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1.
Almost all of the soybean cultivars (Glycine max L. Merr.) form nodules on their roots by infection with Bradyrhizobium japonicum. However, it has been observed that the soybean cultivars harboring nodulation conditioning genes, for instance Rj 2, Rj 3, and Rj 4, do not form effective nodules with some strains of B. japonicum. Ishizuka et al. (1991b) classified the rhizobia isolated from field-grown soybeans into nodulation-types A, B, and C based on the compatibility with these Rj-soybean cultivars. Nodulation-type B is incompatible with Rj 2 Rj 3-cultivars and type C is incompatible with Rj 4-cultivars. Nodulation-type A rhizobia are compatible with both Rj 2 Rj 3 - and Rj 4-cultivars. Furthermore, the Rj-cultivars were found to prefer appropriate nodulation-type rhizobia for nodulation, that is, non-Rj-, Rj 2 Rj 3 -, and Rj 4-cultivars prefer nodulation-types A, C, and B rhizobia, respectively (Ishizuka et al. 1991a). In the present study, in order to examine the mechanisms of preference of Rj-cultivars for rhizobial strains for nodulation, root exudates from Rj-cultivars were examined for their effects on the growth rate and chemotaxis of various types of B. japonicum strains.  相似文献   

2.
For the increase of the occupation ratio of inoculum strain in the competition with indigenous rhizobia, the relationship between Rj-genotypes of soybean and the preference of Rj-cultivars for various types of rhizobia for nodulation was investigated by using the Rj 2 Rj 4-genotype of soybean isolated from the cross between the Rj 2 Rj 3-cultivar IAC-2 and Rj 4-one Hill (Ishizuka et al. 1993: Soil Sci. Plant Nutr., 39, 79-86). Firstly, these Rj 2 Rj 4-genotypes were found to harbor the Rj 3-gene. The Rj 2Rj3Rj4-genotypes of soybean were considered to exhibit a more narrow microsymbiont range for nodulation than the Rj 2 Rj 3-and Rj4-cultivars. Therefore, rhizobia were isolated from the nodules of various Rj-genotypes of soybeans grown in soils, and the preference of the Rj 2 Rj 3 Rj 4-genotype for indigenous rhizobia was examined. The nodule occupancy of serotype 110 was significantly higher in the bacteroids of the nodules from the Rj 2 Rj 3 Rj 4-rgenotypes than in those from the other genotypes, non Rj-, Rj 2 Rj 3-, and Rj 4-cultivars. These results demonstrated that the Rj 2 Rj 3 Rj 4-genotype prefers more actively serogroup USDA110 to the others of rhizobia. Thus, Rj 2 Rj 3 Rj 4-genotype is superior to non- Rj-, Rj 2 Rj 3-, and Rj 4-genotypes for the formation of efficient nodules for nitrogen fixation.  相似文献   

3.
Bradyrhizobium species are symbiotic partners of soybean plants. However, some Bradyrhizobium bacteria do not form functional nodules on the roots of Rj4 genotype soybean cultivars. Our objective was to identify the strains of Bradyrhizobium (i.e., type C strains) that are least competent to form nodules on the roots of this plant genotype. We checked (i) previously isolated type C strains of Myanmar Bradyrhizobium elkanii (MMY6-1, MMY6-2, and MMY6-5), (ii) previously isolated type C strains of Myanmarese Bradyrhizobium spp. (MMY3-5 and MMY3-7), and (iii) strain Is-34 of B. japonicum, for nodule formation when associated with Rj4 and other Rj genotype soybeans. Strains in groups (i), (ii), and (iii) are known to be incompatible with Rj4 soybean genotypes. MMY3-5 and MMY3-7 produced functional nodules when associated with Rj4 and other Rj genotype soybean cultivars, except Hill (Rj4) cultivar. The ratios of ineffective nodule numbers/total nodule numbers (I/T ratios) for MMY6-1, MMY6-2, MMY6-5, and Is-34 in association with Rj4 soybean cultivars were > 0.5, demonstrating incompatibility between these bacterial strains and the Rj4 genotype. Interestingly, the I/T ratios of MMY6-1 and MMY6-2 were higher than that of Is-34 in almost all Rj4 soybean cultivars. Thus, the nodule-forming abilities of the B. elkanii strains MMY6-1 and MMY6-2 were strongly suppressed in Rj4 soybean cultivars; these strains may therefore be useful to identify the Rj4 genotype in soybean cultivars.  相似文献   

4.
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   

5.
The selection of effective rhizobia for higher efficiency nitrogen fixation is one of the most important steps for inoculant production. Therefore, this experiment was conducted to select the most effective type A and type B strains for specific Rj-gene harboring soybean varieties and to test the symbiotic effectiveness of selected strains on different Rj-gene harboring soybean varieties. Screening experiments using the specific soybean varieties were done with a completely randomized design and three replications in this study. Evaluation of the effective Myanmar Bradyrhizobium strains for plant growth, nodulation and N2 fixation were studied in pot experiments using sterilized vermiculite in the Phytotron (controlled-environmental condition). Then, a pot experiment was conducted using Futsukaichi soil in the screen house (natural environmental condition). The N2 fixation ability of soybean was evaluated by acetylene reduction activity (ARA) and the relative ureide index method. In the first screening experiment, type A and type B strains with higher nitrogen fixation and proper nodulation on their respective soybean cultivars were selected for the next screening. In the second screening, Bradyrhizobium elkanii AHY3-1 (type A), Bradyrhizobium japonicum SAY3-7 (type A), B. elkanii BLY3-8 (type B) and B. japonicum SAY3-10 (type B) isolates, which showed higher nitrogen fixation and nodulation in Yezin-3 (Rj4) and Yezin-6 (non-Rj), were selected for the next experiment. In the third screening experiment, SAY3-7 and BLY3-8, which had higher nitrogen fixing potential and proper nodulation, were selected as effective isolates. These two isolates were compatible with non-Rj and Rj4 soybean varieties for nodulation and nitrogen fixation. Based on the results of the screening experiment, these two strains were tested for their symbiotic efficacy in Futsukaichi soil. This study shows that inoculation treatment of SAY3-7 and BLY3-8 significantly increased plant growth, nodulation, and N2 fixation at the V6, R3.5 and R8 stages in Yezin-3 (Rj4) and/or Yezin-6 (non-Rj), and the seed yield at R8 stage, in Yezin-3 (Rj4) and Yezin-6 (non-Rj) soybean varieties compared with the control treatment. It can be concluded that SAY3-7 and BLY3-8 are suitable for inoculant production because of their higher nitrogen fixation ability, proper nodulation and better productivity of Myanmar soybean cultivars.  相似文献   

6.
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   

7.
Summary Previous laboratory and greenhouse studies have shown that phages significantly reduce soil populations of homologous rhizobia. Reductions in nodulation and N2 fixation have also been observed. The purpose of the current study was to examine the effect of a phage specific ofBradyrhizobium japonicum USDA 117 on nodulation, nodule occupancy, N2 fixation and soybean growth and yield under field conditions. The phage was inoculated in combination withB. japonicum USDA 117 and/orB. japonicum USDA 110 (resistant strain) into a rhizobia-free sandy loam soil and planted toGlycine max (L.) Merr. Williams. When the phage was applied to soil inoculated withB. japonicum USDA 117 alone, significant reductions in nodule weight and number, shoot weight, foliar N, nitrogenase activity, and seed index were observed. When, however, the soil also contained the non-homologous strain,B. japonicum USDA 110, no significant effects on any of these parameters were found. Nodule occupancy by competing strains ofB. japonicum USDA 110 and USDA 117 was also affected by the phage. In soil which did not contain the phage, 46% and 44% of the identified nodules were occupied by USDA 110 and 117, respectively. When the phage was present in the soil, nodule occupancy byB. japonicum USDA 117 was reduced to 23%, while occupancy byB. japonicum USDA 110 was increased to 71%. These results suggest that nodulation by selected strains of rhizobia can be restricted and nodulation by more effective, inoculated strains can be increased through the introduction of a homologous phage to soils.  相似文献   

8.
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   

9.
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   

10.
Abstract

Genetic diversity and distribution of indigenous soybean-nodulating bradyrhizobia in Japan were investigated based on restriction fragment length polymorphism analysis of PCR product (PCR-RFLP) analysis of the 16S?23S rDNA internal transcribed spacer (ITS) region using Bradyrhizobium USDA strains as reference strains. Soil samples were collected from five field sites in Hokkaido, Fukushima, Kyoto, Miyazaki and Okinawa in Japan. A total of 300 isolates were derived from three Rj-genotype soybean cultivars, Akishirome (non-Rj), CNS (Rj 2 Rj 3) and Fukuyutaka (Rj 4), and five field site combinations. The PCR products of the ITS region were digested with HaeIII, HhaI, MspI and XspI. Electrophoresed patterns were analyzed for phylogenetic relationship using Bradyrhizobium reference strains. Results revealed 22 RFLP patterns and 11 clusters. The RFLP patterns of the seven clusters were similar or identical to Bradyrhizobium japonicum USDA 6, 38, 110, 115, 123 and Bradyrhizobium elkanii USDA 76 and 94. Four minor clusters were independent from the clusters of the reference strains. The isolation ratio revealed the major clusters at each field site. These results suggested that major clusters of indigenous bradyrhizobia might be in the order Bj123, Bj38, Bj110, Bj6 and Be76 from the northern to southern regions in Japan.  相似文献   

11.
Summary A method was developed to improve the colonizing ability of inoculated strains of root-nodule bacteria using aliette (aluminum tris-O-ethyl phosphonate), a basipetally translocated fungicide. Aliette applied to seeds of alfalfa inoculated with an aliette-resistant strain of Rhizobium meliloti increased the numbers of R. meliloti in the rhizosphere after 3 but not 37 days, increased the number of nodules, and with some seed treatments, increased the growth of alfalfa. The enhanced colonization by R. meliloti as a result of seed treatment with aliette lasted for at least 31 days for alfalfa, although plant weights did not increase, Colonization by R. meliloti was further enhanced if seeds and foliage were treated with the fungicide. Coating seeds or sparaying the foliage with aliette also increased the number and weight of nodules and nitrogenase activity in soybeans inoculated with an aliette-resistant strain of Bradyrhizobium japonicum. The stimulation of B. japonicum in the rhizosphere and of nodulation was evident with successive plantings of soybeans if the seeds for each planting were treated with the chemical, but aliette did not increase the yield of inoculated soybeans in the subsequent plantings. With only the seeds of the first planting of inoculated soybeans treated with aliette, the numbers of B. japonicum in the rhizosphere of subsequent plantings were only occasionally greater and the numbers of nodules on the later plantings were not increased. We suggest that root colonization, nodulation, and N2 fixation by Rhizobium and Bradyrhizobium may be enhanced by the use of basipetally translocated antimicrobial compounds together with root-nodule bacteria that are resistant to those compounds.  相似文献   

12.
Several commercial fungicide seed treatments were evaluated for their possible effect on the survival of Bradyrhizobium japonicum on seeds and on the nodulation and yield of soybeans in a greenhouse and a field experiment. quinolate Pro (carbendazim and oxine copper), Vitavax 200FF (carboxin and thiram), and Monceren (pencycuron) had a small effect or no effect on the survival of B. japonicum and on the nodulation and yield of soybeans. They can thus be considered compatible with soybean seed inoculation. Germipro UFB (carbendazim and iprodione), Apron 35J (metalaxyl), and Tachigaren (hymexazol) decreased B. japonicum survival and the nodulation and yield of soybeans and thus cannot be considered compatible with soybean seed inoculation.  相似文献   

13.
Abstract

The diversities of communities of soybean-nodulating indigenous bradyrhizobia in Japan were estimated using mathematical ecology methods based on the results of polymerase chain reaction–restriction fragment length polymorphism analysis of the 16S–23S rDNA internal-transcribed spacer region. Polar ordination analysis indicated a significant correlation between the compositions of bradyrhizobial communities and northern latitudes in Japan. This result suggests that the composition and the geographical distribution of indigenous soybean-nodulating bradyrhizobia might be affected by soil temperature and the associated diversity of the host plants acclimatized to a particular climate. Furthermore, for estimation of the compositional difference in bacterial communities among three different Rj-genotype soybean cultivars, an analysis of the diversity indexes was conducted and this analysis indicated differences in the composition of the communities isolated from the Rj 2 Rj 3-genotype compared with those from the non-Rj- and Rj 4-genotype cultivars. This result suggests that Rj 2 Rj 3-genotype soybeans might affect not only compatibility with particular bradyrhizobia, but also preference to bradyrhizobia for nodulation.  相似文献   

14.
15.
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   

16.
A group of Bradyrhizobium strains isolated from soybean plants in Thailand did not correspond to any known DNA homology groups of Bradyrhizobium japonicum and Bradyrhizobium elkanii reported by Hollis et al. (J. Gen. Microbiol., 123, 215–222, 1981). To clarify the phenotypic characteristics of the group, serological properties and intrinsic antibiotic resistance (IAR) profile of 94 Thai strains were compared with those of USDA and Japanese strains. Indirect ELISA tests for each Thai strain were performed agaiIl.st polyclonal antisera prepared against 15 USDA standard serotype strains of B. japonicum and B. elkanii. Among the 94 Thai strains tested, 36 which were previously identified as B. elkanii, with the exception of one strain, were strongly responsive to an antiserum prepared against USDA 31. The remaining 58 strains, with the exception of two strains, showed multiple cross reactions which were peculiar to the Thai strains. These serological reaction patterns did not correspond to any known serogroups labeled as B. japonicum and B. elkanii. In the IAR test, the taxonomically unknown Thai soybean bradyrhizobia exhibited a high level of resistance to neomycin (50 µg/mL), polymyxin (50 µg/mL), nalidixic acid (15 µg/mL), and kanamycin (15 µg/mL). Kanamycin could thus be useful in combination with neomycine and nalidixic acid for distinguishing between the unknown Thai strains and strains of B. japonicum and B. elkanii. Our results demonstrated that the unknown Thai strains were serologically and IAR-phenotypically remote from both B. japonicum and B. elkanii.  相似文献   

17.
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   

18.
Peanuts (Arachis hypogaea L.) were introduced to China about 500 years ago. However, the diversity of Rhizobial strains in China that can nodulate peanut was poorly understand. Diversity and phylogeny of 50 slow-growing strains, isolated from root nodules of peanut in different geographical regions of China, were studied using polyphasic techniques. All stains were clustered by phenotypic tests into two distinct groups: Group I: 16S rRNA RFLP genotype 3, and Group II, which divided into 16S rRNA RFLP genotypes 1 and 2. Genotype 1 shares the same genotype with USDA110, USDA122 and USDA127 of Bradyrhizobium japonicum, and genotype 2 solely consisted of extra-slow growing bradyrhizobia isolated from Hongan, China. Results of 16S rRNA sequencing revealed that peanut bradyrhizobia were phylogenetically related to B. japonicum and their sequence divergence was less than 1.1%. Based upon the size of the internally transcribed spacer (ITS) between the16S and 23S RNA genes, strains were classified into ITS-I, ITS-II and ITS-III genotypes. Strains could be further divided into sub-clusters IA, IB, IIa, IIb and IIc five sub-clusters through ITS PCR-RFLP and repetitive extragenic palindromic PCR (REP-PCR) analysis. Host specificity test revealed that all peanut bradyrhizobia tested nodulated Phaseolus vulgaris and strains of clusters IIb and IIc nodulated Glycine soja efficiently. Bradyrhizobia isolated from peanut were related, but still exhibited phylogenetical divergence with B. japonicum.  相似文献   

19.
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   

20.
In order to select appropriate Bradyrhizobium USDA reference strains for primary grouping of indigenous soybean bradyrhizobia, we systematically constructed phylogenetic trees of 20 USDA strains based on DNA sequence analysis and PCR-restriction fragment length polymorphism (RFLP) targeted to 16S rDNA and the internal transcribed spacer (ITS) region between 16S and 23S rDNAs. The phylogenetic trees of 16S rDNA showed 3 major groups, cluster USDA 110 (USDA 62, 110, 122, 125, and 129), cluster USDA 6 (USDA 4, 6T, 38, 115, 123, 127, 135, and 3622T) and cluster B. elkanii (USDA 31, 46, 61, 76T, 94, and 130), as well as the phylogenetically independent strain USDA 124. The topology of the ITS trees was almost similar to that of 16S rDNA, although the positions of two extra-slow-growing strains, USDA 135 and USDA 3622T were variable among the ITS sequences, PCR-RFLP of the ITS region and 16S rDNA. Only two strains, USDA 110 and USDA 122, harbored hup genes and they fell into the USDA 110 cluster. These results suggest that PCR-RFLP analysis of 16S rDNA and the 16S-23S rDNA ITS region may be useful for the grouping of bradyrhizobia and for the first screening of hup-positive strains. Based on the above results, we propose a minimum set of USDA strains reflecting Bradyrhizobium diversity that includes B. japonicum USDA 6T, B. japonicum USDA 110, B. japonicum USDA 124, and B. elkanii USDA 76T. In addition, an extra-slow-growing strain with the serotype USDA 135 might be necessary for genomic diversity analysis of bradyrhizobia, because their phylogenetic positions were variable.  相似文献   

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