Soil fumigation and compost amendment alter soil microbial community composition but do not improve tree growth or yield in an apple replant site

Apple replant disease (ARD) is a disease complex that reduces survival, growth and yield of replanted trees, and is often encountered in establishing new orchards on old sites. Methyl bromide (MB) has been the fumigant used most widely to control ARD, but alternatives to MB and cultural methods of control are needed. In this experiment, we evaluated the response of soil microbial communities and tree growth and yield to three pre-plant soil treatments (compost amendment, soil treatment with a broad-spectrum fumigant, and untreated controls), and use of five clonal rootstock genotypes (M.7, M.26, CG.6210, G.30 and G.16), in an apple replant site in Ithaca, New York. Polymerase chain reaction (PCR)—denaturing gradient gel electrophoresis (DGGE) analysis was used to assess changes in the community composition of bacteria and fungi in the bulk soil 8, 10, 18 and 22 months after trees were replanted. PCR-DGGE was also used to compare the community composition of bacteria, fungi and pseudomonads in untreated rhizosphere soil of the five rootstock genotypes 31 months after planting. Tree caliper and extension growth were measured annually in November from 2002 to 2004. Apple yield data were recorded in 2004, the first fruiting year after planting. Trees on CG.6210 rootstocks had the most growth and highest yield, while trees on M.26 rootstocks had the least growth and lowest yield. Tree growth and yield were not affected by pre-plant soil treatment except for lateral extension growth, which was longer in trees growing in compost-treated soil in 2003 as compared to those in the fumigation treatment. Bulk soil bacterial PCR-DGGE fingerprints differed strongly among the different soil treatments 1 year after their application, with the fingerprints derived from each pre-plant soil treatment clustering separately in a hierarchical cluster analysis. However, the differences in bacterial communities between the soil treatments diminished during the second year after planting. Soil fungal communities converged more rapidly than bacterial communities, with no discernable pattern related to pre-plant soil treatments 10 months after replanting. Changes in bulk soil bacterial and fungal communities in response to soil treatments had no obvious correlation with tree performance. On the other hand, rootstock genotypes modified their rhizosphere environments which differed significantly in their bacterial, pseudomonad, fungal and oomycete communities. Cluster analysis of PCR-DGGE fingerprints of fungal and pseudomonad rhizosphere community DNA revealed two distinct clusters. For both analyses, soil sampled from the rhizosphere of the two higher yielding rootstock genotypes clustered together, while the lower yielding rootstock genotypes also clustered together. These results suggest that the fungal and pseudomonad communities that have developed in the rhizosphere of the different rootstock genotypes may be one factor influencing tree growth and yield at this apple replant site.     

Rhizosphere bacterial community composition in natural stands of Carex arenaria (sand sedge) is determined by bulk soil community composition

The relative importance of specific plant properties versus soil characteristics in shaping the bacterial community structure of the rhizosphere is a topic of considerable debate. Here, we report the results of a study on the bacterial composition of the rhizosphere of the wild plant Carex arenaria (sand sedge) growing at 10 natural sites in The Netherlands. The soil properties of the sandy soils at these sites were highly disparate, most notably in pH, chloride and organic matter content. Rhizosphere and bulk soil bacterial communities were examined by culture-independent means, namely, 16S rDNA-directed PCR-DGGE profiling. Large differences were observed between the bacterial communities of the different sites for both bulk and rhizosphere soil. Cluster analysis of bacterial profiles revealed that the rhizosphere community of each site was generally more closely related to the bulk soil community of that site rather than to rhizosphere communities of other sites. Hence, bacterial community structure within the rhizosphere of C. arenaria appeared to be determined to a large extent by the bulk soil community composition. This conclusion was supported by a reciprocal planting experiment, where C. arenaria shoots of different sites yielded highly similar rhizosphere communities when planted in the same soil.     

Comparison of root system architecture and rhizosphere microbial communities of Balsas teosinte and domesticated corn cultivars

The progenitor of maize is Balsas teosinte (Zea mays subsp. parviglumis) which grows as a wild plant in the valley of the Balsas river in Mexico. Domestication, primarily targeting above-ground traits, has led to substantial changes in the plant's morphology and modern maize cultivars poorly resemble their wild ancestor. We examined the hypotheses that Balsas teosinte (accession PI 384071) has a) a different root system architecture and b) a structurally and functionally different rhizosphere microbial community than domesticated cultivars sweet corn (Zea mays subsp. mays accession PI 494083) and popping corn (Zea mays subsp. mays accession PI 542713). In a greenhouse experiment, five plants from each corn variety were grown in individual pots containing a Maury silt loam – perlite (2:1) mixture and grown to the V8 growth stage at which rhizosphere bacterial and fungal community structure was assessed using terminal restriction fragment length polymorphism and fatty acid methyl ester analysis. Functional characteristics of the rhizosphere were assayed by examining the potential activity of seven extracellular enzymes involved in carbon, nitrogen and phosphorus cycling. Root system architecture was characterized by root scans of sand grown plants at the V5 growth stage. Compared to the control the sweet corn rhizosphere had different bacterial and fungal community structure, decreased fungal diversity and increased bacterial abundance. Teosinte caused a significant change in the rhizosphere bacterial and fungal community structure and increased bacterial abundance, but no significant decrease in bacterial or fungal diversity where the former was found to be significantly greater than in the sweet corn rhizosphere. Popping corn did not trigger significant changes in the bacterial or fungal diversity and bacterial abundance in the soil. The individual popping corn plants changed the bacterial and fungal communities in different directions and the overall effect on community structure was significant, but small. Of the enzymes analyzed, potential N-acetylglucosaminidase (NAG) activity was found to contributed most to the differentiation of teosinte rhizosphere samples from the other corn varieties. The teosinte root system had proportionally more very fine (diameter < 0.03 mm) roots than popping corn and sweet corn and it developed the highest root to shoot dry weight ratio, followed by popping corn. Sweet corn had significantly lower average root diameter than popping corn and teosinte and grew proportionally the least below-ground dry mass. The results allude to functional and structural differences in the rhizosphere microbial communities of the corn varieties that, with additional research, could lead to useful discoveries on how corn domestication has altered rhizosphere processes and how plant genotype influences nutrient cycling.     

Rhizosphere fungal communities are influenced by Senecio jacobaea pyrrolizidine alkaloid content and composition

The contents and the compositions of the pyrrolizidine alkaloid (PA) complex of ragwort (Senecio jacobaea L.) were examined as potential drivers of fungal community structure in the rhizosphere. S. jacobaea plants within the coastal sand dune reserve of Meijendel (the Netherlands) were assayed for concentration and composition of PAs in roots. Rhizosphere soil was collected from pre-flowering plants, which differed up to 8-fold in PA production, and represented both jacobine and senecionine/seneciphylline chemotypes. Bulk soil samples from the same site were also collected for comparative examination. A culture-independent approach, involving direct DNA isolation, PCR of fungal 18S rRNA genes, and denaturing gradient gel electrophoresis (DGGE), was applied to compare the fungal communities of plants with different PA contents, as well as differences between bulk and rhizosphere samples. Cluster analysis of PCR-DGGE profiles revealed no clear evidence for PA-induced selection of specific fungal communities. However, canonical variance analysis showed that fungal communities associated with high-PA jacobine chemotypes could be discriminated from low PA samples and from the senecionine/seneciphylline chemotypes. The diversity of DGGE banding patterns, both in terms of band number and evenness, showed a trend toward lower diversity in the rhizosphere of high-PA plants as compared to low-PA plants and bulk soil. These results indicate that PA chemotypes of S. jacobaea differ in their influence on soil-borne fungal communities, with jacobine-containing plants exerting a greater selection in the rhizosphere than plants containing senecionine/seneciphylline.     

<Emphasis Type="Italic">p</Emphasis>-Coumaric can alter the composition of cucumber rhizosphere microbial communities and induce negative plant-microbial interactions

Phenolics from root exudates or decaying residues are usually referred as autotoxins of several plant species. However, how phenolics affect soil microbial communities and their functional significances are poorly understood. Rhizosphere bacterial and fungal communities from cucumber (Cucumis sativus L.) seedlings treated with p-coumaric acid, an autotoxin of cucumber, were analyzed by high-throughput sequencing of 16S rRNA gene and internal transcribed spacer amplicons. Then, feedback effects of the rhizosphere biota on cucumber seedlings were evaluated by inoculating non-sterilized and sterilized rhizosphere soils to sterilized background soils. p-Coumaric acid decreased the bacterial diversity of rhizosphere but increased fungal diversity and altered the compositions of both the bacterial and fungal communities. p-Coumaric acid increased the relative abundances of microbial taxa with phenol-degrading capability (such as Chaetomium, Humicola, and Mortierella spp.) and microbial taxa which contained plant pathogens (such as Fusarium spp.). However, p-coumaric acid inhibited the relative abundances of Lysobacter, Haliangium, and Gymnoascus spp., whose species can have pathogen-antagonistic and/or plant-growth-promoting effects. The positive effect of cucumber rhizosphere microbiota on cucumber seedling growth was reduced by p-coumaric acid. Overall, our results showed that, besides its direct phytotoxicity, p-coumaric acid can inhibit cucumber seedling growth through generating negative plant-soil microbial interactions.     

The influence of tillage on the structure of rhizosphere and root-associated arbuscular mycorrhizal fungal communities

Soil environmental factors affect the structure of arbuscular mycorrhizal (AM) fungal communities present in soil. However, it is not understood to which degree management practices such as tillage lead to dissimilarities between intra- and extraradical AM fungal communities. This study aims to assess the influence of two different soil management practices (conventional tillage and no-till) on the diversity of AMF communities, both in rhizosphere soil and inside corn roots. We hypothesized that under no-till, roots are colonized as they grow through the undisturbed fungal mycelia left from the previous crop whereas under conventional tillage they are colonized by those propagules that survived disturbance and can re-establish in their new relocated and mixed environment. We predicted that the degree of similarity of AM fungal communities inside versus outside the roots would be greater under no-till than under tillage. Using terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis we observed a different AM fungal community present in roots under no-till than under conventional tillage. Moreover, the communities present in the rhizosphere soil were different than in the roots of the corn plants. These results suggest that soil management does alter the diversity of AM fungal communities associated with corn roots and that plants influence the structure of the AMF community colonizing their roots. Sequencing results indicated that the majority of AMF species found in this agricultural soil was Glomus spp. However, further work is required to determine the extent to which AM fungal genotypic alterations by soil management influences competitive relationships.     

Diversity and antagonistic potential of Pseudomonas spp. associated to the rhizosphere of maize grown in a subtropical organic farm

Pseudomonas spp. are one of the most important bacteria inhabiting the rhizosphere of diverse crop plants and have been frequently reported as biological control agents (BCAs). In this work, the diversity and antagonistic potential of Pseudomonas spp. in the rhizosphere of maize cultivars Nitroflint and Nitrodent grown at an organic farm in Brazil was studied by means of culture-dependent and -independent methods, respectively. Sampling of rhizosphere soil took place at three different stages of plant development: 20, 40 and 106 days after sowing. A PCR-DGGE strategy was used to generate specific Pseudomonas spp. fingerprints of 16S rRNA genes amplified from total community rhizosphere DNA. Shifts in the relative abundance of dominant populations (i.e. PCR-DGGE ribotypes) along plant development were detected. A few PCR-DGGE ribotypes were shown to display cultivar-dependent relative abundance. No significant differences in diversity measures of DGGE fingerprints were observed for different maize cultivars and sampling times. The characterisation and assessment of the antagonistic potential of a group of 142 fluorescent Pseudomonas isolated from the rhizosphere of both maize cultivars were carried out. Isolates were phenotypically and genotypically characterised and screened for in vitro antagonism towards three phytopathogenic fungi and the phytopathogenic bacterium Ralstonia solanacearum. Anti-fungal activity was displayed by 13 fluorescent isolates while 40 isolates were antagonistic towards R. solanacearum. High genotypic and phenotypic diversity was estimated for antagonistic fluorescent Pseudomonas spp. PCR-DGGE ribotypes displayed by antagonists matched dominant ribotypes of Pseudomonas DGGE fingerprints, suggesting that antagonists may belong to major Pseudomonas populations in the maize rhizosphere. Antagonists differing in their genotypic and phenotypic characteristics shared the same DGGE electrophoretic mobility, indicating that an enormous genotypic and functional diversity might be hidden behind one single DGGE band. Cloning and sequencing was performed for a DGGE double-band which had no corresponding PCR-DGGE ribotypes among the antagonists. Sequences derived from this band were affiliated to Pseudomonas stutzeri and P. alcaligenes 16S rRNA gene sequences. As used in this study, the combination of culture-dependent and -independent methods has proven to be a powerful tool to relate functional and structural diversity of Pseudomonas spp. in the rhizosphere.     

Seasons differently impact the structure of mineral weathering bacterial communities in beech and spruce stands

Bacterial communities play an essential role in the sustainability of forest ecosystems by releasing from soil minerals the nutritive cations required not only for their own nutrition but also for that of trees. If it is admitted that the nutritional needs of trees vary during seasons, the seasonal dynamics of the mineral weathering bacterial communities colonizing the tree rhizosphere remain unknown. In this study, we characterized the mineral weathering efficacy of bacterial strains, from the rhizosphere and the adjacent bulk soil at four different seasons under two different tree species, the evergreen spruce and the deciduous beech, using a microplate assay that measures the quantity of iron released from biotite. We showed that the functional and taxonomic structures of the mineral weathering bacterial communities varied significantly with the tree species as well as with the season. Notably, the Burkholderia strains from the beech stand appeared more efficient to weather biotite that the one from the spruce stand. The mineral weathering efficacy of the bulk soil isolates did not vary during seasons under the beech stand whereas it was significantly higher for the spring and summer isolates from the spruce stand. The weathering efficacy of the rhizosphere isolates was significantly higher for the autumn isolates compared to the isolates sampled in the other seasons under the beech stand and in summer compared to the other seasons under spruce. These results suggest that seasonal differences do occur in forest soil bacterial communities and that evergreen and deciduous trees do not follow the same dynamic.     

Influence of grass species and soil type on rhizosphere microbial community structure in grassland soils

A number of studies have reported species specific selection of microbial communities in the rhizosphere by plants. It is hypothesised that plants influence microbial community structure in the rhizosphere through rhizodeposition. We examined to what extent the structure of bacterial and fungal communities in the rhizosphere of grasses is determined by the plant species and different soil types. Three grass species were planted in soil from one site, to identify plant-specific influences on rhizosphere microbial communities. To quantify the soil-specific effects on rhizosphere microbial community structure, we planted one grass species (Lolium perenne L.) into soils from three contrasting sites. Rhizosphere, non-rhizosphere (bulk) and control (non-planted) soil samples were collected at regular intervals, to examine the temporal changes in soil microbial communities. Rhizosphere soil samples were collected from both root bases and root tips, to investigate root associated spatial influences. Both fungal and bacterial communities were analysed by terminal restriction fragment length polymorphism (TRFLP). Both bacterial and fungal communities were influenced by the plant growth but there was no evidence for plant species selection of the soil microbial communities in the rhizosphere of the different grass species. For both fungal and bacterial communities, the major determinant of community structure in rhizospheres was soil type. This observation was confirmed by cloning and sequencing analysis of bacterial communities. In control soils, bacterial composition was dominated by Firmicutes and Actinobacteria but in the rhizosphere samples, the majority of bacteria belonged to Proteobacteria and Acidobacteria. Bacterial community compositions of rhizosphere soils from different plants were similar, indicating only a weak influence of plant species on rhizosphere microbial community structure.     

Impact of fresh root material and mature crop residues of oilseed rape (Brassica napus) on microbial communities associated with subsequent oilseed rape

Glasshouse bioassays were conducted to assess the impact of different inputs of oilseed rape plant material on soil and rhizosphere microbial diversity associated with subsequently grown oilseed rape (Brassica napus) plants. The first bioassay focussed on the effect of oilseed rape rhizodeposits and fresh detached root material on microbial communities, in a rapid-cycling experiment in which oilseed rape plants were grown successively in pots of field soil for 4 weeks at a time, with six cycles of repeated vegetative planting in the same pot. Molecular analyses of the microbial communities after each cycle showed that the obligate parasite Olpidium brassicae infected the roots of oilseed rape within 4 weeks after the first planting (irrespective of the influence of rhizodeposits alone or in the presence of fresh detached root material), and consistently dominated the rhizosphere fungal community, ranging in relative abundance from 43 to 88 % when oilseed rape was grown more than once in the same soil. Fresh detached root material also led to a reduction in diversity within the soil fungal community, due to the increased relative abundance of O. brassicae. In addition, rhizosphere bacterial communities were found to have a reduced diversity over time when fresh root material was retained in the soil. In the second glasshouse experiment, the effect of incorporating mature, field-derived oilseed rape crop residues (shoots and root material) on microbial communities associated with subsequently grown oilseed rape was investigated. As before, molecular analyses revealed that O. brassicae dominated the rhizosphere fungal community, despite not being prevalent in either the residue material or soil fungal communities.     

Changes in microbial communities and carbohydrate profiles induced by the mycorrhizal fungus (Glomus intraradices) in rhizosphere of olive trees (Olea europaea L.)

The influence of inoculation of olive trees with arbuscular mycorrhizal (AM) fungi, Glomus (G) intraradices, on microbial communities and sugar concentrations, were examined in rhizosphere of olive trees (Olea europaea L.). Analyses of phospholipid and neutral lipid fatty acids (PLFA and NLFA, respectively) were then used to detect changes in microbial community structure in response to inoculation of plantlets with G. intraradices.Microscopic observations studies revealed that the extraradical mycelium of the fungus showed formation of branched absorbing structures (BAS) in rhizosphere of olive tree. Root colonization with the AM fungi G. intraradices induced significant changes in the bacterial community structure of olive tree rhizosphere compared to non-mycorrhizal plants. The largest proportional increase was found for the fatty acid 10Me18:0, which indicated an increase in the number of actinomycetes in mycorrhizal rhizosphere soil, whereas the PLFAs i15:0, a15:0, i16:0, 16:1ω7 and cy17:0 which were used as indicators of bacteria decreased in mycorrhizal treatment compared to non-mycorrhizal control treatment. A highest concentration of glucose and trehalose and a lowest concentration of fructose, galactose, sucrose, raffinose and mannitol were detected in mycorrhizal rhizosphere soil. This mycorrhizal effect on rhizosphere communities may be a consequence of changes in characteristics in the environment close to mycorrhizal roots.     

Nematode assemblages in the rhizosphere of spring barley (Hordeum vulgare L.) depended on fertilisation and plant growth phase

Nematodes from rhizosphere soil of barley grown at three fertiliser treatments (control (0), NK and NPK) were studied in a field experiment. Sampling was done twice, during vegetative growth and flowering, respectively, to determine how fertiliser effects on nematode assemblages depended on plant growth phase. At the growth stage the proportion of fungal feeding nematodes (dominated by Aphelenchoides spp. and Aphelenchus sp.) was highest in NK. During flowering, the abundance and proportion of fungal feeders in the 0 and NPK plots had increased and reached a level similar to the NK plot. Overall densities of bacterial feeders (mainly Cephalobidae and Rhabditidae) were similar, but opportunistic bacterial feeders constituted a higher proportion in the fertilised plots compared to the unfertilised. Ectoparasitic plant feeders (Tylenchorhynchus sp.) were more numerous in NK and NPK than in the control at both sampling dates. Endoparasite (Pratylenchus spp.) numbers were lower in the NPK plot at the growth stage. Numbers of Tylenchidae increased between samplings. The classification of Tylenchidae as epidermal cell and root hair feeders as opposed to hyphal feeders is discussed. Results thus indicate that: (i) bacterial and especially fungal feeding nematodes are stimulated by unbalanced fertilisation; (ii) ectoparasitic plant feeders are stimulated by N-fertilisation, while migratory endoparasites are inhibited at high and balanced fertilisation; (iii) nutrient effects diminish after plants reach the flowering stage.     

In situ dynamics of soil fungal communities under different genotypes of potato, including a genetically modified cultivar

Fungi are key to the functioning of soil ecosystems, and exhibit a range of interactions with plants. Given their close associations with plants, and importance in ecosystem functioning, soil-borne fungi have been proposed as potential biological indicators of disturbance and useful agents in monitoring strategies, including those following the introduction of genetically modified (GM) crops. Here we report on the impact of potato crop varieties, including a cultivar that was genetically modified for its starch quality, on the community composition of the main phyla of fungi in soils, i.e. Ascomycota, Basidiomycota and Glomeromycota in rhizosphere and bulk soil. Samples were collected at two field sites before sowing, at three growth stages during crop development and after the harvest of the plants, and the effects of field site, plant growth stage and plant cultivar (genotype) on fungal community composition assessed using three phylum-specific T-RFLP profiling strategies and multivariate statistical analysis (NMDS ordinations with ANOSIM test). In addition, fungal biomass, arbuscular mycorrhizal colonization of roots and activities of extracellular fungal enzymes (laccases, Mn-peroxidases and cellulases) involved in degradation of lignocelluloses-rich organic matter were determined. Fungal community compositions, densities and activities were observed to differ significantly between the rhizosphere and bulk soil. The most important factors determining fungal community composition and functioning were plant growth stage for the rhizosphere communities and location and soil properties for the bulk soil communities. The basidiomycetes were the most numerous fungal group in the bulk soils and in the rhizosphere of young plants, with a shift toward greater ascomycete numbers in the rhizosphere at later growth stages. There were no detectable differences between the GM cultivar and its parental cultivar in terms of influence on fungal community structure of function. Fungal community structure and functioning of both GM- and parental cultivars fell within the range of other cultivars at most sampling moments.     

Structural differences in the rhizosphere communities of legumes are not equally reflected in community-level physiological profiles

The relationship of structural diversity and differences in the functional potentials of rhizosphere communities of alfalfa, common bean and clover was investigated in microcosms. PCR-SSCP (single strand conformation polymorphism) analysis of 16S rRNA genes revealed significant differences in the composition of the leguminous rhizosphere communities at the shoot stage of plants grown in the same soil. Sequencing of dominant SSCP-bands indicated the presence of plant specific organisms. The partial rRNA gene sequences were related to members of the α- and γ-Proteobacteria, Bacteroidetes and Actinobacteria. Besides the plant species, the soil also affected the structural diversity in rhizospheres. The dominant bacterial populations of alfalfa grown in soils with different agricultural histories were assigned to different taxonomic groups. Addressing the functional potentials, community-level physiological profiles (CLPP) were generated using BIOLOG GN^®. The three leguminous rhizosphere communities could be differentiated by principle component analysis, though the overall analysis indicated that the metabolic potential of all rhizosphere samples was similar. The functional variation examined in rhizospheres of alfalfa was minor in response to the soil origin and was found not to be significant different at different growth stages. The results indicate that similar functional potentials may be provided by structurally different bacterial communities.     

Microbial communities in forest floors under four tree species in coastal British Columbia

We compare forest floor microbial communities in pure plots of four tree species (Thuja plicata, Tsuga heterophylla, Pseudotsuga menziesii, and Picea sitchensis) replicated at three sites on Vancouver Island. Microbial communities were characterised through community level physiological profiles (CLPP), and profiling of phospholipid fatty acids (PLFA).Microbial communities from cedar forest floors had higher potential C utilisation than the other species. The F layer of the forest floor under cedar contained significantly higher bacterial biomass (PLFA) than the F layer under the other three tree species. There were differences in microbial communities among the three sites: Upper Klanawa had the highest bacterial biomass and potential C utilisation; this site also had the highest N availability in the forest floors. Forest floor H layers under hemlock and Douglas-fir contained greater biomass of Gram positive, Gram negative bacteria and actinomycetes than F layers based on PLFA, and H layers under spruce contained greater biomass of Gram negative bacteria than F layers. There were no significant differences in bacterial biomass between forest floor layers under cedar. Fungal biomass displayed opposite trends to bacteria and actinomycetes, being lowest in cedar forest floors, and highest in the F layer and at the site with lowest N availability. There were also differences in community composition among species and sites, with cedar forest floors having a much lower fungal:bacterial ratio than spruce, hemlock and Douglas-fir. The least fertile Sarita Lake site had a much greater fungal:bacterial ratio than the more fertile San Juan and Upper Klanawa sites. Forest floor layer had the greatest effect on microbial community structure and potential function, followed by site, and tree species. The similarity in trends among measures of N availability and microbial communities is further evidence that these techniques provide information on microbial communities that is relevant to N cycling processes in the forest floor.     

Brassica napus seed meal soil amendment modifies microbial community structure, nitric oxide production and incidence of Rhizoctonia root rot

A low glucosinolate content (21.8 μmol g⁻¹) Brassica napus seed meal (RSM) applied to orchard soils altered communities of both pathogenic and saprophytic soil micro-organisms. RSM amendment reduced infection by native and introduced isolates of Rhizoctonia spp. and recovery of Pratylenchus spp. from apple roots. Root infection by Rhizoctonia solani AG-5 was also suppressed in split-root assays where a portion of the root system was cultivated in RSM-amended soils and the remainder grown in the presence of the pathogen but lacking RSM. R. solani hyphal growth was not inhibited by RSM amendment. Suppression of Pratylenchus was attained to an equivalent extent by amending soils with either RSM or soybean meal (SM) when applied to provide a similar N content. Thus, glucosinolate hydrolysis products did not appear to have a significant role in the suppression of Rhizoctonia spp. or Pratylenchus spp. obtained via RSM amendment. RSM amendment elevated populations of Pythium spp. and of ammonia-oxidizing bacteria that release nitric oxide but suppressed fluorescent pseudomonad numbers. Streptomyces spp. soil populations increased significantly in response to RSM but not SM amendment. The vast majority of Streptomyces spp. recovered from the apple rhizosphere produced nitric oxide and possessed a nitric oxide synthase homolog. We propose that transformations in the bacterial community structure are associated with the observed control of Rhizoctonia root rot, with NO production by soil bacteria potentially having a role in the induction of plant systemic resistance.     

苹果树根际促生细菌种群分析

利用选择性培养基, 对多年生苹果树根际与连作幼树根际促生细菌进行了分离和测数, 并采用BOX-PCR技术进行聚类分析。结果表明: 多年生苹果树根际细菌总量及固氮细菌、解磷细菌、硅酸盐细菌、拮抗细菌4类根际促生细菌的数量均高于连作幼树根际。在多年生苹果树根际, 硅酸盐细菌的数量最大, 解磷细菌和固氮细菌的数量次之, 拮抗细菌的数量最小; 在连作幼树根际, 解磷细菌的数量最大, 硅酸盐细菌和固氮细菌的数量次之, 拮抗细菌的数量最小。从两种土壤中获得的促生细菌分离株的BOX-PCR图谱最大的相异百分数都在1.25以上, 说明这些细菌分离株的遗传进化距离比较接近。在细菌BOX-PCR图谱相异百分数为0.25的水平上, 多年生苹果树根际促生细菌分为79个聚类群, 其中固氮细菌18个聚类群, 解磷细菌29个聚类群, 硅酸盐细菌19个聚类群, 拮抗细菌18个聚类群; 连作幼树根际促生细菌分为46个聚类群, 其中固氮细菌15个聚类群, 解磷细菌19个聚类群, 硅酸盐细菌8个聚类群, 拮抗细菌9个聚类群。多年生苹果树4类根际促生细菌的多样性、丰富度和均匀度指数均高于连作幼树根际, 而优势度指数低于连作幼树根际。与连作幼树相比, 多年生苹果树根际促生细菌具有丰富的种属多样性。     

连作对再植枸杞根际细菌群落多样性和群落结构的影响研究

受枸杞道地产区土地资源等因素限制,连作障碍已成为影响枸杞产业发展的重要原因之一,导致严重的经济损失.研究连作条件下枸杞农田土壤生态系统微生物群落的演替规律对枸杞产业的可持续发展具有重要的理论意义.以宁夏银川市南梁农场连作多年的枸杞地为研究对象,利用Illumina MiSeq测序技术分析了连作对再植枸杞根际/非根际细菌群落的影响.结果表明,连作地显著抑制再植枸杞苗地径的增加,且其土壤pH较对照样地显著降低(p<0.05).测序结果证实,与对照样地相比,连作地再植枸杞根际土壤细菌物种数显著降低(p<0.05),细菌群落α多样性下降(p>0.05).主坐标分析表明,连作和对照样地间枸杞非根际细菌群落结构无明显差异,但连作显著改变再植枸杞根际细菌的群落结构.对细菌群落丰度的统计分析发现,连作地枸杞根际浮霉菌门、非根际假单胞菌门的相对丰度较对照样地显著降低(p<0.05).此外,冗余分析结果表明:枸杞园土壤pH和有效磷含量是影响枸杞非根际土壤细菌群落结构变化的主要因素,分别解释了41.8%和35.4%的群落结构变化(p<0.05),其他土壤因子无统计学意义,但土壤理化因子对再植枸杞根际细菌群落结构变化的影响均未达显著水平.这些结果证实连作能够显著抑制再植枸杞生长、影响再植枸杞根际细菌群落结构和多样性,干扰枸杞与土壤细菌群落间的互作关系.这些研究结果将为解析枸杞连作障碍机制提供理论基础.     

Principal component analysis and discriminant analysis (PCA-DA) for discriminating profiles of terminal restriction fragment length polymorphism (T-RFLP) in soil bacterial communities

To assess the impact of a transgenic crop on soil environment, we compared soil bacterial communities from the rhizospheres of cucumber green mottle mosaic virus (CGMMV)-resistant transgenic watermelon (Citrullus vulgaris [Twinser] cv. Gongdae) and non-transgenic parental line watermelon at an experimental farm in Miryang, Korea. Soil microbial community structure was studied using terminal restriction fragment length polymorphism (T-RFLP) using HaeIII and HhaI enzymes on products from polymerase chain amplification reactions (PCR) of total DNA from rhizosphere. We used principal component analyses (PCA) to reduce dimensionality of T-RFLP profiles before comparison. On these PCA scores, we conducted discrimination analyses to compare soil microbial communities from the rhizosphere of transgenic and non-transgenic. Discriminant analyses indicate that microbial communities from rhizosphere of transgenic and non-transgenic watermelon did not differ with significance at 95% level. Our study could be used as a model case to assess the environmental risk assessment of transgenic crops on soil microbial organisms.     

Structure and function of microbial communities in the rhizosphere of Scots pine after tree-felling

We evaluated changes occurring in the rhizosphere microbial communities of Scots pine (Pinus sylvestris L.) due to tree-felling and decrease of the photosynthetic C flow into the soil under field conditions over one growing season. Samples were taken from tree rhizospheres, freshly felled stump rhizospheres and bulk soil. We used culture dependent (CFU counts, community level physiological profiles, CLPPs) and independent methods (fluorogenic MUF-substrates, PLFA pattern and PCR-DGGE) to monitor the microbial communities in soil samples. The numbers of cultivable bacteria and amounts of phosphatase activity in the rhizosphere of trees were significantly higher compared with those in the bulk soil. The organic C consuming community measured by CLPP was stimulated directly after the tree-felling in stump rhizospheres; utilization of the disintegration components of cellulose, hemicellulose and chitin increased. Furthermore, bacterial and fungal biomass as well as chitin decomposers (CFU) increased in the stump rhizosphere. After 11 weeks of tree-felling the stump rhizosphere soluble PO₄-P and NH₄-N as well as amounts of total C and N began to resemble the concentrations measured in the bulk soil. However, the stump rhizosphere community structure detected by PLFA and PCR-DGGE still resembled that of the tree rhizosphere.