Sulfate-reducing bacteria in rice field soil and on rice roots.

Rice plants that were grown in flooded rice soil microcosms were examined for their ability to exhibit sulfate reducing activity. Washed excised rice roots showed sulfate reduction potential when incubated in anaerobic medium indicating the presence of sulfate-reducing bacteria. Rice plants, that were incubated in a double-chamber (phylloshpere and rhizosphere separated), showed potential sulfate reduction rates in the anoxic rhizosphere compartment. These rates decreased when oxygen was allowed to penetrate through the aerenchyma system of the plants into the anoxic root compartment, indicating that sulfate reducers on the roots were partially inhibited by oxygen or that sulfate was regenerated by oxidation of reduced S-compounds. The potential activity of sulfate reducers on rice roots was consistent with MPN enumerations showing that H2-utilizing sulfate-reducing bacteria were present in high numbers on the rhizoplane (4.1 x 10(7) g-1 root fresh weight) and in the adjacent rhizosperic soil (2.5 x 10(7) g-1 soil dry weight). Acetate-oxidizing sulfate reducers, on the other hand, showed highest numbers in the unplanted bulk soil (1.9 x 10(6) g-1 soil dry weight). Two sulfate reducing bacteria were isolated from the highest dilutions of the MPN series and were characterized physiologically and phylogenetically. Strain F1-7b which was isolated from the rhizoplane with H2 as electron donor was related to subgroup II of the family Desulfovibrionaceae. Strain EZ-2C2, isolated from the rhizoplane on acetate, grouped together with Desulforhabdus sp. and Syntrophobacter wolinii. Other strains of sulfate-reducing bacteria originated from bulk soil of rice soil microcosms and were isolated using different electron donors. From these isolates, strains R-AcA1, R-IbutA1, R-PimA1 and R-AcetonA170 were Gram-positive bacteria which were affiliated with the genus Desulfotomaculum. The other isolates were members of subgroup II of the Desulfovibrionaceae (R-SucA1 and R-LacA1), were related to Desulforhabdus sp. (strain BKA11), Desulfobulbus (R-PropA1), or culstered between Desulfobotulus sapovorans and Desulfosarcina variabilis (R-ButA1 and R-CaprA1).     

Fluorescent pseudomonads in the rhizosphere of plants and their relation to root exudates

Fluorescent pseudomonads were present in chernozem soil not influenced by plant roots (10(3)-10(4) per g dry soil) in the rhizosphere soil of various plants (10(4)-10(5) per g soil) and on roots (10(3) to 10(7) per g fresh roots), depending on the species and age of the plant. Relative species representation of fluorescent pseudomonads changed on the roots and in the plant rhizosphere as compared with free soil. Pseudomonas fluorescens, representing 60-93% of the population of fluorescent pseudomonads predominated on the roots of all plants investigated. Somewhat different results were obtained in rhizosphere soil. Relatively higher numbers of P. fluorescens were detected in the rhizosphere soil of cucumber and maize, numbers in the rhizosphere soil of wheat were practically the same as in free soil and higher numbers of P. putida were found in the rhizosphere soil of barley. Almost all components contained in the root exudates of the plants studied, including beta-pyrazolylalanine from the root exudates of cucumbers were utilized as carbon and energy sources. Root exudates of wheat and maize were utilized by the strain P. putida K2 with an efficiency of 73-91%, depending on species and age of the plant.     

Sulfate-reducing Bacteria and Mercury Methylation in the Water Column of the Lake 658 of the Experimental Lake Area

Sulfate-reducing bacteria (SRB) appear to be the main mediators of mercury methylation in sediments, which are deemed to be major sites of methylmercury (MMHg) production. However, recent studies have also found significant MMHg formation in the water column of lakes across North America. To investigate the potential involvement of SRB in mercury methylation in the water column of a stratified oligotrophic lake, two of the main families of SRB (Desulfobacteraceae and Desulfovibrionaceae) were quantified by Real-Time Polymerase Chain Reaction of the 16S rRNA gene. MMHg production was measured applying a stable isotope technique using ¹⁹⁸HgCl. Methylation assays were conducted at different water depths and under stimulation with lactate, acetate or propionate and inhibition with molybdate. Desulfobacteraceae and Desulfovibrionaceae16S rRNA gene copies in control samples accounted for 0.05% to 33% and <0.01% to 1.12% of the total bacterial 16S rRNA, respectively. MMHg formation was as high as 0.3 ng L^?1 day^?1 and largest in lactate amended samples. Strain isolation was only achieved in lactate amended media with all isolated strains being SRB belonging to the Desulfovibrio genus according to their 16S rRNA gene sequence. Isolated strains methylated between 0.06 and 0.2% of ¹⁹⁸HgCl per day. Acetate and propionate did not stimulate mercury methylation as much as lactate. Two strains were identified as Desulfovibrio sp. 12ML1 (FJ865472) and Desulfovibrio sp. 12ML3 (FJ865473), based on partial sequences of their 16S rRNA and DSR gene. Methylation assays and bacteria characterization suggest that Desulfovibrionaceae is an important mercury methylators in Lake 658. Supplemental materials are available for this article. Go to the publisher's online edition of Geomicrobiology Journal to view the free supplemental file.     

Spatial distribution of methane-oxidizing activity in a flooded rice soil

In a study on spatial distribution of methane oxidation in an unplanted flooded field, methane-oxidizing activity, analysed in soil samples under laboratory conditions, decreased with increasing depth (25 cm and beyond). In a flooded field planted to rice, rates of methane oxidation followed the order : rhizosphere (collected from roots at 10-20 cm depth) > surface soil at (0-1 cm) > subsurface soil at 10-20 cm depth, diagonally 10-15 cm away from the centre of hill. Application of ammonium sulfate and, to a lesser extent, urea to surface, rhizosphere and subsurface soil samples from flooded field planted to rice effected a distinct inhibition of methane oxidation. Nitrification inhibitors (thiourea, sodium thiosulfate and dicyandiamide) were also effective in inhibiting methane oxidation. Both surface and rhizosphere soil samples harbored higher populations of methane-oxidizing bacteria than the subsurface soil. Inhibition of methane oxidation in surface and rhizosphere soil samples concomitant with the suppression of autotrophic ammonium oxidizers by nitrification inhibitors implicates an active involvement of autotrophic ammonium oxidizers in methane oxidation.     

The rhizosphere microflora of wheat grown under controlled conditions II. Influence of the stage of growth of the plant,soil fertility and leaf treatment with urea on the rhizosphere soil microflora

Summary The rhizosphere effect of seminal roots of seedlings and of nodal roots of tillering plants of spring wheat ‘Kaspar’ was investigated under controlled conditions. The total number of micro-organisms recorded in the rhizosphere soil were significantly higher than for the non-rooted soil when investigated with the soil dilution plate method, but lower when fluorescence microscopy was used. Additions of inorganic fertilizer (NPK) decreased their numbers especially in rhizosphere soil of seminal roots and in non-rhizosphere soil, but did not change the ratio between bacteria and actinomycetes (B/A). In the rhizosphere soil the B/A ratio was higher than in the non-rhizosphere soil. An effect of urea leaf treatment was found with the dilution-plate method only in the rhizosphere soil of nodal roots, 3 to 7 days after the first treatment. Increased numbers of actinomycetes were found in this period in NPK fertilized soil, whereas increased numbers of bacteria were found at both fertility levels.     

Evaluation of the sulfate-reducing bacterial population associated with stored swine slurry

Hydrogen sulfide, produced by sulfate-reducing bacteria (SRB), is one of the most potent malodors emitted from anaerobic swine waste storage systems. However, little is known about the prevalence and diversity of SRB in those systems. The goals of this study were to evaluate the SRB population in swine manure storage systems and to develop quantitative, real-time PCR (QRT-PCR) assays to target four of the SRB groups. Dissimilatory sulfite reductase (DSR) gene sequences were obtained from swine slurry stored in underground pits (43 clones) or in lagoons (34 clones). QRT-PCR assays were designed to target the dsrA gene of four novel groups of SRB. Sequences of dsrA clones from slurry samples grouped with those from three different cultured SRB: Desulfobulbus sp. (46 clones), Desulfovibrio sp. (24 clones and 5 isolates), and Desulfobacterium sp. (7 clones). However, DsrA sequences from swine slurry clones were generally less than 85% similar to those of cultured organisms. SRB from all four targeted SRB groups were detected in underground waste storage pits (6.6 x 10(3)-8.5 x 10(7) dsrA copies mL(-1) slurry), while only two groups of SRB were detected in lagoons (3.2 x 10(5)-2.5 x 10(6) dsrA copies mL(-1) slurry). To date, this is the only study to evaluate the phylogeny and concentration of SRB in any livestock waste storage system. The new QRT-PCR assays should facilitate sensitive, specific detection of the four novel groups of SRB in livestock waste storage systems.     

Structure and activity of bacterial community inhabiting rice roots and the rhizosphere

Root-derived carbon provides a major source for microbial production and emission of CH4 from rice field soils. Therefore, we characterized the structure and activity of the bacterial community inhabiting rice roots and the rhizosphere. In the first experiment, DNA retrieved from rice roots was analysed for bacterial 16S rRNA genes using cloning, sequencing and in situ hybridization. In the second experiment, rice plants were pulse-labelled with 13CO2 (99% of atom 13C) for 7 days, and the bacterial RNA was isolated from rhizosphere soil and subjected to density gradient centrifugation. RNA samples from density fractions were analysed by terminal restriction fragment length polymorphism fingerprinting, cloning and sequencing. The experiments showed that the dominant bacteria inhabiting rice roots and the rhizosphere particularly belonged to the Alphaproteobacteria, Betaproteobacteria and Firmicutes. The RNA stable isotope probing revealed that the bacteria actively assimilating C derived from the pulse-labelled rice plants were Azospirillum spp. (Alphaproteobacteria) and members of Burkholderiaceae (Betaproteobacteria). Both anaerobic (e.g. Clostridia) and aerobic (e.g. Comamonas) degraders were present at high abundance, indicating that root environments and degradation processes were highly heterogeneous. The relative importance of iron and sulfate reducers suggested that cycling of iron and sulfur is active in the rhizosphere.     

Composition of Archaeal Community in a Paddy Field as Affected by Rice Cultivar and N Fertilizer

Methanogenesis in paddy fields is significantly influenced by environmental and field management factors such as rice cultivar and nitrogenous fertilizer. However, it has been unclear whether such effects are reflected in the structure of methanogenic archaeal populations. In the present study, molecular analyses including cloning and sequencing and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of archaeal 16S rRNA genes were used to characterize the methanogenic archaeal assemblages and to identify the effect of environmental variables including rice cultivar and N fertilizer on archaeal community compositions in a Chinese paddy field soil. The correlation between methanogenic archaeal composition and environmental variables was explored by correspondence analysis. The results showed that the spatial or niche factor (rice roots versus rhizosphere, surface, and the deeper layer soils) had the greatest influence on the archaeal community composition. There was an obvious enrichment or selection of hydrogenotrophic as opposed to acetoclastic methanogens by rice roots. The archaeal community also changed, though slightly, between the rhizosphere and bulk soils and between the surface soil and the deeper layer soil. However, rice cultivar and N fertilizer appear to have an effect only on methanogens tightly associated with rice roots.     

利用发光酶基因标记技术跟踪棉花根圈中的绿针假单胞菌PL9L

采用细菌转化和杂交的方法,成功地将全套发光酶基因标记系统Tn7luxCDABE引入绿针假单胞菌(Pseudomonas chlororaphis)PL9,得到稳定的发光标记菌PL9L。采用发光菌落平板计数法和X射线胶片自显影法,通过盆栽试验和盒裁试验,研究了发光标记菌PL9L在棉花根圈的定殖动态和分布规律。盆栽试验结果表明,在灭菌土盆栽中,播种后6d左右PL9L在棉花根圈的定殖水平达最高(31×109cfu/g根土),播种后56d左右趋向稳定,PL9L数量为17×102cfu/g根土;未灭菌土盆载中,播种后8d左右PL9L的定殖水平达最高(11×109cfu/g根土),46d左右趋向稳定,菌数为14×102cfu/g根土。盒栽试验结果表明,PL9L可从种子向根尖方向扩散,但并不与根的伸长生长同步,播种后36d,灭菌土盒栽中PL9L可扩散至种子下方120cm以内,而未灭菌土盒栽中PL9L扩散至110cm以内。在棉花根尖区域均未检测到PL9L。     

Population dynamics of a motile and a non-motile Azospirillum lipoferum strain during rice root colonization and motility variation in the rhizosphere

Abstract: Azospirillum lipoferum 4B and non-motile A. lipoferum 4T have been simultaneously isolated from rice rhizosphere at the same frequency. A. lipoferum 4T showed stable morphological and metabolic traits which are atypical for A. lipoferum species such as lack of motility, carbohydrate metabolism and laccase activity. Inoculation experiments showed that A. lipoferum 4T, but not A. lipoferum 4B, needed rice roots to stabilize in sterile soil. Both strains were able to colonize efficiently rice roots (10⁸ cfu g⁻¹ fresh roots) but motile form 4B remained dominant. In spite of their phenotypical differences, A. lipoferum 4B and 4T co-existed without exclusion in sterile soil (planted or not) and rice rhizosphere. Inoculation of rice roots with A. lipoferum 4B showed that rice rhizosphere enhanced the frequency of appearance of stable non-motile forms (40%). This percentage was weaker in plantlet growth medium (4%). However, these non-motile bacteria kept the same biochemical traits than the motile parental strain 4B (carbohydrates metabolism, laccase activity).     

稻、麦根系H~ 的分泌与介质磷水平的关系

水稻、小麦根系H~ 的分泌量随供磷水平的降低而增加,并存在明显的昼夜变化。在自然光照下H~ 分泌量随光强度增加而增多,同时强光比黑暗时H~ 分泌对磷供应水平更为敏感。磷供应不足还诱导水稻根系柠檬酸分泌量增加,而苹果酸则差异不明显。难溶性磷的溶解率与根系H~ 和柠檬酸分泌所导致的根际pH下降有密切联系。因此,在有效磷不足的条件下可明显提高稻、麦根际土壤中难溶性磷的利用率,其中丰产型小麦和粳稻品种对土壤中磷利用的根际效应更为显著。     

硅酸盐细菌NBT菌株在小麦根际定殖的初步研究

对硅酸盐细菌NBT菌株进行了耐药性标记,得到稳定的链霉素抗性标记菌NBT菌株,采用选择性培养基分离计数,通过琼脂平板和盆栽试验,研究了标记菌NBT在小麦根际的定殖动态及影响因素。结果表明,在灭菌土盆栽中,播种后9d左右NBT菌株在小麦根际的定殖水平达最高(1．4×10^8cfu·g^-1根土),播种后54d左右趋向稳定,NBT菌株细胞数量为2．4×10^3cfu·g^-1根土;未灭菌土盆栽中,播种后9d左右NBT菌株的定殖水平达最高(3．8×10^8cfu·g^-1根土),60d左右趋向稳定,菌数为3．1×10^3cfu·g^-1根土,牛物和非牛物因素对NBT菌株定殖小麦根系有影响。     

Rates of root and organism growth, soil conditions, and temporal and spatial development of the rhizosphere

BACKGROUND: Roots growing in soil encounter physical, chemical and biological environments that influence their rhizospheres and affect plant growth. Exudates from roots can stimulate or inhibit soil organisms that may release nutrients, infect the root, or modify plant growth via signals. These rhizosphere processes are poorly understood in field conditions. SCOPE AND AIMS: We characterize roots and their rhizospheres and rates of growth in units of distance and time so that interactions with soil organisms can be better understood in field conditions. We review: (1) distances between components of the soil, including dead roots remnant from previous plants, and the distances between new roots, their rhizospheres and soil components; (2) characteristic times (distance(2)/diffusivity) for solutes to travel distances between roots and responsive soil organisms; (3) rates of movement and growth of soil organisms; (4) rates of extension of roots, and how these relate to the rates of anatomical and biochemical ageing of root tissues and the development of the rhizosphere within the soil profile; and (5) numbers of micro-organisms in the rhizosphere and the dependence on the site of attachment to the growing tip. We consider temporal and spatial variation within the rhizosphere to understand the distribution of bacteria and fungi on roots in hard, unploughed soil, and the activities of organisms in the overlapping rhizospheres of living and dead roots clustered in gaps in most field soils. CONCLUSIONS: Rhizosphere distances, characteristic times for solute diffusion, and rates of root and organism growth must be considered to understand rhizosphere development. Many values used in our analysis were estimates. The paucity of reliable data underlines the rudimentary state of our knowledge of root-organism interactions in the field.     

Spatial variation of active microbiota in the rice rhizosphere revealed by in situ stable isotope probing of phospholipid fatty acids

This report is part of a serial study applying stable isotope labelling to rice microcosms to track the utilization of recently photosynthesized carbon by active microbiota in the rhizosphere. The objective of the present study was to apply phospholipid fatty acid-based stable isotope probing (PLFA-SIP) to detect the spatial variation of active microorganisms associated with rhizosphere carbon flow. In total, 49 pulses of 13CO2 were applied to rice plants in a microcosm over a period of 7 days. Rhizosphere soil was separated from bulk soil by a root bag. Soil samples were taken from rhizosphere and bulk soil, and the bulk soil samples were further partitioned both vertically (up layer and down layer) and horizontally with increasing distance to the root bag. Incorporation of 13C into PLFAs sharply decreased with distance to the roots. The labelling of 16:1omega9, 18:1omega7, 18:1omega9, 18:2omega6,9 and i14:0 PLFAs was relatively stronger in the rhizosphere while that of i15:0 and i17:0 increased in the bulk soil. The microorganisms associated with 16:1omega9 were active in both up- and down-layer soils. The microorganisms represented by i14:0, 18:1omega7 and 18:2omega6,9 exhibited a relatively higher activity in up-layer soil, whereas those represented by i15:0 and i17:0 were more active in down-layer soil. These results suggest that in the rhizosphere Gram-negative and eukaryotic microorganisms were most actively assimilating root-derived C, whereas Gram-positive microorganisms became relatively more important in the bulk soil. The active populations apparently differed between up- and down-layer soil and in particular changed with distance to the roots, demonstrating systematic changes in the activity of the soil microbiota surrounding roots.     

Root-induced changes in radial oxygen loss, rhizosphere oxygen profile, and nitrification of two rice cultivars in Chinese red soil regions

Aims

To evaluate the external and internal morphological differences of roots that might influence rice root radial oxygen loss (ROL) and the corresponding rhizosphere nitrification activity, growth characteristics and nitrogen nutrition of rice.

Methods

The root ROL and rhizosphere oxygen profile were determined using a miniaturised Clark-type oxygen microelectrode system, and the rhizosphere nitrification activity was studied with a short-term nitrification activity assay.

Results

The rice biomass, nitrogen accumulation and nitrogen use efficiency (NUE) of ZH (high yield) were significantly higher than those of HS (low yield). The root biomass, number, diameter and porosity of ZH were also much greater than those of HS. The inner and surface oxygen concentrations of the root of ZH were significantly higher than those of HS. The order of paddy soil oxygen penetration depth was ZH?>?HS?>?CK, and the order of the oxygen concentrations detected in the water layer and rhizosphere soil was the same. The rhizosphere nitrification activity and nitrate concentration of ZH were significantly higher than those of HS.

Conclusions

More porous and thicker roots improved the individual root ROL, and more adventitious root numbers enhanced the entire plant ROL and correspondingly improved the rhizosphere nitrification activity, which might influence the growth and nitrogen nutrition of rice.     

Influence of Soil Type,Cultivar and Verticillium dahliae on the Structure of the Root and Rhizosphere Soil Fungal Microbiome of Strawberry

Sustainable management of crop productivity and health necessitates improved understanding of the ways in which rhizosphere microbial populations interact with each other, with plant roots and their abiotic environment. In this study we examined the effects of different soils and cultivars, and the presence of a soil-borne fungal pathogen, Verticillium dahliae, on the fungal microbiome of the rhizosphere soil and roots of strawberry plants, using high-throughput pyrosequencing. Fungal communities of the roots of two cultivars, Honeoye and Florence, were statistically distinct from those in the rhizosphere soil of the same plants, with little overlap. Roots of plants growing in two contrasting field soils had high relative abundance of Leptodontidium sp. C2 BESC 319 g whereas rhizosphere soil was characterised by high relative abundance of Trichosporon dulcitum or Cryptococcus terreus, depending upon the soil type. Differences between different cultivars were not as clear. Inoculation with the pathogen V. dahliae had a significant influence on community structure, generally decreasing the number of rhizosphere soil- and root-inhabiting fungi. Leptodontidium sp. C2 BESC 319 g was the dominant fungus responding positively to inoculation with V. dahliae. The results suggest that 1) plant roots select microorganisms from the wider rhizosphere pool, 2) that both rhizosphere soil and root inhabiting fungal communities are influenced by V. dahliae and 3) that soil type has a stronger influence on both of these communities than cultivar.     

Analogous wheat root rhizosphere microbial successions in field and greenhouse trials in the presence of biocontrol agents Paenibacillus peoriae SP9 and Streptomyces fulvissimus FU14

Two Pythium-infested soils were used to compare the wheat root and rhizosphere soil microbial communities from plants grown in the field or in greenhouse trials and their stability in the presence of biocontrol agents. Bacteria showed the highest diversity at early stages of wheat growth in both field and greenhouse trials, while fungal diversity increased later on, at 12 weeks of the crop cycle. The microbial communities were stable in roots and rhizosphere samples across both soil types used in this study. Such stability was also observed irrespective of the cultivation system (field or greenhouse) or addition of biocontrol coatings to wheat seeds to control Pythium disease (in this study soil infected with Pythium sp. clade F was tested). In greenhouse plant roots, Archaeorhizomyces, Debaryomyces, Delftia, and unclassified Pseudeurotiaceae were significantly reduced when compared to plant roots obtained from the field trials. Some operational taxonomic units (OTUs) represented genetic determinants clearly transmitted vertically by seed endophytes (specific OTUs were found in plant roots) and the plant microbiota was enriched over time by OTUs from the rhizosphere soil. This study provided key information regarding the microbial communities associated with wheat roots and rhizosphere soils at different stages of plant growth and the role that Paenibacillus and Streptomyces strains play as biocontrol agents in supporting plant growth in infested soils.     

Occurrence and dynamics of phototrophic purple nonsulphur bacteria compared with other asymbiotic nitrogen fixers in ricefields of Egypt

The occurrence and the dynamics of phototrophic purple nonsulphur bacteria (PPNSB) as well as Azospirillum, Azotobacter, Clostridium, and cyanobacteria at different rice growth stages were studied in two ricefields, at Kafr-El-Shiekh and Al-Fayoum in Egypt.The PPNSB existed in the both rice fields examined, but their numbers varied according to field conditions, habitat and rice growth stage. After transplanting, the number of PPNSB increased gradually, reached its maximum at maximum tillering stage, and thereafter declined toward harvest time. Numbers of PPNSB were generally comparable with that of the heterotrophic N₂-fixers namely Azospirillum, Azotobacter, Clostridium and cyanobacteria, while that of phototrophic purple and green sulphur bacteria were relatively lower.The highest PPNSB numbers were generally found in rhizosphere (10³–10⁶ per g^–1 dw soil) followed by soil (10³–10⁵ per g^–1 dw soil) and floodwater (10–10² per ml). Rice plants showed a positive rhizosphere effect on PPNSB, clostridia, Azotobacter and Azospirillum, negative rhizosphere effect on cyanobacteria and green sulphur bacteria, and no effect on purple sulphur bacteria.     

Laboratory acetylene reduction assay for relative measurement of N2-fixing activities associated with field-grown wetland rice plants

Summary A short-term laboratory acetylene reduction assay using cut plant-soil samples incubated in the dark was developed for measuring relative N₂-fixing activities associated with field-grown rice plants. The assay sample consists of rhizosphere soil, root, and cut stem and leaf sheath. The cut plant-soil assay is relatively simple, rapid, and convenient; it reduces, if not eliminates, the problems encountered in whole-plant (field, pot, and water culture) and excised roots assays. Varietal differences in N₂-fixing activity were detected with the new assay technique.     

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.