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1.
A study was undertaken to determine the free-living culturable diazotrophic bacteria of paddy soils from a long-term fertilizer management experiment. Long-term application of different fertilizers significantly affected the population of free-living diazotrophs. Out of 165 distinct bacterial morphotypes observed during the isolation process, only 32 were positive for both acetylene reduction assay (ARA), and nifH gene screening. The ARA activity of the isolates ranged from 1.8 to 2,844.7 nmol ethylene h?1 mg protein?1. The 16S rRNA analysis identified the isolates to be members of 13 different genera viz. Bacillus, Pseudomonas, Paenibacillus, Serratia, Ochrobactrum, Lysinibacillus, Burkholderia, Brevundimonas, Herbaspirillum, Novosphingobium, Sphingomonas, Xanthomonas, and Azorhizobium. Though partial nifH gene sequencing of diazotrophic isolates showed good consistency with that of 16S rRNA-based identification, some nifH sequences were similar to a variety of uncultured nitrogen-fixing bacteria. The diversity of free-living diazotrophic bacteria and the wide distribution of nifH sequences indicate the potential contribution of these microorganisms to nitrogen input to paddy fields.  相似文献   

2.
The intercellular colonization of rice roots by Azorhizobium caulinodans and other diazotrophic bacteria has been studied using strains marked with the lacZ reporter gene. A. caulinodans were able to enter the roots of rice at emerging lateral roots (lateral root cracks) by crack entry and this was observed by light microscopy. After colonization of lateral roots, bacteria moved into intercellular space within the cortical cell layer of roots. Naringenin at 1×10-5 and 5×10-5 M concentration significantly enhanced root colonization. The role of nodABC and regulatory nodD genes was also studied; lateral root crack (LRC) colonization of rice was shown to be Nod factor and NodD independent. Lateral root crack colonization of rice was also observed with similar frequency following inoculation with Azospirillum brasilense and the colonization by A. brasilense was stimulated by naringenin and other flavonoid molecules.  相似文献   

3.
Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.  相似文献   

4.
Micropropagated sugarcane plants have been used in Brazil for almost three decades. Besides the improvement in plant health, micropropagated sugarcane carries no endophytic plant growth-promoting bacteria. The Brazilian inoculation technology to reintroduce diazotrophic bacteria in micropropagated sugarcane plantlets revealed a synergistic-like effect in PGP-bacteria mixed inoculations. The infection model of single diazotrophic bacteria species in sugarcane was studied in detail, but still many questions remain open. In this study we used a combined fluorescence in situ hybridization (FISH) and a cultivation based approach (MPN) to evaluate the colonization of sugarcane plantlets by mixed inocula. The highest colonization for three out of the five species studied was obtained with a mixed inoculum, when the Azospirillum amazonense showed an increase by almost 100 times in colonization and Herbaspirillum spp. and Burkholderia tropica was determined at 107 cells per gram root fresh weight. All of the inoculated bacterial species could be detected using the FISH probes 12 h after bacterial inoculation. The FISH results confirmed the MPN counts and showed differences in the population numbers and colonization behavior of particular bacterial inoculum strains in the different mixed inocula. A putative antagonistic effect among the inoculated H. seropedicae and H. rubrisubalbicans strains was observed using FISH, as well as the better competitiveness of B. tropica as compared to the A. amazonense strain. The observed data probably reflect also specific interactions with the sugarcane variety used in this particular inoculation system, and may not be generalized as a rule. This is the first study about the competition for sugarcane colonization in a mixed bacterial inoculum.  相似文献   

5.
The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the 15N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.  相似文献   

6.
The largest numbers of the Brazilian traditional upland rice varieties are found in the Maranhão state, Northeast region of Brazil. However, no information is available on the diazotrophic bacterial population associated as well as the plant growth promoting potential when these traditional genotypes are inoculated with native strains. Here, we evaluated the response of ten traditional rice varieties to inoculation with ten diazotrophic strains, previously isolated from rice soil of this region and screened for their ability to produce indole-3-acetic acid (IAA) in vitro. The procedure for selection of the best diazotrophic strain/rice variety interaction involved three steps: gnotobiotic conditions, soil pot and field experiments. The gnotobiotic experiment showed that the Azospirillum amazonense strain AR3122 increased the biomass of the traditional varieties Cana Roxa and Cana Forte (28 and 48%, respectively) while this effect was less evident for the other combination of strains/rice varieties. The soil pot experiment showed that the combination of Burkholderia vietnamiensis strain AR 1122 and traditional variety Arroz 70 was superior to the other strains/varieties and the treatment fertilized with 100 kg N ha−1. The best performance of the Burkholderia vietnamiensis strain AR1122/variety Arroz 70 was confirmed in the field experiment. There was an increase of up 10 and 29% in the grain yield in comparison to both the N fertilization and Herbaspirillum seropedicae ZAE 94 strain treatments, respectively. In contrast, the response of the commercial variety Bonança to inoculation with strain AR1122 was much lower, suggesting that a biofertilizer inoculation program for traditional rice varieties should consider the genetic interaction between strain and rice variety. The diazotrophic B. vietmaniensis strain AR1122 was a good biofertilizer candidate for inoculation of traditional rice varieties and therefore should be used for further studies to confirm the strain-genotype effect envisaging a sustainable rice crop system mainly in the Northeast region of Brazil.  相似文献   

7.
Take-all of wheat caused by Gaeumannomyces graminis var. tritici was less when soils in glasshouse pots were fertilized with NH4+-N than with NO3?-N. The form of N did not alter countable populations of microorganisms in the rhizosphere or rhizoplane, but altered the numbers of bacteria and streptomycetes that inhibited the pathogen's growth in vitro. The pH of the medium used to isolate these microorganisms, whether similar or dissimilar to the pH of the rhizosphere, had some influence both upon countable populations and upon the proportions of antagonists. Highest counts of the rhizoplane microflora were on agar media with a pH similar to that of the soil. Most antagonists were isolated from a soil that is physically and chemically conducive to parasitism of wheat roots by Gaeumannomyces, but which contains a microflora suppressive toward the parasitic colonization of the roots. Isolates of the general bacterial flora, of Pseudomonas spp. and of streptomycetes, but not of Bacillus spp. inhibited the in vitro growth of G. graminis.  相似文献   

8.
A total of 45 cyanobacterial strains isolated from rice fields near Loktak Lake in Manipur, India were tested for their rice root colonization capacity under light and under darkness. Twenty-one of these strains showed significant colonization of rice roots. The average colonization values were 637 and 381?μg chl a g?1 root dry wt in N2 medium and 792 and 451?μg chl a g?1 root dry wt in NO 3 ? medium under light and darkness, respectively. Thus, while the colonization was higher under light and in NO 3 ? medium, there was significant level of colonization under darkness in N2 medium (381?μg chl a g?1 root dry wt). A 16S rRNA gene fragment-based denaturing gradient gel electrophoresis analysis revealed difference in the competence of individual strains to colonize rice roots exposed to individual or mixed population. The colonization pattern of seven strains used in competition experiments was found to be biphasic. A 16S rRNA gene-based phylogenetic analysis revealed high level of molecular similarity among strains of Nostoc and Anabaena.  相似文献   

9.
 Four experiments were performed under gnotobiotic conditions to select strains of the endophytic diazotrophs Herbaspirillum seropedicae and Burkholderia spp. as inocula of rice plants. Eighty strains of H. seropedicae originally isolated from rice, sorghum and maize plants, were tested in test tube cultures with N-free agar as the substrate. Rice plants showed medium and high increases in their fresh weight in response to inoculation with nineteen strains. These strains were tested again, and six strains were then selected to evaluate their contribution to the N of the plant via biological N2 fixation (BNF) using an agar growth medium containing 5 mg N l–1of 15N-labelled (NH4)2SO4. The contribution of the strains to plant N via BNF varied from 54% when rice plants were inoculated with strain ZAE94, to 31% when strain ZAE67 was used. These results were confirmed in the fourth gnotobiotic experiment, which also included strains of the new N-fixing bacteria belonging to the genus Burkholderia, isolated from rice, as well as a strain of Burkholderia vietnamiensis, isolated from rice rhizosphere. Burkholderia spp. strains showed similar effects to those observed for H. seropedicae strains, while B. vietnamiensis fixed only 19% of plant total N. The best four strains were tested in a pot experiment where pre-germinated, inoculated rice seedlings were grown in soil labelled with 15N. The results confirmed the gnotobiotic experiments, although the levels of N in the rice plants derived from BNF of the selected H. seropedicae and Burkholderia spp. strains were lower. Nevertheless, there was an increase in N content in grains of inoculated plants, and the results showed that the method used for strain selection is very useful and can be applied to other strains of N2-fixing bacteria and plants. Received: 4 May 1999  相似文献   

10.
Biological nitrogen(N) fixation(BNF) plays a significant role in maintaining soil fertility in paddy field ecosystems. Rice variety influences BNF, but how different rice varieties regulate BNF and associated diazotroph communities has not been quantified. Airtight,field-based ~(15)N_2-labelling growth chamber experiments were used to assess the BNF capacity of different rice varieties. In addition,both the 16 S rRNA and nifH genes were sequenced to assess the influence of different rice varieties on bacterial and diazotrophic communities in paddy soils. After subjecting a rice-soil system to 74 d of continuous airtight, field-based ~(15)N_2 labelling in pots in a growth chamber, the amounts of fixed N were 22.3 and 38.9 kg ha~(-1) in inbred japonica(W23) and hybrid indica(IIY) rice cultivars planted in the rice-soil systems, respectively, and only 1%–2.5% of the fixed N was allocated to the rice plants and weeds. A greater abundance of diazotrophs was found in the surface soil(0–1 cm) under IIY than under W23. Sequencing of the 16 S rRNA gene showed significantly greater abundances of the cyanobacterial genera Nostoc, Anabaena, and Cylindrospermum under IIY than under W23.Sequencing of the nifH gene also showed a significantly greater abundance of Nostoc under IIY than under W23. These results indicate that the hybrid rice cultivar(IIY) promoted BNF to a greater extent than the inbred rice cultivar(W23) and that the increase in BNF might have been due to the enhanced heterocystous cyanobacteria Nostoc.  相似文献   

11.
《Applied soil ecology》2007,35(1):25-34
Rice is usually grown in N-deficient soils, demanding that the element be supplied to the field by commercially available N fertilizers. Unfortunately, a substantial amount of the urea-N or NO3-N applied as fertilizers is lost through different mechanisms, causing environmental pollution problems. Utilization of biological N2 fixation (BNF) technology can decrease the application of N fertilizers, reducing environmental risks. This study evaluated the effects of four free-living N-fixing bacterial species, isolated from oligotrophic soil conditions, as single inoculants or combined with arbuscular mycorrhizal fungi (Glomus clarum), on the development of rice plants grown as flooded or upland rice, in the greenhouse. Upland rice roots were inoculated with Methylobacterium sp., Burkholderia sp. and Sphingomonas sp., whereas the species Burkholderia sp., Pseudomonas sp. and Sphingomonas sp., were inoculated on flooded rice. Inoculants consisted of individual bacterial species or their mixtures, with or without G. clarum. Controls included non-bacteria/non-AM fungi, and AM fungi alone. Experiments were carried out in five replicates. The presence of G. clarum decreased or did not significantly affect plant growth under the different culture conditions. The presence of AM fungi stimulated the N-fixing bacterial population of upland rice. Bacterial species had different effects, under both culture conditions, and some genera of N-fixing bacteria increased root and shoot growth at different plant growth stages. The level of mycorrhiza colonization had no influence on plant growth  相似文献   

12.
Summary Sweet potatoes were micropropagated and then transplanted from axnic conditions to fumigated soil in pots in the greenhouse. Spores of Glomus clarum were obtained from Brachiaria decumbens or from sweet potatoes grown in soil infected with this fungus and with an enrichment culture of Acetobacter diazotrophicus. Three experiments were carried out to measure the beneficial effects of vesicular-arbuscular mycorrhizal (VAM) fungi-diazotroph interactions on growth, nutrition, and infection of sweet potato by A. diazotrophicus and other diazotrophs obtained from sweet potato roots. In two of these experiments the soils had been mixed with 15N-containing organic matter. The greatest effects of mycorrhizal inoculation were observed with co-inoculation of A. diazotrophicus and/or mixed cultures of diazotrophs containing A. diazotrophicus and Klebsiella sp. The tuber production was dependent on mycorrhization, and total N and P accumulation were increased when diazotrophs and G. clarum were applied together with VAM fungal spores. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM fungi or when present within G. clarum spores. More pronounced effects on root colonization and intraradical sporulation of G. clarum were observed when A. diazotrophicus was co-inoculated. In non-fumigated soil, dual inoculation effects, however, were of lower magnitude. 15N analysis of the aerial parts and roots and tubers at the early growth stage (70 days) showed no statistical differences between treatments except for the VAM+Klebsiella sp. treatment. This indicates that the effects of A. diazotrophicus and other diazotrophs on sweet potato growth were caused by enhanced mycorrhization and, consequently, a more efficient assimilation of nutrients from the soil than by N2 fixation. The possible interactions between these effects are discussed.  相似文献   

13.
Summary This study is an attempt to describe the dominant N2-fixing microflora associated with the roots of wetland rice. Rice cultivar Giza 171 was grown in a phytotron on two alluvial Egyptian soils for 8 days, a stage when the nitrogenase activity of undisturbed plants reached a level of 245 × 10–6 mol C2H4 h–1 g–1 dry weight of leaf. The roots and rhizosphere soils were then used for counting and isolating dominant diazotrophs. Counts and initial enrichment steps were carried out on a selective medium made of an axenic rice plantlet, the spermosphere model, incubated under 1 % acetylene. The counts were very high, exceeding 108 bacteria g–1 dry weight of rhizosphere soil. Enterobacteriaceae were dominant; most isolates were Enterobacter cloacae belonging to different biotypes in the two soils. Enterobacter agglomerans, Citrobacter freundii and Klebsiella planticola were also present as members of the dominant microflora. Azospirillum brasilense and Azospirillum lipoferum were present as well, but less abundant.  相似文献   

14.
The numbers of culturable diazotrophic endophytic bacteria (CDEB) from roots, stems and leaves of sugarcane submitted to organic, inorganic or no fertilization were compared. In order to determine the size of the N2 fixing populations, the Most Probable Number technique (MPN) was used. The quantification of diazotrophic bacteria by using the acetylene reduction assay (ARA) was more accurate than observing the bacterial growth in the vials; to confirm N2 fixing capability, the detection of gene nifH was performed on a sample of 105 isolated bacteria. The production of extracellular enzymes involved in the penetration of the plants by the bacteria was also studied. The results showed that organic fertilization enhances the number of CDEB when compared with conventional fertilization used throughout the growing season. The maximum number of bacteria was detected in the roots. Roots and stems presented the greatest number of CDEB in the middle of the cropping season and in leaves numbers varied according to the treatment. Using two pairs of primers and two different methods, the nifH gene was found in 104 of the 105 tested isolates. Larger amounts of pectinase were released by isolates from sugarcane treated with conventional fertilizers (66%), whereas larger amounts of cellulase were released by strains isolated from sugarcane treated with organic fertilizers (80%).  相似文献   

15.
Climate change and human activity have led to the degradation of desert wetlands. Free-living diazotrophs are vital for soil nitrogen input. However, a comprehensive understanding of how soil free-living diazotrophic communities and their co-occurrence patterns respond to desert wetland degradation is lacking. Here, quantitative polymerase chain reaction (qPCR), amplicon sequencing targeting nitrogenase gene (nifH), and network analysis were used to investigate the abundance, diversity, community composition, and co-occurrence patterns of soil free-living diazotrophs along the wetland degradation gradient, i.e., non-degraded (ND), lightly degraded (LD), moderately degraded (MD), and severely degraded (SD), in the southeastern Mu Us Desert, northern China. The abundance and Shannon, Simpson, Chao 1, and ACE indexes decreased (P < 0.05) by 14.6%, 20.7%, 2.1%, 46.5%, and 45.0%, respectively, in SD wetland, whereas no significant difference (P > 0.05) was observed between ND and LD wetlands. The relative abundance of Proteobacteria generally decreased (by 53.5%–19.7%) across the different degradation levels, while the relative abundance of Cyanobacteria increased (by 6.2%–40.1%) from ND to MD levels. The abundance, diversity, and community composition of diazotrophs were most strongly related to soil organic carbon, followed by total nitrogen, moisture, and pH. The least number of network nodes and edges and the lowest density were observed for MD and SD wetlands, indicating that the complexity of free-living diazotrophic networks was reduced by continued degeneration. Overall, severe desert wetland degradation affected the abundance, diversity, and network complexity of soil free-living diazotrophs more negatively than light degradation. This degradation promoted the growth of autotrophic diazotrophs and inhibited the growth of heterotrophic diazotrophs. These changes were mostly related to the loss of soil organic carbon.  相似文献   

16.
Rhizosphere bacteria may enhance plant uptake of Fe by producing siderophores that chelate sparingly soluble Fe3+ in calcareous soils. To evaluate the extent to which plants benefit from colonization of the roots by prolific siderophore-producing bacteria, we inoculated two oat cultivars with six strains of bacteria that produced high concentrations of siderophores under Felimiting conditions in vitro. Oat cv Coker 227, an Fe-efficient cultivar, which produces the phytosiderophore avenic acid, and cv TAM 0-312, and Fe-inefficient cultivar, which does not produce the phytosiderophore, were grown in a calcareous soil (Weswood silt loam) on a light bench in the laboratory. Half of the plants were fertilized with a nutrient solution containing 5 mM Fe and half with a nutrient solution containing no Fe. After 6 weeks of growth, we compared colonization of the roots by the inoculant bacteria and the dry weight and Fe content of roots and shoots. Three species of Pseudomonas colonized the roots of both oat cultivars in high numbers (106 cells g-1 root dry weight), whereas the remaining bacteria colonized the roots in substantially lower numbers (104 cells g-1 root dry weight). Plants fertilized with 5 mM Fe were larger and supported greater numbers or rhizosphere bacteria per gram of root than plants not supplied with Fe. Comparisons of the Fe content and dry weight of roots and shoots revealed few significant differences between inoculated and uninoculated plants, or among the plants inoculated with the different strains of siderophore-producing bacteria. The differences that were observed revealed no consistent response to inoculation. We conclude that inoculation of the roots of the two oat cultivars with bacteria that produce high concentrations of siderophores in response to an Fe deficiency had little or no effect on Fe acquisition by the plants.  相似文献   

17.
Intercropping with aerobic rice or arbuscular mycorrhizal fungi (AMF) colonization alleviated watermelon wilt disease, which is likely attributed to rice root exudates or AMF depressing watermelon wilt pathogen. However, it is unclear whether rice root exudates transfers to watermelon rhizosphere soil and whether AMF affects the transfer of rice root exudates to watermelon rhizosphere soil. A rhizobox experiment, with aerobic rice under 14?CO2, was conducted to investigate the effect of AMF colonization on carbon (C) transfer from rice to watermelon and on phosphorus (P) uptake by both watermelon and rice. The rhizobox was separated into labelling side (L side) and sampling side (S side) by inserting nylon mesh in the middle of the box. The L side was planted with aerobic rice, and the S side was aerobic rice (monocropping) or watermelon (intercropping). When 14?CO2 was added to rice canopy at the L side, 14?C activities of rice roots and rhizosphere soils in the L side were increased by intercropping with watermelon or AMF colonization. The 14?C was detected in roots and rhizosphere soils of rice and watermelon in the S side, but no differences were found among different treatments. 14?C activities in leaves were improved by AMF inoculation in the S side, regardless of rice or watermelon. Mycorrhizal colonization stimulated P absorption and translocation to rice in intercropping system. These findings suggest that AMF colonization could increase C transfer from rice to watermelon while intercropping with watermelon could promote AMF colonization and P uptake by rice.  相似文献   

18.
The active release of amino acids by diazotrophic rhizobacteria into the natural environment or under in vitro conditions is poorly described in the literature. This capacity could be an important trait in the plant–bacteria interaction and in plant growth promotion. The ability of releasing amino acids into a medium free of N-combined was studied in cultures of five diazotrophic genera, Beijerinckia, Burkholderia, Enterobacter, Klebsiella, and Pseudomonas, isolated from the sugarcane rhizosphere. Eleven different amino acids were excreted into the culture media by 40% of the isolates. The highest amino acid diversities were found in Beijerinckia (ICBR 177) and Enterobacter (ICBR 200). The highest quantities were excreted by Beijerinckia (ICBR 177), Enterobacter (ICBR 200), Pseudomonas (ICBR 56), and Klebsiella (ICBR 183). The production of amino acids by rhizobacteria may play an important role in the growth of plants and might also have a direct application in agricultural technologies. The release of amino acids could explain one of the positive effects of plant growth-promoting bacteria and encourage further studies of this subject.  相似文献   

19.
Colonization behaviour of two enterobacterial strains on cereals   总被引：4，自引：0，他引：4  
 Two diazotrophic enterobacterial strains, Pantoea agglomerans D5/23 and Klebsiella pneumoniae CC12/12, were observed in sterile and non-sterile hydroponic and soil experiments in order to determine, by means of an immunological detection method (double antibody sandwich enzyme linked immunosorbent assay), their colonization sites, their migration within individuals of different plant species, and their ability to compete with indigenous organisms. To investigate the interaction between bacteria and plants, root and shoot samples were analysed using electron microscopy. Field experiments were performed to determine the growth-promoting abilities of the bacterial strains. In field experiments, inoculation with P. agglomerans led to an increase in the grain yield of different wheat (Triticum aestivum) cultivars. The same strain was also able to colonize the rhizosphere and the phyllosphere of different cereals due to its ability to migrate within the plant. Roots and media were colonized 10–100 times more intensively than shoots, with about 106 cells g–1 root and 104 cells g–1 shoot. We found that P. agglomerans colonized the root and plant-growth medium of wheat to a greater extent than those of rye (Secale cereale) and barley (Hordeum vulgare), whereas the colonization of shoots was higher in rye and barley compared to wheat. Furthermore, while cell numbers of K. pneumoniae in media and roots were 10 times higher than cell numbers of P. agglomerans, only the latter markedly increased root growth. We were able to detect significant differences in colonization numbers between treatments even if the data were not normally or log-normally distributed or the variances were not homogenous. Received: 14 June 1999  相似文献   

20.
Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.  相似文献   

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