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1.
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   

2.
Lotus uliginosus is generally nodulated by rhizobia of the genus Bradyrhizobium when used for improvement of Uruguayan pastures. The genetic diversity and phylogenetic relationships of 111 isolates from nodules of L. uliginosus collected from four fields with or without prior inoculation history were analyzed in this study. Genetic diversity estimated by ERIC-PCR revealed 75 different genomic fingerprints, and showed a relatively greater value compared with other methods and varied by soil type. 16S ribosomal RNA gene RFLP analysis revealed three different ribogroups, A, B and C, with 71 isolates in ribogroup A, three isolates in ribogroup B and only one in ribogroup C. Phylogenetic analyses based on 16S RNA gene sequences, ITS, as well as atpD, recA and glnII gene sequences indicated that ribogroup A strains were affiliated with B. japonicum bv. genistearum strains. The three isolates in ribogroup B did not clearly associate with any Bradyrhizobium species described previously and could represent a novel species within this genus. Unlike B. japonicum strains these isolates were able to nodulate and fix nitrogen with other Lotus species as well as with Spartium, a leguminous shrub. The unique isolate in ribogroup C clustered with Mesorhizobium and appeared genetically and phenotypically related to broad host-range Mesorhizobium sp. NZP2037. Our data suggest that Uruguayan soils contain native or naturalized bradyrhizobia that are able to nodulate L. uliginosus as efficiently as the commercial strain NZP2309 but could have adaptive advantages making them more suitable for inoculant purposes.  相似文献   

3.
We investigated the taxonomic position and symbiotic capabilities of two root-nodule bacterial strains isolated from the South African herbaceous, papilionoid legume Rhynchosia ferulifolia. The 16S rRNA gene sequence of the two strains was determined along with intragenic sequences of nodA and nifH, together with their symbiotic capabilities when inoculated onto the papilionoid legumes R. ferulifolia, Rhynchosia caribaea, Rhynchosia minima and Macroptilium atropurpureum (Siratro). Burkholderia phymatum STM815T, Cupriavidus taiwanensis LMG 19424T and root-nodule bacteria isolated from R. minima and Rhynchosia totta were included in the study. Root-nodule bacteria isolated from R. ferulifolia, WSM3937 and WSM3930, belong to the genus Burkholderia and are most closely related to Burkholderia terricola (98.8% similarity). The phylogenetic analysis of nodA and nifH revealed substantial similarity of the novel strains with Burkholderia tuberum STM678T, a β-rhizobium also originated from South Africa, and only a distant relationship with South American Mimosa-nodulating β-rhizobia. R. ferulifolia was effectively nodulated only by Burkholderia sp. WSM3937 and WSM3930 and not by bradyrhizobia isolated from Rhynchosia minima and Rhynchosia totta or STM815 and LMG 19924. Nodules induced by the novel strains were determinate and hosted well organized symbiosomes within infected cells. In this study we describe a new symbiotic N-fixing relationship between Burkholderia sp. and the South African legume R. ferulifolia. This is the first report of N-fixation between β-rhizobia and an herbaceous, papilionoid legume from which the strains were originally isolated. The level of N-fixation in this symbiosis approached that achieved by effectively nodulated Medicago sativa and suggests that the β-rhizobia may have a role in N-fixation in agricultural systems.  相似文献   

4.
The nodulation of Lotus pedunculatus and the multiplication of three Rhizobium loti (fast-growing, acid-producing) and two Bradyrhizobium (slow-growing, alkali-producing) strains was investigated in acidified rooting solution. R. loti strains multiplied at pH 4.5 but Bradyrhizobium strains failed to multiply. No difference in growth rate between R. loti and Bradyrhizobium strains was apparent in rooting solution at pH 6.7. Similar responses to pH were observed in yeast extract-mannitol broth except that Bradyrhizobium strains multiplied more slowly than R. loti at pH 6.7. All strains nodulated L. pedunculatus growing in acid (pH 4.5) rooting solution when presented as single cultures. Following inoculation with 1:1 mixtures of R. loti and Bradyrhizobium strains, R. loti formed 93% of nodules at pH 4.5 and significantly fewer nodules (66%) at pH 6.7. These results demonstrate a competitive advantage for acid-tolerant strains over acid-sensitive strains in nodulation of their lost legume at pH 4.5.  相似文献   

5.
《Applied soil ecology》2007,35(2):441-448
The size of the background rhizobial population can often determine the success of field nodulation and persistence of inoculant rhizobia. Field experiments were conducted to determine the nodulation response of annual medics (Medicago spp.) in a pasture-wheat-pasture rotation when grown in soils of contrasting pH and rhizobial populations. Medicago truncatula Gaertn. and M. polymorpha L. were inoculated with one of three different strains of Sinorhizobium medicae (WSM540, WSM688) or S. meliloti (NA39) or left uninoculated and sown in two fields of pH (CaCl2) 5.9 and 7.2 of differing soil rhizobial backgrounds (11 and 7.1 × 104 cells/g soil, respectively). Nodulation was assessed in years 1 and 3 of the rotation. At the site with a small rhizobial background, M. polymorpha nodulated poorly when inoculated with the acid-sensitive strain NA39 but nodulated well when inoculated with acid-tolerant strains WSM688 and WSM540. M. truncatula had a similar extent of nodulation with each of the rhizobial inoculants. At the site with a large rhizobial background all treatments had greater than 85% of plants nodulated. Nodule occupancies, assessed by PCR, provided further insight: at the site with a small rhizobial background both medic species successfully nodulated with the acid-tolerant strains WSM540 and WSM688 and these strains persisted to year 3. However, at the site with large rhizobial background, only one strain, WSM688, was identified from M. truncatula nodules in year 3. This study highlights the importance of edaphic constraints and plant–rhizobia interactions to the successful development of nodulation in a field environment.  相似文献   

6.
Twenty-eight Rhizobium strains were isolated from the root nodules of faba bean (Vicia faba L.) collected from 11 governorates in Egypt. A majority of these strains (57%) were identified as Rhizobium leguminosarum bv. viciae (Rlv) based on analysis of a nodC gene fragment amplified using specific primers for these faba bean symbionts. The strains were characterized using a polyphasic approach, including nodulation pattern, tolerance to environmental stresses, and genetic diversity based on amplified ribosomal DNA-restriction analysis (ARDRA) of both 16S and 23S rDNA. Analysis of tolerance to environmental stresses revealed that some of these strains can survive in the presence of 1% NaCl and a majority of them survived well at 37 °C. ARDRA indicated that the strains could be divided into six 16S rDNA genotypes and five 23S rDNA genotypes. Sequence analysis of 16S rDNA indicated that 57% were Rlv, two strains were Rhizobium etli, one strain was taxonomically related to Rhizobium rubi, and a group of strains were most closely related to Sinorhizobium meliloti. Results of these studies indicate that genetically diverse rhizobial strains are capable of forming N2-fixing symbiotic associations with faba bean and PCR done using nodC primers allows for the rapid identification of V. faba symbionts.  相似文献   

7.
This study tested the competitive ability of three locally isolated Cyclopia rhizobia and strain PPRICI3, the strain currently recommended for the cultivation of Cyclopia, a tea-producing legume. Under sterile glasshouse conditions, the three locally isolated strains were equally competitive with strain PPRICI3. In field soils, the inoculant strains were largely outcompeted by native rhizobia present in the soil, although nodule occupancy was higher in nodules growing close to the root crown (the original inoculation area). In glasshouse experiments using field soil, the test strains again performed poorly, gaining less than 6% nodule occupancy in the one soil type. The presence of Cyclopia-compatible rhizobia in field soils, together with the poor competitive ability of inoculant strains, resulted in inoculation having no effect on Cyclopia yield, nodule number or nodule mass. The native rhizobial population did not only effectively nodulate uninoculated control plants, they also out-competed introduced strains for nodule occupancy in inoculated plants. Nonetheless, the Cyclopia produced high crop yields, possibly due to an adequate supply of soil N.  相似文献   

8.
The Medicago sativa-Sinorhizobium symbiosis is challenged by acidity, resulting in generally poor nodulation and production. Medicago murex, however, can nodulate and grow at low pH. The effect of low pH on signal exchange in the Sinorhizobium-Medicago symbiosis was studied to gain a greater understanding of the basis for poor nodulation of M. sativa compared to M. murex. Root exudates from M. sativa and M. murex grown in buffered nutrient solution at pH 4.5, 5.8 and 7.0, were collected to measure the expression of nodB induction in Sinorhizobium. A nodB-gusA fusion was constructed and inserted into Sinorhizobium medicae strains WSM419 (acid tolerant) and CC169 (acid sensitive). We identified greater induction by root exudates from both Medicago spp. collected at pH 4.5 than at pH 5.8 and 7.0, less induction by M. murex than M. sativa and less induction of WSM419 than CC169. The same major inducing compounds, 4′,7-dihydroxyflavanone (liquiritigenin), 4′,7-dihydroxyflavone, and 2′,4′,4-trihydroxychalcone (isoliquiritigenin), were identified in exudates of M. murex and M. sativa at all pH values, although in increasing amounts at lower pH. Poor nodulation of M. sativa relative to M. murex under acid conditions is not the consequence of decreased induction of Sinorhizobium nodB by chemical inducers present in the root exudates of both species at low pH.  相似文献   

9.
Root-nodule bacteria that nodulate the legume genus Lotononis are being investigated to develop new forage species for agriculture. Bacteria isolated from nodules of Lotononis angolensis were fast-growing, highly mucoid and pink-pigmented, and on the basis of 16S rRNA phylogeny <94% related to other genera in the Alphaproteobacteria. Root-nodule bacteria isolated from other Lotononis species (L. bainesii, L. solitudinis and L. listii) resembled the more common dry, slow-growing, pink-pigmented rhizobia previously described for L. bainesii. These isolates could be attributed to the Methylobacterium genus, although not to the type species Methylobacterium nodulans. Further differences were uncovered with nodulation studies revealing that nodule isolates from L. angolensis were effective at nitrogen fixation on their host plant, but could nodulate neither L. bainesii nor L. listii. Reciprocal tests showed isolates from L. bainesii, L. listii and L. solitudinis were incapable of nodulating L. angolensis effectively. Nodule morphology for L. bainesii, L. angolensis and L. listii was characteristically lupinoid, with little structural divergence between the species, and with nodules eventually enclosing the entire root.  相似文献   

10.
Lime pelleting of the inoculated seed is recommended for most pasture legume species to improve survival of the rhizobia on the seed and to counter deleterious effects of soil or fertiliser acidity on rhizobial numbers. Except for New South Wales, lime pelleting is specifically not recommended for serradella (Ornithopus spp.). Our objectives were to evaluate effects of lime pelleting on bradyrhizobial numbers on seed, and nodulation and growth of the serradella plants. Three experiments are reported at two acid-soil sites in northern New South Wales involving four cultivars of yellow serradella (Ornithopus compressus) and Bradyrhizobium sp. (Lupinus) strains WSM471 (current inoculant strain) and WU425 and WSM480. Lime pelleting increased bradyrhizobial numbers on seed, 24 h after inoculation, by an average of 90%. Similarly, lime pelleting increased nodulation and shoot dry matter of the inoculated plants by an average of 57 and 28%, respectively. The three strains were similar in effects on plant growth. Relative values for shoot dry weight, averaged over sites, were 100 for WSM471 and 98 for both WU425 and WSM480. Our results confirmed previous research that lime pelleting inoculated serradella seed was not deleterious to survival of the bradyrhizobial inoculum, and showed that it could result in enhanced symbiotic activity of the inoculum in some instances. We recommend lime pelleting of serradella and that WSM471 remain the inoculant strain.  相似文献   

11.
In 1993 and 1994, 12 bacterial isolates were isolated from root nodules of cicer milkvetch (Astragalus cicer). In the tests for nodulation of A. cicer by these bacterial isolates, five were found to form hypertrophic structures, while only two formed true nodules. These true nodules were formed in a sterilized soil system. This system might be able to act as a DNA donor to provide residual DNA to other microbes in the soil. The rhizobial isolates were thought to have lost genetic material crucial to nodulation during the isolation process. This hypothesis was supported by an experiment in which isolate B2 was able to nodulate A. cicer in vermiculite culture after being mixed with heat-killed rhizobia, Rhizobium leguminosarum bv. trifolii and R. loti. The nodulation would not occur in vermiculite culture system without the heat-killed rhizobia. Based on the biochemical data, the B2 and 9462L, which formed true nodules with A. cicer, were closely related. The rhizobia type cultures that nodulate A. cicer include Bradyrhizobium japonicum, Rhizobium leguminosarum bv. trifolii, R. leguminosarum bv. viceae, and R. loti. All of these rhizobia were from different cross-inoculation groups. The B2 and 9462L isolates could only nodulate Medicago sativa, Phaseolus vulgaris, and Melilotus officinalis, but not these species within the genus from which they were isolated: Astragalus. The traditional cross-inoculation group concept obviously does not fit well in the classification of rhizobia associated with Astragalus. The rhizobia isolated from A. cicer can be quite different, and the rhizobia able to renodulate A. cicer also quite diverse. Received: 27 June 1996  相似文献   

12.
Diversity of rhizobia nodulating Lotus corniculatus grown in geographical regions with different rainfall regimes in northern and southern Uruguay, was estimated using 168 root nodule isolates. ERIC-PCR analysis revealed no correlation between observed fingerprints and the geographical origin of isolates. Despite the commercial strain U510 has been used for decades to inoculate L. corniculatus, none of the isolates corresponded to this strain. Phylogenetic analyses using 16S rRNA and atpD genes, and ITS sequences clustered all the isolates within genus Mesorhizobium. A great majority of the isolates likely belong to the species M. huakuii, as does the commercial strain U510. The remaining isolates were closely related to either M. septentrionale or M. caraganae. Although no M. loti-like bacteria were identified, all isolates carried symbiotic genes closely related to M. loti and other narrow host range Lotus rhizobia. A significant portion of the Uruguayan isolates were as efficient as the reference strain U510 in symbiosis with L. corniculatus. A few of the isolates were also capable of nitrogen fixation in symbiosis with L. uliginosus, albeit with lower efficiency than reference strains. Our results indicate that rhizobia nodulating L. corniculatus in Uruguay are genetically and phenotypically diverse, and that the commercial strain U510 is probably not adapted to survive the Uruguayan edaphoclimatic conditions.  相似文献   

13.
《Journal of plant nutrition》2013,36(9):1367-1375
A study was conducted to compare the responses to acid pH of Medicago sativa and Lotus glaber, two forage legumes with different environmental requirements, either supplied with inorganic nitrogen (N) or inoculated with different strains of their nodule bacteria. Medicago sativa showed, in both treatments, a significant reduction in total dry weight at pH below 6.0. In contrast, Lotus glaber grew equally well at all the pHs assayed in the presence of adequate N. Under inoculated conditions, in the absence of N supply, plant growth was dependent on the bacterial strain used. When the ability of each strain to multiply in culture medium was examined, it was observed that Sinorhizobium meliloti strains showed a pH-sensitive response that inhibited growth at pH 4.0, whereas Mesorhizobium loti strains showed normal growth at this pH. These results suggest that for the effectiveness of Mesorhizobium loti–Lotus glaber symbiosis in acid soils the major factor to be considered is the tolerance of the bacterial strain to acid conditions, while the limiting factors for the Sinorhizobium meliloti–Medicago sativa symbiosis are the sensitivity to low pH of both the plant and its bacteria.  相似文献   

14.
Bacteria were isolated from root-nodules collected from indigenous legumes at 38 separate locations in the Gascoyne and Pilbara regions of Western Australia. Authentication of cultures resulted in 31 being ascribed status as root-nodule bacteria based upon their nodulation of at least one of eight indigenous legume species. The authenticated isolates originated from eight legume genera from 19 sites. Isolates were characterised on the basis of their growth and physiology; 20 isolates were fast-growing and 11 were slow-growing (visible growth within 3 and 7 d, respectively). Fast-growers were isolated from Acacia, Isotropis, Lotus and Swainsona, whilst slow-growers were from Muelleranthus, Rhynchosia and Tephrosia. Indigofera produced one fast-growing isolate and seven slow-growing isolates. Three indigenous legumes (Swainsona formosa, Swainsona maccullochiana and Swainsona pterostylis) nodulated with fast-growing isolates and four species (Acacia saligna, Indigofera brevidens, Kennedia coccinea and Kennedia prorepens) nodulated with both fast- and slow-growing isolates. Swainsona kingii did not form nodules with any isolates. Fast-growing isolates were predominantly acid-sensitive, alkaline- and salt-tolerant. All slow-growing isolates grew well at pH 9.0 whilst more than half grew at pH 5.0, but all were salt-sensitive. All isolates were able to grow at 37 °C. The fast-growing isolates utilised disaccharides, whereas the slow-growing isolates did not. Symbiotic interactions of the isolates were assessed on three annual, one biennial and nine perennial exotic legume species that have agricultural use, or potential use, in southern Australia. Argyrolobium uniflorum, Chamaecytisus proliferus, Macroptilium atropurpureum, Ononis natrix, Phaseolus vulgaris and Sutherlandia microphylla nodulated with one or more of the authenticated isolates. Hedysarum coronarium, Medicago sativa, Ornithopus sativus, Ornithopus compressus, Trifolium burchellianum, Trifolium polymorphum and Trifolium uniflorum did not form nodules. Investigation of the 31 authenticated isolates by polymerase chain reaction with three primers resulted in the RPO1 primer distinguishing 20 separate banding patterns, while ERIC and PucFor primers distinguished 26 separate banding patterns. Sequencing the 16S rRNA gene for four fast- and two slow-growing isolates produced the following phylogenetic associations; WSM1701 and WSM1715 (isolated from Lotus cruentus and S. pterostylis, respectively) displayed 99% homology with Sinorhizobium meliloti, WSM1707 and WSM1721 (isolated from Sinorhizobium leeana and Indigofera sp., respectively) displayed 99% homology with Sinorhizobium terangae, WSM1704 (isolated from Tephrosia gardneri) shared 99% sequence homology with Bradyrhizobium elkanii, and WSM1743 (isolated from Indigofera sp.) displayed 99% homology with Bradyrhizobium japonicum.  相似文献   

15.
The 15N natural abundance technique is one of those most easily applied ‘on farm’ to evaluate the contribution of biological N2 fixation (BNF) to legume crops. When proportional BNF inputs are high, the accuracy of this technique is highly dependent on an accurate estimate of the 15N abundance of the N derived from N2 fixation (the ‘B’ value). The objective of this study was to determine the influence of soybean variety on ‘B’ value. Plants of five soybean varieties were inoculated separately with two Bradyrhizobium strains (one Bradyrhizobium japonicum and one Bradyrhizobium elkanii) grown in pots of soil virtually free of bradyrhizobia capable of nodulating soybean. The proportion of N derived from BNF (%Ndfa) was estimated in separate pots where a small quantity of enriched 15N ammonium sulphate was added. The %Ndfa was then used with the 15N natural abundance data of the nodulated soybean and non-N2-fixing reference plants, to determine the ‘B’ value for each soybean variety/Bradyrhizobium association. The varieties nodulated by the B. japonicum strain showed significantly greater N content and %Ndfa than those nodulated by the B. elkanii strain, and in all cases the ‘B’ value of the shoot tissue (‘Bs’) was higher. The differences in ‘Bs’ values between varieties nodulated by the same Bradyrhizobium strain were insignificant, indicating that this parameter is influenced much more by the Bradyrhizobium strain than by the variety of the host plant.  相似文献   

16.
We have previously reported significant strain–host incompatibility between the microsymbiont Rhizobium leguminosarum biovar trifolii and Trifolium spp. related to geographic and phenological barriers. Additionally, we have shown that an effective symbiosis between strains of R. l. trifolii and clover was established despite the soil harbouring ineffective R. l. trifolii capable of nodulating the host. We termed this “selection” for effective symbiosis. This paper reports glasshouse-based experiments to validate and further explore this in situ selection phenomenon. The effect of cell density and strain ratio at the time of inoculation, as well as soil pH, were investigated on two hosts (Trifolium purpureum and Trifolium polymorphum) that were each exposed to one microsymbiont capable and one incapable of N2 fixation. In co-inoculation experiments at a cell density of 104 cells/mL, each host nodulated solely with its effective strain, even when this strain was outnumbered 100-fold by the ineffective strain. However, the selection process ceased when the effective strain was outnumbered 1000-fold. At higher basal cell concentrations of 105–108 cells/mL, selection for WSM1325 to form effective nodulation on T. purpureum was evident, but was significantly reduced as the ratio of ineffective cells in the inoculum increased above 4-fold. The results indicate that the selection mechanism is highly dependent on the basal rhizobial cell density. Soil pH did not significantly alter the process, and both strains grew at similar rates and formed nodules at similar rates. A preliminary investigation into the genetic backgrounds of WSM1325 and WSM2304 revealed that although their 16S rRNA sequences were identical, they have considerable differences in their symbiotic and chromosomal replicons through examination of atpD, GSII and nodD sequences.  相似文献   

17.
Physiological responses to acid stress in two strains of Rhizobium leguminosarum bv trifolii of differing acid-soil tolerance were compared. Acidity affected the size and morphology of the acid-tolerant strain, WSM409, but not of the acid-sensitive strain, TA1. Acid grown cells of WSM409 and TA1 had less cell-associated Ca and Mg and more P than cells grown at pH 7.0. Potassium content was lower in acid grown cells; WSM409 was less affected by pH than that in TA1. WSM409 was more tolerant of pH shock at pH 3.5 when grown at pH 4.8 than when grown at pH 7.0. TA1 was more sensitive to pH shock when grown at pH 4.8 than when grown at pH 7.0. WSM409 shows a characteristic adaptive acid tolerance response, whereas TA1 shows an acid sensitive response.  相似文献   

18.
Elaeagnus compatible Frankia isolates from Tunisian soil have been previously clustered with Frankia, colonizing Elaeagnaceae and Rhamnaceae in two different phylogenetic subgroups, while strain BMG5.6 was described as a new lineage closely related to Frankia and Micromonospora genera. In this study we further assess the diversity of captured Frankia and the relationship with BMG5.6-like actinobacteria, by using nifH gene sequences. Using PCR-RFLP screening on DNA extracted from lobe nodules, additional microsymbionts sharing BMG5.6 features have been detected proving a widespread occurrence of these actinobacteria in Elaeagnus root nodules. Neighbour-Joining trees of Frankia nifH sequences were consistent with previously published 16S rRNA and GlnII phylogenetic trees. Although four main clades could be discerned, actinobacterial strain BMG5.6 was clustered with Frankia strains isolated from Elaeagnus. The present study underscored the emanation of new diazotrophic taxon isolated from actinorhizal nodules occupying intermediate taxonomic position between Frankia and Micromonospora. Moreover, its aberrant position in nifH phylogeny should open network investigations on the natural history of nitrogen-fixing gene among actinobacteria.  相似文献   

19.
Although rhizobia for common bean (Phaseolus vulgaris L.) are established in most Brazilian soils, understanding of their genetic diversity is very poor. This study characterized bean strains from two contrasting ecosystems in Brazil, the Northeast Region, with a semi-arid climate and neutral soils and the South Region, with a humid subtropical climate and acid soils. Seedlings of the cultivars Negro Argel and Aporé were used to trap 243 rhizobial isolates from 12 out of 14 sites. An analysis of ERIC-PCR products revealed enormous variability, with 81% of the isolates representing unique strains considering a level of 70% of similarity. In general, there was no effect of either the bean cultivar, or the ecosystem on rhizobial diversity. One-hundred and one strains showing genetic relatedness (ERIC-PCR) less than 70% were further analyzed using restriction fragment length polymorphism (RFLP) of the 16 S rDNA cleaved with five restriction enzymes. Twenty-five different profile combinations were obtained. Rhizobium etli was the predominant species, with 73 strains showing similar RFLP profiles, while 12 other strains differed only by the profile with one restriction enzyme. Fifty strains were submitted to sequencing of a 16 S rDNA fragment, and 34 clustered with R. etli, including strains with RFLP-PCR profiles similar to those species or differing by one restriction enzyme. However, other strains differing by one or two enzymes were genetically distant from R. etli and two strains with identical profiles showed higher similarity to Sinorhizobium fredii. Other strains showed higher similarity of bases with R. tropici, R. leguminosarum and Mesorhizobium plurifarium, but some strains were quite dissimilar and may represent new species. Great variability was also verified among the sequenced strains in relation to the ability to grow in YMA at 40 °C, in LB, to synthesize melanin in vitro, as well as in symbiotic performance, including differences in relation to the described species, e.g. many R. etli strains were able to grow in LB and in YMA at 40 °C, and not all R. tropici were able to nodulate Leucaena.  相似文献   

20.
Common bean (Phaseolus vulgaris) is native to the Americas, and Rhizobium etli is the dominant microsymbiont in both the Mesoamerican and the Andean centers of genetic diversification. Wild common beans are not found in Brazil, although the legume has been cropped in the country throughout time and all but one of the rhizobial species that nodulate it (Rhizobium gallicum) have been broadly detected in Brazilian soils. However, the majority of the effective rhizobial strains isolated so far from field-grown plants belong to R. tropici. This study describes the analysis of symbiotic and non-symbiotic genes of 15 effective R. tropici strains, isolated from four geographically distant regions in Brazil. With RFLP-PCR of the 16S and 23S rRNA genes and sequence analysis of 16S rRNA, two clusters were observed, one related to R. tropici type A and another to type B strains. Diversity in ribosomal genes was high, indicating that type A strains might represent a new species. High intraspecies diversity was also observed in the rep-PCR analysis with BOX, ERIC and REP primers. However, in the RFLP-PCR analysis of nifH and nodC genes, all R. tropici showed unique combinations of profiles, which might reflect an evolutionary strategy to maximize N2 fixation.  相似文献   

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