Rapid cycling of nitrogen and phosphorus from dying roots of Lolium perenne

Summary Previous experiments, using ³²P pulse labelling, showed that when roots of Lolium perenne were detached from the shoot, a substantial proportion of the phosphorus in the roots could within a few weeks be released and be captured by another, living plant. This paper describes experiments designed to confirm and further investigate this rapid nutrient transfer. Roots from plants grown with ample N and P were detached and placed in litter bags in soil. They lost up to 60% of their initial N and up to 70% of their P in three weeks. Even when roots were grown with deficient P supply, resulting in C:P ratios of 300–400, they lost 20–30% of their initial P. Time-courses of ³²P loss from roots suspended in solution gave results which agreed with these figures. The initially rapid rate of ³²P loss had declined greatly within three weeks. In a pot experiment small L. perenne plants showed a marked increase in their N and P content during 30 days after a neighbouring large plant's shoot was removed, supporting rapid capture of nutrients lost from the detached roots. To investigate P loss from roots while attached to the shoot, L. perenne shoots were clipped every four days and ³²P loss from the roots measured. After the third clip the rate of loss increased, eventually to more than four times that from the control plants.     

Mobilization and transfer of nutrients from litter to tree seedlings via the vegetative mycelium of ectomycorrhizal plants

The ability of the mycorrhizal fungus Paxillus involutus to mobilize nitrogen and phosphorus from discrete patches of beech ( Fagus sylvatica ), birch ( Betula pendula ) and pine ( Pinus sylvestris ) litter collected from the fermentation horizon of three forest soils, and to transfer the nutrients to colonized B. pendula Roth seedlings, was investigated in transparent observation chambers. The mycelium of P. involutus foraged intensively in all three types of litter, leading to a significant decline in their phosphorus contents after 90 d. Over the same period only one of the litter types, beech, showed more than a 10% loss of its N contents. Exploitation of the litter led to invigoration of the vegetative mycelium of the fungus throughout the chambers as well as to significant increases of biomass production and leaf area in seedlings grown in the plus litter (+L) relative to those in minus litter (−L) systems. The yield increases were associated with gains in whole plant tissue content and concentration of P, but in content only in the case of N. Calculations suggest that a major proportion of the phosphorus lost from litter originated in its organic fraction. The possible basis of the discrepancy between values of N loss from litter and gain by the plant is discussed and the extent to which the distinctive pattern of nutrient mobilization is a feature peculiar to this fungus-plant combination is considered. It is concluded that nutrient mobilization from natural organic substrates in the fermentation horizon of forest soils may be a key function of the vegetative mycelium of mycorrhizal systems. The need for experimental analyses of a greater range of fungus-plant partnerships is stressed.     

Specificity of interplant cycling of phosphorus: The role of mycorrhizas

The cycling of nutrients from dying roots of one plant (the donor) to other intact plants (the receivers) was examined in a series of pot experiments. In each pot receiver plants formed either the same or a different type of mycorrhiza as the donor plant and was therefore respectively either capable or incapable of forming mycorrhizal hyphal links to the donor. There was a preferential transfer of ³²P from the dying roots of the vesicular-arbuscular mycorrhizal (VAM) Lolium perenne to VAM-infected trees Acer pseudoplatanus and Fraxinus excelsior compared to the ectomycorrhizal (ECM) Larix eurolepis, this despite an apparently greater competitive ability of L. eurolepis to obtain ³²P from the soil. Following the death of L. perenne roots there was also an increase in total P in the VAM tree receiver. These findings could not be explained by similarities in rooting distribution of the VAM-infected plants.In a similar study of the transfer of ³²P between heathland plants there was a preferential cycling of ³²P from one ericoid mycorrhizal Calluna vulgaris to another rather than to the VAM Molinia caerulea. In contrast, when ³²P was supplied directly to the soil, M. caerulea obtained significantly more ³²P than C. vulgaris. These results are discussed in relation to the potential role of interplant mycorrhizal links in the cycling of nutrients within partially closed cycles and the implications that this might have for species balance in plant communities.     

Loss of exudates from the roots of perennial ryegrass inoculated with a range of micro-organisms

To determine the effect of microbial metabolites on the release of root exudates from perennial ryegrass, seedlings were pulse labelled with [¹⁴C]-CO₂ in the presence of a range of soil micro-organisms. Microbial inoculants were spatially separated from roots by Millipore membranes so that root infection did not occur. Using this technique, only microbial metabolites affected root exudation. The effect of microbial metabolites on carbon assimilation and distribution and root exudation was determined for 15 microbial species. Assimilation of a pulse label varied by over 3.5 fold, dependent on inoculant. Distribution of the label between roots and shoots also varied with inoculant, but the carbon pool that was most sensitive to inoculation was root exudation. In the absence of a microbial inoculant only 1% of assimilated label was exuded. Inoculation of the microcosms always caused an increase in exudation but the percentage exuded varied greatly, within the range of 3–34%.     

林地覆盖经营对雷竹鞭根主要养分内循环的影响

为了给林地覆盖经营雷竹(Phyllostachys violascens)林可持续经营提供理论参考,探讨了休养式覆盖经营(覆盖3a后休养3a)、长期覆盖经营(覆盖6a)和不覆盖雷竹林(CK)2年生壮龄竹鞭及其1级、2级根N、P、K、Mg、Ca、Fe浓度和养分迁移、内循环率的差异。结果表明:不同覆盖经营年限雷竹林N、P、K、Mg、Ca和Fe浓度总体上1级根显著高于2级根。1级根和2级根中均存在N、P、K、Mg的养分内循环,且1级根养分内循环率大于2级根,Fe、Ca内循环不明显。N、P、K、Mg养分浓度与养分迁移速率随时间的推延,1级根为持续降低,2级根为先升高后降低。与不覆盖雷竹林相比,休养式林地覆盖经营总体上提高了1级、2级根的N、P、K、Ca的浓度和P、K、Mg的迁移速率、N、P、K的迁移量、P、K的养分内循环率以及1级根Mg的浓度和迁移量、2级根N的迁移速率和Mg的内循环率;长期林地覆盖经营虽提高了雷竹1级根N、K的浓度和N的迁移量及2级根N的浓度和内循环率,但总体上降低了1级根P、K、Mg和2级根N、P、Mg的迁移量与1级、2级根P、Mg的迁移速率及P、K、Mg的养分内循环率。研究表明:雷竹林鞭根中存在明显的养分内循环,且1级根对养分内循环的贡献较大。休养式林地覆盖经营利于雷竹林对养分的循环利用,而长期覆盖经营阻碍了根系对养分的平衡吸收,减弱了根系养分的内循环,不利于雷竹林的生长更新。     

Soluble carbohydrates in roots of leek (Allium porrum) plants in relation to phosphorus supply and VA mycorrhizas

Leek plants (Allium porrum L.) inoculated with Glomus mosseae were raised on sterilized soil/sand medium amended with Ca(H₂PO₄)₂.H₂O to test the hypothesis that high concentration of soil P inhibits formation of vesicular-arbuscular (VA) mycorrhizas by reducing concentration of soluble carbohydrate in the root. When P supply was increased, from either P addition or VA mycorrhizal infection, there was initially also an increase in concentration of soluble carbohydrate in the root. At the concentration of soil P at which infection was reduced, concentration of soluble carbohydrate was at its maximum. Therefore the above hypothesis is discounted. An increased delay in infection establishment and a greater number of abortive entry points would suggest that high concentration of soil P reduces VA mycorrhizal infection by changing the anatomy of the root to make it resistant to fungal penetration.     

Effects of nutrients and arbuscular mycorrhizal colonization on the growth of Salix gracilistyla seedlings in a nutrient-poor fluvial bar

A field survey and a pot culture experiment were conducted to examine the effects of nutrients (N and P) and arbuscular mycorrhizal (AM) fungi on the growth of Salix gracilistyla, a pioneer plant in riparian habitats. The plants growing in the field were colonized by AM and/or ectomycorrhizal fungi. However, the direct effect of AM colonization on seedling growth was not detected in the pot culture experiment. In contrast, N application significantly promoted plant growth, suggesting that the growth of S. gracilistyla seedlings is largely limited by the availability of N in the field.     

Formation of structures resembling ericoid mycorrhizas by the root endophytic fungus Heteroconium chaetospira within roots of Rhododendron obtusum var. kaempferi

A resynthesis study was conducted to clarify the relationship between the root endophyte, Heteroconium chaetospira and the ericaceous plant, Rhododendron obtusum var. kaempferi. The host plant roots were recovered 2 months after inoculation, and the infection process and colonization pattern of the fungus were observed under a microscope. The hyphae of H. chaetospira developed structures resembling ericoid mycorrhizas, such as hyphal coils within the host epidermal cells. These structures were morphologically the same as previously reported ericoid mycorrhizal structures. The frequencies of hyphal coils within the epidermal cells of host roots ranged from 13 to 20%. H. chaetospira did not promote or reduce host plant growth. This is the first reported study that H. chaetospira is able to form structures resembling mycorrhizas within the roots of ericaceous plants.     

The effect of soil pH on rhizosphere carbon flow of Lolium perenne

Perennial rye-grass plants were grown at 15°C in microcosms containing soil sampled from field plots that had been maintained at constant pH for the last 30 years. Six soil pH values were tested in the experiment, with pH ranging from 4.3–6.5. After 3 weeks growth in the microcosms, plant shoots were exposed to a pulse of ¹⁴C-CO₂. The fate of this label was determined by monitoring ¹⁴C-CO₂ respired by the plant roots/soil and by the shoots. The ¹⁴C remaining in plant roots and shoots was determined when the plants were harvested 7 days after receiving the pulse label. The amount of ¹⁴C (expressed as a percentage of the total ¹⁴C fixed by the plant) lost from the plant roots increased from 12.3 to 30.6% with increasing soil pH from 4.3 to 6. Although a greater percentage of the fixed ¹⁴C was respired by the root/soil as soil pH increased, plant biomass was greater with increasing soil pH. Possible reasons for observed changes in the pattern of ¹⁴C distribution are discussed and, it is suggested that changes in the soil microbial biomass and in plant nitrogen nutrition may, in particular be key factors which led to increased loss of carbon from plant roots with increasing soil pH.     

Morphological types of arbuscular mycorrhizas in pioneer woody plants growing in an oil palm farm in Sumatra, Indonesia

Morphological types of arbuscular mycorrhizas (AM) in pioneer woody plants were examined in an oil palm farm 1 year after reclamation. In total, 18 plant species belonging to 15 genera in 9 families were examined, and the Arum type was found in 12 species belonging to 9 genera in 5 families. In contrast, the Paris type was found in 5 species belonging to 5 genera in 4 families. The number of plant species with Arum-type AM was always higher than that with Paris-type AM in all four sampling plots. All the examined species in Euphorbiaceae, most of which are fast-growing pioneer trees and were dominant in forming the canopy in secondary forests, were found to have Arum-type AM. These results suggest that Arum-type colonization is beneficial for fast-growing woody plant species.     

Effect of white clover cultivar on apparent transfer of nitrogen from clover to grass and estimation of relative turnover rates of nitrogen in roots

The apparent transfer of N from clover to associated grass was evaluated over a four year period both on the basis of harvested herbage and by taking account of changes in N in stubble and root (to 10 cm depth) in swards with perennial ryegrass and three different white clover cultivars differing in leaf size. The large leaved Aran transferred 15% of its nitrogen while Huia transferred 24% and the small leaved Kent Wild White transferred 34%. When changes in stubble and root N were taken into account the percentage of N transferred was calculated to be 5% less than in harvested herbage only, as the small leaved types had proportionately more N in the roots and stolons, but the large leaved type was probably more competitive towards the grass.Loss of N from clover roots from July to October was compared to that from grass roots in a grass/white clover sward continuously stocked with steers using a method which incorporated tissue turnover and ¹⁵N dilution techniques. Less than 1 mg N m^-2 d^-1 was lost from the grass roots. In contrast 8 mg m^-2 d^-1 were estimated to be lost from clover roots while 12 mg N m^-2 d^-1 were assimilated.It is concluded that clover cultivar and competitive ability on grass have to be taken into account together with the relationship between N turnover in roots and N available for grass growth when modelling N transfer in grass/clover associations.     

A rapid staining procedure for arbuscules of living arbuscular mycorrhizas using neutral red as acidotropic dye

Arbuscules are the most conspicuous structures of arbuscular mycorrhizas. Due to their large surface area, they are regarded as putative sites of nutrient exchange between host and symbiont. A new staining technique for arbuscules is presented in which arbuscules are selectively stained by acidotropic (i.e. based on the ion-trap mechanism) accumulation of neutral red (activity stain). Criteria to distinguish acidotropic staining from staining due to chemical affinity are presented together with approaches to minimize background staining and to check for complete penetration of the dye through the root tissues. The non-toxic staining technique allows detection of arbuscules in living roots within two to six hours. The histochemical properties of neutral red, microscopic techniques, potential pitfalls, the arbuscular life cycle, and aspects of future research are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Association of a fungal endophyte in perennial ryegrass with antibiosis to larvae of the southern armyworm, Spodoptera eridania

Larvae of the Southern armyworm Spodoptera eridania Cramer (Lepidoptera:Noctuidae), feeding on perennial ryegrasses (Lolium perenne L.) infected with an endophytic fungus (Acremonium loliae Latch, Christensen and Samuels), had a much lower survival rate (7–13%) than larvae feeding on endophyte-free ryegrasses (82–90%). Death of the larvae on endophyte-infected entries occurred rapidly between 144 h and 168 h of feeding. This corresponded with armyworms feeding on the base of the plant, where endophyte concentration is highest. Twenty-four hours after the start of the bioassay the larval mass and rate of larval development were significantly higher on endophytic entries. From 48–144 h no differences were seen, but after 144 h the mass of larvae on endophyte-infected grasses sharply declined. Observations from this bioassay further substantiate the association between A. loliae-infected ryegrass and antibiosis to several lepidopterous and coleopterous insect pests.

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Résumé Le ray-grass vivace (Lolium perenne), contaminé par le champignon endophyte Acremonium loliae Latch, Christensen & Samuels, a présenté une augmentation de la resistance à de nombreux coléoptères et lépidoptères nuisibles. Cette note examine les réactions de Spodoptera eridania Cramer (Lépido., Noctuidae) alimenté sur trois lignées de ray-grass contaminées par le champignon et trois lignées saines. Après 168 h d'alimentation sur ray-grass contaminé, les chenilles presentent une très forte mortalité; la survie n'est que de 7 à 13% contre 82 à 90% pour le ray-grass sain. Le décès brutal des chenilles correspond à leur alimentation sur la base de la plante ou la concentration du champignon est la plus forte. Les chenilles consomment constamment, broutant plus des 2/3 du feuillage du ray-grass; les broutements des six séries ne différaient pas significativement.Au bout de 24 h, la nombre de chenilles passées du 3ème au 4ème stade, et l'augmentation de poids sont plus élevés pour les séries sur plante contaminée, ce qui suggère un avantage initial pour les chenilles en présence de champignon endophyte, l'analyse en poids sec a montré que l'augmentation de poids initial est réel. Entre 48 et 144 h, cependant, le nombre de 4ème stade et le poids des chenilles sont les mêmes dans les deux séries. Après 144 h, le poids des chenilles sur ray-grass contaminé diminue significativement; aucune n'était parvenue au 5ème stade, contre 11% sur ray-grass sain. Nous n'avons pas observé de signes apparents de neurotoxicité. Au lieu de cela, il ya a eu interaction avec un processus physiologique fondamental, ce qui a provoqué une forte perte de poids larvaire et la mort, indiquant l'intervention d'inhibiteurs métaboliques.

     

Does the Response of Perennial Ryegrass to Elevated CO2 Concentration Depend on the Form of the Supplied Nitrogen?

To test whether different nitrogen form (nitrate or ammonium) in substrate can alter the response to elevated partial pressure of CO₂ (pCO₂) plants of perennial ryegrass (Lolium perenne cv. Bastion) were grown from seeds in growth chambers under pCO₂ of either 35 Pa (ambient, CA) or 70 Pa (elevated, CE) in a hydroponic system (with nutrient and pH control) for 24 d. Nitrogen was supplied as ammonium, nitrate or an equimolar mixture of both N forms. Under CE plants grew faster than their counterparts under CA during the first 14 d but after 23 d of cultivation stimulation disappeared. Despite the strong positive effect of mixed forms of N on plant growth, the beneficial effect of CE was similar to that in the other two N treatments. However, the almost alike final growth response to CE had different underlying mechanisms in different N treatments. Plants supplied with nitrate as a sole source of nitrogen had lower leaf mass ratio but much higher specific leaf area compared to plants supplied with ammonium. The decrease in the content of leaf organic N (per unit of structural dry mass) under CE was found only in leaves of plants supplied with ammonium on day 14. Nevertheless, the available form of N evidently contributes to changes of leaf N content under CE. The high levels of N and non-structural saccharides in plants supplied with ammonium at CE suggest that the CO₂ response of these plants was controlled by factors other than amount of available carbon and nitrogen.     

A novel life cycle arising from leaf segments in plants regenerated from horseradish hairy roots

Summary Horseradish (Armoracia rusticana) hairy root clones were established from hairy roots which were transformed with the Ri plasmid in Agrobacterium rhizogenes 15834. The transformed plants, which were regenerated from hairy root clones, had thicker roots with extensive lateral branches and thicker stems, and grew faster compared with non-transformed horseradish plants. Small sections of leaves of the transformed plants generated adventitious roots in phytohormone-free G (modified Gamborg's) medium. Root proliferation was followed by adventitious shoot formation and plant regeneration. Approximately twenty plants were regenerated per square centimeter of leaf. The transformed plants were easily transferable from sterile conditions to soil. When leaf segments of the transformed plants were cultured in a liquid fertilizer under non-sterile conditions, adventitious roots were generated at the cut ends of the leaves. Adventitious shoots were generated at the boundary between the leaf and the adventitious roots and developed into complete plants. This novel life cycle arising from leaf segments is a unique property of the transformed plants derived from hairy root clones.     

The role of the blue mussel Mytilus edulis in the cycling of nutrients in the Oosterschelde estuary (The Netherlands)

The fluxes of particulate and dissolved material between bivalve beds and the water column in the Oosterschelde estuary have been measured in situ with a Benthic Ecosystem Tunnel. On mussel beds uptake of POC, PON and POP was observed. POC and PON fluxes showed a significant positive correlation, and the average C:N ratio of the fluxes was 9.4. There was a high release of phosphate, nitrate, ammonium and silicate from the mussel bed into the water column. The effluxes of dissolved inorganic nitrogen and phosphate showed a significant correlation, with an average N:P ratio of 16.5. A comparison of the in situ measurements with individual nutrient excretion rates showed that excretion by the mussels contributed 31–85% to the total phosphate flux from the mussel bed. Ammonium excretion by the mussels accounted for 17–94% of the ammonium flux from the mussel bed. The mussels did not excrete silicate or nitrate. Mineralization of biodeposition on the mussel bed was probably the main source of the regenerated nutrients.From the in situ observations net budgets of N, P and Si for the mussel bed were calculated. A comparison between the uptake of particulate organic N and the release of dissolved inorganic N (ammonium + nitrate) showed that little N is retained by the mussel bed, and suggested that denitrification is a minor process in the mussel bed sediment. On average, only 2/3 of the particulate organic P, taken up by the mussel bed, was recycled as phosphate. A net Si uptake was observed during phytoplankton blooms, and a net release dominated during autumn. It is concluded that mussel beds increase the mineralization rate of phytoplankton and affect nutrient ratios in the water column. A comparison of N regeneration by mussels in the central part of the Oosterschelde estuary with model estimates of total N remineralization showed that mussels play a major role in the recycling of nitrogen.     

Factors affecting formation of cluster roots in Myrica gale seedlings in water culture

The effect of nutrient deficiency, aeration, phosphorus supply, and nitrogen source on the formation of cluster (proteoid) roots was examined in Myrica gale seedlings growing in water culture. Only the omission of phosphorus resulted in the formation of significant numbers to cluster roots when plants were grown in a number of 1/4 strength Hoagland's solutions, each lacking one mineral nutrient. Aeration shortened the time required for cluster root formation and increased the percentage of plants forming cluster roots. The proportion of the root system comprised of cluster roots decreased as the phosphorus concentration in the solution increased and no cluster roots formed in solutions containing 8 mg P/L. Phosphorus supply also affected total plant biomass, proportion of biomass comprising nitrogen-fixing nodules, shoot:root ratio, phosphorus concentration in the leaves and phosphorus content of the plants. The plants showed luxury consumption of phosphorus and were able to produce large amounts of biomass utilizing only stored phosphorus.Nitrogen source also affected cluster root formation. Urea-fed plants produced cluster roots more quickly and devoted a substantially larger proportion of root growth to cluster roots than did nitrate-fed plants. The longest cluster root axes were produced in nitrate-fed plants supplied with no phosphorus and the shortest were in urea-fed plants at 4 mg P L^–1.Four methods for expressing the extent of cluster root formation were examined and it was concluded that cluster roots as a proportion of total fine root dry weight is preferable in many cases. Formation of cluster roots in response to phosphorus deficiency coupled with previously demonstrated traits allows Myrica gale to adapt to a wide range of soil conditions.