Effect of low pO2 on colonization of maize roots by a genetically altered Pseudomonas putida [PH6(L1019)]

Among the factors which may affect colonization of roots by soil bacteria is that of rhizosphere oxygen partial pressure (pO₂). The oxygen concentration in the root zone influences both microbes and roots. Roots exposed to low pO₂, as might occur during flooding and waterlogging of the soil, become more leaky and loss of soluble carbon increases. To determine whether periods of low pO₂ increased root colonization by a genetically altered pseudomonad we inoculated 3- to 4-week-old maize plants, grown in soil and transferred to a hydroponic system or grown in fritted clay, with Pseudomonas putida PH6(L1019)(lacZY+) following exposure of the roots to air or cylinder N₂. Numbers of heterotrophs and the marked pseudomonad were determined by dilution plating. Low pO₂ generally increased the numbers of bacteria associated with roots exposed to the treatments in solution or in undisturbed fritted clay rooting medium. Under low pO₂ in a hydroponic system, roots of intact maize plants tended also to have higher soluble organic C and hexose (anthrone-detectable sugars) than roots exposed to air. The effect of low pO₂ was most pronounced in the fritted clay where low pO₂ favored colonization by the marked strain; numbers were 3- to 96-fold greater than those on roots flushed with air but accounted for only 0.06–0.61% of the total population. Roots exposed to low pO₂ tended to accumulate more C. Results suggest that in the fritted clay, the pseudomonad was able to exploit the increased C supply and to achieve greater numbers on roots exposed to low pO₂, whereas the dilution of carbon released from roots in the hydroponic apparatus did not allow for the same magnitude of increase on roots. Received: 2 December 1996     

Phosphatase activity in the rhizosphere and its relation to the depletion of soil organic phosphorus

Summary The distribution of phosphatase activity and of phosphate fractions of the soil in the proximity of roots was studied in order to evaluate the significance of phosphatases in P nutrition of various plants (Brassica oleracea, Allium cepa, Triticum aestivum, Trifolium alexandrinum). A considerable increase in both acid and alkaline phosphatase activity in all the four soil-root interfaces was observed. Maximum distances from the root surface at which activity increases were observed ranged from 2.0 mm to 3.1 mm for acid phosphatase and from 1.2 mm to 1.6 mm for alkaline phosphatase. The increase in phosphatase activity depended upon plant age, plant species and soil type. A significant correlation was noticed between the depletion of organic P and phosphatase activity in the rhizosphere soil of wheat (r = 0.99^**) and clover (r = 0.97^**). The maximum organic P depletion was 65% in clover and 86% in wheat, which was observed within a distance from the root of 0.8 mm in clover and 1.5 mm in wheat. Both the phosphatases in combination appear to be responsible for the depletion of organic P.     

黄瓜根部施用化学物质对其霜霉病诱导抗性的研究

分别用几种化学物质从黄瓜幼苗子叶期开始进行根部诱导，结果表明，在自然病原激发病害试验和在人工接种病原激发病害试验中，经苯酚、脲素、无水对氨基苯磺酸和氯化钾诱导的黄瓜植株病株率和病叶率下降显著。对可产生诱导免疫效果的化学物质分析发现，氨类和苯类物质及钾的氯化物与黄瓜叶片对霜霉病的抗性有极为密切的关系。     

Organic acid concentrations in soil solution: effects of young coniferous trees and ectomycorrhizal fungi

Organic acids may play a key role in rhizosphere and pedogenic processes. The effects of young trees and ectomycorrhizas on the soil solution concentrations of low molecular weight organic acids (LMWOAs) were studied in soil columns (E horizon) in the presence or absence of Pinus sylvestris and Picea abies with or without three ectomycorrhizal fungi. Several LMWOAs were identified at concentrations ranging from <0.1 to 11 μM. Compared to soil columns without tree seedlings, the presence of non-mycorrhizal or mycorrhizal tree seedlings sometimes resulted in small but statistically significant increases in citrate, formate, malonate and oxalate concentration. The general nutrient concentration and low P had little short-term effect on soil solution organic acid concentrations. The results suggest that biodegradation rather than production may be the major factor regulating soil solution organic acid concentrations.     

The influence of nitrogen fertilisation on bacterial activity in the rhizosphere of barley

The effect of nitrogen addition on the activity of rhizosphere bacteria was studied using barley seedlings. Three different nitrogen sources were added to the soil (nitrate, ammonium and ammonium+nitrate) at four different concentrations (0, 100, 300 and 500 mg N kg⁻¹ soil) and the plants were allowed to grow for 6 weeks. The bacterial activity was estimated by measuring thymidine and leucine incorporation into bacteria extracted using homogenisation-centrifugation. Bulk soil bacterial activity was low compared with that of rhizosphere bacteria. Nitrogen addition did not affect the activity of the bulk soil bacteria, indicating that the activity was not nitrogen limited. The thymidine and leucine incorporation rates of rhizosphere bacteria decreased when ammonium or ammonium+nitrate was applied compared with the non-amended controls. No effect on bacterial activity was found following nitrate addition. There was a significant positive correlation between rhizosphere bacterial activity and rhizosphere pH. Shoot length following ammonium treatment was significant lower than in the non-amended control, while nitrate and ammonium+nitrate addition had no effect. This indicates that the varying effects due to nitrogen sources on rhizosphere bacterial activity were not due to effects on plant growth.     

Relative importance of motility and antibiosis in the rhizoplane competence of a biocontrol agent Pseudomonas fluorescens MelRC2Rif

Mutants defective in motility or antibiotics production were obtained by Tn5 mutagenesis of a biocontrol agent Pseudomonas fluorescens MelRC2Rif (wt). Tomato or melon seeds were co-inoculated with a Tn5 mutant and wt in a 1:1 ratio and then grown in soil for 10 days. There was no change in ratios of Tn5 mutants defective in antibiosis to wt in the process of rhizoplane colonization, suggesting little contribution of in vitro antibiosis to the rhizoplane competence of P. fluorescens MelRC2Rif. Similar results were also obtained when seeds treated with bacteria were planted in soil artificially infested with fungal pathogens. In contrast, ratios of Tn5 mutants defective in motility to wt significantly decreased, suggesting the contribution of motility to the rhizoplane competence of this bacterium. When a non-motile Tn5 mutant and wt were co-inoculated into soil at a matric potential of pF 2.3 (–20 kPa) and plants were then grown, there was no change in the ratio in rhizoplane colonization, suggesting that motility might have a role in the movement along roots but an insignificant role in the movement from bulk soil towards roots. When they were co-inoculated into 0.2% water agar (WA) instead of soil, a remarkable decline in ratios was detected. Thus it was soil structure that hindered the efficiency of motility. Time course enumeration of rhizoplane colonization of tomatoes grown in WA revealed that motility was an important means of movement towards and/or along roots rather than the multiplication on roots. Received: 8 July 1996     

Gram-negative bacterial flora on the root surface of wheat (Triticum aestivum) grown under different soil conditions

We identified 161 Gram-negative bacterial strains isolated from the root surface of wheat grown under different soil conditions. The strains were divided into seven groups based on major morphological and physiological properties. Taxonomic allocation of the groups was verified by guanine+cytosine contents of DNA. Except for one group, which may be assumed to include bacteria belonging to the genera Flavobacterium and Cytophaga, the various groups were taxonomically united. The distribution of the groups changed with soil improvement. Pseudomonads predominated in unimproved soil, but Flavobacterium and Cytophaga spp. were predominant in the most improved soil. As all the strains were non-fermentative by Hugh and Leifson's test, API 20NE identification was applied. However, many strains were misidentified by this system, especially in the Flavobacterium and Cytophaga spp. group. For ecological studies, the strains were classified to species level by the API 20 NE system and by the results of a combination of guanine+cytosine (mol%) and isoprenoid quinone data. The pattern of distribution of the bacteria on the root surface of wheat varied at species level within one genus depending on soil conditions.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Nitrogen fixation by Azospirillium brasilense in soil and the rhizosphere under controlled environmental conditions

Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O₂ tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C₂H₄ h^-1 per 2kg fresh soil, increasing to 300 nmol C₂H₄ h^-1 when malic acid was added as a C source via a dialyse tubing system. The N₂ fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O₂ than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O₂, with a relatively high level of activity remaining even at an O₂ concentration of 20 kPa. Both NO _inf3 ^sup- and NH _inf4 ^sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N₂-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the ¹⁵N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N₂ fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This ¹⁵N dilution was comparable to that seen in plants inoculated with non-N₂-fixing Psudomonas fluorescens.     

根-土界面的微生态过程与有机污染物的环境行为研究

土壤-植物系统是地球生态系统中与人类生存与健康关系最为密切的亚系统。该系统中有机污染物的运移必须历经根-土界面多层次的微生态过程的控制。这些微生态过程涉及到系统中许多生物、生物化学和物理化学反应机理,与土壤中污染物迁移转化及其归宿等环境行为具有密切的关系。理解这些微生态过程及其对有机污染物环境行为的影响,对提高作物生长、改善土壤环境质量和提高农产品品质安全具有直接的理论和实践指导意义。     

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.