Bacterial associations with the mycorrhizosphere and hyphosphere of the arbuscular mycorrhizal fungus Glomus mosseae

Roots and mycorrhizal fungi may not associate with soil bacteria randomly, but rather in a hierarchical structure of mutual preferences. Elucidation of such structures would facilitate the management of the soil biota to enhance the stability of the plant-soil system. We conducted an experiment utilizing two isolates of soil bacteria to determine their persistence in distinct mycorrhizal regions of the root zone, and their effects on general rhizosphere populations of fluorescent pseudomonads (FP). Split-root sorghum (Sorghum bicolor L.) plants were grown in four-compartment containers, constructed so that the soils in individual compartments held either (1) roots colonized by the arbuscular-mycorrhizal (AM) fungus Glomus mosseae (M), (2) nonAM roots only (R), (3) hyphae of G. mosseae (H), or (4) no mycorrhizal structures (S). The soils were inoculated (10⁷ cells g^-1 dry soil) with antibiotic-resistant (rifampicin, rif; streptomycin, sm) strains of the soil bacteria, Alcaligenes eutrophus (rif^r50) or Arthrobacter globiformis (sm^r250), or were left uninoculated as control. A. eutrophus had been isolated from a specific source (hyphosphere soil of G. mosseae), and A. globiformis from mycorrhizosphere soils of two AM fungi. After 10 wk of growth, the presence of A. eutrophus was barely detectable (<10 cfu g^-1 dry soil) in nonAM (R and S) soils, but persisted well (10⁴ cfu g^-1 dry soil) in AM (H and M) soils. Numbers of A. globiformis were more evenly distributed between all soils, but were highest in the presence of AM roots (M soil). There were varied bacterial effects on root and AM-hyphal development: A. eutrophus decreased hyphal length in H soil, while A. globiformis stimulated root length in M soil. The two bacterial inoculants did not affect numbers of FP in H, R, and M soils, but the AM status of the soils did: the numbers of FP increased in the order M>R>H>S. There was a positive correlation of FP numbers with both bacterial inoculants in M and H soils. Numbers of FP changed with root or hyphal lengths, an effect that was related to changes in the numbers of the inoculated bacteria. The results indicate that the hyphosphere-specific A. eutrophus depended on the presence of G. mosseae, but that the nonspecific A. globiformis did not. The mycorrhizal status of soils may selectively influence persistence of bacterial inoculants as well as affecting the numbers of other native bacteria.     

镉和铅污染对烤烟根区土壤微生物及烟叶品质的影响

通过盆栽试验,研究了Cd和Pb污染对烤烟根区微生物和烟叶品质的影响.结果表明,随着Cd、Pb处理浓度的增加,烤烟根区4种微生物数量均呈减少趋势;低浓度Cd(≤10 mg·kg^-1)和Pb(≤300 mg·kg^-1)下,烤烟根区4种微生物数量降幅最大;烤烟烟叶中品质指标糖/碱比和氮/碱比升高,化学成分组成趋于不协调,不利于烟叶香味的形成.烟叶K含量与Cd和Pb污染浓度呈负相关,与Cd污染下根区钾细菌数量呈显著(或极显著)正相关,与Pb污染根区钾细菌数量相关不显著.     

放线菌制剂对人参生长及根域土壤微生物区系的影响

以小兴安岭地区人参为研究对象,探索放线菌制剂对人参的促生效应及对人参根区、根表土壤微生物区系的影响.结果表明： 经放线菌制剂Streptomyces pactum（Act12）处理后,人参药用部分产量增加,品质改善;叶片诱导酶活性提高,根系活力增强;土壤中细菌、放线菌的数量和比例显著增加,真菌的数量和比例减少.与对照相比,土壤微生物区系结构改变:优势菌荧光假单胞菌、韩国假单胞菌和氧化微杆菌在根区、根表土壤中的数量大幅提高;病原真菌烟色织孢霉在根区土壤中减少,在根表土壤中消失.表明施用放线菌制剂Act12能够改善土壤微生物区系,提高人参植株的抗性和根系活力,增加产量并改善品质.     

Numbers and locations of native bacteria on field-grown wheat roots quantified by fluorescence in situ hybridization (FISH)

Native bacteria, Pseudomonas and filamentous bacteria were quantified and localized on wheat roots grown in the field using fluorescence in situ hybridization (FISH). Seminal roots were sampled through the season from unploughed soil in a conservation farming system. Such soils are spatially heterogeneous, and many roots grow slowly through hard soil with cracks and pores containing dead roots remnant from previous crops. Root and rhizosphere morphology, and contact with soil particles were preserved, and autofluorescence was avoided by observing sections in the far-red with Cy5 and Cy5.5 fluorochromes. Spatial analyses showed that bacteria were embedded in a stable matrix (biofilm) within 11 microm of the root surface (range 2-30 microm) and were clustered on 40% of roots. Half the clusters co-located with axial grooves between epidermal cells, soil particles, cap cells or root hairs; the other half were not associated with visible features. Across all wheat roots, although variable, bacteria averaged 15.4 x 10(5) cells per mm(3) rhizosphere, and of these, Pseudomonas and filaments comprised 10% and 4%, respectively, with minor effects of sample time, and no effect of plant age. Root caps were most heavily colonized by bacteria along roots, and elongation zones least heavily colonized. Pseudomonas varied little with root development and were 17% of bacteria on the elongation zone. Filamentous bacteria were not found on the elongation zone. The most significant factor to rhizosphere populations along a wheat root, however, was contact with dead root remnants, where Pseudomonas were reduced but filaments increased to 57% of bacteria (P < 0.001). This corresponded with analyses of root remnants showing they were heavily colonized by bacteria, with 48% filaments (P < 0.001) and 1.4%Pseudomonas (P = 0.014). Efforts to manage rhizosphere bacteria for sustainable agricultural systems should continue to focus on root cap and mucilage chemistry, and remnant roots as sources of beneficial bacteria.     

Metabolic activity of bacteria isolated from soil, rhizosphere and mycorrhizosphere of pine (Pinus silvestris L.)

Casamino acids were found to be the best substrate for bacteria isolated from soil, rhizosphere and mycorrhizosphere of pine. Most active with this substrate were however the root zone bacteria. Glucose was oxidized similarly by the bacteria isolated from the three sources.     

The role of endophytic bacteria during seed piece decay and potato tuberization

Healthy potato tubers (Solanum tuberosum L.) cv. Kennebec were found to be internally colonized by non-pathogenic bacterial populations originating from root zone soil. These endophytic bacteria were categorized, on the basis of bioassays, as plant growth promoting (PGP), plant growth retarding (PGR) and plant growth neutral (PGN). Genera isolated from tubers included Pseudomonas, Bacillus, Xanthomonas, Agrobacterium, Actinomyces and Acinetobacter. The PGP and PGR isolates were similarly distributed throughout these genera. Bacterial populations increased in the root zone soil directly adjacent to the seed piece during and immediately following seed piece decay. Bacteria sampled at this time were capable of promoting tuber number and weight. The proportions of PGP, PGR and PGN bacteria in the root zone were altered as endophytic bacteria were released from the decaying seed piece. The study indicates that endophytic bacteria present in the seed tubers may play an important role in seed piece decay, tuberization and plant growth.     

3株芽孢杆菌对番茄的促生作用及对番茄根域微生物的影响

通过种子萌发和盆栽促生试验研究3株芽孢杆菌Bs10、Ba12和Bl10对番茄的促生作用及其对番茄根域微生物区系的调节作用.结果表明: 3株芽孢杆菌对番茄种子的胚轴、胚根和番茄植株的生长有明显的促进作用,处理后番茄根系的总长度、总表面积和总体积均显著增加;处理后土壤中细菌数量和比例显著增加,真菌数量和比例明显减少.与对照相比,土壤微生物区系优势菌数量发生改变:优势甲基营养型芽孢杆菌在番茄根区、根表土壤中和根内的数量大幅提高;病原真菌腐皮镰刀菌和尖孢镰刀菌在根区和根表土壤中的数量显著减少.推知芽孢杆菌对根系微生物区系的调节作用是其发挥防病促生作用的重要机制之一.     

Dynamic of the volume and structure of the soil bacterial complex in the presence of azobenzene

Azobenzene exerted no significant effect on the dynamics and the species composition of the saprophytic soil bacterial complex, which remained almost the same as in the control and was characterized by the predominance of Curtobacterium sp., Arthrobacter globiformis, and Bacillus megaterium in all stages of succession. Some heterotrophic bacteria were found to be able to accumulate azobenzene. Bac. cereus and Bac. polymyxa degraded azobenzene during their cultivation in nutrient media.     

Bacterial succession on n-alkanes under the conditions of sulfate reduction

The dynamics of species composition of a hydrocarbon-oxidizing bacteriocenosis of a ground suspension of Mozhaisk Reservoir has been studied. The bacteriocenosis was undergoing development in a paraffin film (model association composed of sulfate-reducing bacteria and hydrocarbon-oxidizing bacteria). The type of bacterial succession did not depend on the depth, from which ground samples were collected. Two microbial species (Pseudomonas sp. and Arthrobacter globiformis) were absolutely dominant. Pseudomonas sp. was dominant at the early and intermediate stages of the succession, whereas A. globiformis was present in the hydrocarbon-oxidizing bacteriocenosis throughout the whole period of the succession. There was a trend toward a gradual increase in the ratio of A. globiformis, and, by the end of the experiment, Pseudomonas sp. was replaced by A. globiformis almost completely. The bacterial species Micrococcus sp. and Rhodococcus erythropolis were minor components of the hydrocarbon-oxidizing bacteriocenosis under the conditions of sulfate reduction. The succession of species of hydrocarbon-oxidizing bacteria in the paraffin film of the model association reflects both the life strategy of the bacterial species under study and the degree of their tolerance to products of sulfate reduction.     

石油污染土壤修复植物的根-土界面微生物特征

选取沈抚灌区的主要修复植物蓖麻为实验材料,分析了蓖麻根区土壤、根际土壤、根面、根内4个层面上细菌和真菌的数量,优势菌种的生理生化特征,及细菌菌株生长营养类型,揭示了根-土界面微生物区系特征与石油污染土壤生物修复的关系.结果发现：修复植物根-土界面上,细菌数量为根际>根面>根内,真菌数量为根内>根面>根际,根面细菌与真菌数量均处于根际与根内区域微生物数量之间;修复植物蓖麻根面区域优势细菌种类最多,根内区域优势真菌种类最多;根际与根内的优势细菌具有较强的降解大分子物质的能力;根面细菌在营养需求分类上可归为氨基酸需求菌群.     

Effect of dicumarol on the growth of some soil microorganisms.

The effect of dicumarol on growth of selected soil bacteria: Azotobacter chroococcum, Arthrobacter globiformis, A. citreus and Bacillus megaterium was studied. The following minimum concentrations were inhibitory in vitro: Arthrobacter citreus--20 mug/ml., Bacillus megaterium--40 mug/ml., Azotobacter chroococcum--40 mug/ml. Arthrobacter globiformis--70 mug/ml. Cells of all microorganisms studied grown in the presence of dicumarol developed aberrant morphological forms.     

Production of B--vitamins by heterotrophic planktonic bacteria isolated from littoral zone of lake Jeziorak.

Different genera and groups of heterotrophic planktonic bacteria of the littoral zone of the lake Jeziorak produced different B-group vitamins. Most numerous among the planktonic bacteria of this zone of the lake were biotin producers and least numerous were organisms synthesizing riboflavin and nicotinic acid. Most bacteria produced one or two vitamins. Three or more vitamins were produced by only a few strains. In autumn no strain produced all five vitamins studied in this work.     

The abundance of pink-pigmented facultative methylotrophs in the root zone of plant species in invaded coastal sage scrub habitat

Pink-pigmented facultative methylotrophic bacteria (PPFMs) are associated with the roots, leaves and seeds of most terrestrial plants and utilize volatile C(1) compounds such as methanol generated by growing plants during cell division. PPFMs have been well studied in agricultural systems due to their importance in crop seed germination, yield, pathogen resistance and drought stress tolerance. In contrast, little is known about the PPFM abundance and diversity in natural ecosystems, let alone their interactions with non-crop species. Here we surveyed PPFM abundance in the root zone soil of 5 native and 5 invasive plant species along ten invasion gradients in Southern California coastal sage scrub habitat. PPFMs were present in every soil sample and ranged in abundance from 10(2) to 10(5) CFU/g dry soil. This abundance varied significantly among plant species. PPFM abundance was 50% higher in the root zones of annual or biennial species (many invasives) than perennial species (all natives). Further, PPFM abundance appears to be influenced by the plant community beyond the root zone; pure stands of either native or invasive species had 50% more PPFMs than mixed species stands. In sum, PPFM abundance in the root zone of coastal sage scrub plants is influenced by both the immediate and surrounding plant communities. The results also suggest that PPFMs are a good target for future work on plant-microorganism feedbacks in natural ecosystems.     

Polychlorinated biphenyl (PCB)-degrading bacteria associated with trees in a PCB-contaminated site

The abundance, identities, and degradation abilities of indigenous polychlorinated biphenyl (PCB)-degrading bacteria associated with five species of mature trees growing naturally in a contaminated site were investigated to identify plants that enhance the microbial PCB degradation potential in soil. Culturable PCB degraders were associated with every plant species examined in both the rhizosphere and root zone, which was defined as the bulk soil in which the plant was rooted. Significantly higher numbers of PCB degraders (2.7- to 56.7-fold-higher means) were detected in the root zones of Austrian pine (Pinus nigra) and goat willow (Salix caprea) than in the root zones of other plants or non-root-containing soil in certain seasons and at certain soil depths. The majority of culturable PCB degraders throughout the site and the majority of culturable PCB degraders associated with plants were identified as members of the genus Rhodococcus by 16S rRNA gene sequence analysis. Other taxa of PCB-degrading bacteria included members of the genera Luteibacter and Williamsia, which have not previously been shown to include PCB degraders. PCB degradation assays revealed that some isolates from the site have broad congener specificities; these isolates included one Rhodococcus strain that exhibited degradation abilities similar to those of Burkholderia xenovorans LB400. Isolates with broad congener specificity were widespread at the site, including in the biostimulated root zone of willow. The apparent association of certain plant species with increased abundance of indigenous PCB degraders, including organisms with outstanding degradation abilities, throughout the root zone supports the notion that biostimulation through rhizoremediation is a promising strategy for enhancing PCB degradation in situ.     

蒙古栎根内及根围土壤中共生真菌和细菌群落组成分析

为探究中国东北地区森林根系和根围土壤中共生真菌的分布状况,分析了黑龙江省五大连池市蒙古栎Quercus mongolica纯林中根内及根围土壤中共生真菌和细菌群落的组成。结果表明,根内真菌的1 295个OTUs中有209个OTUs为共生真菌,隶属于36属,相对丰度25.46%;根围土壤真菌1 513个OTUs中有285个OTUs为共生真菌,隶属于40属,相对丰度59.91%;根内与根围土壤共同拥有共生真菌33属,根内特有3属,根围土壤特有7属。其中,外生菌根真菌为根内和根围土壤中共生真菌的主要类群,分别占共生真菌的98.82%和99.80%。定殖根内的细菌共获得5 550个OTUs,隶属于400属;根围土壤细菌获得8 406个OTUs,隶属于436属,根内细菌群落的Shannon指数和Chao1指数均低于根围土壤的。PICRUSt功能预测分析结果表明,根内的信号转导通路与信号分子和相互作用通路（包括CAM配体、ECM-受体相互作用等通路）的丰度低于根围土壤,而膜运输通路与信号分子和相互作用通路（包括细菌毒素、细胞抗原等通路）的丰度高于根围土壤。根内与根围土壤中菌根辅助细菌组成差异分析结果表明,除慢生根瘤菌属 Bradyrhizobium外,根内其余9属的相对丰度均高于根围土壤,尤其假单胞菌属 Pseudomonas的相对丰度远高于根围土壤。     

Root architecture and wind-firmness of mature Pinus pinaster

This study aims to link three-dimensional coarse root architecture to tree stability in mature timber trees with an average of 1-m rooting depth. Undamaged and uprooted trees were sampled in a stand damaged by a storm. Root architecture was measured by three-dimensional (3-D) digitizing. The distribution of root volume by root type and in wind-oriented sectors was analysed. Mature Pinus pinaster root systems were organized in a rigid 'cage' composed of a taproot, the zone of rapid taper of horizontal surface roots and numerous sinkers and deep roots, imprisoning a large mass of soil and guyed by long horizontal surface roots. Key compartments for stability exhibited strong selective leeward or windward reinforcement. Uprooted trees showed a lower cage volume, a larger proportion of oblique and intermediate depth horizontal roots and less wind-oriented root reinforcement. Pinus pinaster stability on moderately deep soils is optimized through a typical rooting pattern and a considerable structural adaptation to the prevailing wind and soil profile.     

Metabolic Status of Bacteria and Fungi in the Rhizosphere of Ponderosa Pine Seedlings

We determined the quantity and metabolic status of bacteria and fungi in rhizosphere and nonrhizosphere soil from microcosms containing ponderosa pine seedlings. Rhizosphere soil was sampled adjacent to coarse, fine, or young roots. The biovolume and metabolic status of bacterial and fungal cells was determined microscopically and converted to total and active biomass values. Cells were considered active if they possessed the ability to reduce the artificial electron acceptor 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride (INT) to visible intracellular deposits of INT formazan. A colorimetric assay of INT formazan production was also used to assess dehydrogenase activity. INT-active microorganisms made up 44 to 55% of the microbial biomass in the soils studied. The proportion of fungal biomass that exhibited INT-reducing activity (40 to 50%) was higher than previous estimates of the active proportion of soil fungi determined by using fluorescein diacetate. Comparison between soils from different root zones revealed that the highest total and INT-active fungal biomass was adjacent to fine mycorrhizal roots, whereas the highest total and active bacterial biomass was adjacent to the young growing root tips. These observations suggest that fungi are enhanced adjacent to the fine roots compared with the nonrhizosphere soil, whereas bacteria are more responsive than fungi to labile carbon inputs in the young root zone. Colorimetric dehydrogenase assays detected gross differences between bulk and rhizosphere soil activity but were unable to detect more subtle differences due to root types. Determination of total and INT-active biomass has increased our understanding of the role of spatial compartmentalization of bacteria and fungi in rhizosphere carbon flow.     

Cyclic tetrasaccharide-synthesizing enzymes from Arthrobacter globiformis A19

A bacterial strain Arthrobacter globiformis A19 producing cyclic tetrasaccharide (CTS) was isolated from soil. The enzymes, 6-alpha-glucosyltransferase (6GT) and 3-alpha-isomaltosyltransferase (IMT), involved in the synthesis of CTS were purified to homogeneity. The molecular and enzymatic properties of IMT from A. globiformis were similar to those of enzymes from Bacillus globisporus C11 and N75. Arthrobacter 6GT had a smaller molecular mass of 108 kDa and a higher optimum pH of 8.4 than the enzymes from strains of B. globisporus. The genes for IMT (ctsY) and 6GT (ctsZ) were cloned from the genome of A. globiformis A19. The two genes linked together in tandem and formed a gene cluster, ctsYZ. Both of the gene products showed similarities to alpha-glucosidases belonging to glycoside hydrolase family 31, and conserved two aspartic acids corresponding to the putative catalytic residues of the family enzymes. The enzymatic system for the production of CTS consisting of 6GT and IMT might be widespread among bacteria.     

Development of rhizosphere and rhizoplane microflora of Aristida coerulescens in the Libyan desert

Summary Microflora of rhizosphere soil, rhizoplane and macerated root-portions of Aristida coerulescens, naturally occurring in the Libyan desert, were different in count and isolates, in the different root zones. A rhizosphere effect characteristic of each zone is shown. The root base contained the lowest numbers of microflora (bacteria and fungi) whilst the root tip included the highest counts. Distribution of most of the individual fungal species in the different root zones and root-surfaces is given in text.     

Production of organic acids by bacteria isolated from soil, rhizosphere and mycorrhizosphere of pine (Pinus sylvestris L.)

The bacteria studied released into the medium ten to eleven organic acids. Soil organisms excreted mainly pyruvic and alpha-ketoglutaric acids, while strains from the root zone--gluconic acid and unidentified uronic acid (y2). Mean indices of total production of the organic acids by bacteria were in the following order: rhizosphere less than soil less than mycorrizosphere. Bacteria from the root zone released into the medium very large amounts of pyruvic, gluconic, and uronic (y2) acids--in some instances several times higher than bacterial dry mass.