Mycorrhizal acquisition of inorganic phosphorus by the green-leaved terrestrial orchid Goodyera repens

BACKGROUND AND AIMS: Mycorrhizal fungi play a vital role in providing a carbon subsidy to support the germination and establishment of orchids from tiny seeds, but their roles in adult orchids have not been adequately characterized. Recent evidence that carbon is supplied by Goodyera repens to its fungal partner in return for nitrogen has established the mutualistic nature of the symbiosis in this orchid. In this paper the role of the fungus in the capture and transfer of inorganic phosphorus (P) to the orchid is unequivocally demonstrated for the first time. METHODS: Mycorrhiza-mediated uptake of phosphorus in G. repens was investigated using spatially separated, two-dimensional agar-based microcosms. RESULTS: External mycelium growing from this green orchid is shown to be effective in assimilating and transporting the radiotracer (33)P orthophosphate into the plant. After 7 d of exposure, over 10 % of the P supplied was transported over a diffusion barrier by the fungus and to the plants, more than half of this to the shoots. CONCLUSIONS: Goodyera repens can obtain significant amounts of P from its mycorrhizal partner. These results provide further support for the view that mycorrhizal associations in some adult green orchids are mutualistic.     

Resolving the ‘nitrogen paradox’ of arbuscular mycorrhizas: fertilization with organic matter brings considerable benefits for plant nutrition and growth

Arbuscular mycorrhizal fungi (AMF) can transfer nitrogen (N) to host plants, but the ecological relevance is debated, as total plant N and biomass do not generally increase. The extent to which the symbiosis is mutually beneficial is thought to rely on the stoichiometry of N, phosphorus (P) and carbon (C) availability. While inorganic N fertilization has been shown to elicit strong mutualism, characterized by improved plant and fungal growth and mineral nutrition, similar responses following organic N addition are lacking. Using a compartmented microcosm experiment, we determined the significance to a mycorrhizal plant of placing a ¹⁵N‐labelled, nitrogen‐rich patch of organic matter in a compartment to which only AMF hyphae had access. Control microcosms denied AMF hyphal access to the patch compartment. When permitted access to the patch compartment, the fungus proliferated extensively in the patch and transferred substantial quantities of N to the plant. Moreover, our data demonstrate that allowing hyphal access to an organic matter patch enhanced total plant N and P contents, with a simultaneous and substantial increase in plant biomass. Furthermore, we demonstrate that organic matter fertilization of arbuscular mycorrhizal plants can foster a mutually beneficial symbiosis based on nitrogen transfer, a phenomenon previously thought irrelevant.     

Nutrient transfer from soil nematodes to plants: a direct pathway provided by the mycorrhizal mycelial network

A pathway for the transfer of nutrients from dead nematodes to mycorrhizal plants is described for the first time. Plants of Betula pendula were grown in transparent microcosms in the mycorrhizal (M) or non‐mycorrhizal (NM) condition, either with or without nematode necromass of known nitrogen (N) and phosphorus (P) contents as the major potential source of these elements. Plants colonized by the mycorrhizal fungus Paxillus involutus produced greater yields and had larger N and P contents in the presence of nematodes than did their NM counterparts. The symbiotic systems were shown to exploit the N and P originally contained in necromass more effectively, and to transfer the nutrients to the plants in quantities approximately double those seen in NM systems. Even so, NM plants obtained sufficient N and P from dead nematodes to enable some enhancement of growth. Our observations confirm that mycorrhizal fungi provide the potential for the recycling of nutrients contained in this quantitatively important component of the soil mesofauna and demonstrate that the symbiotic pathway is considerably more effective than that provided by saprotrophs alone. The consequences of this nutrient transfer pathway for nutrient recycling in temperate forest ecosystems are considered.     

Effects of soil fertility and mycorrhizal infection on pollen production and pollen grain size of Cucurbita pepo (Cucurbitaceae)

The effects of soil fertility (two levels of soil nitrogen and two levels of soil phosphorus) and mycorrhizal infection on pollen production and pollen grain size were studied in two cultivars of the common zucchini (Cucurbita pepo). Overall, soil fertility and mycorrhizal infection had significant effects on traits affecting the male function of plants (staminate flower production, pollen production per flower and pollen grain size). There were also differences between the cultivars for these male traits in all three experiments. In addition, pollen grain size decreased toward the end of the growing season. In the mycorrhiza experiment, both phosphate concentration per pollen grain and total phosphate content per anther were greater but not significantly greater in the mycorrhizal plants than in the non-mycorrhizal plants. A significant negative relationship between pollen production and pollen grain size was found in the mycorrhiza and soil phosphorus experiments, indicating that there was a trade-off between pollen production and pollen size. This study is the first to show that mycorrhizal infection has an effect on male function (pollen production and size) in addition to the well-documented effects on female function (fruit/seed production and size).     

Resource allocation to growth or luxury consumption drives mycorrhizal responses

Highly variable phenotypic responses in mycorrhizal plants challenge our functional understanding of plant‐fungal mutualisms. Using non‐invasive high‐throughput phenotyping, we observed that arbuscular mycorrhizal (AM) fungi relieved phosphorus (P) limitation and enhanced growth of Brachypodium distachyon under P‐limited conditions, while photosynthetic limitation under low nitrogen (N) was exacerbated by the fungus. However, these responses were strongly dependent on host genotype: only the faster growing genotype (Bd3‐1) utilised P transferred from the fungus to achieve improved growth under P‐limited conditions. Under low N, the slower growing genotype (Bd21) had a carbon and N surplus that was linked to a less negative growth response compared with the faster growing genotype. These responses were linked to the regulation of N : P stoichiometry, couples resource allocation to growth or luxury consumption in diverse plant lineages. Our results attest strongly to a mechanism in plants by which plant genotype‐specific resource economics drive phenotypic outcomes during AM symbioses.     

拉杰沙希大学校园药用植物菌根研究

Forty different medicinal plants were investigated for arbuscular mycorrhizal association in the Rajshahi University Campus in Bangladesh. The results indicated that 35 different plants were infected by AM (arbuscular mycorrhizal) fungi as found by trypan blue staining procedure. The percentage of root colonization by AM fungi varied from 13.3% to 100%. Mangifera indica and Morus indica have maximum percentage of colonization (100%). The intensity of root colonization were abundant in the plants belonging to the families Anacardiaceae, Asclepiadaceae, Moraceae, Leguminosae and Apocynaceae whereas the intensity of colonization of crop roots were moderate and poor belonging to Gramineae and Leguminosae. The presence of greater number of spore in soil was always associated with the incidence of abundant mycelia. In plant roots the formation of spore and mycelia was restricted by low pH. Number of mycorrhizal fungus spores ranged between 35 to100 per 100g air dried soil in different family respective soils. The frequency of mycorrhizal fungus infection showed positive correlation with soil pH, moisture, water holding capacity, texture, total nitrogen, organic carbon, phosphorus, calcium, potassium, and magnesium. Especially phosphorus and nitrogen in the soil greatly influenced the plant root infection by AM fungi.     

Plant-driven weathering of apatite - the role of an ectomycorrhizal fungus

Ectomycorrhizal (EcM) fungi are increasingly recognized as important agents of mineral weathering and soil development, with far‐reaching impacts on biogeochemical cycles. Because EcM fungi live in a symbiotic relationship with trees and in close contact with bacteria and archaea, it is difficult to distinguish between the weathering effects of the fungus, host tree and other micro‐organisms. Here, we quantified mineral weathering by the fungus Paxillus involutus, growing in symbiosis with Pinus sylvestris under sterile conditions. The mycorrhizal trees were grown in specially designed sterile microcosms in which the supply of soluble phosphorus (P) in the bulk media was varied and grains of the calcium phosphate mineral apatite mixed with quartz, or quartz alone, were provided in plastic wells that were only accessed by their fungal partner. Under P limitation, pulse labelling of plants with ¹⁴CO₂ revealed plant‐to‐fungus allocation of photosynthates, with 17 times more ¹⁴C transferred into the apatite wells compared with wells with only quartz. Fungal colonization increased the release of P from apatite by almost a factor of three, from 7.5 (±1.1) × 10^?10 mol m^?2 s^?1 to 2.2 (±0.52) × 10^?9 mol m^?2 s^?1. On increasing the P supply in the microcosms from no added P, through apatite alone, to both apatite and orthophosphate, the proportion of biomass in roots progressively increased at the expense of the fungus. These three observations, (i) proportionately more plant energy investment in the fungal partner under P limitation, (ii) preferential fungal transport of photosynthate‐derived carbon towards patches of apatite grains and (iii) fungal enhancement of weathering rate, reveal the tightly coupled plant–fungal interactions underpinning enhanced EcM weathering of apatite and its utilization as P source.     

Mycorrhizal fungi suppress aggressive agricultural weeds

Plant growth responses to arbuscular mycorrhizal fungi (AMF) are highly variable, ranging from mutualism in a wide range of plants, to antagonism in some non-mycorrhizal plant species and plants characteristic of disturbed environments. Many agricultural weeds are non mycorrhizal or originate from ruderal environments where AMF are rare or absent. This led us to hypothesize that AMF may suppress weed growth, a mycorrhizal attribute which has hardly been considered. We investigated the impact of AMF and AMF diversity (three versus one AMF taxon) on weed growth in experimental microcosms where a crop (sunflower) was grown together with six widespread weed species. The presence of AMF reduced total weed biomass with 47% in microcosms where weeds were grown together with sunflower and with 25% in microcosms where weeds were grown alone. The biomass of two out of six weed species was significantly reduced by AMF (?66% & ?59%) while the biomass of the four remaining weed species was only slightly reduced (?20% to ?37%). Sunflower productivity was not influenced by AMF or AMF diversity. However, sunflower benefitted from AMF via enhanced phosphorus nutrition. The results indicate that the stimulation of arbuscular mycorrhizal fungi in agro-ecosystems may suppress some aggressive weeds.     

Carbon transfer associated with assimilation of organic nitrogen sources by silver birch (Betula pendula Roth.)

Summary Qualitative and quantitative aspects of heterotrophic carbon assimilation by mycorrhizal plants of birch (Betula pendula) were examined. Plants were grown aseptically from seed in the mycorrhizal condition with the fungus Hebeloma crustuliniforme and in the non-mycorrhizal condition, with protein as their sole exogenous nitrogen source. Yields and nitrogen contents were determined in some of the plants, while the roots of others were supplied with ¹⁴C-labelled protein and their shoots exposed for up to 72 h to different irradiance regimes. Only mycorrhizal plants utilised the organic nitrogen. Uptake of carbon associated with this utilisation and its translocation to the leaves was demonstrated directly by means of autoradiography. Amounts of activity transferred to shoots were greatest in low irradiance regimes. Calculation of net carbon gain from the heterotrophic source, based upon the assumption that breakdown products of protein are assimilated as amino-acids, indicates that over a 55-day growth period up to 9% of plant C may be derived from protein. The physiological and ecological significance of these findings are discussed.     

丛枝菌根真菌对枳根净离子流及锌污染下枳苗矿质营养的影响

盆栽实验研究了不同施Zn水平(0、300 mg/kg和600 mg/kg)下,接种丛枝菌根真菌Glomus intraradices对枳苗生长、Zn、Cu、P、K、Ca、Mg分布的影响,并采用非损伤微测技术测定分析了菌根化与非菌根化枳根净Ca²⁺、H⁺、NO₃^-离子流动态。结果表明:(1)在不同施Zn水平下,接种菌根真菌显著提高了枳苗地上部及根部鲜重;随着施Zn水平的提高,菌根侵染率呈降低趋势,枳苗地上部与根部Zn含量呈增加趋势,且接种株根部Zn含量显著高于未接种株。(2)接种株未施Zn处理的地上部Cu、P、K、Mg和根部Cu含量、施600 mg/kg Zn处理的根部Cu及施300 mg/kg Zn处理的根部P含量均显著高于对照,而菌根真菌侵染对枳苗Ca含量并无显著性影响。(3)接种株未施Zn处理的根部距根尖端0 μm和600 μm处净Ca²⁺流出速率、600 μm处净H⁺流入速率、2400 μm处净NO₃^-流入速率均显著高于未接种株。     

Cycling of nutrients from dying roots to living plants,including the role of mycorrhizas

This paper presents information about the release of nitrogen and phosphorus from dying grass roots and the capture of phosphorus by other, living plants. We have paid particular attention to the part played by mycorrhizas in this phosphorus capture, and the possible importance of mycorrhizal links between dying and living roots.WhenLolium perenne plants were grown with ample nutrients and their roots then detached and buried in soil, about half the nitrogen and two-thirds of the phosphorus was lost in three weeks, but only one-fifth of the dry weight. The C:N and C:P ratios suggest that microbial growth in the roots would at first be C-limited but would become N- and P-limited within three weeks.Rapid transfer of³²P can occur from dying roots to those of a living plant if the two root systems are intermingled. The amount transferred was substantially increased in two species-combinations that are known to form mycorrhizal links between their root systems. In contrast, in a species-combination where only the living (receiver) plant could become mycorrhizal no significant increase of³²P transfer occurred. This evidence, although far from conclusive, suggests that mycorrhizal links between dying and living roots can contribute to nutrient cycling. This research indicates a major difference in nutrient cycling processes between perennial and annual crops.     

Effect of diazotrophic bacteria isolated from a mycelium of arbuscular mycorrhizal fungi on colonization of maize roots by Glomus fistulosum

The inoculation of mycorrhizal maize plants with three isolates of microaerophilic diazotrophic bacteria obtained from the mycelium of arbuscular mycorrhizal fungi associated with three grasses (Arrhenatherum elatius - bacterial isolate ARR, Agropyrum repens - isolate AGR and Poa annua - isolate POA) caused no increase in nitrogen content in plant biomass. The inoculation with bacterial isolate ARR resulted in the decreased plant growth. Bacterial isolate AGR decreased the percentage of the root length colonized by arbuscular mycorrhizal fungus Glomus fistulosum. The inoculation with both mycorrhizal fungus and isolate POA increased significantly the concentration of phosphorus in plant shoots compared to uninoculated control. This revised version was published online in July 2006 with corrections to the Cover Date.     

Root symbioses ofAlnus glutinosa (L.) gaertn. and their possible role in alder decline: A preliminary study

During the last few years alder has declined in South Bohemia. The possible role of mycorrhizal and actinorhizal symbioses is reviewed and some of the preliminary results from experiments testing the influence of these symbioses on alder growth and the influence of eutrophication on the development of these symbioses are reported. Seedlings ofAlnus glutinosa were inoculated with arbuscular mycorrhizal (AM) fungi and the actinomyceteFrankia in experiment 1, and with rhizosphere soil collected from field sites with different degrees of alder damage in experiment 2. In both experiments, a solution containing nitrate, ammonia and phosphorus in concentrations simulating eutrophic waters, was applied. Both symbioses markedly promoted the growth of the seedlings in experiment 1. The plants inoculated with the rhizosphere soil microflora in experiment 2 were larger than the control plants. Response of the seedlings to the inoculation with the soil from the rhizosphere of damaged alder trees from six field sites differs, even though no correlation was found relating growth to the health status of the trees. Nutrient treatment did not have any effect on the growth of seedlings in either experiment. The dry weight ofFrankia was greater in mycorrhizal plants compared to nonmycorrhizal plants and mycorrhizal colonization is reduced inFrankia inoculated plants supplemented with phosphorus in experiment 1. Nitrogen enhanced mycorrhizal colonization in nodulated plants which were not supplemented with phosphorus no effect of nitrogen on actinorhiza was observed.     

Studies of iron transport by arbuscular mycorrhizal hyphae from soil to peanut and sorghum plants

 The influence of an arbuscular mycorrhizal (AM) fungus on phosphorus (P) and iron (Fe) uptake of peanut (Arachis hypogea L.) and sorghum (Sorghum bicolor L.) plants was studied in a pot experiment under controlled environmental conditions. The plants were grown for 10 weeks in pots containing sterilised calcareous soil with two levels of Fe supply. The soil was inoculated with rhizosphere microorganisms only or with rhizosphere microorganisms together with an AM fungus (Glomus mosseae [Nicol. & Gerd.] Gerdemann & Trappe). An additional small soil compartment accessible to hyphae but not roots was added to each pot after 6 weeks of plant growth. Radiolabelled P and Fe were supplied to the hyphae compartment 2 weeks after addition of this compartment. After a further 2 weeks, plants were harvested and shoots were analysed for radiolabelled elements. In both plant species, P uptake from the labelled soil increased significantly more in shoots of mycorrhizal plants than non-mycorrhizal plants, thus confirming the well-known activity of the fungus in P uptake. Mycorrhizal inoculation had no significant influence on the concentration of labelled Fe in shoots of peanut plants. In contrast, ⁵⁹Fe increased in shoots of mycorrhizal sorghum plants. The uptake of Fe from labelled soil by sorghum was particularly high under conditions producing a low Fe nutritional status of the plants. These results are preliminary evidence that hyphae of an arbuscular mycorrhizal fungus can mobilise and/or take up Fe from soil and translocate it to the plant. Accepted: 6 March 1998     

Plant responsiveness to mycorrhizas differs from dependence upon mycorrhizas

Soil phosphorus response curves of plants with and without mycorrhizas reflect two different, but complementary, phenomena. The first, plant responsiveness to mycorrhizas, is represented by the difference in growth between plants with and without mycorrhizas at any designated level of phosphorus availability. This is also a measure of mycorrhizal fungus effectiveness. The second, the lowest level of phosphorus availability at which plants can grow without mycorrhizas, is here termed dependence upon mycorrhizas. The latter definition differs from conventional usage which fails to distinguish dependence from responsiveness. Sigmoid curves generated by the three-parameter, logistic equation generally can model the responses of plants to mycorrhizas and phosphorus addition and can be used to assess responsiveness, effectiveness, and dependence. Such curves reveal that plant responsiveness or fungus effectiveness determined at a single level of phosphorus availability may be misleading when used to compare different host species' intrinsic capacities to respond to different mycorrhizal fungus species. Instead, the same relative position should be evaluated among phosphorus response curves for different species combinations. Dependence of a plant species known to benefit from mycorrhizas can be assessed with reference to only the phosphorus response curve of plants without mycorrhizas. Dependence is a constitutive property of plant species that can be used to classify them as facultatively or obligately mycotrophic. Dependence is a plant attribute upon which natural selection can act, but responsiveness and effectiveness cannot be selected directly because they are emergent properties of the interaction between plant and fungus species.     

Nitrogen delivery to maize via mycorrhizal hyphae depends on the form of N supplied

Arbuscular mycorrhizal fungi can enhance nutrient acquisition by a plant via their extraradical hyphae. This is particularly true for phosphorus, but the case for nitrogen (N) has been less clear. In our growth systems there was a small air-gap between root and hyphal compartments, which eliminated diffusion of nutrients between compartments. Moreover, our methods allowed us to distinguish between nitrate and ammonium. We found that N transfer to Zea maize L. depends on the sources fed to the hyphae of Glomus aggregatum Schenck & Smith. In experiment 1, despite the fact that plant demand for N was already met, plants received 10 times as much ¹⁵N from ammonium than from nitrate. In experiment 2, 74% of shoot-N was derived from the slow-release urea added to the hyphal compartment while only 2.9% was derived from the nitrate-N. Intraradical hyphae isolated from roots contained a considerable amount of ¹⁵N in the cell wall even when ¹⁵N-nitrate was the source. We conclude that the mycorrhizal fungus can rapidly deliver ammonium-N to the plants, and that while the fungus can absorb nitrate, it apparently lacks the capacity to transfer it to the plant.     

Carbon and nitrogen flow in silver birch and Norway spruce connected by a common mycorrhizal mycelium

 Spruce and birch seedlings were grown together in boxes filled with unsterile peat. Both seedlings were colonized by the ectomycorrhizal fungus Scleroderma citrinum. The two plants thus shared a common external mycelium. ¹⁵N-labelled ammonium was supplied exclusively to the fungus, while the birch or the spruce plant was continuously fed with ¹³C-labelled CO₂ for 72 h. The carbon and nitrogen transfer rates were strikingly different for birch and spruce seedlings. The mycorrhizal mycelium received carbohydrates mainly from the birch plant and the nitrogen transfer by the fungus to the plants was largely directed towards the birch. Carbon assimilates were also transferred in both directions between birch and spruce; however, there was no conclusive evidence for a net transfer of carbon between the plants. Accepted: 20 September 1996     

Functional diversity of arbuscular mycorrhizal fungal isolates in relation to extraradical mycelial networks

We investigated the functional significance of extraradical mycorrhizal networks produced by geographically different isolates of the arbuscular mycorrhizal fungal (AMF) species Glomus mosseae and Glomus intraradices. A two-dimensional experimental system was used to visualize and quantify intact extraradical mycelium (ERM) spreading from Medicago sativa roots. Growth, phosphorus (P) and nitrogen (N) nutrition were assessed in M. sativa plants grown in microcosms. The AMF isolates were characterized by differences in extent and interconnectedness of ERM. Phenotypic fungal variables, such as total hyphal length, hyphal density, hyphal length per mm of total or colonized root length, were positively correlated with M. sativa growth response variables, such as total shoot biomass and plant P content. The utilization of an experimental system in which size, growth rate, viability and interconnectedness of ERM extending from mycorrhizal roots are easily quantified under realistic conditions allows the simultaneous evaluation of different isolates and provides data with a predictive value for selection of efficient AMF.     

接种AM真菌对玉米和油菜种间竞争及土壤无机磷组分的影响

在温室盆栽条件下,分别模拟单作、间作和尼龙网分隔种植,比较接种丛枝菌根(arbuscular mycorrhizal, AM)真菌Glomus intraradices和Glomus mosseae对菌根植物玉米和非菌根植物油菜生长和磷吸收状况的影响,并分析土壤中各无机磷组分的变化。结果发现,接种AM真菌可以促进土壤中难溶性磷(Ca₁₀-P和O-P)向有效态磷转化,并显著降低总无机磷含量 (P<0.05),显著提高菌根植物玉米的生物量和磷吸收量(P<0.05),特别是在间作体系中使玉米的磷营养竞争比率显著提高了45.0%-104.1% (P<0.05),显著降低了油菜的生物量和磷吸收量(P<0.05),从而增强了了菌根植物的竞争优势,降低了非菌根植物与菌根植物的共存能力。揭示了石灰性土壤中AM真菌对植物物种多样性的影响,有助于更加全面地理解AM真菌在农业生态系统中的作用。     

Glycine-Glomus-Rhizobium Symbiosis : VI. Photosynthesis in Nodulated, Mycorrhizal, or N- and P-Fertilized Soybean Plants

Soybean (Glycine max [L.] Merr. cv Hobbit) plants were grown in a growth chamber for 56 days in a phosphorus- and nitrogen-deficient soil and were colonized by the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae (Nicol. & Gerd) Gerd. and Trappe and Rhizobium japonicum strain USDA 136, or by either organism alone, or by neither. Non-VAM plants received supplemental phosphorus and nonnodulated plants supplemental nitrogen to achieve the same rate of growth in all treatments. Plants of all four treatments had the same (P > 0.05) dry weights at harvest, but VAM plants had higher rates of CO₂ exchange (CER, P < 0.05) and lower leaf P concentrations (P < 0.01). Leaf nitrogen concentrations were lower in nodulated than in nitrogen-supplemented plants (P < 0.01) while starch concentrations were higher (P < 0.01). There was a significant negative relationship between nitrogen and starch (r = −0.989). Statistical evaluation of the data showed that some parameters (CER, leaf area and phosphorus content) were associated with phosphorus nutrition (or the presence of the VAM fungus), others (leaf fresh weight and root dry weight) with nitrogen nutrition (or the presence of Rhizobium), and some (leaf nitrogen and starch content) by both factors. The development of microsymbiont structures and nodule activity were significantly lower in the tripartite association than in plants colonized by one endophyte only. The findings suggest that endophyte effects go beyond those of simple nutrition and associated source-sink relationships.