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1.
Seventeen Sinorhizobium meliloti strains from seven provinces in China were used to screen highly effective strains for alfalfa cultivar in a greenhouse study and their symbiotic relationship and competitive ability were studied in the field. CCBAU30138 was the most effective strain, as evidenced by increase in dry weights. A field experiment showed that the inoculation of alfalfa with CCBAU30138 resulted in increases of 11.9% and 19.6% of dry matter production and crude protein production, respectively, in forage of monocultured plants. The total dry matter yields of alfalfa and tall fescue in binary culture were increased by 16.3% by inoculation of alfalfa with this strain. These results showed that S. rneliloti strain CCBAU30138 was an effective inoculant both in the greenhouse and in the field. The analysis of randomly amplified polymorphic DNA （RAPD） by polymerase chain reaction （PCR） from nodule extracts showed that the strain CCBAU30138 had high competitiveness in the field. It occupied 47.5% of nodules in alfalfa monoculture and 44.4% of nodules in alfalfa-tall fescue binary culture after 20 weeks of growth. In conclusion, a simple system to select highly effective and competitive symbiotic strains specific to alfalfa was established. Using this system, a s.train suitable for the alfalfa cultivar ‘Vector＇ grown in Wuqiao County of Hebei Province was obtained.  相似文献   

2.
The shoot dry weight of alfalfa inoculated with an effective strain of Rhizobium meliloti mixed with an ineffective strain in different ratios was found to be directly proportional to the log of the number of effective nodules. Consequently the comparison of the shoot dry weight of plants inoculated with a mixture of effective and ineffective strains with the shoot dry weight of plants inoculated with the effective strain should allow the estimation of the relative competitiveness of the effective strains. To check this. the competitiveness of 14 antibiotic-resistant strains of R. leguminosarum was evaluated in this way and compared with the ability of the strains to form nodules when inoculated to seeds of Vicia faba planted in a soil containing indigenous R. leguminosarum. The percentage of recovery of the inoculum strains in the nodules of field-grown fababeans was positively correlated with the competitiveness of the strains as estimated by the greenhouse test. This simple way of evaluating the nodulating competitiveness of strains of rhizobia being indicative of their competitive behaviour with indigenous rhizobia in the field could therefore be useful for screening a large number of strains for competitiveness.  相似文献   

3.
《Applied soil ecology》2003,22(1):55-66
The ability of antibiotic-producing streptomycetes to colonize alfalfa (Medicago sativa L.) plants and influence the activities of a fungal plant pathogen (Phoma medicaginis var. medicaginis) and a mutualistic symbiont (Sinorhizobium meliloti) was investigated. Streptomyces strains were introduced around seeds at the time of planting. Hyphal filaments and spore chains were observed by scanning electron microscopy on roots of alfalfa seedlings receiving the streptomycete amendment. Streptomyces strain densities on leaves decreased 10–100-fold over an 8-week period, while densities on roots remained constant over time. The Streptomyces strains also colonized alfalfa root nodules. We then tested the ability of 15 antibiotic-producing strains of Streptomyces to inhibit in vitro growth of Phoma medicaginis var. medicaginis Malbr. & Roum., the causal agent of spring blackstem and leaf spot of alfalfa. The majority of the Streptomyces strains inhibited growth of three diverse strains of P. medicaginis. In a detached leaf assay, one Streptomyces strain decreased leaf spot symptoms caused by P. medicaginis when inoculated onto leaves 24 h before the pathogen. Two Streptomyces strains decreased defoliation caused by P. medicaginis when the streptomycetes were introduced around seeds at the time of planting. We also examined inhibitory activity of Streptomyces strains against 11 strains of S. meliloti. Eight of the 15 Streptomyces strains inhibited in vitro growth of five or more of the S. meliloti strains, while four Streptomyces strains had no effect on growth of any test strains. In a growth chamber assay, two of six Streptomyces strains, when inoculated into the planting mix, significantly reduced plant dry weight compared to the treatment with S. meliloti alone, but did not significantly reduce the number of nodules. These results suggest that careful selection of Streptomyces isolates for use in biological control of plant diseases will limit the potential negative impacts on rhizobia.  相似文献   

4.
Strigolactones have recently been suggested to be phytohormones that are present in all plants. Strigolactones are released by roots into the rhizosphere, stimulating the seed germination of parasitic plants such as Striga spp. and Orobanche spp. and play a crucial role in the interaction between plants and symbiotic arbuscular mycorrhizal fungi.By applying different concentrations of the synthetic strigolactone analogue GR24 to alfalfa (Medicago sativa) inoculated with Sinorhizobium meliloti we could show that in alfalfa nodulation is positively affected by the presence of the strigolactone analogue GR24. Moreover, we could show that this increased nodulation cannot be linked with a stimulatory effect of GR24 on the growth or the expression of nod genes of S. meliloti.Putative mechanisms operating in the plant in response to the addition of GR24 and leading to increased nodule formation by rhizobia are discussed.  相似文献   

5.
The acid tolerance of Sinorhizobium meliloti in culture media and in soils is considered a useful criteria to select for strains with improved survival in agricultural acidic soils. Using a glass tube system with gamma-irradiated soil at different pH values, we analysed the survival of two different alfalfa-nodulating rhizobia: S. meliloti (pHlimit for growth 5.6–6.0) and the acid-tolerant Rhizobium sp. LPU83, closely related to the strain Rhizobium sp. Or191 (pHlimit for growth below 5.0). Although the acid-tolerant rhizobia showed a slightly better survival during the first months in acid soil (pH=5.6), none of the strains could be detected 2 months after inoculation (bacterial counts were below 103 colony-forming units (cfu)/30 g of soil). The inclusion of two alfalfa plants/glass tube with soil, however, supported the persistence of both types of rhizobia at pH 5.6 for over 2 months with counts higher than 9×106 cfu/30 g of soil. Remarkably, in the presence of alfalfa the cell densities reached by S. meliloti were higher than those reached by strain LPU83, which started to decline 1 week after inoculation. Although more acid-sensitive in the culture medium than the Or191-like rhizobia, in the presence of the host plant the S. meliloti strains showed to be better adapted to the free-living condition, irrespective of the pH of the soil.  相似文献   

6.
The symbiotic nitrogen-fixing soil bacterium, Sinorhizobium meliloti, is well known for its ability to interact with the leguminous plant Medicago sativa L. It has, however, not been reported that this species possesses the capability to degrade toxic nitroaromatic compounds, such as 2,4-dinitrotoluene (DNT) which is commonly associated with the degradation of the explosive trinitrotoluene (TNT). In this study, the pJS1 DNT-biodegradative plasmid was genetically transferred to S. meliloti strain USDA 1936, which was confirmed by plasmid profile analysis. Several standard analytical and chemical tests including high performance liquid chromatography (HPLC), nitrite (NO2) release assays, rhizosphere population and plant greenhouse studies were conducted to test the ability of S. meliloti to degrade 2,4-DNT. The possible presence of 2,4-DNT remaining in the treated soil was tested, and no 2,4-DNT had been absorbed by the soil. The pJS1-carrying recombinant strain DHK1 produced ‘ARC’ alfalfa plants that were almost 2-fold higher in shoot dry weight than that produced by the parent strain on soil containing 0.14 mM 2,4-DNT. The transconjugant strain DHK1 reduced significantly one-third more 2,4-DNT in both 0.14 and 0.28 mM contaminated soil, and in 0.55 mM contaminated soil it degraded 94% of the 2,4-DNT present. In liquid cultures, however, only about 4% reduction in 2,4-DNT concentrations was obtained in 10 days. We interpret the results as clearly establishing that genetic modification was successfully used, for the first time, to improve the capability of the symbiotic nitrogen-fixing soil bacterium S. meliloti DHK1 to bioremediate in situ 2,4-DNT-contaminated soil in the presence of alfalfa plants.  相似文献   

7.
Ineffective mutants of four effective strains of Rhizobium meliloti were isolated and tested for their ability to compete with effective parents or antibiotic resistant mutants in the formation of nodules on Medicago sativa. In 5 out of the 6 cases studied, ineffective mutants were no different to effective strains of the same origin in their competitive ability. A difference in selection for infection by the host plant was observed between equally effective strains as well as between ineffective strains. Except for one pair of strains, the more-competitive effective strains (resistant or not to antibiotics) had the same origin as the more-competitive ineffective strains. For such strains the ability to compete with other strains to form nodules was a characteristic of each parent strain. Competitiveness was independent of effectiveness and had been retained during mutation.  相似文献   

8.
Using Tn5 mutagenesis of lysogenic Sinorhizobium meliloti strain L5–30 (L54), three neomycin-resistant transconjugants differing in phage-resistance profiles were isolated. Two of them increased the dry weight of plants and were capable of establishing root nodules, whereas the third one was ineffective. The bacterium-phage interaction did not have observable consequences in the Medicago sativa - S. meliloti symbiosis because it did not affect the number of nodules on M. sativa or plant dry weight.  相似文献   

9.
In this paper the interactive effects of different Sinorhizobium meliloti strains and alfalfa cultivars on the host plant growth and nodulation were evaluated. In the experiments, six alfalfa cultivars (Giza-1, Ismailia-94, Kometa, Radius, Luzella and Legend) as well as five S. meliloti strains (ARC-1, ARC-2, A2, TAL and L5-30) were included. The alfalfa cultivars, Giza-1 and Ismailia-94, exhibited the highest average dry forage weight with all studied strains. Generally, plants inoculated with the S. meliliti ARC-2 strain produced significantly higher shoot and root dry weights over all evaluated cultivars. The strain L5-30 of S. meliloti produced the highest nodule number with all the studied alfalfa cultivars whereas, among tested cultivars, N2-fixing Luzella showed the highest nodulation with the majority of rhizobial strains. Analysis of correlation coefficients for the studied alfalfa-rhizobium associations revealed positive and significant relationships of shoot dry weight with root dry weight. In contrast, nodule number per plant correlated negatively with forage dry weight of plants. It was also found that nodule number per plant is not a promising trait with which to select the most effective N2-fixing rhizobium-legume systems; selection of such symbiotic associations should be done on the basis of shoot and root dry weights.  相似文献   

10.
Alfalfa (Medicago sativa L.) is cultivated in arid and semi-arid regions where salinity is one of the main limiting factors for its production. Thus, this experiment was conducted to evaluate the efficacy of arbuscular mycorrhizal fungus (AMF), Glomus mosseae, alfalfa rhizobia Sinorhizobium meliloti (R) seed inoculation in the development of salinity tolerance of different alfalfa cultivars (Rehnani, Pioneer and Bami) under a variety of salinity levels. The results revealed that under non-stress condition, root mycorrhizal infection, nodulation (the number and weight of nodules per plant), potassium (K), calcium (Ca), phosphorus (P), zinc (Zn), copper (Cu) and magnesium (Mg) contents of the root and shoot, the value of the K/Na ratio, protein [calculated from the nitrogen (N) content] and proline contents of the shoot and the alfalfa yield were found to be the highest while Na contents of the root and shoot were seen to be the lowest when seeds were double inoculated followed by mycorrhizae, rhizobium and control treatments, respectively. Similarly, under salinity condition, the greatest amounts of mycorrhizal infection, nodulation, root and shoot P contents, the value of K/Na ratio, the shoot proline content and the root Ca content were enhanced with the least amount of leaf Na content related to the cases of seeds which were double inoculated, followed by mycorrhizae, rhizobium and control treatments respectively. The results suggested that inoculation of alfalfa seed with AMF or R, especially double inoculation, causes a considerable increase in alfalfa yield under both saline and non-saline conditions by increasing colonization, nodulation and nutrient uptake.  相似文献   

11.
《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   

12.
The soil bacterium Sinorhizobium meliloti forms a symbiotic relationship with alfalfa (Medicago sativa) roots, which results in the formation of intracellular root nodules. This symbiosis increases nitrogen (N) in the soil; however, to establish such a synergistic relationship, a complex communication system is required between the bacterium and its legume host. Rhizobacteria are known to respond to plant root exudates and produce signal molecules known as “Nod” factors. Research suggests that the brown seaweed (Ascophyllum nodosum) extract (ANE) stimulates both root nodulation and growth of alfalfa (Khan et al. 2011 Khan, W., Palanisamy, R., Critchley, A. T., Smith, D. L., Papadopoulos, Y. and Prithiviraj, B. 2011. Ascophyllum nodosum (brown seaweed) extract and its organic fractions stimulate root nodulation and growth of alfalfa (Medicago sativa). Communications in Soil Science and Plant Analysis (Accepted),  [Google Scholar]). To elucidate the mechanism of action, the effects of ANE on the early stages of root–rhizobia interactions were examined. A. nodosum extract (ANE) and its organic fractions were prepared and alfalfa roots were treated. After 2 days, the treated roots were inoculated with S. meliloti. The roots from treated plants were excised and observed for colony-forming units. To verify whether ANE elicited the synthesis and secretion of factors similar to those induced by luteolin, S. meliloti cultures were treated with ANE and the bacterial components were analyzed by high-pressure liquid chromatography (HPLC). To study Nod factor induction by S. meliloti due to ANE treatment, a root hair deformation assay was performed. A translational fusion of S. meliloti NodC:LacZ (strain JM57) was used to observe the effect of ANE on bacterial gene expression. When S. meliloti culture medium was supplemented with ANE, no effect on bacterial growth was observed. However, it was observed that the attachment of S. meliloti to the root hairs was improved. Similarly in vitro ANE root treatments, followed by S. meliloti inoculation, increased bacterial colonies. HPLC profiles and a root hair deformation assay suggested that ANE elicits production of compounds similar to the Nod factor, which are normally induced by the plant signaling molecule luteolin. The results suggest that ANE may contain compound(s) that promote the legume–rhizobia symbiotic relationship and plant signaling.  相似文献   

13.
In this study, rhizobia strains isolated from white clover (Trifolium repens) root nodules were evaluated in an effort to identify an efficient nitrogen-fixing rhizobia strain that can also improve the growth of rice plants (Oryza sativa). White clover plants were collected from seven sites in south Brazil, and 78 native rhizobia isolates were obtained. The genetic diversity analysis of those isolates was carried out by BOX-polymerase chain reaction. Overall, the native rhizobia isolated showed a high genetic diversity, but when the bacterial isolates from the same site were compared, the diversity was lower. One native rhizobia, POA3 (isolated from the Porto Alegre locality), was able to promote the growth of both plants and is therefore a good candidate for new inoculant formulation. Finally, we can conclude that the community of native rhizobia symbiont of white clover plants in southern Brazil is highly diverse and the growth promotion effect of rhizobia inoculation on rice plants was more pronounced in a poor nutrient substrate condition than in a rich nutrient substrate condition.  相似文献   

14.
The distribution and symbiotic efficiency of nodule bacteria Rhizobium leguminosarum_bv. trifolii F., Sinorhizobium meliloti D., Rhizobium galegae L., and Rhizobium leguminosarum bv. viciae F. in Lithuanian soils as dependent on the soil acidity were studied in the long-term field, pot, and laboratory experiments. The critical and optimal pH values controlling the distribution of rhizobia and the symbiotic nitrogen fixation were determined for every bacterial species. The relationship was found between the soil pH and the nitrogen-fixing capacity of rhizobia. A positive effect of liming of acid soils in combination with inoculation of legumes on the efficiency of symbiotic nitrogen fixation was demonstrated.  相似文献   

15.
When a bacteriocin-sensitive and a bacteriocin-resistant strain of Rhizobium were added together to plant-tubes, containing Trifolium subterraneum cv. Dwalganup seedlings, the bacteriocinresistant strain occupied less than 10% of the nodules sites. But when a bacteriocin-producer was added with the inoculum, the bacteriocin-resistant strain occupied 75% of the nodule sites. In a replicate experiment, using a different bacteriocin-sensitive strain, the proportion of the nodules formed by the bacteriocin-resistant strain was increased from 17%, in the absence of the bacteriocin-producer, to 100% in its presence.Similar experiments were set up in non-sterile soil that contained ineffective, bacteriocin-sensitive rhizobia. When an effective bacteriocin-resistant strain was inoculated with a bacteriocin-producing strain, the proportion of nodules formed by the effective strain was greater than in the absence of the bacteriocin-producer. This was because nodulation by the indigenous rhizobia was reduced in the presence of the bacteriocin-producer.Such experiments suggest that bacteriocin-producing strains of R. trifolii that reduce the nodulation success of competing bacteriocin-sensitive strains could be used to increase the proportion of nodules formed by a desired bacteriocin-resistant strain.  相似文献   

16.
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   

17.
Studies were conducted to evaluate the survival and persistence of Sinorhizobium meliloti 104A14 and two acid phosphatase-negative mutants in Kirkland (fine, mixed, thermic Udertic Paleustolls) silt loam soils with various fertility levels, and to assess the impact of inoculation on nodule occupancy and soil microbial community structure in the inoculated alfalfa (Medicago sativa L.) rhizosphere. Recovery of the inoculated strains was 100% (in the order of 108 cells g−1 soil) immediately following inoculation to soils, but decreased from 108 cells g−1 soil to undetectable levels in a nutrient-poor soil within 32 days. In a nutrient-rich soil, approximately 2–3% (4.7–7.43×106 cells g−1 soil) of the mutants and 23% (5.84×107 cells g−1 soil) of the wild-type inocula persisted for more than 64 days. Survivability and persistence of the wild-type S. meliloti were significantly greater than that of the genetically modified acid phosphatase negative mutants in all the soils tested. The persistence and nodule occupancy of the introduced S. meliloti in sterile and non-sterile soils were also tested for two repeated alfalfa growth periods in the same plant growth units, with a 1 month interval in between and no additional inoculation for the second period. Nodule occupancy of the introduced S. meliloti in non-sterile soils ranged from 30 to 60% for the first period and 85 to 100% for the second period. Our results suggest that survival and persistence of S. meliloti was enhanced by alfalfa cultivation and increased soil fertility, but impaired by mutation of acid phosphatase genes regardless of phosphorus nutritional levels. Moreover, inoculation with genetically modified S. meliloti strain 104A14 promoted indigenous bacterial growth in soil (increased bacterial population from 1.4×106 to 4.3×106 cells g−1 soil), but not the growth of fungi and yeast. However, inoculation of the wild-type S. meliloti or genetically modified mutants did not result in significant changes in microbial community structure as indicated by EP indices and ratios of r/K strategists.  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.
《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   

20.
A collection of 299 isolates of rhizobia nodulating Medicago truncatula was isolated from 10 Tunisian soils and was characterized by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR/RFLP) of 16S rRNA gene. Results showed that 227 and 72 isolates were assigned, respectively, to Sinorhizobium meliloti and Sinorhizobium medicae. In 9 out of 10 soils S. meliloti was detected, whereas S. medicae was recovered from only 5 out of 10 soils. The cross-nodulation of three populations of M. truncatula grown on Bulla Regia soil, which contained naturally the two Sinorhizobium species, showed that M. truncatula population collected from Amra site was selective to S. meliloti at least in soil conditions. Forty-eight isolates of each Sinorhizobium species trapped by M. truncatula populations collected from Bulla Regia, Soliman and Rhayet sites on Bulla Regia soil were characterized by repetitive extragenic palindromic-PCR (REP-PCR) and showed a clear distinction between the two Sinorhizobium species and a higher diversity for S. meliloti.  相似文献   

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