Leaching of genetically modified Pseudomonas fluorescens through organic soils: Influence of temperature,soil pH,and roots

Summary The effects of soil temperature and bulk soil pH on the vertical translocation of a genetically modified Pseudomonas fluorescens inoculum were studied in reconstituted soil microcosms, in the presence and absence of growing Lolium perenne roots. The inoculated microcosms received one rainfall event per day (5 mm h^-1 for 6 h) for 5 days and the resulting leachate was quantitatively assayed for the presence of the modified pseudomonad. Soil temperature affected the total number of modified pseudomonads detected in the leachate over the 5 days, with significantly lower numbers detected at 25°C compared to 5°C. The bulk soil pH also affected leaching of the inoculum, with significantly greater numbers detected in the effluent at pH 7.5 than at pH 4.5. In the absence of L. perenne, greater numbers of the modified pseudomonads were detected in the pH 7.5 soil after 5 days of leaching compared to soil at pH 4.5. L. perenne roots decreased the number of cells of the inoculum that were leached and detected in the soil after 5 days of leaching. In the soil microcosms used for the pH study the distribution of the inoculum remaining with the soil was altered by L. perenne roots. At each pH value the proportion of cells detected within the soil below the surface 2 cm of the microcosms was greater in the presence of L. perenne roots. The results of this study indicate that soil temperature, bulk soil pH, and the presence of root systems are important factors in determining the extent of inoculum translocation, and should be considered in the design and interpretation of field experiments.     

Responses by iron-efficient and inefficient oat cultivars to inoculation with siderophore-producing bacteria in a calcareous soil

Rhizosphere bacteria may enhance plant uptake of Fe by producing siderophores that chelate sparingly soluble Fe³⁺ in calcareous soils. To evaluate the extent to which plants benefit from colonization of the roots by prolific siderophore-producing bacteria, we inoculated two oat cultivars with six strains of bacteria that produced high concentrations of siderophores under Felimiting conditions in vitro. Oat cv Coker 227, an Fe-efficient cultivar, which produces the phytosiderophore avenic acid, and cv TAM 0-312, and Fe-inefficient cultivar, which does not produce the phytosiderophore, were grown in a calcareous soil (Weswood silt loam) on a light bench in the laboratory. Half of the plants were fertilized with a nutrient solution containing 5 mM Fe and half with a nutrient solution containing no Fe. After 6 weeks of growth, we compared colonization of the roots by the inoculant bacteria and the dry weight and Fe content of roots and shoots. Three species of Pseudomonas colonized the roots of both oat cultivars in high numbers (10⁶ cells g^-1 root dry weight), whereas the remaining bacteria colonized the roots in substantially lower numbers (10⁴ cells g^-1 root dry weight). Plants fertilized with 5 mM Fe were larger and supported greater numbers or rhizosphere bacteria per gram of root than plants not supplied with Fe. Comparisons of the Fe content and dry weight of roots and shoots revealed few significant differences between inoculated and uninoculated plants, or among the plants inoculated with the different strains of siderophore-producing bacteria. The differences that were observed revealed no consistent response to inoculation. We conclude that inoculation of the roots of the two oat cultivars with bacteria that produce high concentrations of siderophores in response to an Fe deficiency had little or no effect on Fe acquisition by the plants.     

Possible influence of Rhizobium on VA mycorrhiza metabolic activity in double symbiosis of alfalfa plants (Medicago sativa L.) grown in a pot experiment

Summary Alfalfa (Medicago sativa, L. cv Aragón) plants were grown under greenhouse conditions in pots of inert sand and vermiculite. The plants were inoculated with Rhizobium meliloti strain 102F28, with Glomus fasciculatus or with a mixture of both microorganisms. Plants inoculated with both Rhizobium and Glomus had the highest shoot dry weight and the lowest root-to-shoot ratio. Roots from dually inoculated plants also had a higher oxygen uptake and nodule nitrogenase activity than those from plants inoculated with either of the two microsymbionts alone. However, the dry weight of the roots from only VAM-infected plants was higher than those from Rhizobium or from Rhizobium plus Glomus-inoculated ones. These differences did not correlate with succinate dehydrogenase activity, which was similar between treatments. Nutrient element concentrations were increased in dually infected plants in comparison with those of plants inoculated with only Rhizobium or Glomus. These data suggest that Rhizobium may affect fungal metabolism and that the effect is not achieved via the tricarboxylic acid pathway.     

The effect of vesicular-arbuscular mycorrhizal inoculation on nutrient uptake and yield of alley-cropped cassava in a degraded Alfisol of southwestern Nigeria

Leaf and root (tuber) nutrient uptake patterns of cassava (Manihot esculenta Crantz) alley-cropped with gliricidia (Gliricidia sepium), leucaena (Leucaena leucocephala), and senna [(Senna (syn. Cassia) siamea] as influenced by vesicular-arbuscular mycorrhizal (VAM) inoculation in a degraded Alfisol were investigated in consecutive years. The cassava plants were mulched with fresh prunings of each hedgerow tree species at 2-month intervals in the second and third years of alley cropping. While VAM inoculation significantly influenced the root uptake of nutrients, the leaf uptake was not affected except for the uptake of P. In most cases, there was no difference in the nutrient concentration between inoculated and uninoculated plants, either in the leaf or in the root, indicating that the productivity of cassava was regulated by the amount of nutrients the roots could absorb. In spite of similar total soil N in all inoculated and uninoculated alley-cropped cassava plots and similar exchange-able soil K contents in inoculated and uninoculated alley-cropped cassava plots with leucaena and senna, greater uptake of N, P, and K and greater concentrations of K were observed in roots of inoculated alley-cropped cassava with gliricidia and leucaena than with senna. These results indicated that greater mineralization and availability of nutrients to cassava roots from prunings of nodulating gliricidia and leucaena than from non-nodulating senna may be important, particularly with efficient VAM inoculation, in these alley-cropping systems. Also, for similar nutrients in the inoculated and uninoculated cassava soils alley-cropped with each hedgerow species, VAM inoculation significantly enhanced cassava root dry weights, indicating that an effective VAM fungus can be an agent of greater nutrient uptake in a competitive environment.     

油菜苗期根系缺硼对保护酶的变化及多胺积累的影响

The effects of boron deficiency on the membrane permeability,the lipid peroxidation of membrane ,the activities of the protective enzymes and the accumulation of polyamines in the roots of rape (Brassica napus L.cv Zhongyou 821)plants were examined using solution culture experiment .Compared to the 20 mmol B m^-3 treatment ,boron-deficient treatment( 2 mmol B m^-3) decreased root dry weight and increased the rate of solute leakage and malondialdehyde (MDA) concentration in the roots of rape.Similar patterns of change were observed in the level of lipid peroxidation (MDA concentration) and the rate of solute leakage under boron deficiency.Results suggested that a signifcant alteration of membrane composition had occurred under B deficiency.The concentration of putrescin and the ration of putrescine to spermidine spermine in the roots of rape increaed significantly,and the activities of superoxide dismutase and catalase decreased in the roots of rape during B deficiency,But peroxidase activity in root of B-defcient plant was higher than that of control plant.     

模拟酸雨对龙眼幼果多胺含量的影响研究

以8年生乌龙岭龙眼为试验材料,研究不同pH的模拟酸雨对龙眼幼果多胺含量的影响。结果表明,酸雨胁迫下龙眼幼果多胺(PAs)、腐胺(Put)、亚精胺 精胺(Spd Spm)含量上升,并随果实的发育而逐渐下降。易脱落幼果的PAs、Put、Spd Spm含量均显著高于正常幼果。     

Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.), and sweet sorghum (Sorghum vulgare)

Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.     

Effects of polyamines on the ion content and solutes excretion of isolated embryonic axes and the germination rate of seeds of chickpea

The effect that three polyamines, spermine, spermidine and cadaverine, have on the growth rate and the endogenous ion content of isolated, chickpea (Cicer arietinum, L.) embryonic axes and the germination of whole seeds has been studied. Both the growth rate and ion content are in the same direction, but depending upon each individual polyamine and the concentration. In moderate quantities, spermine increases growth rate, spermidine less so and cadaverine not at all; while at high quantities, spermine greatly reduces growth rate, spermidine less so and cadaverine has no effect. Cadaverine has more effect on the germination of whole seeds. The excretion of aminoacids and sugars into the medium in response to these polyamines was significantly augmented.     

Measurement of pH at the root surface of red clover (Trifolium pratense) grown in soils differing in proton buffer capacity

Summary Measurements of pH were made at the root surface of Trifolium pratense, using Sb electrodes. Nodulated plants were grown in rhizotrones on a sandy soil free of carbonate and on a clay soil rich in carbonate. In the sandy soil, pH at the surface of root laterals was about 1 unit lower than in the bulk soil. The lowest pH values were found at the root tips. In the calcareous soil, pH measured at the root surface did not differ from pH in the bulk soil. This soil had a much higher H⁺ buffer capacity than the sandy soils. It seems likely that H⁺ ions excreted from the roots grown in the calcareous soil were directly neutralized by soil carbonate.     

Response of maize to mycorrhizal colonization at varying levels of zinc and phosphorus

A greenhouse experiment was conducted in a red sandy loam soil (Alfisol) to study the responses of arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith inoculated (M+) and uninoculated (M−) maize (Zea mays L) plants exposed to various levels of P (15 and 30 mg kg⁻¹) and Zn (0, 1.25, and 2.5 mg kg⁻¹). Roots and shoots were sampled at 55 and 75 days after sowing and assessed for their nutritional status, root morphology, and root cation exchange capacity (CEC) besides grain quality. Mycorrhizal plants had longer and more extensive root systems than nonmycorrhizal plants, indicating that M+ plants are nutritionally rich, especially with P, which directly assisted in the proliferation of roots. Further, root CEC of M+ plants were consistently higher than those of M− plants, suggesting that mycorrhizal colonization assists in the acquisition of nutrients from soil solution. Mycorrhizal inoculated plants had significantly (P ≤ 0.01) higher P and Zn concentrations in roots, shoots, and grains, regardless of P or Zn levels. The available Zn and P status of AM fungus-inoculated soils were higher than unioculated soils. The data suggest that mycorrhizal symbiosis improves root morphology and CEC and nutritional status of maize plants by orchestrating the synergistic interaction between Zn and P besides enhancing soil available nutrient status that enables the host plant to sustain zinc-deficient conditions.     

Toxicity of copper and zinc in seedlings of Mung bean and inducing accumulation of polyamine

A solution culture was conducted to investigate the effects of copper (Cu) and zinc (Zn) toxicity on growth of mung bean (Phaseolus aures Roxb. cv VC‐3762) and accumulation of polyamine. Eight‐day‐old seedlings were grown in diluted nutrient solution with different concentrations of Cu and Zn for 6 days. Results showed that elongation of epicotyl and fresh weight of plants were decreased by 10 μM Cu and 100 μM Zn significantly compared to control (0.03 μMCu and 0.1 μMZn). Accumulation of polyamine, especially putrescine (Put) was found in the epicotyl of mung bean seedlings. Addition of 5 mM calcium (Ca) into nutrient solution improved the growth of 10 μM Cu‐treated seedling, and decreased the concentration of Put and increased concentrations of spermine and spermidine in epicotyl of plants. Moreover, supplying Put did not increase tolerance of plant to Cu or Zn. It was suggested that Put accumulation resulting from toxicity of Cu and Zn might merely be a symptom of stress injury.     

Development of stable isotope dilution assays for the simultaneous quantitation of biogenic amines and polyamines in foods by LC-MS/MS

Microbial amino acid metabolism may lead to substantial amounts of biogenic amines in either spontaneously fermented or spoiled foods. For products manufactured with starter cultures, it has been suggested that certain strains may produce higher amounts of such amines than others; however, to support efforts of food manufacturers in mitigating amine formation, reliable methods for amine quantitation are needed. Using 10 isotopically labeled biogenic amines as the internal standards, stable isotope dilution assays were developed for the quantitation of 12 biogenic amines and of the 2 polyamines, spermine and spermidine, in one LC-MS/MS run. Application of the method to several foods revealed high concentrations of, for example, tyramine and putrescine in salami and fermented cabbage, whereas histamine was highest in Parmesan cheese and fermented cabbage. On the other hand, ethanolamine was highest in red wine and Parmesan cheese. The results suggest that different amino acid decarboxylases are active in the respective foods depending on the microorganisms present. The polyamine spermine was highest in salami and tuna.     

Dual inoculation of pretransplant stage Oryza sativa L. plants with indigenous vesicular-arbuscular mycorrhizal fungi and fluorescent Pseudomonas spp.

Summary This study examined the response of rice (Oryza sativa L.) plants at the pretransplant/nursery stage to inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi and fluorescent Pseudomonas spp., singly or in combination. The VAM fungi and fluorescent Pseudomonas spp. were isolated from the rhizosphere of rice plants. In the plants grown in soil inoculated with fluorescent Pseudomonas spp. alone, I found increases in shoot growth, and in root length and fine roots, and decreases in root growth, and P and N concentrations. In contrast, in the plants colonized by VAM fungi alone, the results were the reverse of those of the pseudomonad treatment. Dual inoculation of soil with VAM fungi and fluorescent Pseudomonas spp. yielded plants with the highest biomass and nutrient acquisition. In contrast, the plants of the control treatment had the lowest biomass and nutrient levels. The dual-inoculated plants had intermediate root and specific root lengths. The precentages of mycorrhizal colonization and colonized root lengths were significantly lower in the dual-inoculated treatment than the VAM fungal treatment. Inoculation of plants with fluorescent Pseudomonas spp. suppressed VAM fungal colonization and apparently reduced photosynthate loss to the mycorrhizal associates, which led to greater biomass and nutrient levels in dual-inoculated plants compared with plants inoculated with VAM fungi alone. Dual inoculation of seedlings with fluorescent Pseudomonas spp. and VAM fungi may be preferable to inoculation with VAM alone and may contribute to the successful establishment of these plants in the field.     

Effect of Azospirillum brasilense inoculation on root morphology and respiration in tomato seedlings

Summary The level of Azospirillum brasilense strain Cd colonization in the rhizosphere of some vegetables was 10⁴–10⁵ colony-forming units (CFU) per root of one plant in 2-week-old plants inoculated with 5 × 10⁸ Azospirillum cells. Significant increases in root length (35%) and in top (90%) and root (50%) dry weight and total leaf area (90%) were observed in 18-day-old inoculated tomato plants compared with non-inoculated controls. An inoculum concentration of 1 × 10⁸ to 5 × 10⁸ CFU/ml stimulated the appearance of root hairs. Large numbers of bacteria (1 × 10⁹ CFU/ml) caused asymmetrical growth of the root tip. In a petri dish system, Azospirillum (1 × 10⁸ CFU/ml) increased root dry weight (150%), protein content (20%), respiration rate per root (70%) and the specific activity of malate dehydrogenase (45%–65%) over non-inoculated controls. The specific respiration rate, expressed as micromol of O₂ per minute per milligram of dry weight of roots, was significantly lower in inoculated roots, suggesting that less energy was spent for accumulation of more dry material.     

丛枝菌根对酸枣实生苗耐盐性的影响

本文研究了在土中加入不同量NaCl条件下 (0、1 5、3 0、4 5gkg-1干土 )接种丛枝菌根真菌 (AMF)Glomusmosseae对盆栽酸枣 (ZizyphusspinosusHu)实生苗生长及耐盐性的影响。结果表明 ,无论接种与否 ,植株的高度、根茎叶的干鲜重均随土壤NaCl浓度的增加而降低 ,而根、茎、叶和整株的Na浓度及Na全量均随土壤NaCl浓度的增加而增大。在土壤盐浓度相同的条件下 ,接种AMF植株的生长量 (株高、鲜重、干重等 )和叶片的叶绿素含量显著高于不接种植株。接种AMF的植株茎、叶中Na浓度低于不接种植株 ,而根中Na浓度、植株Na总量显著高于不接种植株。盐浓度最大的接种处理 ,其植株生长量和叶片叶绿素含量均高于不加盐不接菌处理。播种时进行盐胁迫处理和播种后 4 0d开始进行盐胁迫处理对菌根的侵染率、植株生长的影响差异不显著。上述四种盐浓度播种时进行盐处理的接种AMF植株的总干重比不接种植株分别提高 16 4 %、14 9%、4 8%、35 % ,在播种后 4 0d进行盐处理的接种AMF的植株比不接种植株分别提高 194 %、12 7%、72 %、4 6 %。结果证明 ,酸枣实生苗具有较强的耐盐性 ,其生长对菌根真菌有很强的依赖性 ,接种菌根真菌提高了其耐盐能力。     

Effect of soil pH on plant growth and nodulation of cowpea

Abstract

A study was conducted to evaluate the effect of soil pH on rhizobium inoculation, plant growth and nodulation of cowpea (Vigna unguiculata). Both inoculated and non‐inoculated seeds of the cultivar ‘California Blackeye No. 5’ were grown in the greenhouse in plastic pots with growth medium being a Norfolk sandy loam (Fine, loamy siliceous, thermic, Typic Palendult) soil under different pH levels. Both soil pH and rhizobium inoculation significantly affected root length, plant height, nodule and pod number per plant. Within the pH range of 6.6 to 7.6, these growth parameters generally were at their maximum, decreasing above or below this pH range. Non‐inoculated plants produced some nodules, indicating failure of the methyl bromide to totally destroy all residual soil rhizobta before inoculation treatment.

The inoculated plants produced more seeds and the increased number of nodules of treated plants was directly related to increased seed weight. Since nodule number was highest at the approximate pH range of 6.6 to 7.6, this range was considered optimum for nodulation of cowpea by this strain of rhizobium under greenhouse conditions. At pH 7.5 and above, roots tended to be more fibrous and nodules were generally smaller in size.     

Peculiar effect of Azospirillum inoculation on growth and nitrogen balance of winter wheat (Triticum aestivum)

Summary Wheat plants (Triticum aestivum) grown in pots and in the field under the Mediterranean climate of the south of France were inoculated with a strain of Azospirillum brasilense. Comparisons with non-inoculated plants grown under the same conditions showed significant responses to inoculation with an increase in the number of fertile tillers, shoot and root dry weight, and root to shoot biomass ratio. The roots of inoculated plants attracted relatively more assimilates than those of the control plants until a late stage of growth (heading stage) but the rhizosphere respiration expressed per unit of root growth was not increased by inoculation. Nitrogen yield, both total and in grains, was also enhanced; however, N percentages of all aerial parts of the plants grown in pots were always statistically lower after inoculation than in the control. At maturity, the N % in seeds was 1.81 and 2.45, respectively. The possible mechanisms of this effect of inoculation under the experimental conditions of this study are discussed.     

Colonization of wheat seedling (Triticum aestivum) roots by Pseudomonas fluorescens and Bacillus subtilis

Summary Colonization patterns of Pseudomonas fluorescens and Bacillus subtilis on roots of wheat seedlings growing on water agar were studied qualitatively by replica printing and quantitatively by the plate count method. The results indicated a stronger colonization potential for P. fluorescens (up to 10⁷ cfu/cm root) than for B. subtilis (up to 10⁵ cfu/cm root). Although the numbers of both species were lower when inoculated together, the observed colonization patterns on the roots were comparable to those found with single inoculations. For none of these bacteria was active migration along the root surface in any direction observed, indicating that distal positions are reached mainly by a passive displacement on the root tip and elongating cells. Ecological implications of the observed phenomena are discussed.     

烟草配方肥增效剂对烤烟根系内源激素和多胺含量的影响

本试验研究了烟草配方肥增效剂对进入成熟期后烟株根系内源激素含量和多胺含量以及烟叶产量和产值的影响.结果表明,施用配方肥增效剂可提高烟株根系内赤霉素(GA3)、生长素(IAA)和精胺含量,降低脱落酸(ABA)和腐胺含量,其中精胺含量随着肥料用量的减少而增加.前期,配方肥增效剂对根系内亚精胺含量的影响很小,但后期会导致亚精胺含量迅速增高.增效剂对根系内源激素含量和多胺含量的影响以每株施用45.45g配方肥和0.4545g增效剂处理效果最好.施用增效剂能提高烟叶的产量和产值,减少肥料用量的增效剂处理虽降低了烟叶产量,但因改善了烟叶品质而提高其产值,增效剂对烟叶产量和产值的影响以每株施用36.36g配方肥和0.4545g增效剂处理效果最好.     

Population densities of clover rhizobia in Texas pastures and response to liming

Summary Clovers are widely used forage legumes on acidic soils in Texas and need inoculation with appropriate rhizobia when first introduced. Acidic soils are not conducive to survival of clover rhizobia. A survey of pastures was undertaken to determine the number of rhizobia present. The effect of liming acidic soils on the survival of clover rhizobia was also evaluated in the laboratory. The number of clover rhizobia was more than 100 cells g^-1 soil in 70% of the pastures surveyed but populations within pastures varied by more than two orders of magnitude. The number of years of clover production beyond 1 year did not affect the rhizobial population density. The soil pH of twelve samples was below 5.0 and six samples had populations of rhizobial lower than 100 g^-1 soil. Eleven out of sixteen samples from fields that had grown clover and had pH values above 6.0 had populations exceeding 1000 g^-1 soil and only three samples had populations lower than 100 g^-1 soil. Incubating indigenous or inoculated rhizobia in well-mixed soils having pH values of 5.1 or below resulted in populations declining to below 10 g^-1 soil in 6 weeks. Mixing of soils with pH values of up to 5.4 induced reduction of rhizobial numbers, possibly by destroying microsites. Liming of soils to increase pH values above 5.5 improved survival of native or inoculated rhizobia in most cases.