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     

Endomycorrhizae in a newly cultivated acidic meadow: Effects of three years of barley cropping,tillage, lime,and phosphorus on root colonization and soil infectivity

The dynamics of mycorrhizae under disturbance created by crop production is not well understood. A 3-year experiment was undertaken on a nutrient-poor and acidic land that had last been cultivated in the early 1970s. We observed the effects of cropping spring barley (Hordeum vulgare L.) under four P-fertilizer levels and four levels of lime, in a minimum (rototillage), a reduced (chisel), or a conventional tillage system, on the mycorrhizal receptiveness of the host (maximum level of mycorrhizal colonization, as measured at harvest) and soil infectivity most probable number method. The host receptiveness decreased with time, while crop yields and soil infectivity increased simultaneously with time. Liming increased mycorrhizal colonization of barley roots and soil infectivity. P additions decreased root colonization but did not significantly affect the most probable number valuse. Slightly higher soil infectivity estimates were found under reduced tillage.     

Effects of elevated CO2 concentration on rhizodeposition from Lolium perenne grown on soil exposed to 9 years of CO2 enrichment

The effects of enriched CO₂ atmosphere on partitioning of recently assimilated carbon were investigated in a plant-soil-microorganism system in which Lolium perenne seedlings were planted into cores inserted into the resident soil within a sward that had been treated with elevated CO₂ for 9 consecutive years, under two N fertilisation levels (Swiss FACE experiment). The planted cores were excavated from the ambient (35 Pa pCO₂) and enriched (60 Pa pCO₂) rings at two dates, in spring and autumn, during the growing season. The cores were brought back to the laboratory for ¹⁴C labelling of shoots in order to trace the transfer of recently assimilated C both within the plant and to the soil and microbial biomass. At the spring sampling, high N supply stimulated shoot and total dry matter production. Consistently, high N enhanced the allocation of recently fixed C to shoots, and reduced it to belowground compartments. Elevated CO₂ had no consequences for DM or the pattern of C allocation. At the autumn sampling, at high N plot, yield of L. perenne was stimulated by elevated CO₂. Consistently, ¹⁴C was preferentially allocated aboveground and, consequently belowground recent C allocation was depressed and rhizodeposition reduced. At both experimental periods, total soil C content was similar in all treatments, providing no evidence for soil carbon sequestration in the Swiss Free Air CO₂ Enrichment experiment (FACE) after 9 years of enrichment. Recently assimilated C and soil C were mineralised faster in soils from enriched rings, suggesting a CO₂-induced shift in the microbial biomass characteristics (structure, diversity, activity) and/or in the quality of the root-released organic compounds.     

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.     

不同樱桃砧木根际微生物和养分的动态变化

研究樱桃年生长周期内不同樱桃砧木(大青叶、山樱、考特和马哈利)对根际微生物和养分动态变化的影响。结果表明,樱桃年生长周期内,各砧木根际微生物数量呈现先上升再下降的变化趋势,且均以细菌最多,放线菌次之,真菌最少。根际有机质和全氮呈先上升后下降又上升的变化趋势,全磷和全钾呈先上升后下降的变化趋势,碱解氮和速效磷呈逐步上升的变化趋势,速效钾呈先降低后上升的变化趋势,且各砧木根际各养分差异显著。4种樱桃砧木土壤营养水平基本能够满足樱桃生长的需要,但根际养分亏缺。因此,选择合适的樱桃砧木,有利于土壤微生物数量和养分的提高,从而为樱桃树生长创造良好的土壤生态环境。     

Distribution of phosphatase activity and various bacterial phyla in the rhizosphere of Hordeum vulgare L. depending on P availability

Despite the importance of the rhizosphere for nutrient turnover, little is known about the spatial patterns of organic phosphorus mineralization by plants and by microorganisms in the rhizosphere. Therefore, the distribution of acid and alkaline phosphatase activity and the abundance of bacteria belonging to various bacterial phyla were investigated in the rhizosphere of barley (Hordeum vulgare L.) as dependent on the availability of inorganic P. For this purpose, we conducted a greenhouse experiment with barley growing in inclined boxes that can be opened to the bottom side (rhizoboxes), and applied soil zymography and fluorescence-in situ-hybridization (FISH). Acid phosphatase activity was strongly associated with the root and was highest at the root tips. Due to P fertilization, acid phosphatase activity decreased in the bulk soil, and less strongly in the rhizosphere. Alkaline phosphatase activity, i.e., microbial phosphatase activity was high throughout the soil in the control treatment and was reduced due to inorganic P fertilization especially in the rhizosphere and less strongly in the bulk soil. P-fertilization slightly increased the total number of bacteria in the rhizosphere. Moreover, P-fertilization decreased the abundance of Firmicutes and increased the abundances of Beta- and Gamma-Proteobacteria. The total number of bacterial cells was significantly higher at the root surface than at the root tip and at a distance of 30 μm from the root surface. Our results show that alkaline phosphatase activity decreased more strongly in the rhizosphere than in the bulk soil due to P fertilization, which might be because of greater C deficiency in the bulk soil compared to the rhizosphere. Furthermore, the results indicate a spatial separation between hotspots of acid phosphatase activity and hotspots of bacteria in the rhizosphere of H. vulgare. Taken together, our study shows that bacteria and phosphatase activity were very heterogeneously distributed in soil, and that the effects of P fertilization on phosphatase activity differed strongly between bulk soil and rhizosphere as well as between various zones of the rhizosphere.     

Organic acid induced release of nutrients from metal-stabilized soil organic matter – The unbutton model

Processes of soil organic matter (SOM) stabilization and the reverse, destabilization of SOM resulting in subsequent release and mobilization of nutrients from SOM, remain largely unresolved. The perception of SOM as supramolecular aggregates built of low molecular mass biomolecules is currently emerging. Polyvalent metal cations contribute to SOM tertiary structure by bridging functional groups of such molecules (Simpson et al., 2002). The strong bond to metals protects high quality organic material from being immediately accessed and decomposed. Here we propose a three-step process by which low molecular mass organic acids (LMMOAs) and hydrolytic enzymes act in series to destabilize SOM supramolecules to release organic nitrogen (N) and phosphorus (P) for local hyphal and root uptake. Complexation of the stabilizing metals by fungal-released LMMOA gives fungal-root consortia direct access to organic substrates of good quality. Because of their small sizes and carboxyl group configuration, citric and oxalic acids are the most effective LMMOAs forming stable complexes with the main SOM bridging metals Ca and Al in SOM. Citrate, forming particularly strong complexes with the trivalent cations Al and Fe, is dominant in soil solutions of low-productive highly acidic boreal forest soils where mycorrhizal associations with roots are formed predominantly by fungi with hydrophobic hyphal surfaces. In these systems mycelia participate in the formation of N-containing SOM with a significant contribution from strong Al bridges. In less acidic soils of temperate forests, including calcareous influenced soils, SOM is stabilized predominantly by Ca bridges. In such systems mycorrhizal fungi with more hydrophilic surfaces dominate, and oxalic acid, forming strong bidentate complexes with Ca, is the most common LMMOA exuded. A plant-fungus driven biotic mechanism at the supramolecular aggregate level (10³–10⁵ Da) resolves micro-spatial priming of SOM, where the destabilization step is prerequisite for subsequent release of nutrients.     

Enhanced bioremediation of soil containing 2,4-dinitrotoluene by a genetically modified Sinorhizobium meliloti

The symbiotic nitrogen-fixing soil bacterium, Sinorhizobium meliloti, is well known for its ability to interact with the leguminous plant Medicago sativa L. It has, however, not been reported that this species possesses the capability to degrade toxic nitroaromatic compounds, such as 2,4-dinitrotoluene (DNT) which is commonly associated with the degradation of the explosive trinitrotoluene (TNT). In this study, the pJS1 DNT-biodegradative plasmid was genetically transferred to S. meliloti strain USDA 1936, which was confirmed by plasmid profile analysis. Several standard analytical and chemical tests including high performance liquid chromatography (HPLC), nitrite (NO₂) release assays, rhizosphere population and plant greenhouse studies were conducted to test the ability of S. meliloti to degrade 2,4-DNT. The possible presence of 2,4-DNT remaining in the treated soil was tested, and no 2,4-DNT had been absorbed by the soil. The pJS1-carrying recombinant strain DHK1 produced ‘ARC’ alfalfa plants that were almost 2-fold higher in shoot dry weight than that produced by the parent strain on soil containing 0.14 mM 2,4-DNT. The transconjugant strain DHK1 reduced significantly one-third more 2,4-DNT in both 0.14 and 0.28 mM contaminated soil, and in 0.55 mM contaminated soil it degraded 94% of the 2,4-DNT present. In liquid cultures, however, only about 4% reduction in 2,4-DNT concentrations was obtained in 10 days. We interpret the results as clearly establishing that genetic modification was successfully used, for the first time, to improve the capability of the symbiotic nitrogen-fixing soil bacterium S. meliloti DHK1 to bioremediate in situ 2,4-DNT-contaminated soil in the presence of alfalfa plants.     

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。     

盐碱胁迫对枸杞生长及根际适应能力的影响

通过水培试验研究不同盐碱胁迫条件对枸杞生长以及根际pH值和电导率的影响。结果表明:枸杞在含有NaCl的营养液中培养,随着NaCl浓度的升高,枸杞的株高、鲜质量和干质量减少,根系对根际的pH值和电导率调节能力减弱;此外,枸杞能够在1.4%NaCl水培条件下生长,1.6%NaCl以上水培条件会导致枸杞落叶直至死亡。