两株解磷细菌的解磷活性及作用机制研究

解磷细菌在增加土壤可溶性磷含量、提高磷肥利用效率方面具有重要作用。为选筛高效解磷菌、探讨其解磷机制,本文利用平板溶磷圈法筛选解磷细菌,采用钼锑抗比色法研究其解磷活性,苯磷酸二钠法研究其磷酸酶活性,利用薄层层析分析其产生的有机酸,根据生理生化特征和16S r RNA基因序列系统发育分析,确定其分类学地位。结果表明,菌株JXJ-11和JXJ-15对植酸钙的降解活性很强,3 d后培养液中可溶性磷浓度分别增加219 mg·L~(-1)和216 mg·L~(-1);对磷酸钙降解活性较弱,最高可溶性磷浓度仅为植酸钙的21.79%~30.37%;解磷细菌可分泌酸性、中性和碱性磷酸酶,降解不溶性磷,可能产生丙酸和琥珀酸等有机酸,降低培养液p H,增加可溶性磷浓度。两株细菌均为革兰氏阴性杆菌,无芽孢,产生硫化氢,其中菌株JXJ-11的16S rRNA基因序列与Sphingomonas melonis DAPP-PG 224T和S.aquatilis JSS7T相似性最高(99.79%),菌株JXJ-15的16S rRNA基因序列与Klebsiella pneumoniae subsp.pneumoniae DSM 30104T相似性最高(99.73%),根据以上信息,确定菌株JXJ-11和JXJ-15分别是鞘氨醇单胞菌属和克雷白氏杆菌属的成员。菌株JXJ-11和JXJ-15的解磷机制包括分泌有机酸和磷酸酶,其中JXJ-11在微生物磷肥研制方面具有潜在应用价值。     

Effect of extracellular-enzyme activities on solubilization rate of soil organic nitrogen

In a sandy soil containing ¹⁵N-labeled active (soluble and easily degradable) and non-labelled passive (recalcitrant) fractions of soil organic matter, the rate of net N mineralization (solubilization) was determined during a 55-day incubation at 25°C, 63% water-holding capacity and different levels of soil extracellular-enzyme activities. The active fraction of soil N was labelled by preincubation (at 5°C and 74% water-holding capacity for 6 months) of soil amended with ¹⁵N-labeled plant material. Increases in the activity of extracellular-enzymes in soil were induced by the addition of glucose and KH₂PO₄ at the beginning of the incubation. The results show that the contents of total soluble N (NO ₃ ^– –N+NH ₄ ⁺ –N + soluble organic N) were significantly higher in glucose-amended soil compared to the unamended soil. The increases in soluble N in soil amended with 1 and 2 mg glucose g^-1 dry soil corresponded to a mean rate of net solubilization of 7.9±1.4 and 18.8±0.7 nmol N g^-1 dry soil day^-1, respectively. The mean rate of net N solubilization (3.6±1.0 nmol N g^-1 dry soil day^-1) in unamended soil was significantly lower than those of glucose amended soils. The content of ¹⁵N in total soluble N in soil amended with 2 mg glucose, for example, was diluted from 3.11±0.08 atom% before the incubation to 2.77±0.03 atom% after 55 days. This indicates that 89% of soluble-N accumulated in soil by the end of the incubation originated from the active fraction of soil N and the rest, estimated at 11%, originated from the passive fraction. The activities of soluble and total proteases as well as the rate of N solubilization in the soil increased with the application of glucose. The activity of these extracellular enzymes was highly correlated with the rates of net N solubilization. Thus, increases in extracellular-enzyme activities in glucose-amended soils had a priming effect on the solubilization of ¹⁵N-labeled active and non-labeled passive fractions of soil organic N. It seems that the activity of extracellular-enzymes expressed in terms of total and soluble protease activities could be a rate-limiting factor in the processes of soil organic N solubilization.     

Phosphate solubilization by a wild type strain and UV-induced mutants of Aspergillus tubingensis

Phenotypic mutants of Aspergillus tubingensis were obtained by UV irradiation and phosphate solubilization ability of these mutants were studied and compared with the wild type strain. Low phosphate solubilizing mutant was also selected in this study. Among the different mutants, AtM-5 and AtM-2 showed highest P solubilization when tri-calcium phosphate and rock phosphate were used as P source compared to the wild type strain and other mutants. These mutants also showed maximum acid phosphatase and phytase activity. These results suggest that P solubilization by these isolates is due to lowering of pH of the culture filtrate and also the activity of acid phosphatase and phytase.     

Vitamin production in relation to phosphate solubilization by soil bacteria

Studies were conducted to determine the relationship of vitamin production and ability to dissolve bicalcium phosphate in bacteria isolated from rhizosphere, rhizoplane and control soil. Production of vitamin B₁₂, riboflavin, niacin, pantothenate and biotin was determined using a bioassay procedure that may be useful for large-scale screening of bacteria for vitamin production.Among isolates producing one or more vitamins, 76.4, 37.5 and 57.5% of control, rhizosphere and rhizoplane isolates, respectively, solubilized phosphate. However, phosphate-solubilizing isolates from rhizosphere and rhizoplane were more active vitamin producers than solubilizing isolates from control soil, and non-solubilizing isolates from any of the three regions.Production of vitamin B₁₂, riboflavin and niacin by rhizosphere isolates and of riboflavin by rhizoplane isolates was also correlated with ability to solubilize phosphate, but similar relationships were not observed for control isolates.     

土壤中耐铅微生物的筛选

通过在培养基中加入一定浓度的Pb(300mg／kg),从土壤中分离到12株耐Pb微生物菌株。其中细菌菌株为1～5号,放线菌为6～9号,真菌为10～12号。经过形态学观察、拮抗试验以及液体培养和土壤模拟试验中降低Pb效果测试,发现12株菌对有效态铅都有一定的降低作用。在液体培养中3号、4号、6号、7号、8号和10号菌株的降低效果最好,降低率超过70％,在土壤培养中3号和7号的降低作用最明显,降低率分别为63．8％和66％。     

Phosphate fertilizer enhancing soil erosion: effects and mechanisms in a variably charged soil

Purpose

The extensive application of phosphate fertilizers could produce a series of environmental problems by adsorbing on the surface of soil particle and migrating into water during soil erosion. Therefore, this study is to explore the effects of phosphate on soil aggregate stability and soil erosion and to analyze the mechanisms of phosphate enhancing soil erosion from the scope of soil charge density, electric field, and particle interaction.

Materials and methods

A variable charged soil (0–20 cm) was pre-treated firstly by KCl, K₂HPO₄, and KH₂PO₄, respectively. Under the conditions of KCl, K₂HPO₄, and KH₂PO₄ solutions with concentrations of 0.0001, 0.001, 0.01, 0.1, and 1 mol L^?1, (1) the amounts of soil particle transport and erosion intensities were measured using rainfall simulation with electrolyte solutions instead of rainwater; (2) the aggregate stabilities were measured by weighing the particles and micro-aggregates of <2, 5, and 10 μm after soil aggregate breakdown in electrolyte solutions; and (3) the zeta potentials of soil particles were measured in electrolyte solutions.

Results and discussion

The application of K₂HPO₄ and KH₂PO₄ in soil strongly enhanced soil aggregate breakdown and soil erosion, while in KCl application, soil aggregates were stable and erosion did not occur. Moreover, the intensities of soil aggregate breakdown and soil erosion for K₂HPO₄ application were much stronger than that for KH₂PO₄ application. Phosphate, especially K₂HPO₄, strongly increased surface negative charge density of soil particles and thus increased the electrostatic repulsive pressure between adjacent soil particles in aggregates, and as a result, the soil erosion intensity increased. However, the surface charge density was not increased by the increased pH, specific adsorption, and dispersion force adsorption but possibly attributed to a non-classic induction force adsorption arising from the anionic non-classic polarization in the strong electric field around soil particle surface.

Conclusions

The application of phosphate decreased aggregate stability and stimulated soil erosion through increasing charge density of particle surface by a new non-classic induction adsorption of phosphate.

     

Abiotic solubilization of soil organic matter,a less-seen aspect of dissolved organic matter production

Dissolved organic matter (DOM) is a small but reactive pool of the soil organic matter (SOM) that contributes to soil dynamics including the intermediary pool spanning labile to resistant SOM fractions. The solubilization of SOM (DOM production) is commonly attributed to both microbially driven and physico-chemically mediated processes, yet the extent to which these processes control DOM production is highly debated. We conducted a series of experiments using ¹³C-ryegrass residue or its extract (¹³C-ryegrass-DOM) separately under sterile and non-sterile conditions to demonstrate the importance of DOM production from microbial and physico-chemical processes. Soils with similar properties but differing in parent material were used to test the influence of mineralogy on DOM production. To test the role of the source of C for DOM production, one set of soils was leached frequently with ¹³C-ryegrass-DOM and in the other set of soils ¹³C-ryegrass residue was incorporated at the beginning of the experiment into the soil and soils were leached frequently with 0.01 mol L⁻¹ CaCl₂ solution. Leaching events for both treatments occurred at 12-d intervals over a 90-day period. The amount of dissolved organic C and N (DOC and DON) leached from residue-amended soils were consistently more than 3 times higher in sterile than non-sterile soils, decreasing with the time. Despite changes in the concentration of DOC and DON and the production of CO₂, the proportion of DOC derived from the ¹³C-ryegrass residue was largely constant during the experiment (regardless of microbial activity), with the majority (about 70%) of the DOM originating from native SOM. In ¹³C-residue-DOM treatments, after successive leaching events and regardless of the sterility conditions i) the native SOM consistently supplied at least 10% of the total leached DOM, and ii) the contribution of native SOM to DOM was 2–2.9 times greater in ¹³C-residue-DOM amended soils than control soils, suggesting the role of desorption and exchange reactions in DOM production in presence of fresh DOM input. The contribution of the native SOM to DOM resulted in higher aromaticity and humification index. Our results suggest that physico-chemical processes (e.g. exchange or dissolution reactions) can primarily control DOM production. However, microbial activity affects SOM solubilization indirectly through DOM turnover.     

Phosphate solubilization by Chryseobacterium sp. and their combined effect with N and P fertilizers on plant growth promotion

The capabilities of soil microorganisms to solubilize phosphate have been known for many years, but their isolation and use as crop inoculants have met with little success. Thirty-five bacterial isolates were screened for their phosphate-solubilizing ability, and two of them, PSR10 and RGR13, identified through 16S rDNA sequencing as Chryseobacterium sp. PSR10 and Escherichia coli RGR13, respectively, screened for plant growth promotion in the greenhouse. Seed inoculation of Macrotyloma uniflorum (horsegram) by Chryseobacterium sp. PSR10 showed better plant growth promotion in sterilized and unsterilized soil under greenhouse conditions and was selected for a field experiment with 100, 50 and 30% of recommended doses of nitrogen and phosphorus fertilizer. Seed inoculation with 50% of the recommended dose of nitrogen and phosphorus increased plant growth (agronomical parameters, chlorophyll content, nitrate reductase activity, phosphorus content and crop yield). We conclude that effective plant growth-promoting bacterium Chryseobacterium sp. PSR10 broadens the spectrum of phosphate solubilizers available for field applications and might be used together with 50% dose of nitrogen and phosphate.     

Environmental influences on Acinetobacter sp. strain BD413 transformation in soil

The ability of various environmental factors (root exudate from silver tussock, blue tussock, flax, wheat, ryegrass and lupin; simulated-root exudate; moisture; temperature; soil density; salinity; sewage sludge; fertiliser; pesticide) to promote or inhibit transformation of the soil-dwelling bacterium Acinetobacter baylyi BD413 (pFG4ΔnptII) was investigated using soil microcosm studies. A marker-rescue system was used to monitor the transfer of a functional nptII gene from exogenous chromosomal DNA to A. baylyi BD413 (pFG4ΔnptII). Significant differences were detected in A. baylyi BD413 (pFG4ΔnptII) transformation rates in three sterile New Zealand agricultural soils. Addition of simulated-root exudate to the sterile soil was essential for transformation of A. baylyi BD413 (pFG4ΔnptII) in the soil types tested, but addition of plant exudates collected from a variety of New Zealand cropping and native plants did not promote transformation rates to above detectable limits. Increases in soil temperature and bulk density increased the transformation rate but this effect was not consistent across all three soil types. Application of sewage sludge to sterile soils significantly increased transformation in the sandy soil but not in the silt loam and fine sandy loam soil types. Fertiliser (superphosphate) and herbicide (glyphosate) applied at agronomic rates did not affect transformation rates; however, when used at 5× and 50× the agronomic rate respectively, transformation was significantly reduced in all three sterile soils. These results suggest that competence and transformation of the A. baylyi BD413 (pFG4ΔnptII) in soils is highly dependent on the presence of nutrients and is also influenced by the soil texture.     

垃圾堆肥对难溶性磷转化及土壤磷素吸附特性影响

在城市生活垃圾进行工厂化堆肥过程中,加入难溶性磷矿粉,探讨堆肥对难溶性磷的转化能力及堆肥产品培肥后对土壤磷素吸附特性的影响。结果表明,加入磷矿粉可使堆肥中活性有机磷、中等活性有机磷、中稳性有机磷、高稳性有机磷及速效磷含量均有不同程度的提高,与对照相比分别增加212.69%、80.36%、61.21%、62.74%、157.89%。通过电镜观察表明,堆肥后磷矿粉典型的矿物特征消失,表面呈蜂窝状。将堆肥后的产品进行培肥试验表明,富磷垃圾肥处理可明显改善土壤磷素的吸附特性,与施化肥相比,最大吸附量(Q_m)下降8.76%,最大缓冲容量(Q_m·K)下降13.58%,而磷素的吸附饱和度(DPS)、零净吸附浓度磷(EPC₀)则呈不同程度的增加,幅度依次为98.52%、7.13%。试验结果显示,通过堆肥生产富磷垃圾肥可为解决中国磷素资源缺乏、化学磷肥利用率低等问题提供一条生物学途径。     

添加紫云英对稻田土壤颗粒吸附磷酸盐的影响

【目的】研究添加紫云英(Chinese milk vetch, CMV)对土壤颗粒表面磷素吸附特征的影响机制,为绿肥高效利用提供理论依据。【方法】采用土培试验方法,设置4个紫云英翻压量梯度：CMV0 (0)、CMV1(15000 kg/hm²)、CMV2 (22500 kg/hm²)和CMV3 (30000 kg/hm²),淹水培养30天后,取土样,过250μm筛后,分为细砂粒(48～250μm)、粉粒(2～48μm)和粘粒(<2μm),分别测定土壤有机质、全氮、全磷和有效磷含量,并分别进行磷酸盐的等温吸附和动力学吸附试验。【结果】与CMV0相比,添加紫云英显著提高了土壤颗粒中有机质、全氮、全磷和有效磷含量,尤以砂粒中的提高幅度最大,分别达到33.42%～81.04%、4.83%～15.17%、45.45%～51.52%和40.76%～60.70%;添加紫云英降低了砂粒和粘粒的比表面积,但是提高了粉粒的比表面积。土壤颗粒对磷素的吸附可用Langmuir吸附等温线方程很好地拟合,添加紫云英提高了各粒径土壤对磷的理论最...     

Phosphate sorption curves as a soil testing technique: A simplified approach

Abstract

Single point phosphate sorption curves effectively estimated phosphate fertilizer requirements while requiring less work than multiple point curves. Correlation coefficients of 0.961 and 0.981 were obtained when phosphate rates, estimated using single point curves, were compared with those from multiple point curves. Reducing the ionic strength of the supporting electrolyte solution by using 0.001 M CaCl₂ or water rather than 0.01 M CaCl₂ during equilibration increased dissolved inorganic phosphate 73% and 141% on the average thus improving analytical precision. The relative effects of salt concentration were sufficiently independent of pH and phosphorus concentration to suggest that a simple conversion factor can be used to convert external P requirements from one set of equilibrium conditions (salt concentration) to another.     

Phosphate eesoeption isotherms in four selected tropical soils and one temperate soil

Abstract

Some problem areas connected with soil testing and with reporting soil test and plant analysis results are explored, A uniform system for reporting soil test results is suggested as a step towards further standardization of soil test methods within and among states. Four categories are suggested as a common terminology for soil test and plant analysis results. These are: deficient, probable response, adequate and excessive.     

Phosphate desorption from the surface of soil mineral particles by a phosphate-solubilizing fungus

High phosphate (Pi) sorption in soils is a serious limiting factor for plant productivity and Pi fertilization efficiency, particularly in highly weathered and volcanic ash soils. In these soils, the sorbed Pi is so strongly held on the surfaces of reactive minerals that it is not available for plant root uptake. The use of phosphate-solubilizing microorganisms (PSM) capable of Pi desorption seems to be a complementary alternative in the management of these soils. The aim of this study was to evaluate the effectiveness of the soil fungus Mortierella sp., a known PSM, to desorb Pi from four soil minerals differing in their Pi sorption capacity. The fungus was effective in desorbing Pi from all tested minerals except from allophane, and its desorption depended on the production of oxalic acid. The effectiveness of the fungus to desorb Pi was ranked as montmorillonite > kaolinite > goethite > allophane. The quantity of desorbed Pi increased by increasing the amount of sorbed Pi. The Pi sorption capacity expressed as P_0.2 value (amount of P required to increase a solution P concentration up to 0.2 mg L^?1) was a good indicator of the effectiveness of Mortierella sp. to desorb Pi from soil minerals.     

Estimating soil C loss potentials from the C to N ratio

Thirty-seven arable soil samples (0–20 cm depth) ranging in clay content from 4 to 40% and in C to N ratio from 8 to 20 were incubated at ?100 hPa and 20 °C for 238 days. The CO₂-C evolved during days 105–147 and days 0–238 was related to the soil C to N ratio by a power function. We hypothesized the existence of a threshold value for the soil C to N ratio below which CO₂-C evolution remains constant. A soil C to N threshold value of 9.7 was estimated for both incubation periods and the function predicted that the CO₂-C evolution was halved when the soil C to N ratio approached 14. Data obtained by Springob and Kirchmann [Springob, G., Kirchmann, H., 2002. C-rich sandy Ap horizons of specific historical land-use contain large fractions of refractory organic matter. Soil Biology & Biochemistry 34, 1571–1581.] showed a similar relationship, but with a C to N threshold value of 10.8. We suggest that the relationship established between the C to N ratio of a soil and its CO₂-C loss during incubation represents a simple but useful measure for predicting soil C loss potentials on a more general level.     

Implications of iron solubilization on soil phosphorus release in seasonally flooded forests of the lower Orinoco River, Venezuela

The microbial reduction of Fe oxides is thought to contribute with the release of P in sedimentary environments. However, secondary reactions of the bioproduced Fe(II) with P in solution, can lead to a decrease in the soluble P concentration. In this study, we examined how the reduction of Fe(III) affects the soluble P concentration, when the soils of a seasonally flooded forest gradient are subjected to anaerobic conditions. Soil samples were collected during the dry season from two zones subjected to different flooding intensity: MAX and MIN zones that were inundated 8 and 2 months per year, respectively. When anaerobic conditions were applied to soils from both zones, a clear stimulatory effect on the Fe(III) reduction was observed. However, bioproduced Fe(II) underwent secondary chemical reactions, masking the extend of Fe(III) reduction of these soils. Iron was reduced mainly during the first 15 days of the anaerobic incubation and it was stimulated by a pulse of labile carbon. Iron dissolution did not lead to an increase of the soluble P content. However, in both zones P was high and positively correlated with Fe(II), implying that soil P mobilization was linked to Fe dissolution. In the MIN zone, soluble P concentration decreased, probably as a consequence of the secondary reactions of solubilized P with other non-redox sensitive soils elements. Fe solubilization also had an effect on the activity of acid phosphatase and consequently in the solubilization of P from the organic pool. In conclusion, the P cycle in these soils is strongly coupled to C and Fe cycles.     

Competitiveness of native Bradyrhizobium japonicum strains in two different soil types

Interstrain competitiveness is a key factor affecting the performance of rhizobium inoculant. In the present study five native strains of Bradyrhizobium japonicum, namely SSF 4, SSF 5, SSF 6, SSF 7 and SSF 8, were assessed for their competitiveness in nodulating soybean using serological methods. The strains were inoculated individually or with the type strain USDA 110 at a 1:1 ratio. Nodule occupancy determined by immunofluorescence and dot immunoblot assay revealed that under in vitro conditions SSF 8 is more competitive than USDA 110 whereas the others were less competitive. The competitive ability of these strains was also estimated in pot culture in the field. In red soil both SSF 8 and USDA 110 were equally competitive whereas in black soil SSF 8 competed better than USDA 110 and produced more nodules. In a black soil field trial using a randomized block design, USDA 110 or SSF 8, when inoculated alone, occupied the majority of the nodules and enhanced nodule dry weight and shoot biomass. SSF 8 was more competitive when the strains were co-inoculated. Received: 1 November 1996     

Phosphate solubilization by <Emphasis Type="Italic">Penicillium</Emphasis> spp. closely associated with wheat roots

In this study we found that Penicillium spp. exhibiting P-solubilizing activity are common both on and in the roots of wheat plants grown in southern Australian agricultural soils. From 2,500 segments of washed and surface-disinfested root pieces, 608 and 223 fungi were isolated on a selective medium, respectively. All isolates were screened for P solubilization on solid medium containing hydroxyapatite (HA); 47 isolates (5.7%) solubilized HA and were identified as isolates of Penicillium or its teleomorphs. These isolates were evaluated for solubilization of Idaho rock phosphate (RP) in liquid culture. Penicillium bilaiae strain RS7B-SD1 was the most effective, mobilizing 101.7 mg P l^–1 after 7 days. Other effective isolates included Penicillium simplicissimum (58.8 mg P l^–1), five strains of Penicillium griseofulvum (56.1–47.6 mg P l^–1), Talaromyces flavus (48.6 mg P l^–1) and two unidentified Penicillium spp. (50.7 and 50 mg P l^–1). A newly isolated strain of Penicillium radicum (KC1-SD1) mobilized 43.3 mg P l^–1. RP solubilization, biomass production and solution pH for P. bilaiae RS7B-SD1, P. radicum FRR4718 or Penicillium sp. 1 KC6-W2 was determined over time. P. bilaiae RS7B-SD1 solubilized the greatest amount of RP (112.7 mg P l^–1) and had the highest RP-solubilizing activity per unit of biomass produced (up to 603.2 g P l^–1 mg biomass^–1 at 7 days growth). This study has identified new isolates of Penicillium fungi with high mineral phosphate solubilizing activity. These fungi are being investigated for the ability to increase crop production on strong P-retaining soils in Australia.     

萌芽开花期梨枣对土壤水势的响应

本试验选取4年生梨枣(Ziziphus jujuba Mill.)为试验材料,设置-31~-51 kPa、-41~-84 kPa、-59~-132kPa和-161 kPa(不灌溉)4个土壤水势水平,结合茎直径微变化、营养生长与生殖生长量,分析了梨枣在萌芽开花期对土壤水势的响应。结果表明:1)在梨枣萌芽期,土壤水势在-41~-84 kPa范围波动时,茎直径日最大收缩量最小,营养生长与生殖生长最旺盛,是该时期适宜的土壤水势范围;花芽分化适宜的土壤水势范围为-41~-132 kPa。梨枣萌芽期适当的水分亏缺不影响花芽分化,但减弱了营养生长量,优化了营养生长与生殖生长的比例。2)在梨枣开花期,土壤水势范围为-54~-78 kPa时,茎直径日最大收缩量最小,坐果率较高,为开花期适宜的土壤水势范围;土壤水势为-79~-114 kPa时坐果率最高。开花期轻度的水分亏缺可显著提高坐果率。3)土壤水势过高或过低都抑制梨枣的营养生长、生殖生长和坐果,但水涝能促进茎直径生长,水分缺亏则抑制茎直径生长。4)在平水年,梨枣的萌芽期无需灌水,自然降雨即能满足其萌芽展叶、花芽分化的需要,但开花期,降雨不能满足梨枣高坐果率的需要。