected for serogroup distribution only at site 2, with N fertilizer decreasing nodule occupancy by the inoculated strain and increasing the occurrence of indigenous strains. Consequently, although no benefits were obtained by the combination of inoculation and N fertilizer, this practice may be feasible with the selection of appropriate N-tolerant strains from the indigenous rhizobial population. Received: 26 May 1999  相似文献   

12.
 In a previous study soybean Bradyrhizobium strains, used in Brazilian studies and inoculants over the last 30 years, and strains adapted to the Brazilian Cerrados, a region frequently submitted to environmental and nutritional stresses, were analyzed for 32 morphological and physiological parameters in vivo and in vitro. A cluster analysis allowed the subdivision of these strains into species Bradyrhizobium japonicum, Bradyrhizobium elkanii and a mixed genotype. In this study, the bacteria were analyzed for nodulation, N2 fixation capacity, nodule occupancy and the ability to increase yield. The goal was to find a relationship between the strain groups and the symbiotic performance. Two strains of Brazilian B. japonicum showed higher rates of N2 fixation and nodule efficiency (mg of N mg–1 of nodules) under axenic conditions. These strains also showed greater yield increases in field experiments when compared to B. elkanii strains. However, no differences were detected between B. japonicum and B. elkanii strains when comparing nodule occupancy capacity. The adapted strains belonging to the serogroup B. elkanii SEMIA 566, most clustered in a mixed genotype, were more competitive than the parental strain, and some showed a higher capacity of N2 fixation. Some of the adapted strains, such as S-370 and S-372, have shown similar N2 fixation rates and nodulation competitiveness to two Brazilian strains of B. japonicum. This similarity demonstrates the possibility of enhancing N2 fixing ability, after local adaptation, even within B. elkanii species. Differences in the DNA profiles were also detected between the parental SEMIA 566 and the adapted strains by analyses with the ERIC and REP-PCR techniques. Consequently, genetic, morphological and physiological changes can be a result of adaptation of rhizobia to the soil. This variability can be used to select strains capable of increasing the contribution of N2 fixation to soybean nutrition. Received: 28 May 1997  相似文献   

13.
The competition with established soil populations of Bradyrhizobium able to nodulate soybean has been one of the major constraints to the introduction of more efficient strains in Cerrados soils. The effects of nodulation establishment and persistence of four serologically distinct strains of Bradyrhizobium japonicum (CPAC 15 and CPAC 7, belonging to serogroups USDA 123 and CB 1,809) and B. elkanii (29 W and SEMIA 587, belonging to serogroups 29 W and 587) were examined. These strains were introduced in a dark-red oxisol, without indigenous populations of soybean bradyrhizobia, and were evaluated for 6 years. The experimental design was a completely randomized block with four replicates. In the first year, besides the inoculation treatments, there was also an uninoculated control. In the second year, the main plots were split into three sub-plots and treatments consisted of an uninoculated control, CPAC 7 and CPAC 15. In the third year, the entire area was inoculated with CPAC 7. In the fourth and sixth years, the plots were planted with soybean without inoculation, and in the fifth year the plots were left fallow. The strains introduced in the first year influenced nodule occupancy by strain CPAC 7 until the third successive growing season. By the fourth and sixth years, as a consequence of the dispersal of strains serologically related to serocluster 123 in the entire experimental area, this serogroup dominated the nodulation, occurring, on average, in more than 50% of the nodules of the treatments where it had never been inoculated.  相似文献   

14.
A glasshouse study was undertaken to investigate the effects of the copper fungicide Kocide 101 and its residues in soil on the growth, nodulation and nitrogen fixation of beans (Phaseolus vulgaris L.). The soil used was a sandy clay loam classified as Typic Rhodustalf. The bean variety SUA 90 was used as test crop. The bean rhizobia strains CIAT 899, PV, and a local isolate were used. Kocide 101 applied at the recommended rate (equivalent to 1.7 mg kg‐1 soil) had no significant negative effects on the growth, nodulation or nitrogen fixation of bean plants. Higher levels of Kocide 101 significantly (P < 0.05) reduced plant growth, nodulation and nitrogen fixation. The bean plants inoculated with the “local isolate”; rhizobia had the highest dry matter weights, nodule numbers and nodule dry weights, and also had more N fixation. They were followed by those inoculated with the PV, strain and, lastly, those inoculated with CIAT 899. The growth and nodulation of bean plants were still curtailed by the Kocide 101 residues four months after the fungicide was first applied to the soil. Therefore, occurrence of high levels of Kocide 101 in soils can have long‐term effects on the performance of the bean‐rhizobia symbiosis.  相似文献   

15.
Abstract

Hydrophobic mutants of the Bradyrhizohium japonicum strain 138NR were obtained by transposon Tn5 mutagenesis followed by replica-plating on polystyrene plates. Fifteen mutants were isolated at a frequency of 106. Gel-filtration analysis of the exopolysaccharides revealed that the hydrophobic mutants produced a significantly smaller amount of low-molecular-weight polysaccharides than the parent. Four of the isolated mutants formed a larger number of nodules on soybean (Glycine max L. Merr. cv. Tamahomare) than the parent, two were superior in symbiotic nitrogen fixation, and two were symbiotically defective on soybean. Competitive nodulation abilities of the mutants were examined by inoculating them to soybean with B. japonicum strain 123ET as a competitor and determining the nodule occupancy based on the antibiotic resistance. All the mutants tested except for the symbiotically defective ones were superior in their competitive nodulation ability to the parent strain. When inoculated 24 h before the inoculation with 123ET, the mutants exhibited an increased nodule occupancy (44–93% compared to 9% by the parent).  相似文献   

16.
The compatibility between rhizobia and host plants for nodulation was determined based on the genetic and physiological properties of both symbionts. It has been observed that soybean varieties carrying the Rj-gene were not nodulated effectively by certain strains or groups of rhizobia. Soybeans carrying the Rj 2-gene, Rj 2-varieties, were found to nodule ineffectively by the rhizobial strains belonging to the 3-24-44 and 122 serogroups (Caldwell 1966). In the same way, Rj 3- and Rj 4-varieties were found to nodule ineffectively by strains USDA 33 (Vest 1970) and USDA 61 (Vest and Caldwell 1972), respectively.  相似文献   

17.
Cropping in low fertility soils, especially those poor in N, contributes greatly to the low common bean (Phaseolus vulgaris L.) yield, and therefore the benefits of biological nitrogen fixation must be intensively explored to increase yields at a low cost. Six field experiments were performed in oxisols of Paraná State, southern Brazil, with a high population of indigenous common bean rhizobia, estimated at a minimum of 103 cells g–1 soil. Despite the high population, inoculation allowed an increase in rhizobial population and in nodule occupancy, and further increases were obtained with reinoculation in the following seasons. Thus, considering the treatments inoculated with the most effective strains (H 12, H 20, PRF 81 and CIAT 899), nodule occupancy increased from an average of 28% in the first experiment to 56% after four inoculation procedures. The establishment of the selected strains increased nodulation, N2 fixation rates (evaluated by total N and N-ureide) and on average for the six experiments the strains H 12 and H 20 showed increases of 437 and 465 kg ha–1, respectively,in relation to the indigenous rhizobial population. A synergistic effect between low levels of N fertilizer and inoculation with superior strains was also observed, resulting in yield increases in two other experiments. The soil rhizobial population decreased 1 year after the last cropping, but remained high in the plots that had been inoculated. DGGE analysis of soil extracts showed that the massive inoculation apparently did not affect the composition of the bacterial community.  相似文献   

18.
For examining the probability of increase in the occupation ratio of inoculated rhizobium in nodules, various Rj-soybean cultivars including the Rj 2 Rj 3 Rj 4-lines of soybean were grown in a field of the Kyushu University Farm. Bradyrhizobium japonicum USDA110 that carries uptake hydrogenase (Hup+) was used as an inoculum. The relative efficiency of nitrogen fixation generally increased by the inoculation. However, there were no significant differences in the effects among the genotypes of the host plants. The occupation ratio of serogroup USDA110 in the nodules on the taproot of the inoculated plants was in the range of 77–100%, suggesting that the B. japonicum strain USDA110 infected taproots immediately after inoculation. The occupation ratios in the nodules on the lateral roots were 53–67, 40–86, 63–83, and 62–77% in inoculated plants of the non-Rj-, Rj 2 Rj 3-, Rj 4-, and Rj 2 Rj 3 Rj 4-genotypes, respectively, and they decreased in all the genotypes with the progression of growth. At the time of the first sampling, the occupation ratios on the lateral roots of these Rj 2 Rj 3 Rj 4-genotypes showed values intermediate between those of IAC-2 (Rj 2 Rj 3) and Hill (Rj 4) , which were the parent cultivars of the Rj 2 Rj 3 Rj 4-lines, B340, B349, and C242. The reduction in the occupation ratio of the serogroup USDA110 for about 1 month after the first sampling was the lowest (0.13–0.16) in the Rj 2 Rj 3 Rj 4-genotypes, excluding B349, followed by the non-Rj- and Rj 2 Rj 3-genotypes and highest (0.52–0.69) in the Rj 4-genotypes, excluding Hill. Therefore, it was considered that the population of compatible rhizobia with host soybean plants increased in the rhizosphere with the progression of the development and growth. The results showed that with the expansion of the root area of host plants, the occupation ratio of type A rhizobia including the serogroup USDA110 was high. Therefore, the Rj 2 Rj 3 Rj 4-genotypes were superior to other Rj-genotypes in terms of the inoculation effects of nodulation type A rhizobium, B. japonicum USDA110. However, the preference of the Rj 2 Rj 3 Rj 4-genotype for serogroup USDA110 is not sufficient to rule out the competition with the other serogroups in this study. Therefore, the study should be centered on the isolation of more efficient (Hup+) and highly compatible rhizobial strains with the Rj 2 Rj 3 Rj 4- genotypes.  相似文献   

19.
Compatible rhizobia strains are essential for nodulation and biological nitrogen fixation (BNF) of hairy vetch (Vicia villosa Roth, HV). We evaluated how past HV cultivation affected nodulation and BNF across host genotypes. Five groups of similar HV genotypes were inoculated with soil dilutions from six paired fields, three with 10-year HV cultivation history (HV+) and three with no history (HV?), and used to determine efficiency of rhizobia nodulation and BNF. Nodulation was equated to nodule number and mass, BNF to plant N and Rhizobium leguminosarum biovar viceae (Rlv) soil cell counts using qPCR to generate an amplicon of targeted Rlv nodD genes. Both HV cultivation history and genotype affected BNF parameters. Plants inoculated with HV+ soil dilutions averaged 60 and 70 % greater nodule number and mass, respectively. Such plants also had greater biomass and tissue N than those inoculated with HV? soil. Plant biomass and tissue N were strongly correlated to nodule mass (r 2?=?0.80 and 0.50, respectively), while correlations to nodule number were low (r 2?=?0.50 and 0.31, respectively). Although hairy vetch rhizobia occur naturally in soils, past cultivation of HV was shown in this study to enhance nodulation gene-carrying Rlv population size and/or efficiency of rhizobia capable of nodulation and N fixation.  相似文献   

20.
Interstrain competitiveness is a key factor affecting the performance of rhizobium inoculant. In the present study five native strains of Bradyrhizobium japonicum, namely SSF 4, SSF 5, SSF 6, SSF 7 and SSF 8, were assessed for their competitiveness in nodulating soybean using serological methods. The strains were inoculated individually or with the type strain USDA 110 at a 1:1 ratio. Nodule occupancy determined by immunofluorescence and dot immunoblot assay revealed that under in vitro conditions SSF 8 is more competitive than USDA 110 whereas the others were less competitive. The competitive ability of these strains was also estimated in pot culture in the field. In red soil both SSF 8 and USDA 110 were equally competitive whereas in black soil SSF 8 competed better than USDA 110 and produced more nodules. In a black soil field trial using a randomized block design, USDA 110 or SSF 8, when inoculated alone, occupied the majority of the nodules and enhanced nodule dry weight and shoot biomass. SSF 8 was more competitive when the strains were co-inoculated. Received: 1 November 1996  相似文献   

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