Forms of phosphorus in bacteria and fungi isolated from two Australian soils

To understand the origin of organic and condensed forms of phosphorus (P) in soils, detailed information about P forms in microorganisms is required. We isolated 7 bacteria and 8 fungi from two Australian soils and analyzed the P forms in their pure cultures by extraction with NaOH-EDTA followed by ³¹P solution nuclear magnetic (NMR) spectroscopy. The bacteria belonged to the actinobacteria and the fungi to the ascomycota, as determined by rDNA sequencing. The proportions of broad forms of P were significantly different between the bacterial and fungal isolates (analysis of similarities, p = 0.001). Ortho-, pyro- and polyphosphate were present in higher proportions in fungi, while monoester and diester P were present in higher proportions in bacteria. Spectral deconvolution of the monoester region revealed 15 distinct resonances. The three major ones, which were identified by spiking experiments as glycerol 1-phosphate, glycerol 2-phosphate and adenosine-5′-monophosphate (AMP), comprised 56–74% of P in the monoester region. Ordination by principal component analysis and testing for treatment effects using analysis of similarities showed significant separation of P distribution in the monoester region between bacterial and fungal isolates (p = 0.007). However, neither group of microorganisms had a specific single P form which might be considered characteristic. As such, it may be difficult to distinguish soil P from bacterial or fungal origins, with the possible exception of a predominantly fungal origin of pyro- and polyphosphate. The identification of three major resonances in the monoester region of microorganisms is important, since the same resonances are found in ³¹P NMR spectra of soil extracts.     

土壤分离的细菌溶磷和解磷能力研究

Microorganisms capable of solubilizing and mineralizing phosphorus （P） pools in soils are considered vital in promoting P bioavallability. The study was conducted to screen and isolate inorganic P-solubilizing bacteria （IPSB） and organic P-mineralizing bacteria （OPMB） in soils taken from subtropical flooded and temperate non-flooded soils, and to compare inorganic P-solubilizing and organic P-solubilizing abilities between IPSB and OPMB. Ten OPMB strains were isolated and identified as Bacillus cereus and Bacillus megaterium, and five IPSB strains as B. megaterium, Burkholderia caryophyUi, Pseudomonas cichorii, and Pseudomonas syringae. P-solubilizing and -mineralizing abilities of the strains were measured using the methods taking cellular P into account. The IPSB strains exhibited inorganic P-solubilizing abilities ranging between 25.4-41.7 μg P mL^-1 and organic P-mineralizing abilities between 8.2-17.8μg P mL^-1. Each of the OPMB strains also exhibited both solubilizing and mineralizing abilities varying from 4.4 to 26.5 μg P mL^-1 and from 13.8 to 62.8 μg P mL^-1, respectively. For both IPSB and OPMB strains, most of the P mineralized from the organic P source was incorporated into the bacterial cells as cellular P. A significantly negative linear correlation （P 〈 0.05） was found between culture pH and P solubilized from inorganic P by OPMB strains. The results suggested that P solubilization and mineralization could coexist in the same bacterial strain.     

Contributions by fungi and bacteria to aggregate stability of cultivated soils

Stability changes during ageing (sterile) or incubation (non-sterile) of both natural field aggregates and remoulded aggregates from five soils were studied for periods up to 30 days. Growth of fungal hyphae, estimated by ergosterol measurement, corresponded to temporary stability increases in both types of aggregates during the first 15 days. Thereafter, fungal hyphae disappeared and were replaced by actinomycetes and bacteria. Increased stability due to entanglement by hyphae was comparable to that due to thixotropy in remoulded aggregates. Bacterial growth accompanied the fall in stability associated with fungal decline, but had little direct effect in stabilizing soil aggregates. Destruction of polysaccharides by periodate oxidation greatly diminished aggregate stability. The role of bacterial polysaccharides in soil aggregate stability is discussed.     

Mineralization of bacteria and fungi in chloroform-fumigated soils

It has been suggested by others that the size of the flush of mineralization caused by CHC1₃ fumigation can be used to estimate the amount of microbial biomass in soils. Calculation of biomass from the flush requires that the proportion of CHCl₃-killed cell C mineralized be known. To determine this proportion, 15 species of [¹⁴C]labelled fungi and 12 species of [¹⁴C]labelled bacteria were added to four types of soil and these were fumigated for 24 h with CHC1₃, reinoculated with unfumigated soil, and incubated at 22°C for 10 days. The average percentage mineralization of the fungi was 43.7 ± 5.3, while the average for the bacteria was 33.3 ± 9.9. Using a 1:3 ratio for distribution of total biomass between the bacterial and fungal populations, respectively, it was calculated that the average mineralization of both types of cells was 41.1%. In experiments conducted to determine if CHC1₃ vapour alters stabilized microbial metabolites or dead microbial cells in a manner which makes them more susceptible to degradation, it was found that both fumigated and unfumigated dead fungal materials mineralized to the same extent in soil during 10 days of incubation.     

外生菌根真菌对土壤无机磷迁移的影响

Ectomycorrhizal(EM) fungi could form symbiosis with plant roots and participate in nutrient absorption; however, many EM species commonly found in forest soils, where phosphorus(P) concentration and availability are usually very low, particularly in tropical and subtropical areas, have not yet been investigated for their efficiencies to mobilize soil P. In this study, fungal growth, P absorption,efflux of protons and organic acids, and soil P depletion by four isolates of EM fungi isolated either from acidic or calcareous soils were compared in pure liquid culture using soil as a sole P source. Boletus sp. 7(Bo 7), Lactarius deliciosus 3(Ld 3), and Pisolithus tinctorius 715(Pt 715) from acidic and P-deficient soils of southwestern China showed higher biomass and P concentration and accumulation than Cenococcum geophilum 4(Cg 4) from a calcareous soil of Inner Mongolia, northern China, after 4 weeks of liquid culture. Oxalate, malate, succinate, acetate, and citrate concentrations in the culture solutions varied significantly with fungal species,and oxalate accounted for 51.5%–91.4% of the total organic acids. Organic acids, particularly oxalate, in the culture solutions may lead to the solubilization of iron-bound P(Fe-P), aluminum-bound P(Al-P), and occluded P(O-P) from soil phosphates. Fungal species also varied greatly in proton efflux, which decreased the culture solution pH and may dissolve calcium-bound P(Ca-P) in soil.This could be the reason for the increment of both inorganic P in the culture solutions and Olsen P in the soil when EM fungi were present. Total inorganic P, the sum of Al-P, Fe-P, O-P, and Ca-P, in the culture solutions was positively correlated with the total concentration of organic acids in the culture solutions(r = 0.918*, n = 5), but negatively with both the total inorganic P in soil(r =-0.970**, n = 5) and the culture solution pH(r =-0.830*, n = 5). These suggested variable efficiencies of EM fungal species to mobilize inorganic P fractions from soil, which could make EM trees to utilize inorganic P in the same way like EM fungi and adapt to the soils with various P concentrations and availabilities.     

Biodegradation of ochratoxin A by fungi isolated from grapes

Ochratoxin A is a mycotoxin present in several food products for which levels should be reduced. Chemical, physical, and biological methods have been proposed for the detoxification of mycotoxins, biological methods being the more promising ones. In this report, filamentous fungi isolated from Portuguese grapes were assessed for ochratoxin A degradation capabilities. It was observed that 51 of the 76 tested strains, predominantly aspergillus species, were able to degrade more than 80% of ochratoxin A added to the culture medium and that the most potent species (more than 95% of initial amount) were the black aspergilli, A. clavatus, A. ochraceus, A. versicolor, and A. wentii. Other fungi frequently isolated from grapes, such as Alternaria, Botrytis, Cladosporium, and Penicillium, also showed significant degradation capabilities. It was observed that the compounds obtained from the degradation of ochratoxin A by black aspergilli and by A. ochraceus and A. wentii strains were different.     

Assessing the tolerance to heavy metals of arbuscular mycorrhizal fungi isolated from sewage sludge-contaminated soils

Different fungal ecotypes were isolated from soils which had received long-term applications of metal-contaminated sewage sludge with the aim of studying the degree of tolerance and adaptation to heavy metals of arbuscular mycorrhizal (AM) fungi. The development and structural aspects of AM colonization produced by the different fungal isolates were studied using two host plants, Allium porrum and Sorghum bicolor, which were grown in either contaminated or non-contaminated soils. Four different AM fungi were successfully isolated from the experimental field plots: (i) Glomus claroideum, isolated from plots receiving only inorganic fertilizer; (ii) another apparently similar ecotype of Glomus claroideum, but isolated from plots with 300 m³ ha⁻¹ year⁻¹ of contaminated sludge added, (iii) an unidentified Glomus sp., present only in the less contaminated plots (100 m³ ha⁻¹ year⁻¹ of unamended sludge) and (iv) Glomus mosseae, isolated from plots receiving 100 or 300 m³ ha⁻¹ year⁻¹ of amended or unamended sludge (intermediate rates of contamination). There were consistent differences in behaviour among the four AM fungi tested with regard to the colonization levels they produced in non-contaminated and contaminated soils. Both total and arbuscular colonization were affected by heavy metal contamination. The main conclusions of this study are that Glomus sp. and G. mosseae isolates are strongly inhibited by heavy metals, which acted mainly by interfering with the growth of the external mycelium, and also by limiting the production of arbuscules. Our results suggest that G. claroideum isolates, particularly the ecotype which was isolated from the plots receiving the highest dose of metal-contaminated sludge, shows a potential adaptation to increased metal concentration in soil.     

Utilization of rock phosphate in alkaline soils by plants inoculated with mycorrhizal fungi and phosphate-solubilizing bacteria

Interactions between vesicular-arbuscular (VA) mycorrhizal fungi and phosphate-solubilizing bacteria were studied in a low-phosphate alkaline soil amended with 0, 0.1% and 0.5% rock phosphate. Endogone (E3 and yellow vacuolate spore types) and two bacteria able to solubilize rock phosphate in vitro and produce plant growth regulating substances were used as inocula. Lavender (Lavandula spica var. vera L.) plants with mycorrhiza plus bacteria (either E3 plus bacteria or “yellow vacuolate” plus bacteria treatments) took up more total P than plants with either Endogone or bacteria separately at each concentration of rock phosphate. Plants not inoculated with bacteria or Endogone derived no benefit from the rock phosphate.     

PAH biodegradation by telluric saprotrophic fungi isolated from aged PAH-contaminated soils in mineral medium and historically contaminated soil microcosms

Journal of Soils and Sediments - Remediation of contaminated soils is of high relevance considering losses of this limited resource in most countries through erosion or through destruction for...     

Humic acid bleaching by white-rot fungi isolated from biosolids compost

A screening assay for isolating humic-substances degrading fungi from biosolids compost at the thermophillic phase employed using plates containing 2,2′-azino-bis(3-ethylbenzothiazoline-sulfonic acid) (ABTS), MnCl₂ or the monoazo dye Acid Red 183. Two of the most active fungi out of 70 fungal strains isolated were identified based on rDNA sequences and designated Trametes sp. M23 and Phanerochaete sp. Y6 (accessions no. DQ408582 and DQ438910). These isolates, when compared to a model white-rot fungi T. versicolor and P. chrysosporium, showed the ability to bleach humic acids extracted from biosolids compost while growing under solid-state conditions using perlite as a solid support. T. versicolor and Trametes sp. M23 also exhibited the ability to bleach humic acids from a peat source. Interestingly, only Trametes sp. M23 bleached leonardite humic acid, which is considered to be a highly aromatic and stable type of natural organic matter. To the best of our knowledge, this is the first report of white-rot fungi being isolated and identified from thermophilic composts. Since these fungi are capable of degrading lignin and humic acid and were found active in organic-matter-degradation processes, we suggest that they may play a significant role in the degradation and transformation of these refractory substances during composting.     

Acetylene reduction by bacteria isolated from the rhizosphere of wheat

An asymbiotic bacillus possessing N₂-ase activity and capable of growth on low-N media was consistently isolated from the rhizosphere of a chromosome substitution line of spring wheat in which a pair of chromosomes 5D from the cultivar Rescue had been substituted for those of ‘Cadet’. None of the bacilli were isolated from either of the two parent cultivars, Rescue and Cadet, or from the corresponding substitution line involving a homoeologous chromosome 5B. Maximum C₂H₂ reduction to C₂H₄ by pure culture isolates was found to occur at a partial pressure of 0.2 atm C₂H₂. C₂H₂ reduction was diminished in the presence of O₂ and nearly ceased at a partial pressure of 0.08 atm O₂.The data presented suggest that altering the genetics of the wheat plant can change the root environment to favor the establishment of bacilli that exhibit N₂-ase activity in pure culture.     

A preliminary study of the growth of fungi and bacteria from temperate and antarctic soils in relation to temperature

The growth of fungi isolated from a lowland temperate site (Roudsea Wood National Nature Reserve), an upland temperate moorland (Moor House National Nature Reserve) and an oceanic Antarctic island (Signy, S. Orkneys) was compared at 1, 14 and 25°C. This showed that low temperatures caused greatest retardation of growth in fungi from the warmest site (Roudsea) and least from the coldest site (Signy Island). At Moor House, fungi which were isolated most frequently in winter were able to grow better at 1°C than summer forms. The fungal flora of Signy Island was restricted and consists of cold tolerant cosmopolitan species which have been selected by or become adapted to the prevailing low temperatures. Of fungi isolated from any two of the sites, Mortierella alpina and Mucor hiemalis showed temperature adaptation correlated with prevailing site temperature, while Trichoderma viride, Penicillium thomii, and P. frequentans showed no adaptation.     

Phosphate solubilizing bacteria and fungi in desert soils: species,limitations and mechanisms

Desert soils are infertile, and the ability to improve them by P-fertilization is limited by the solubility of phosphate. We aimed to understand the function of phosphate solubilizing bacteria and the mechanisms behind phosphate solubilization in desert soils. Vegetated and barren desert soils, mine spoil and a fertile temperate grassland loam were sampled. Bacteria and fungi were isolated and identified, and their phosphate-solubilizing abilities were measured in vitro. The release of plant available PO₄, SO₄, NO₃ and NH₄ from desert soils did not compare with that of a grassland soil. Desert soils had substantially lower solubilization than grassland, 162 and 99–121 µg PO₄-P g^?1 dry soil, respectively. Phosphate-solubilizing bacteria and fungi were inhabiting the soils. Si addition increased phosphate solubilization of fungi by 50%. The isolated microbes were shown, using ³¹P nuclear magnetic resonance (NMR) analysis, to rapidly take-up both intracellular and extracellular phosphate during the phosphate solubilizing process. Desert soil had potentially active microbial populations that are capable to solubilize inorganic phosphorus; S and Si as the limiting factors. Acidification as the main mechanism to solubilize mineral phosphate was not as evident in our desert soils as in former studies dealing more fertile soils.     

Growth measurements of saprotrophic fungi and bacteria reveal differences between canopy and forest floor soils

Canopy-held organic matter develops into a distinct soil system separate from the forest floor in wet temperate coniferous forests, creating a natural microcosm. We distinguished between fungal and bacterial components of the decomposer community in one site with Maple (Acer macrophyllum) and one site with Alder (Alnus rubra) by using direct measurements of growth; acetate incorporation into ergosterol, and leucine incorporation for fungi and bacteria, respectively. The higher organic matter content of the canopy soils correlated with higher fungal growth. The relative importance of fungi, indicated by fungal:bacterial growth ratio, was higher in the canopy soil of the Maple site, while there was no difference in the Alder site. The high C:N ratio of the Maple canopy soil likely contributed to this difference. These results demonstrate a divergence between canopy and forest floor that should be explored to gain insights in decomposer ecology using the natural microcosms that the canopy soils provide.     

Solubilization of iron and calcium phosphates by soil fungi isolated from coffee plantations

Almost 900 fungal isolates were obtained from eight coffee plantations in Colombia and Mexico. Of these, 76 isolates showed activity to solubilize Ca₃(PO₄)₂ (PCa) and FePO₄·H₂O (PFe), which had been added to agar in a plate test. Generally, PCa was better solubilized than PFe. Colombian isolates were generally somewhat less effective than Mexican isolates. The two most effective isolates from each country with apparent highest PFe, solubilization potential were selected and cultivated in liquid medium containing PFe, which is more prevalent in tropical soils. The pH value, solubilized P in the medium and P uptake in fungal biomass were determined. After 24 days, Cylindrocarpon didymum and C. obtusisporum (both from Colombia) had solubilized 9.9 and 6.4 mg PO₄ ^3--P L^?1 and took up 8.6 and 11.6 mg P in biomass. Penicillium janthinellum and Paecilomyces marquandii (both from Mexico) solubilized 7.0 and 1.9 mg PO₄ ^3--P L^?1 and took up 11.3 and 17.3 mg P in biomass. The potential practical use of the four fungal isolates for different strategies in making more P available for coffee growth is discussed.     

Mineral transformations by fungi in culture and in soils

• 1 Although micro-organisms are regarded as the principal agents of mineral cycling in soils, the role of bacteria has generally been emphasized, while that of fungi has been neglected.
• 2 Fungi are able to transform the majority of elements in vitro but whether they play an important role in soil is as yet unknown.
• 3 There is sufficient circumstantial evidence from soil studies to suggest that fungi may under certain conditions nitrify and oxidize reduced forms of sulphur. Their role in denitrification and sulphate reduction process is more speculative. While they appear incapable of nitrogen fixation, fungi undoubtedly play a major role in the mineralization of organic N,P, and S in soils, in the solubilization of insoluble phosphates, and participate in oxidation of manganese. Perhaps one of their most important roles is in the dissolution of silica and rocks thus releasing ions into the soil solution during weathering.
• 4 The ability of fungi to oxidize elements in vitro does not compare with that of the chemoautotrophic bacteria. On the other hand in vitro activity tells us little about the activity of an organism in the soil.
• 5 At present our appreciation of the part played by fungi in mineral cycling in soils is limited by the techniques available, but there is little doubt that they have a major role to play in the cycling of elements other than carbon.

     

Isolation and identification of iron-reducing bacteria from gley soils

Seventy-one facultative anaerobic bacteria, capable of reducing iron oxide in pure culture, were isolated from three differently gleyed subsoils. The bacteria were picked at random from poured plates (10⁻⁵ and 10⁻⁶) inoculated with serially diluted soil samples. An attempt was made to identify these strains by morphological and biochemical tests. Among these 71 iron-reducing bacteria, all except three were capable of reducing nitrate to nitrite and 35 reduced nitrite further into gaseous compounds (denitrification), but only one strain (Bacillus subtilis) produced H₂S. Based upon their physiological and morphological properties, 38 strains were allotted to the genus Pseudomonas, 31 sporeformers to the genus Bacillus and two were regarded to be coryneform (Arthrobacter?) bacteria. Species identified were Ps. denitrificans (23), ps. stutzeri (8) ps. fluorescens-putida (5), Bacillus cereus (6), B. cereus var. mycoides (14) and Bacillus subtilis (9). Two spore-forming bacilli, two non-pigmented pseudomonads and two coryneform type of bacteria could not be identified. The significance of the enzyme nitrate reductase (nitratase) of these bacteria for anaerobic respiration and as a mechanism of iron reduction is discussed.     

Earthworms increase the ratio of bacteria to fungi in northern hardwood forest soils, primarily by eliminating the organic horizon

The exotic earthworm invasion in hardwood forests of the northern United States is associated with many ecosystem-level changes. However, less is known about the effects of the invasion on the composition of the soil microbial community through which ecosystem-level changes are mediated. Further, earthworm effects on soil microbial community composition have not been well studied in the field. To evaluate changes in bacterial and fungal abundance associated with the earthworm invasion we quantified bacterial and fungal biomass by microscopic counts in paired earthworm-invaded (earthworm) and earthworm-free (reference) plots in five forest stands in central New York (USA). Earthworms significantly increased the ratio of bacteria to fungi on an area basis (per m²), by more than two times in mid-summer and early autumn. While this effect was associated primarily with the lack of the fungal-dominated organic horizon in earthworm plots, a higher ratio of bacteria to fungi in the surface 5 cm mineral soil also contributed as it developed between spring and mid-summer. Earthworm reduction of fungal biomass was confirmed by substantially lower growth of fungal hyphae into mesh sand bags in earthworm compared to reference plots. Burrowing activity by the earthworm Lumbricus terrestris increased the ratio of bacteria to fungi over the short-term within earthworm plots, introducing small-scale spatial heterogeneity associated with burrows. Our study suggests that the exotic earthworm invasion in these northern hardwood forests markedly increased the ratio of bacteria to fungi by eliminating the fungal-rich organic horizon, and was associated localized increases in bacterial vs. fungal abundance in mineral soil, setting the stage for future research into linkages between the earthworm invasion, bacterial and fungal abundance, and ecosystem processes.     

光合细菌对鱼病原细菌生长的影响

从发病的棒花鱼（Abbottina rivularis）和鲫鱼（Carassius auratus）分离得到7株菌,经人工感染证实其中的B3、J1和J2等3株菌均为病原细菌。B3和J1鉴定为嗜水气单胞菌（Aeromonas hydrophila）,J2鉴定为豚鼠气单胞菌（Aeromonas caviae）。采用平板扩散法测定了光合细菌嗜酸红假单胞菌Y7（Rhodopseudomonas acidophila Y7）、球形红假单胞菌X3（R．spheroids X3）和桃红荚硫菌S（Tniocapsa roseopersicina S）对病原细菌B3和J2生长的影响。结果表明：在营养琼脂平板上3种光合细菌对两种病原细菌（B3和J2）无明显拮抗作用,但光合细菌能覆盖在病原细菌上生长;3种光合细菌对病原细菌生长有明显的抑制作用,其中球形红假单胞菌X3抑制作用最大,但X3代谢产物对病原细菌生长无明显抑制作用,说明X3对病原细菌的抑制作用来自菌体细胞。X3还能明显抑制水体中的病原菌B3和J2的生长。     

Susceptibility of Pythium spp and other fungi to antagonism by the mycorparasite Phytium Oligandrum

Phytium spp and isolates within species differed in susceptibility to the mycoparasite Pythium oligandrum Drechs., as evidenced by their degree of inhibition by it on cellulose and ability to support its growth across their colonies on agar. Yet no Phythium sp. was highly susceptible to it, and P. graminicola Subramanian was highly resistant.No evidence was found that P. oligandrum produces toxins active against other fungi. In liquid culture, P. oligandrum grew on undisturbed colonies of Phialophora sp. (highly susceptible) but not P. ultimum Trow or Fusarium culmorum (W. G. Sm.) Sacc. (moderately resistant) and not Rhizoctonia solani Kuhn (highly resistant). It grew in culture filtrates of Phialophora sp., P. ultimum and F. culmorum, utilizing organic nitrogen and thiamine released by these fungi, but not in culture filtrates of R. solani. It grew on mycelial macerates of all these fungi, though poorly on those of R. solani. Resistance to parasitism by P. oligandrum seems to reside, at least partly, in low levels of nutrient release from host hyphae.