Actinomycetes in the Rhizosphere of Barley Grown on Acid Soddy Podzolic Soil

The study of various factors (soil acidity, the variety of barley, and their developmental phases) on the rhizosphere actinomycete complex showed that it is soil acidity that substantially influences the population of rhizosphere actinomycetes. The effect of soil acidity was most likely due to the different tolerance of rhizosphere actinomycetes to high concentrations of the aluminum and hydrogen ions. The developmental phases of barley correlated with the population indices of only one genus of actinomycetes, Micromonospora.     

番茄根际微生物种群动态变化及多样性

采用盆栽试验的方法对番茄根际主要微生物种群在不同生育期的动态变化进行了跟踪研究.结果表明,在番茄整个生育期内,可培养细菌数量在初花期和初果期时最多;放线菌数量从苗期到末期逐渐减少;真菌数量逐渐增多.番茄对细菌根际效应明显.DGGE图谱显示不同生育期番茄根际均具有较高的细菌多样性.根际细菌种类和数量在初花期发生较为显著的变化,初果期根际群落多样性指数(H)和物种丰度(S)值都达到最高,微生物最丰富,是筛选拮抗菌的较好时期.     

Micro-organisms in the rhizosphere of plants inoculated withAzotobacter chroococcum

Summary The hypothesis that inoculation withAzotobacter chroococcum affects the growth of plants indirectly through changing the rhizosphere microflora was investigated. Inoculated and uninoculated wheat and tomato plants were grown in the glasshouse in two different soils, and total bacteria, chitinolytic bacteria, actinomycetes, glucosefermenting bacteria, aerobic cellulose-decomposing bacteria, and anaerobes were determined in intervals in the rhizosphere and in the soil. Root-surface fungi were studied using the Harley and Waid's root-washing technique¹⁰. Azotobacter became established in the rhizosphere of wheat and tomato plants and stimulated their growth. All the bacterial groups examined were more abundant in the rhizosphere than in the soil. Inoculation with Azotobacter delayed the colonization of roots by bacteria, actinomycetes, and fungi in the rhizosphere, but had no effect on other organisms. Inoculation did not affect the dominant root-surface fungi, and minor changes were not consistent.Part of a thesis accepted by the University of London for the degree of Ph.D. in Microbiology.     

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.     

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.     

Colonization of plant rhizosphere by actinomycetes of different genera

The survival of environmental isolates of actinomycetes introduced with the seeds of agricultural plants in the root-free soil and in the rhizosphere and rhizoplane was studied. Different strategies of colonization of the rhizosphere were revealed for the representatives of the genera Streptomyces, Micromonospora, and Streptosporangium, the organisms typical for the moderate climate rhizosphere. The plants of winter rye (Secale cereale L.) inoculated with actinomycetes were shown to have growth advantages, while the cow clover plants (Trifolium pratense L.) had no growth advantages compared to uninoculated plants. The role of the plant component in the interaction with mycelial prokaryotes is discussed.     

Effects of organic amendments to soil on the rhizosphere microflora of antirrhinum infected withVerticillium dahliae Kleb.

Summary Organic (e.g. chitin, green manure, cellulose) amendments to soil induced quantitative and qualitative changes in the rhizosphere microflora of antirrhinum plants infected withVerticillium dahliae Kleb. Whereas reduction in disease severity occurred with chitin and green manure amendments, an increase in disease severity was observed with the application of cellulose. The reduction of the disease severity with chitin and green manure may be correlated with the increased population of actinomycetes in the antirrhinum rhizosphere.     

Phenotypic characterization of microbes in the rhizosphere of Alyssum murale

Metal hyperaccumulator plants like Alyssum murale are used for phytoremediation of Ni contaminated soils. Soil microorganisms are known to play an important role in nutrient acquisition for plants, however, little is known about the rhizosphere microorganisms of hyperaccumulators. Fresh and dry weight, and Ni and Fe concentrations in plant shoots were higher when A. murale was grown in non-sterilized compared to sterilized soils. The analysis of microbial populations in the rhizosphere of A. murale and in bulk soils demonstrated that microbial numbers were affected by the presence of the plant. Significantly higher numbers of culturable actinomycetes, bacteria and fungi were found in the rhizosphere compared to bulk soil. A higher percent of Ni-resistant bacteria were also found in the rhizosphere compared to bulk soil. Percentage of acid producing bacteria was higher among the rhizosphere isolates compared to isolates from bulk soil. However, proportions of siderophore producing and phosphate solubilizing bacteria were not affected by the presence of the plant. We hypothesize that microbes in the rhizosphere of A. murale were capable of reducing soil pH leading to an increase in metal uptake by this hyperaccumulator.     

Colonization of plant rhizosphere by actinomycetes of different genera

The survival of environmental isolates of actinomycetes introduced with the seeds of agricultural plants in root-free soil and in the rhizosphere and rhizoplane was studied. Different strategies of colonization of the rhizosphere were revealed for the representatives of the genera Streptomyces, Micromonospora, and Streptosporangium, organisms typical for the moderate climate rhizosphere. The plants of winter rye (Secale cereale L.) inoculated with actinomycetes were shown to have growth advantages, while the cow clover plants (Trifolium pratense L.) had no growth advantages compared to uninoculated plants. The role of the plant component in the interaction with mycelial prokaryotes is discussed.     

Mobilisation of Cu,Mn and Zn in the soil solutions of barley rhizospheres

Summary We report data showing changes during the growing season in the concentrations of manganese, zinc and copper in the rhizosphere in soil solutions of barley plants. Substantial mobilisation of these trace elements occurred during the early stages of rhizosphere development so that soil solution concentrations increased three fold for the copper and zinc and fifteen fold for manganese.     

Distribution of metabolic activity and phosphate starvation response of lux-tagged Pseudomonas fluorescens reporter bacteria in the barley rhizosphere.

The purpose of this study was to determine the metabolic activity of Pseudomonas fluorescens DF57 in the barley rhizosphere and to assess whether sufficient phosphate was available to the bacterium. Hence, two DF57 reporter strains carrying chromosomal luxAB gene fusions were introduced into the rhizosphere. Strain DF57-40E7 expressed luxAB constitutively, making bioluminescence dependent upon the metabolic activity of the cells under defined assay conditions. The DF57-P2 reporter strain responded to phosphate limitation, and the luxAB gene fusion was controlled by a promoter containing regulatory sequences characteristic of members of the phosphate (Pho) regulon. DF57 generally had higher metabolic activity in a gnotobiotic rhizosphere than in the corresponding bulk soil. Within the rhizosphere the distribution of metabolic activity along the root differed between the rhizosphere soil and the rhizoplane, suggesting that growth conditions may differ between these two habitats. The DF57-P2 reporter strain encountered phosphate limitation in a gnotobiotic rhizosphere but not in a natural rhizosphere. This difference in phosphate availability seemed to be due to the indigenous microbial population, as DF57-P2 did not report phosphate limitation when established in the rhizosphere of plants in sterilized soil amended with indigenous microorganisms.     

Nitrogen availability to Pseudomonas fluorescens DF57 is limited during decomposition of barley straw in bulk soil and in the barley rhizosphere.

The availability of nitrogen to Pseudomonas fluorescens DF57 during straw degradation in bulk soil and in barley rhizosphere was studied by introducing a bioluminescent reporter strain (DF57-N3), responding to nitrogen limitation, to model systems of varying complexity. DF57-N3 was apparently not nitrogen limited in the natural and sterilized bulk soil used for these experiments. The soil was subsequently amended with barley straw, representing a plant residue with a high carbon-to-nitrogen ratio (between 60 and 100). In these systems the DF57-N3 population gradually developed a nitrogen limitation response during the first week of straw decomposition, but exclusively in the presence of the indigenous microbial population. This probably reflects the restricted ability of DF57 to degrade plant polymers by hydrolytic enzymes. The impact of the indigenous population on nitrogen availability to DF57-N3 was mimicked by the cellulolytic organism Trichoderma harzianum Rifai strain T3 when coinoculated with DF57-N3 in sterilized, straw-amended soil. Limitation occurred concomitantly with fungal cellulase production, pointing to the significance of hydrolytic activity for the mobilization of straw carbon sources, thereby increasing the nitrogen demand. Enhanced survival of DF57-N3 in natural soil after straw amendment further indicated that DF57 was cross-fed with carbon/energy sources. The natural barley rhizosphere was experienced by DF57-N3 as an environment with restricted nitrogen availability regardless of straw amendment. In the rhizosphere of plants grown in sterilized soil, nitrogen limitation was less severe, pointing to competition with indigenous microorganisms as an important determinant of the nitrogen status for DF57-N3 in this environment. Hence, these studies have demonstrated that nitrogen availability and gene expression in Pseudomonas is intimately linked to the structure and function of the microbial community. Further, it was demonstrated that the activities of cellulolytic microorganisms may affect the availability of energy and specific nutrients to a group of organisms deficient in hydrolytic enzyme activities.     

Nitrogen Availability to Pseudomonas fluorescens DF57 Is Limited during Decomposition of Barley Straw in Bulk Soil and in the Barley Rhizosphere

The availability of nitrogen to Pseudomonas fluorescens DF57 during straw degradation in bulk soil and in barley rhizosphere was studied by introducing a bioluminescent reporter strain (DF57-N3), responding to nitrogen limitation, to model systems of varying complexity. DF57-N3 was apparently not nitrogen limited in the natural and sterilized bulk soil used for these experiments. The soil was subsequently amended with barley straw, representing a plant residue with a high carbon-to-nitrogen ratio (between 60 and 100). In these systems the DF57-N3 population gradually developed a nitrogen limitation response during the first week of straw decomposition, but exclusively in the presence of the indigenous microbial population. This probably reflects the restricted ability of DF57 to degrade plant polymers by hydrolytic enzymes. The impact of the indigenous population on nitrogen availability to DF57-N3 was mimicked by the cellulolytic organism Trichoderma harzianum Rifai strain T3 when coinoculated with DF57-N3 in sterilized, straw-amended soil. Limitation occurred concomitantly with fungal cellulase production, pointing to the significance of hydrolytic activity for the mobilization of straw carbon sources, thereby increasing the nitrogen demand. Enhanced survival of DF57-N3 in natural soil after straw amendment further indicated that DF57 was cross-fed with carbon/energy sources. The natural barley rhizosphere was experienced by DF57-N3 as an environment with restricted nitrogen availability regardless of straw amendment. In the rhizosphere of plants grown in sterilized soil, nitrogen limitation was less severe, pointing to competition with indigenous microorganisms as an important determinant of the nitrogen status for DF57-N3 in this environment. Hence, these studies have demonstrated that nitrogen availability and gene expression in Pseudomonas is intimately linked to the structure and function of the microbial community. Further, it was demonstrated that the activities of cellulolytic microorganisms may affect the availability of energy and specific nutrients to a group of organisms deficient in hydrolytic enzyme activities.     

荒漠草原区土壤粒径组成对柠条根际土壤微生物数量及酶活性的影响

柠条(Caragana korshinskii)是荒漠草原区主要的造林绿化树种,研究其根际土壤微生物和酶活性与不同土壤类型土壤粒径组成的关系有重要意义,然而土壤粒径对荒漠草原柠条根际土壤微生物数量和酶活性的影响知之甚少,探讨土壤颗粒组分与微生物数量、土壤酶活性之间的关系,以及土壤颗粒组成对荒漠草原区固沙灌木植物柠条根际土壤微生物数量及酶活性的影响,可为揭示荒漠草原土壤退化及生态修复提供参考。以宁夏荒漠草原区土壤粒径组成差异显著的灰钙土、红黏土、风沙土环境下栽植的柠条为研究对象,研究不同土壤颗粒组成对根际土壤微生物数量及酶活性的相互关系与影响。结果表明:土壤微生物的数量表现为细菌放线菌真菌。根际土壤中的细菌、真菌数量显著高于非根际,且在3种不同类型的土壤中随着细砂粒的增多,真菌和放线菌数量逐渐降低,而细菌数量呈先增大后减小的趋势;根际与非根际土壤的蔗糖酶、碱性磷酸酶及过氧化氢酶活性均呈现出灰钙土红黏土风沙土的趋势,红黏土根际土壤中的脲酶活性显著高于灰钙土与风沙土;除过氧化氢酶外,土壤酶活性表现为根际高于非根际,在3种不同类型的土壤中随着细砂含量的增加,土壤酶活性均呈递减趋势。土壤颗粒组成与微生物数量之间没有明显的相关性,而与土壤酶活性之间显著相关,土壤酶活性与黏粒、粉粒呈正相关,与细砂、中砂呈负相关关系,根际土壤中酶活性更高,能够为植物及微生物提供更多的营养。     

Bacterial origin and community composition in the barley phytosphere as a function of habitat and presowing conditions

An understanding of the factors influencing colonization of the rhizosphere is essential for improved establishment of biocontrol agents. The aim of this study was to determine the origin and composition of bacterial communities in the developing barley (Hordeum vulgare) phytosphere, using denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes amplified from extracted DNA. Discrete community compositions were identified in the endorhizosphere, rhizoplane, and rhizosphere soil of plants grown in an agricultural soil for up to 36 days. Cluster analysis revealed that DGGE profiles of the rhizoplane more closely resembled those in the soil than the profiles found in the root tissue or on the seed, suggesting that rhizoplane bacteria primarily originated from the surrounding soil. No change in bacterial community composition was observed in relation to plant age. Pregermination of the seeds for up to 6 days improved the survival of seed-associated bacteria on roots grown in soil, but only in the upper, nongrowing part of the rhizoplane. The potential occurrence of skewed PCR amplification was examined, and only minor cases of PCR bias for mixtures of two different DNA samples were observed, even when one of the samples contained plant DNA. The results demonstrate the application of culture-independent, molecular techniques in assessment of rhizosphere bacterial populations and the importance of the indigenous soil population in colonization of the rhizosphere.     

Bacterial diversity of the rhizosphere of maize (Zea mays) grown in tropical soil studied by temperature gradient gel electrophoresis

The bacterial diversity and population dynamics in the rhizosphere of two maize cultivars (Nitroflint and Nitrodent) grown in tropical soils was studied, by traditional cultivation techniques and 16S rRNA gene-based molecular analysis of DNA directly extracted from soil and rhizosphere samples. Rhizosphere and soil samples were taken at three different plant growth stages. Total aerobic bacterial counts were determined. Fingerprints of the most dominant bacterial population were generated by TGGE separation of 16S rRNA gene fragments amplified from total community DNA using eubacterial specific primers. To reduce the complexity of TGGE fingerprints or to analyse less abundant populations, primers specific for different phylogenetic groups have been used. A comparison of the cfu obtained for rhizosphere of both cultivars indicated significant differences only for rhizosphere and soil samples taken 40 days after sowing. However, a comparison of TGGE patterns indicated that the composition of the bacterial community analysed at different plant growth stages for both cultivars was similar. A comparison of -, -proteobacterial and actinomycete TGGE patterns of both cultivars confirmed this observation. The eubacterial TGGE profiles reflected strong seasonal population shifts in the bacterial rhizosphere community of both maize cultivars which could be also observed in the TGGE patterns of - and -proteobacteria and to a lesser extent for actinomycetes. The rhizosphere effect was much more pronounced for young roots compared to samples taken from mature maize plants. The rhizosphere fingerprints showed a reduced complexity for young plants with up to five dominating bands while for mature plants patterns similar to those of soil were observed. Sequencing of dominant clones indicated that the dominant population found at all plant growth stages can be assigned to Arthrobacter populations.     

Predicted microbial biomass in the rhizosphere of barley in the field

Summary Laboratory data for the loss of root material by barley and field data for the growth of barley plants in Syria and in England have been combined to predict the amount of material lost by barley roots during a season, and to predict the resulting microbial biomass in the rhizosphere. The predicted microbial biomass C in the rhizosphere ranged from 10–34% of the total plant biomass C depending mainly upon the value used for rate of loss of root material. Total loss of root material predicted during a season in England constituted 7.7–25.4 percent of C fixed by photosynthesis. The major assumptions made in these calculations are considered, and the predicted values discussed in relation to reported values for soil microbial biomass, CO₂ fluxes from soil and associative nitrogen fixation.     

Succession of indigenous Pseudomonas spp. and actinomycetes on barley roots affected by the antagonistic strain Pseudomonas fluorescens DR54 and the fungicide imazalil

In recent years, the interest in the use of bacteria for biological control of plant-pathogenic fungi has increased. We studied the possible side effects of coating barley seeds with the antagonistic strain Pseudomonas fluorescens DR54 or a commercial fungicide, imazalil. This was done by monitoring the number of indigenous Pseudomonas organisms and actinomycetes on barley roots during growth in soil, harvest after 50 days, and subsequent decomposition. Bacteria were enumerated by traditional plate spreading on Gould's S1 agar (Pseudomonas) and as filamentous colonies on Winogradsky agar (actinomycetes) and by two quantitative competitive PCR assays. For this we developed an assay targeting Streptomyces and closely related genera. DR54 constituted more than 75% of the Pseudomonas population at the root base during the first 21 days but decreased to less than 10% at day 50. DR54 was not successful in colonizing root tips. Initially, DR54 affected the number of indigenous Pseudomonas organisms negatively, whereas imazalil affected Pseudomonas numbers positively, but the effects were transient. Although plate counts were considerably lower than the number of DNA copies, the two methods correlated well for Pseudomonas during plant growth, but after plant harvest Pseudomonas-specific DNA copy numbers decreased while plate counts were in the same magnitude as before. Hence, Pseudomonas was 10-fold more culturable in a decomposition environment than in the rhizosphere. The abundance of actinomycetes was unaffected by DR54 or imazalil amendments, and CFU and quantitative PCR results correlated throughout the experiment. The abundance of actinomycetes increased gradually, mostly in numbers of DNA copies, confirming their role in colonizing old roots.     

Effect of Plant Species on Persistence of Paecilomyces lilacinus Strain 251 in Soil and on Root Colonization by the Fungus

The effect of 12 plant species on the persistence of Paecilomyces lilacinus strain 251 in soil was investigated. After incorporating formulated conidia into non-sterile soil followed by transplanting different test plants, the population dynamic of the fungus was determined over 100 days. At termination of the experiment, the fungal population in the planted soil was compared to the density of P. lilacinus in the rhizosphere and the percent increase or decrease was calculated for each crop. In addition, the potential of P. lilacinus strain 251 to colonize roots endophytically was investigated. Comparison of the slopes describing the population dynamics of the fungus showed no significant differences between soil without plants and soil from the root zone of the majority of the test plants. Bean was the only plant species consistently exerting a negative effect on the persistence of P. lilacinus strain 251 in the soil. For the first time, P. lilacinus strain 251 was isolated in significant numbers from healthy root tissue of barley plants.     

Effects of the Biocontrol Agent Aspergillus flavipes on the Soil Microflora and Soil Enzymes in the Rooting Zone of Pepper Plants Infected with Phytophthora capsici

This study examined the effect of ASD strain (Aspergillus flavipes), isolated from continuous cropping soil for pepper and named by the sampling position, on soil microflora and soil enzymes in rooting zone soil of healthy and diseased (Phytophthora capsici) pepper plants. Results showed that the ASD strain could significantly reduce the number of bacteria and actinomycetes, with a significant increase in fungi in the rhizosphere soil of both healthy and diseased plants. With increasing colonization time of the ASD strain, the number of bacteria and actinomycetes decreased initially and then increased gradually, while the number of fungi was first increased significantly and later decreased slowly. The soil enzyme activities of urease, acid phosphatase, invertase and dehydrogenase were significantly increased by the ASD strain, while the activity of catalase was not significantly increased. As time from inoculation with the ASD strain increased, the activities of various enzymes were higher than controls. Maximum enzyme activities were found on the tenth day after ADS inoculation. The response of soil enzyme activities affected by the ASD strain was as follows: urease > dehydrogenase > invertase > acid phosphatase > catalase. These results suggest that the biocontrol of ASD strain could improve the micro ecology of rhizosphere soil.