Functional aspects of root architecture and mycorrhizal inoculation with respect to nutrient uptake capacity

The aim of this research was to investigate the effect of arbuscular mycorrhizal (AM) colonisation on root morphology and nitrogen uptake capacity of carob ( Ceratonia siliqua L.) under high and low nutrient conditions. The experimental design was a factorial arrangement of presence/absence of mycorrhizal fungus inoculation ( Glomus intraradices) and high/low nutrient status. Percent AM colonisation, nitrate and ammonium uptake capacity, and nitrogen and phosphorus contents were determined in 3-month-old seedlings. Grayscale and colour images were used to study root morphology and topology, and to assess the relation between root pigmentation and physiological activities. AM colonisation lead to a higher allocation of biomass to white and yellow parts of the root. Inorganic nitrogen uptake capacity per unit root length and nitrogen content were greatest in AM colonised plants grown under low nutrient conditions. A better match was found between plant nitrogen content and biomass accumulation, than between plant phosphorus content and biomass accumulation. It is suggested that the increase in nutrient uptake capacity of AM colonised roots is dependent both on changes in root morphology and physiological uptake potential. This study contributes to an understanding of the role of AM fungi and root morphology in plant nutrient uptake and shows that AM colonisation improves the nitrogen nutrition of plants, mainly when growing at low levels of nutrients.     

Modelling the contribution of arbuscular mycorrhizal fungi to plant phosphate uptake

In this paper, we investigate the role of arbuscular mycorrhizal fungi in plant phosphorus nutrition. We develop a mathematical model which quantitatively assesses the contribution of external fungal hyphae to plant phosphate uptake.We derive an equation for solute uptake by a growing fungal mycelium which we couple with a model for root uptake. We analyse the model using nondimensionalization and numerical simulations.Simulations predict that removal of phosphate from soil is dominated by hyphal uptake as opposed to root uptake. Model analysis shows that the depletion zones around hyphae overlap within 8 h and that the transfer between fungus and root is a critical step for the behaviour of phosphorus within the mycelial phase. We also show that the volume fraction of mycelium is negligibly small in comparison to other soil phases.This is the first model to quantify the contribution of mycorrhizal fungi to plant phosphate uptake. A full data set for model parametrization and validation is not currently available. Therefore, more complete sets of experimental measurements are necessary to make this model more applicable.     

菌根真菌与植物共生营养交换机制研究进展

菌根是陆地生态系统普遍存在的、由土壤中的菌根真菌侵染宿主植物根系形成的联合共生体.菌根的建立是以共生体双方的营养交换为基础的:菌根真菌从土壤中吸收氮、磷等营养物质并转运给宿主植物,供其生长;作为交换,植物则以脂质或糖的形式向菌根真菌提供其生长所必需的碳水化合物.近年来,菌根真菌与宿主植物间的营养交换机制一直是研究的热点,国内外对菌根真菌介导的植物营养物质吸收和转运机制的研究也取得了巨大进展.本文综述了丛枝和外生两种菌根真菌与宿主植物间营养交换的最新研究进展,尤其是碳、氮、磷等几种重要营养物质的吸收与双向转运机制,以及营养交换在菌根形成中的潜在调控作用,并对目前存在的关键问题和未来研究方向进行了分析和展望,这对菌根模型的建立及菌根效益的优化具有重要意义.     

Pathways of Phosphorus Absorption and Early Signaling between the Mycorrhizal Fungi and Plants

This review highlights the key role that mycorrhizal fungi play in making phosphorus (Pi) more available to plants, including pathways of phosphorus absorption, phosphate transporters and plant-mycorrhizal fungus symbiosis, especially in conditions where the level of inorganic phosphorus (Pi) in the soil is low. Mycorrhizal fungi colonization involves a series of signaling where the plant root exudates strigolactones, while the mycorrhizal fungi release a mixture of chito-oligosaccharides and liposaccharides, that activate the symbiosis process through gene signaling pathways, and contact between the hyphae and the root. Once the symbiosis is established, the extraradical mycelium acts as an extension of the roots and increases the absorption of nutrients, particularly phosphorus by the phosphate transporters. Pi then moves along the hyphae to the plant root/fungus interface. The transfer of Pi occurs in the apoplectic space; in the case of arbuscular mycorrhizal fungi, Pi is discharged from the arbuscular to the plant’s root symplasm, in the membrane that surrounds the arbuscule. Pi is then absorbed through the plant periarbuscular membrane by plant phosphate transporters. Furthermore, plants can acquire Pi from soil as a direct absorption pathway. As a result of this review, several genes that codify for high-affinity Pi transporters were identified. In plants, the main family is Pht1 although it is possible to find others such as Pht2, Pht3, Pho1 and Pho2. As in plants, mycorrhizal fungi have genes belonging to the Pht1 subfamily. In arbuscular mycorrhizal fungi we found L1PT1, GiPT, MtPT1, MtPT2, MtPT4, HvPT8, ZmPht1, TaPTH1.2, GmosPT and LYCes. HcPT1, HcPT2 and BePT have been characterized in ectomycorrhizal fungi. Each gene has a different way of expressing itself. In this review, we present diagrams of the symbiotic relationship between mycorrhizal fungi and the plant. This knowledge allows us to design solutions to regional problems such as food production in soils with low levels of Pi.

     

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.     

不同林龄杉木人工林菌根侵染特征研究

丛枝菌根真菌是一种通过土壤侵染植物根系,与寄主植物互利共生的重要有益真菌。探究不同林龄杉木林中菌根侵染状况与土壤养分的变化规律,有利于深入认识丛枝菌根真菌—杉木相互作用的养分调控因素,从而为改善杉木人工林土壤肥力、促进杉木林可持续经营提供依据。分别选取10a、25a、45a杉木纯林,分析了不同林分菌根侵染率与孢子密度及部分土壤养分因子(全磷、速效磷、全钾、速效钾)的关系。结果表明:(1)菌根侵染率与孢子密度均呈现出随林龄增大而增大的趋势,pH随林龄增大而减少的趋势;(2)根际土中磷的含量总体偏低,而且受到土壤酸化流失和丛枝菌根真菌积累的双重影响呈现出先减少后增加的趋势;(3)虽然有效钾含量随林龄变化趋势不显著,但丛枝菌根真菌能促进土壤钾的积累。因此,丛枝菌根真菌能有效调控根际土的养分动力学特征,减缓土壤酸化造成的养分流失。     

丛枝菌根真菌与氮添加对不同根形态基因型水稻氮吸收的影响

植物主要依赖自身根系从土壤中获取矿质养分; 具有不同根形态的植物对于养分的吸收能力存在差异。丛枝菌根真菌(AMF)能与陆地植物根系形成共生关系, 帮助植物吸收矿质养分。但是, AMF对于植物根系养分吸收的促进效应是否会受根形态的影响还鲜有研究。该研究选取4种不同根形态基因型水稻(根毛缺陷突变体rhl1、侧根缺陷突变体iaa11、不定根缺失突变体arl1和野生型Kas)为研究对象, 设置2种施氮水平处理(低氮: 20 mg·kg^-1氨氮; 高氮: 100 mg·kg^-1氨氮), 利用稳定同位素¹⁵N示踪标记技术, 探究AMF和氮添加对不同根形态植物氮吸收的影响。研究结果发现, 相比低氮处理, 高氮处理下, rhl1、Kas、iaa11与arl1的茎叶¹⁵N浓度分别提高了60%、72%、128%与118%, 说明氮添加显著促进了水稻氮吸收, 且iaa11与arl1对氮添加的响应更强烈。在低氮水平下, AMF对rhl1、Kas、iaa11与arl1氮吸收的平均效应值分别为17%、31%、42%、51%, 表明AMF对于植物氮吸收的促进效应受根形态影响, iaa11与arl1对AMF的响应明显高于Kas与rhl1; 相较于低氮水平, 高氮水平下AMF对于不同根形态水稻氮吸收的促进效应都会显著降低, 表明氮添加削弱了AMF对植物氮吸收的促进效应。该研究阐明了4种不同根形态基因型水稻氮养分吸收存在显著差异, 其中氮吸收能力较弱的基因型水稻对AMF的响应更强, 该结果补充了植物与AMF在养分吸收上存在功能互补的控制实验证据。     

Plant–microbes interactions in enhanced fertilizer-use efficiency

The continued use of chemical fertilizers and manures for enhanced soil fertility and crop productivity often results in unexpected harmful environmental effects, including leaching of nitrate into ground water, surface run-off of phosphorus and nitrogen run-off, and eutrophication of aquatic ecosystems. Integrated nutrient management systems are needed to maintain agricultural productivity and protect the environment. Microbial inoculants are promising components of such management systems. This review is a critical summary of the efforts in using microbial inoculants, including plant growth-promoting rhizobacteria and arbuscular mycorrhizal fungi for increasing the use efficiency of fertilizers. Studies with microbial inoculants and nutrients have demonstrated that some inoculants can improve plant uptake of nutrients and thereby increase the use efficiency of applied chemical fertilizers and manures. These proofs of concept studies will serve as the basis for vigorous future research into integrated nutrient management in agriculture.     

Quantification of water uptake by arbuscular mycorrhizal hyphae and its significance for leaf growth, water relations, and gas exchange of barley subjected to drought stress

Arbuscular mycorrhizal fungi alleviate drought stress in their host plants via the direct uptake and transfer of water and nutrients through the fungal hyphae to the host plants. To quantify the contribution of the hyphae to plant water uptake, a new split-root hyphae system was designed and employed on barley grown in loamy soil inoculated with Glomus intraradices under well-watered and drought conditions in a growth chamber with a 14-h light period and a constant temperature (15 degrees C; day/night). Drought conditions were initiated 21 days after sowing, with a total of eight 7-day drying cycles applied. Leaf water relations, net photosynthesis rates, and stomatal conductance were measured at the end of each drying cycle. Plants were harvested 90 days after sowing. Compared to the control treatment, the leaf elongation rate and the dry weight of the shoots and roots were reduced in all plants under drought conditions. However, drought resistance was comparatively increased in the mycorrhizal host plants, which suffered smaller decreases in leaf elongation, net photosynthetic rate, stomatal conductance, and turgor pressure compared to the non-mycorrhizal plants. Quantification of the contribution of the arbuscular mycorrhizal hyphae to root water uptake showed that, compared to the non-mycorrhizal treatment, 4 % of water in the hyphal compartment was transferred to the root compartment through the arbuscular mycorrhizal hyphae under drought conditions. This indicates that there is indeed transport of water by the arbuscular mycorrhizal hyphae under drought conditions. Although only a small amount of water transport from the hyphal compartment was detected, the much higher hyphal density found in the root compartment than in the hyphal compartment suggests that a larger amount of water uptake by the arbuscular mycorrhizal hyphae may occur in the root compartment.     

Laboratory and field methods for measurement of hyphal uptake of nutrients in soil

Experimental systems for measuring nutrient transport by arbuscular mycorrhizal (AM) fungi in soil are described. The systems generally include two soil compartments that are separated by fine nylon mesh. Both roots and root-external hyphae grow in one compartment, but only hyphae are fine enough to grow through the mesh into the other compartment. Application of tracer isotopes to the soil of this hyphal compartment can be used to measure nutrient uptake by plants via AM fungal hyphae. Use of compartmented systems is discussed with particular reference to phosphorus, which is the mineral nutrient transported in the largest quantity by AM fungi. Laboratory and field applications of the compartmentation methodology are presented with emphasis on the functioning of native AM fungal communities. Advantages and limitations of the method are considered and future important research directions are discussed in this context. This revised version was published online in June 2006 with corrections to the Cover Date.     

At the Root of the Wood Wide Web: Self Recognition and Non-Self Incompatibility in Mycorrhizal Networks

Arbuscular mycorrhizal (AM) fungi are mutualistic symbionts living in the roots of 80% of land plant species, and developing extensive, below-ground extraradical hyphae fundamental for the uptake of soil nutrients and their transfer to host plants. Since AM fungi have a wide host range, they are able to colonize and interconnect contiguous plants by means of hyphae extending from one root system to another. Such hyphae may fuse due to the widespread occurrence of anastomoses, whose formation depends on a highly regulated mechanism of self recognition. Here, we examine evidences of self recognition and non-self incompatibility in hyphal networks formed by AM fungi and discuss recent results showing that the root systems of plants belonging to different species, genera and families may be connected by means of anastomosis formation between extraradical mycorrhizal networks, which can create indefinitely large numbers of belowground fungal linkages within plant communities.Key Words: arbuscular mycorrhizal symbiosis, extraradical mycelium, anastomosis, plant interconnectedness, self recognition, non-self incompatibility, mycorrhizal networks     

Acquisition of phosphorus and nitrogen in the rhizosphere and plant growth promotion by microorganisms

The rhizosphere is a complex environment where roots interact with physical, chemical and biological properties of soil. Structural and functional characteristics of roots contribute to rhizosphere processes and both have significant influence on the capacity of roots to acquire nutrients. Roots also interact extensively with soil microorganisms which further impact on plant nutrition either directly, by influencing nutrient availability and uptake, or indirectly through plant (root) growth promotion. In this paper, features of the rhizosphere that are important for nutrient acquisition from soil are reviewed, with specific emphasis on the characteristics of roots that influence the availability and uptake of phosphorus and nitrogen. The interaction of roots with soil microorganisms, in particular with mycorrhizal fungi and non-symbiotic plant growth promoting rhizobacteria, is also considered in relation to nutrient availability and through the mechanisms that are associated with plant growth promotion.     

A phosphate transporter gene from the extra-radical mycelium of an arbuscular mycorrhizal fungus Glomus intraradices is regulated in response to phosphate in the environment

The majority of vascular flowering plants are able to form symbiotic associations with arbuscular mycorrhizal fungi. These symbioses, termed arbuscular mycorrhizas, are mutually beneficial, and the fungus delivers phosphate to the plant while receiving carbon. In these symbioses, phosphate uptake by the arbuscular mycorrhizal fungus is the first step in the process of phosphate transport to the plant. Previously, we cloned a phosphate transporter gene involved in this process. Here, we analyze the expression and regulation of a phosphate transporter gene (GiPT) in the extra-radical mycelium of the arbuscular mycorrhizal fungus Glomus intraradices during mycorrhizal association with carrot or Medicago truncatula roots. These analyses reveal that GiPT expression is regulated in response to phosphate concentrations in the environment surrounding the extra-radical hyphae and modulated by the overall phosphate status of the mycorrhiza. Phosphate concentrations, typical of those found in the soil solution, result in expression of GiPT. These data imply that G. intraradices can perceive phosphate levels in the external environment but also suggest the presence of an internal phosphate sensing mechanism.     

Phosphorus and Nitrogen Regulate Arbuscular Mycorrhizal Symbiosis in Petunia hybrida

Phosphorus and nitrogen are essential nutrient elements that are needed by plants in large amounts. The arbuscular mycorrhizal symbiosis between plants and soil fungi improves phosphorus and nitrogen acquisition under limiting conditions. On the other hand, these nutrients influence root colonization by mycorrhizal fungi and symbiotic functioning. This represents a feedback mechanism that allows plants to control the fungal symbiont depending on nutrient requirements and supply. Elevated phosphorus supply has previously been shown to exert strong inhibition of arbuscular mycorrhizal development. Here, we address to what extent inhibition by phosphorus is influenced by other nutritional pathways in the interaction between Petunia hybrida and R. irregularis. We show that phosphorus and nitrogen are the major nutritional determinants of the interaction. Interestingly, the symbiosis-promoting effect of nitrogen starvation dominantly overruled the suppressive effect of high phosphorus nutrition onto arbuscular mycorrhiza, suggesting that plants promote the symbiosis as long as they are limited by one of the two major nutrients. Our results also show that in a given pair of symbiotic partners (Petunia hybrida and R. irregularis), the entire range from mutually symbiotic to parasitic can be observed depending on the nutritional conditions. Taken together, these results reveal complex nutritional feedback mechanisms in the control of root colonization by arbuscular mycorrhizal fungi.     

Phosphate transporters of Medicago truncatula and arbuscular mycorrhizal fungi

Most vascular plants acquire phosphate from their environment either directly, via the roots, or indirectly, via a symbiotic interaction with arbuscular mycorrhizal (AM) fungi. The symbiosis develops in the plant roots where the fungi colonize the cortex of the root to obtain carbon from the plant host, while assisting the plant with acquisition of phosphate and other mineral nutrients from the soil solution. As a first step toward understanding the molecular basis of the symbiosis and phosphate utilization, we have cloned and characterized phosphate transporter genes from the AM fungi Glomus versiforme and Glomus intraradices, and from the roots of a host plant, Medicago truncatula. Expression analyses and localization studies indicate that each of these transporters has a role in phosphate uptake from the soil solution.     

植物菌根共生磷酸盐转运蛋白

大多数植物能和丛枝菌根（arbuscular mycorrhiza, AM）真菌形成菌根共生体。AM能够促进植物对土壤中矿质营养的吸收,尤其是磷的吸收。磷的吸收和转运由磷酸盐转运蛋白介导。总结了植物AM磷酸盐转运蛋白及其结构特征,分析其分类及系统进化,并综述了AM磷酸盐转运蛋白介导的磷的吸收和转运过程及其基因的表达调控。植物AM磷酸盐转运蛋白属于Pht1家族成员,它不仅对磷的吸收和转运是必需的,而且对AM共生也至关重要,为进一步了解菌根形成的分子机理及信号转导途径提供了理论基础。     

A H+-ATPase That Energizes Nutrient Uptake during Mycorrhizal Symbioses in Rice and Medicago truncatula

Most plant species form symbioses with arbuscular mycorrhizal (AM) fungi, which facilitate the uptake of mineral nutrients such as phosphate from the soil. Several transporters, particularly proton-coupled phosphate transporters, have been identified on both the plant and fungal membranes and contribute to delivering phosphate from fungi to plants. The mechanism of nutrient exchange has been studied in plants during mycorrhizal colonization, but the source of the electrochemical proton gradient that drives nutrient exchange is not known. Here, we show that plasma membrane H⁺-ATPases that are specifically induced in arbuscule-containing cells are required for enhanced proton pumping activity in membrane vesicles from AM-colonized roots of rice (Oryza sativa) and Medicago truncatula. Mutation of the H⁺-ATPases reduced arbuscule size and impaired nutrient uptake by the host plant through the mycorrhizal symbiosis. Overexpression of the H⁺-ATPase Os-HA1 increased both phosphate uptake and the plasma membrane potential, suggesting that this H⁺-ATPase plays a key role in energizing the periarbuscular membrane, thereby facilitating nutrient exchange in arbusculated plant cells.     

A critical review on the role of mycorrhizal fungi in the uptake of phosphorus by plants

The beneficial effects of mycorrhizae on plant growth have often been related to the increase in the uptake of immobile nutrients, especially phosphorus (P). In this review the mechanisms for the increase in the uptake of P by mycorrhizae and the sources of soil P for mycorrhizal and non-mycorrhizal plants are examined.Various mechanisms have been suggested for the increase in the uptake of P by mycorrhizal plants. These include: exploration of larger soil volume; faster movement of P into mycorrhizal hyphae; and solubilization of soil phosphorus. Exploration of larger soil volume by mycorrhizal plants is achieved by decreasing the distance that P ions must diffuse to plant roots and by increasing the surface area for absorption. Faster movement of P into mycorrhizal hyphae is achieved by increasing the affinity for P ions and by decreasing the threshold concentration required for absorption of P. Solubilization of soil P is achieved by the release of organic acids and phosphatase enzymes. Mycorrhizal plants have been shown to increase the uptake of poorly soluble P sources, such as iron and aluminium phosphate and rock phosphates. However, studies in which the soil P has been labelled with radioactive ³²P indicated that both mycorrhizal and non-mycorrhizal plants utilized the similarly labelled P sources in soil.     

Nutrient uptake in mycorrhizal symbiosis

The role of mycorrhizal fungi in acquisition of mineral nutrients by host plants is examined for three groups of mycorrhizas. These are; the ectomycorrhizas (ECM), the ericoid mycorrhizas (EM), and the vesicular-arbuscular mycorrhizas (VAM). Mycorrhizal infection may affect the mineral nutrition of the host plant directly by enhancing plant growth through nutrient acquisition by the fungus, or indirectly by modifying transpiration rates and the composition of rhizosphere microflora. A capacity for the external hyphae to take up and deliver nutrients to the plant has been demonstrated for the following nutrients and mycorrhizas; P (VAM, EM, ECM), NH₄ ⁺ (VAM, EM, ECM), NO₃ ^- (ECM), K (VAM, ECM), Ca (VAM, EM), SO₄ ^2- (VAM), Cu (VAM), Zn (VAM) and Fe (EM). In experimental chambers, the external hyphae of VAM can deliver up to 80% of plant P, 25% of plant N, 10% of plant K, 25% of plant Zn and 60% of plant Cu. Knowledge of the role of mycorrhiza in the uptake of nutrients other than P and N is limited because definitive studies are few, especially for the ECM. Although further quantification is required, it is feasible that the external hyphae may provide a significant delivery system for N, K, Cu and Zn in addition to P in many soils. Proposals that ECM and VAM fungi contribute substantially to the Mg, B and Fe nutrition of the host plant have not been substantiated. ECM and EM fungi produce ectoenzymes which provide host plants with the potential to access organic N and P forms that are normally unavailable to VAM fungi or to non mycorrhizal roots. The relative contribution of these nutrient sources requires quantification in the field. Further basic research, including the quantification of nutrient uptake and transport by fungal hyphae in soil and regulation at the fungal-plant interface, is essential to support the selection and utilization of mycorrhizal fungi on a commercial scale.     

Transmembrane electric potential difference of germ tubes of arbuscular mycorrhizal fungi responds to external stimuli

Measurements of the electric potential difference across the hyphal wall and the cell membrane were made on external hyphae of three species of arbuscular mycorrhizal fungus Gigaspora margarita , Scutellospora calospora and Glomus coronatum and on germ tubes of Gi. margarita . The values of transmembrane electric potential difference recorded (∼–40 mV) are less negative than those previously reported from hyphae of arbuscular mycorrhizal fungi closely associated with roots and from filamentous fungi. The external hyphae of arbuscular mycorrhizal fungi grown in soil had similar values of electric potential difference to those grown in soil-less culture, and to germ tubes. Thermodynamic calculations showed that despite these low values of electric potential difference, efficient high-affinity uptake of phosphate is possible. The transmembrane electric potential difference of germ tubes of Gi. margarita became more negative when plant root extract was added to the medium, showing for the first time that the early stages of interaction between plant and fungus occur via direct effects on the plasma membrane rather than via effects on gene expression. Addition of K⁺ reversibly depolarized the transmembrane electric potential difference of germ tubes of Gi. margarita , indicating that despite the low electric potential difference the fungus has control over the permeability of the plasmamembrane to K⁺.