Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization : Hyphal density and mycorrhizal root colonization

The influence of tillage practices on native arbuscular mycorrhizal fungi (AMF) was studied in two, consecutive years in eastern Canada, in two 11 year-old long-term tillage-fertilizer experimental field soils, a sandy loam and a clay, growing corn in monoculture. The three tillage practices were: 1) conventional tillage (CT; fall plowing plus spring disking), reduced tillage (RT; spring disking) and no-till (NT). The corn crop received either inorganic (N and K) or organic (liquid dairy manure) fertilizers. Mycorrhizal hyphal density was estimated from soil samples obtained in early spring (before disking), at the 12–14 leaf stage, at silking, and at harvest. The percentage of corn root colonization by AMF at the 12–14 leaf stage, at silking and at harvest was also determined. The sandy loam was sampled over two consecutive seasons and the clay soil over one season.Densities of total and metabolically active soil hyphae, and mycorrhizal root colonization were significantly lower in CT soil than in RT and NT soil. Lowest soil hyphal densities were observed in early spring. The levels of intra- and extraradical fungal colonization always increased from spring to silking and decreased thereafter. Spring disking had only a small and transient negative effect on hyphal abundance in soil. Fertilization did not influence mycorrhizal colonization of corn or abundance of soil hyphae in the sandy loam soil, but in the clay soil metabolically active hyphae were more abundant with manure application than with mineral fertilization. In 1992, in both soils different tillage systems had same grain yield, however, in 1993, corn yield was higher in NT compared to CT system.     

Influence of arbuscular mycorrhizal fungi on soil structure and aggregate stability of a vertisol

The influence of arbuscular mycorrhizal (AM) fungi on aggregate stability of a semi-arid Indian vertisol was studied in a pot experiment in which Sorghum bicolor (L.) was grown as test plant for 10 weeks. Pasteurized soil inoculated with AM fungi was studied with pasteurized and unpasteurized soils as references. A part of the soil in each pot was placed in nylon mesh bags to separate effects of roots and hyphae. The sorghum plants were planted outside the mesh bags which permitted AM hyphae to enter while excluding roots. Aggregate stability of the soil was determined by wet-sieving and turbidimetric measurements. Development of the AM fungi was quantified as colonized root length and external hyphal length. Soil exposed to growth of roots and hyphae (outside mesh bags) showed aggregates with larger geometric mean diameter (GMD) in pasteurized soil inoculated with AM fungi than in pasteurized uninoculated soil. There was no significant difference in GMD of the inoculated, pasteurized soil and the unpasteurized soil. No significant effects of inoculation or plant growth were found in pasteurized soil exposed to hyphal growth only (inside the mesh bags). However, the unpasteurized soil had significantly higher GMD than the pasteurized soil, irrespective of plants and inoculum. Turbidimetric measurements of soil exposed to roots and hyphae (outside mesh bags) showed the highest aggregate stability for the inoculated pasteurized soil. These results demonstrate that AM fungi contribute to the stabilization of soil aggregates in a vertisol, and that the effect is significant after only one growing season. The effect was associated with both AM hyphae and the stimulation of root growth by AM fungi. The contribution from plant roots and AM hyphae to aggregate stability of different size fractions is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Survival of the external mycelium of a VAM fungus in frozen soil over winter

The present investigation examines (1) whether the external VAM mycelium survives winter freezing to act as a source of inoculum in the spring, and (2) whether soil disturbance reduces the infectivity of the external VAM mycelium following freezing of the soil. Sealed pouches of fine nylon mesh were placed in pots containing soil inoculated with a Glomus species. The mesh was impervious to roots but not to hyphae. Following two 3-week growth cycles of maize in the pots, the pouches were transplanted to the field. Pouches were removed from the field once during the 4 months when the soil was frozen, and once after spring thaw. Measurements were made of VAM spore density, hyphal length and viability in the pouches. Bioassays for infectivity were conducted on all pouches. Some VAM hyphae survived freezing and remained infective following winter freezing, in the absence of plant roots. Soil disturbance did not reduce the infectivity of hyphae following exposure to freezing temperatures. We observed a change in the distribution of viable cytoplasm within hyphae over winter, which we hypothesize represents an adaptation allowing hyphae to survive freezing temperatures. We suggest that the effect of disturbance on hyphal infectivity may be related to this seasonal change in the distribution of hyphal viability.     

Studies on the attractional effect of root exudates on hyphal growth of an arbuscular mycorrhizal fungus in a soil compartment-membrane system

The effects of tomato and bean rhizospheres on hyphal spreading of the arbuscular mycorrhizal (AM) fungus Glomus mosseae were studied using a soil compartment system in combination with hydrophobic polytetrafluorethylene (PTFE) membranes. Both the nylon screen and the PTFE membrane were freely permeable to hyphae but not to roots. Furthermore, the hydrophobic PTFE membrane seemed to be a barrier to the flux of soil solutions containing root exudates. The results show that water soluble exudates of tomato and bean roots greatly stimulate hyphal growth in the soil compartment system used. Moreover, water soluble root exudates of bean exert a clear attractional effect on AM hyphal growth.     

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     

Contribution by two arbuscular mycorrhizal fungi to P uptake by cucumber (Cucumis sativus L.) from32P-labelled organic matter during mineralization in soil

An experiment was set up to investigate the role of arbuscular mycorrhiza (AM) in utilization of P from organic matter during mineralization in soil. Cucumber (Cucumis sativus L.) inoculated with one of two AM fungi or left uninoculated were grown for 30 days in cross-shaped PVC pots. One of two horizontal compartments contained 100 g soil (quartz sand: clay loam, 1:1) with 0.5 g ground clover leaves labelled with³²P. The labelled soil received microbial inoculum without AM fungi to ensure mineralization of the added organic matter. The labelling compartment was separated from a central root compartment by either 37 m or 700 m nylon mesh giving only hyphae or both roots and hyphae, respectively, access to the labelled soil. The recovery of³²P from the hyphal compartment was 5.5 and 8.6% for plants colonized withGlomus sp. andG. caledonium, respectively, but only 0.6 % for the non-mycorrhizal controls. Interfungal differences were not related to root colonization or hyphal length densities, which were lowest forG. caledonium. Both fungi depleted the labelled soil of NaHCO₃-extractable P and³²P compared to controls. A 15–25% recovery of³²P by roots was not enhanced in the presence of mycorrhizas, probably due to high root densities in the labelled soil. The experiment confirms that AM fungi differ in P uptake characteristics, and that mycorrhizal hyphae can intercept some P immobilization by other microorganisms and P-sorbing clay minerals.     

Proton (H+) flux signature for the presymbiotic development of the arbuscular mycorrhizal fungi

Ion dynamics are important for cell nutrition and growth in fungi and plants. Here, the focus is on the relationship between the hyphal H(+) fluxes and the control of presymbiotic growth and host recognition by arbuscular mycorrhizal (AM) fungi. Fluxes of H(+) around azygopores and along lateral hyphae of Gigaspora margarita during presymbiotic growth, and their regulation by phosphate (P) and sucrose (Suc), were analyzed with an H(+)-specific vibrating probe. Changes in hyphal H(+) fluxes were followed after induction by root exudates (RE) or by the presence Trifolium repens roots. Differential sensitivity to P-type ATPase inhibitors (orthovanadate or erythrosin B) suggests an asymmetric distribution or activation of H(+)-pump isoforms along the hyphae of the AM fungi. Concentration of P and Suc affected the hyphal H(+) fluxes and growth rate. However, further increases in H+ efflux and growth rate were observed when the fungus was growing close to clover roots or pretreated with RE. The H(+) flux data correlate with those from polarized hyphal growth analyses, suggesting that spatial and temporal alterations of the hyphal H(+)fluxes are regulated by nutrient availability and might underlie a pH signaling elicitation by host RE during the early events of the AM symbiosis.     

Competition and substrate colonization strategies of three polyxenically grown arbuscular mycorrhizal fungi

Intra- and extraradical colonization competition and hyphal interactions among arbuscular mycorrhizal fungi (AMF) Glomus intraradices, Glomus proliferum and Gigaspora margarita were investigated in two in vitro experimental systems. AMF were polyxenically cultured with a Ri T-DNA transformed carrot root organ culture (ROC) in either big Petri plates containing three culture compartments and a common hyphal compartment (i.e. an independent host root for each AMF) or two by two in the culture compartment of regular bicompartmented Petri dishes (i.e. a common host root and a common hyphal compartment). Maps of the extraradical mycelial development of the three AMF were obtained. Two distinct substrate colonization strategies (Glomus-type and Gigaspora-type) were identified, reflecting intrinsic differences among AMF genera/families. Our data reveal a general lack of antagonism between the isolates when extraradical hyphae explore and exploit the substrate outside the root influence zone; however certain growth restrictions were imposed by Gi. margarita extraradical mycelium when developing near the host root and by G. proliferum intraradical hyphae. This work highlights once more the appropriateness of AM in vitro culture systems to perform in vivo studies on the biology of this symbiosis and opens new avenues to the formulation of in vitro AMF inoculants.     

Comparative structural study ofQuercus serrata andQ. acutissima formed byPisolithus tinctorius andHebeloma cylindrosporum

Ectomycorrhizas were synthesized in pots and growth pouches betweenQuercus serrata, Q. acutissima, and two ectomycorrhizal fungi,Pisolithus tinctorius andHebeloma cylindrosporum. Root morphology and the structure of the mantle and Hartig net were compared using light, fluorescence, scanning and transmission electron microscopy.P. tinctorius initially colonized root cap cells, and eventually produced a highly branched lateral root system with a complete mantle, whereasH. cylindrosporum promoted root elongation with few hyphae on the root apex surface indicating that interaction between roots differs with fungal species. Hartig net structure and hyphal inclusions varied between all the combinations tested. There were structural differences between mycorrhizas ofH. cylindrosporum/Q. acutissima grown in soil and growth pouches, which indicate that the growth pouch environment can induce artefacts in roots. Fruit bodies ofH. cylindrosporum developed in pots withQ. acutissima. AlthoughP. tinctorius has been used to inoculate oak seedlings in the nursery, results of this study indicate thatH. cylindrosporum may also be an effective ectomycorrhizal fungus forQ. serrata andQ. acutissima.     

Chemotropism in the arbuscular mycorrhizal fungus Glomus mosseae

In this work, we report the occurrence of chemotropism in the arbuscular mycorrhizal (AM) fungus Glomus mosseae. Fungal hyphae were able to respond to host-derived signals by reorienting their growth towards roots and to perceive chemotropic signals at a distance of at least 910 microm from roots. In order to reach the source of chemotropic signals, hyphal tips crossed interposed membranes emerging within 1 mm from roots, eventually establishing mycorrhizal symbiosis. The specificity of chemotropic growth was evidenced by hyphal growth reorientation and membrane penetration occurring only in experimental systems set up with host plants. Since pre-symbiotic growth is a critical stage in the life cycle of obligate AM fungal symbionts, chemotropic guidance may represent an important mechanism functional to host root location, appressorium formation and symbiosis establishment.     

Acquisition of N by external hyphae of an arbuscular mycorrhizal fungus and its impact on physiological responses in maize under drought-stressed and well-watered conditions

This study examined the uptake of nitrogen by external hyphae of an arbuscular mycorrhizal (AM) fungus (Glomus intraradices Schenck &; Smith) and its impact on physiological responses in maize plants subjected to well-watered or drought-stressed conditions. Plants were grown in compartmented boxes divided by a nylon mesh (40?μm) into a root compartment and a hyphal compartment. Maize plants (Zea mays cv. 'Tuxpeño sequia' selection cycle C0) were exposed to 2 weeks of drought 56 days after sowing. A ^[15]N tracer was applied as K^[15]NO_[3] to the hyphal compartment at a distance of 5?cm from the root compartment. Root and shoot samples were then analyzed for ^[15]N atom % excess (APE), glutamine synthetase (GS) activity, protein concentration and nutritional status. Evapotranspiration rate and stomatal resistance were monitored daily to determine the degree of drought stress. The APE values for AM shoots and roots were 32% and 33% higher than non-AM shoots and roots, respectively, under drought conditions. This provides clear evidence that the external mycelium of AM fungus transports considerable amounts of ^[15]NO_[3]^[– ]to the host plant under drought conditions. Drought-stressed AM roots had 28% higher GS activity, possibly as a consequence of higher hyphal acquisition of NO_[3]^[–] ions. Mycorrhizal colonization significantly increased the host plant P status regardless of soil moisture regime. In addition, the N status of drought-stressed AM shoots and roots was slightly higher than stressed non-AM shoots and roots. The improved nutritional status may assist AM plants to exploit available soil moisture more efficiently and to maintain higher leaf relative water content under moderate drought conditions.     

Characterisation of arbuscular mycorrhizal fungi colonisation in cluster roots of shape Hakea verrucosa F. Muell (Proteaceae), and its effect on growth and nutrient acquisition in ultramafic soil

Ultramafic soils at Bandalup Hill (Western Australia) are characterised by high concentrations of Ni and low levels of P. Amongst the plant species that can sustain such hostile conditions, Hakea verrucosa F. Muell from a non-mycorrhizal family (Proteaceae) would be expected to rely on cluster roots to access P. However, the acidification of ultramafic soils by cluster roots might increase the dissolution of soil Ni, and therefore its availability to plants. Symbiosis with mycorrhizal fungi, on the other hand, might help to reduce the uptake of Ni by H. verrucosa. Therefore, the aim of this study was to investigate the mycorrhizal status of H. verrucosa, and assess any contribution from mycorrhizal fungi to its growth and nutrient status. Seedlings of H. verrucosa were first grown in undisturbed ultramafic soil cores from Bandalup Hill for 8 weeks to assess the presence of mycorrhizal fungi in their roots. In a second experiment, H. verrucosa seedlings were grown in the same ultramafic soil that was either steamed or left untreated. Seedlings were inoculated with an arbuscular mycorrhizal (AM) fungal consortium from Bandalup Hill. Fungal hyphae, vesicles, as well as intracellular arbuscules and hyphal coils were observed in the cluster roots of H. verrucosa in both experiments. In the first experiment, 57% of the root length was colonized by AM fungi. Seedlings had high (between 1.4 and 1.9) shoot to root ratios and their roots had very few root hairs, despite growing in P-deficient soil. Steaming of the ultramafic soil increased the growth of seedlings and their nutrient uptake. Inoculation with AM fungi reduced the seedling growth in steamed ultramafic soil; however, it increased their shoot P and K concentration and also the shoot K content. The shoot Ni concentration of seedlings was not affected by the presence of AM fungi.     

Extraradical mycelium of the arbuscular mycorrhizal fungus Glomus lamellosum can take up,accumulate and translocate radiocaesium under root-organ culture conditions

Radiocaesium enters the food chain when plants absorb it from soil, in a process that is strongly dependent on soil properties and plant and microbial species. Among the microbial species, arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the root cortex of many plants and develop an extraradical mycelial (ERM) network that ramifies in the soil. Despite the well-known involvement of this ERM network in mineral nutrition and uptake of some heavy metals, only limited data are available on its role in radiocaesium transport in plants. We used root-organ culture to demonstrate that the ERM of the AM fungus Glomus lamellosum can take up, possibly accumulate and unambiguously translocate radiocaesium from a 137Cs-labelled synthetic root-free compartment to a root compartment and within the roots. The accumulation of 137Cs by hyphae in the root-free compartment may be explained by sequestration in the hyphae or by a bottleneck effect resulting from a limited number of hyphae crossing the partition between the two compartments. Uptake and translocation resulted from the incorporation of 137Cs into the fungal hyphae, as no 137Cs was detected in mycorrhizal roots treated with formaldehyde. The importance of the translocation process was indicated by the correlation between 137Cs measured in the roots and the total hyphal length connecting the roots with the labelled compartment. 137Cs may be translocated via a tubular vacuolar system or by cytoplasmic streaming per se.     

Fungal root colonization responses in natural grasslands after long-term exposure to elevated atmospheric CO2

Arbuscular mycorrhizae, ubiquitous mutualistic symbioses between plant roots and fungi in the order Glomales, are believed to be important controllers of plant responses to global change, in particular to elevated atmospheric CO₂. In order to test if any effects on the symbiosis can persist after long-term treatment, we examined root colonization by arbuscular mycorrhizal (AM) and other fungi of several plant species from two grassland communities after continuous exposure to elevated atmospheric CO₂ for six growing seasons in the field. For plant species from both a sandstone and a serpentine annual grassland there was evidence for changes in fungal root colonization, with changes occurring as a function of plant host species. We documented decreases in percentage nonmycorrhizal fungal root colonization in elevated CO₂ for several plant species. Total AM root colonization (%) only increased significantly for one out of the five plant species in each grassland. However, when dividing AM fungal hyphae into two groups of hyphae (fine endophyte and coarse endophyte), we could document significant responses of AM fungi that were hidden when only total percentage colonization was measured. We also documented changes in elevated CO₂ in the percentage of root colonized by both AM hyphal types simultaneously. Our results demonstrate that changes in fungal root colonization can occur after long-term CO₂ enrichment, and that the level of resolution of the study of AM fungal responses may have to be increased to uncover significant changes to the CO₂ treatment. This study is also one of the first to document compositional changes in the AM fungi colonizing roots of plants grown in elevated CO₂. Although it is difficult to relate the structural data directly to functional changes, possible implications of the observed changes for plant communities are discussed.     

Nuclei of symbiotic arbuscular mycorrhizal fungi as revealed by in vivo two-photon microscopy

The present work reports the results obtained from in vivo studies on the distribution and behavior of nuclei of two arbuscular mycorrhizal (AM) fungi growing in symbiosis with tomato root organ cultures (AM monoxenic cultures). Upon staining with 4',6-diamidino-2-phenylindole and two-photon microscopy (2PM) observations, symbiotic thick runner hyphae appeared mostly opaque to 2PM and did not reveal nuclei within them; thin runner hyphae showed dimly stained nuclei along them, whereas nuclei were clearly visible within the branches of the so-called branched absorbing structures. When visible, nuclei appeared anchored laterally at regular intervals along the symbiotic AM extraradical hyphae. Other nuclei migrate through the hyphal central core; this migration occurs in pulses. Simultaneous observations on different areas of extraradical AM mycelium revealed the existence of lysed compartments along the fungal hyphae, containing nuclei remnants and/or chromatin masses. All these results give new insights in (i) the differential permeability of AM hyphae in the symbiotic versus the asymbiotic state; (ii) the behavior and distribution of nuclei along the symbiotic extraradical mycelium; (iii) the occurrence of ageing events within the AM fungal colony; and (iv) the existence of "healing" mechanisms aiming to restrict the damage induced by such ageing or lytic events. An AM fungal strategy for hyphal survival under adverse conditions is also suggested.     

Nitrogen transfer and assimilation between the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith and Ri T-DNA roots of Daucus carota L. in an in vitro compartmented system

Nitrogen metabolism was examined in monoxenic cultures of carrot roots (Daucus carota L.) colonized with the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith. Glutamine synthetase and glutamate dehydrogenase activities were significantly increased in mycorrhizal roots for which only the extraradical mycelium had exclusive access to NH4NO3 in a distinct hyphal compartment inaccessible to the roots. This was in comparison with the water controls but was similar to the enzyme activities of non-arbuscular-mycorrhizal (non-AM) roots that had direct access to NH4NO3. In addition, glutamate dehydrogenase activity was significantly enhanced in AM roots compared with non-AM roots. Carrot roots took up 15NH4+ more efficiently than 15NO3-, and the extraradical hyphae transfered 15NH4+ to host roots from the hyphal compartment but did not transfer 15NO3-. The extraradical mycelium was shown, for the first time, to have a different glutamine synthetase monomer than roots. Our overall results highlight the active role of AM fungi in nitrogen uptake, transfer, and assimilation in their symbiotic root association.     

Interactions between biochar and mycorrhizal fungi in a water-stressed agricultural soil

Biochar may alleviate plant water stress in association with arbuscular mycorrhizal (AM) fungi but research has not been conclusive. Therefore, a glasshouse experiment was conducted to understand how interactions between AM fungi and plants respond to biochar application under water-stressed conditions. A twin chamber pot system was used to determine whether a woody biochar increased root colonisation by a natural AM fungal population in a pasture soil (‘field’ chamber) and whether this was associated with increased growth of extraradical AM fungal hyphae detected by plants growing in an adjacent (‘bait’) chamber containing irradiated soil. The two chambers were separated by a mesh that excluded roots. Subterranean clover was grown with and without water stress and harvested after 35, 49 and 63 days from each chamber. When biochar was applied to the field chamber under water-stressed conditions, shoot mass increased in parallel with mycorrhizal colonisation, extraradical hyphal length and shoot phosphorus concentration. AM fungal colonisation of roots in the bait chamber indicated an increase in extraradical mycorrhizal hyphae in the field chamber. Biochar had little effect on AM fungi or plant growth under well-watered conditions. The biochar-induced increase in mycorrhizal colonisation was associated with increased growth of extraradical AM fungal hyphae in the pasture soil under water-stressed conditions.     

Anatomy and mycotrophy of the achlorophyllous Afrothismia winkleri

Afrothismia winkleri develops fleshy rhizomes, densely covered with small root tubercles, narrowing to filiform roots with age. The exclusively intracellular mycorrhizal fungus has distinct morphologies in different tissues of the plant. In the filiform root the hyphae grow straight and vesicles are borne on short hyphal stalks. The straight hyphae are present in the epidermis of the root tubercles, but change to loosely coiled and swollen hyphae in the rhizome tissue. No penetration from epidermis to root cortex was found. From the rhizome, a separating cell layer permits only one or rarely two hyphal penetrations into the cortex of each root tubercle. The hyphae proceed apically within the root hypodermis in a spiral row of distinctively coiled hyphae, branches of which colonize the inner root cortex. In the inner root cortex the hyphal coils degenerate to amorphous clumps. In older roots the cortex itself also deteriorates, but epidermis, hypodermis, endodermis and central cylinder persist. The mycorrhizal pattern in A. winkleri is interpreted as an elaborate exploitation system whereby the fungus provides carbon and nutrients to the plant and, simultaneously but spatially distinct, its hyphae are used to translocate and store the matter within the plant. Several features indicate that the endophyte is an arbuscular mycorrhizal fungus.     

Phosphorus Effects on the Mycelium and Storage Structures of an Arbuscular Mycorrhizal Fungus as Studied in the Soil and Roots by Analysis of Fatty Acid Signatures

The distribution of an arbuscular mycorrhizal (AM) fungus between soil and roots, and between mycelial and storage structures, was studied by use of the fatty acid signature 16:1(omega)5. Increasing the soil phosphorus level resulted in a decrease in the level of the fatty acid 16:1(omega)5 in the soil and roots. A similar decrease was detected by microscopic measurements of root colonization and of the length of AM fungal hyphae in the soil. The fatty acid 16:1(omega)5 was estimated from two types of lipids, phospholipids and neutral lipids, which mainly represent membrane lipids and storage lipids, respectively. The numbers of spores of the AM fungus formed in the soil correlated most closely with neutral lipid fatty acid 16:1(omega)5, whereas the hyphal length in the soil correlated most closely with phospholipid fatty acid 16:1(omega)5. The fungal neutral lipid/phospholipid ratio in the extraradical mycelium was positively correlated with the level of root infection and thus decreased with increasing applications of P. The neutral lipid/phospholipid ratio indicated that at high P levels, less carbon was allocated to storage structures. At all levels of P applied, the major part of the AM fungus was found to be present outside the roots, as estimated from phospholipid fatty acid 16:1(omega)5. The ratio of extraradical biomass/intraradical biomass was not affected by the application of P, except for a decrease at the highest level of P applied.     

Genetic evidence for auxin involvement in arbuscular mycorrhiza initiation

? Formation of arbuscular mycorrhiza (AM) is controlled by a host of small, diffusible signaling molecules, including phytohormones. To test the hypothesis that the plant hormone auxin controls mycorrhiza development, we assessed mycorrhiza formation in two mutants of tomato (Solanum lycopersicum): diageotropica (dgt), an auxin-resistant mutant, and polycotyledon (pct), a mutant with hyperactive polar auxin transport. ? Mutant and wild-type (WT) roots were inoculated with spores of the AM fungus Glomus intraradices. Presymbiotic root-fungus interactions were observed in root organ culture (ROC) and internal fungal colonization was quantified both in ROC and in intact seedlings. ? In ROC, G. intraradices stimulated presymbiotic root branching in pct but not in dgt roots. pct roots stimulated production of hyphal fans indicative of appressorium formation and were colonized more rapidly than WT roots. By contrast, approaching hyphae reversed direction to grow away from cultured dgt roots and failed to colonize them. In intact seedlings, pct and dgt roots were colonized poorly, but development of hyphae, arbuscules, and vesicles was morphologically normal within roots of both mutants. ? We conclude that auxin signaling within host roots is required for the early stages of AM formation, including during presymbiotic signal exchange.