Functional diversity in arbuscular mycorrhizas: exploitation of soil patches with different phosphate enrichment differs among fungal species

Most terrestrial plant species form associations with arbuscular mycorrhizal fungi (AMF) that transfer soil P to the plant via their external hyphae. The distribution of nutrients in soils is typically patchy (heterogeneous) but little is known about the ability of AMF to exploit P patches in soil. This was studied by growing symbioses of Linum usitatissimum and three AMF (Glomus intraradices, G. mosseae and Gigaspora margarita) in pots with two side-arms, which were accessible to hyphae, but not to roots. Soil in one side-arm was either unamended (P0) or enriched with P; simultaneous labelling of this soil with ³²P revealed that G. intraradices responded to P enrichment both in terms of hyphal proliferation and P uptake, whereas the other AMF did not. Labelling with ³³P of P0 soil in the other side arm revealed that the increased P uptake by G. intraradices from the P-enriched patch was paralleled by decreased P uptake by other parts of the mycelium. This is the first demonstration of variation in growth and nutrient uptake by an AMF as influenced by a localized P enrichment of the soil. The results are discussed in the context of functional diversity of AMF.     

Symbiont identity matters: carbon and phosphorus fluxes between Medicago truncatula and different arbuscular mycorrhizal fungi

Many studies have scrutinized the nutritional benefits of arbuscular mycorrhizal associations to their host plants, while the carbon (C) balance of the symbiosis has often been neglected. Here, we present quantification of both the C costs and the phosphorus (P) uptake benefits of mycorrhizal association between barrel medic (Medicago truncatula) and three arbuscular mycorrhizal fungal species, namely Glomus intraradices, Glomus claroideum, and Gigaspora margarita. Plant growth, P uptake and C allocation were assessed 7 weeks after sowing by comparing inoculated plants with their non-mycorrhizal counterparts, supplemented with different amounts of P. Isotope tracing (³³P and ¹³C) was used to quantify both the mycorrhizal benefits and the costs, respectively. G. intraradices supported greatest plant P acquisition and incurred high C costs, which lead to similar plant growth benefits as inoculation with G. claroideum, which was less efficient in supporting plant P acquisition, but also required less C. G. margarita imposed large C requirement on the host plant and provided negligible P uptake benefits. However, it did not significantly reduce plant growth due to sink strength stimulation of plant photosynthesis. A simple experimental system such as the one established here should allow quantification of mycorrhizal costs and benefits routinely on a large number of experimental units. This is necessary for rapid progress in assessment of C fluxes between the plants and different mycorrhizal fungi or fungal communities, and for understanding the dynamics between mutualism and parasitism in mycorrhizal symbioses.     

Phosphorus Acquisition Strategies within Arbuscular Mycorrhizal Fungal Community of a Single Field Site

Diversity in phosphorus (P) acquisition strategies was assessed among eight isolates of arbuscular mycorrhizal fungi (AMF) belonging to three Glomus species, all obtained from the same field site. Maize (Zea mays L. cv. Corso) was used as a test plant. Compartmented cultivation containers coupled with ³³P radioisotope labeling of soil P were employed to estimate (1) the distance from the roots that AMF were able to acquire soil P from, (2) the rate of soil colonization, (3) the efficiency of uptake of soil P by AMF, (4) benefits provided to maize in terms of P acquisition and growth. Glomus mosseae and G. intraradices took up P 10 cm from roots, whereas G. claroideum only up to 6 cm from the roots. G. mosseae most rapidly colonized the available soil volume and transported significant amounts of P to maize from a distance, but provided no net P uptake benefit to the plants. On the other hand, both G. intraradices and three out of four G. claroideum isolates significantly improved net P uptake by maize. These effects seem to be related to variability between and to a limited extent also within AMF species, in mycelium development, efficiency of hyphal P uptake and effects on plant P acquisition via the root pathway. In spite of absence of maize growth responses to inoculation with any of the AMF isolates, this study indicates remarkable functional diversity in the underground component of the studied field site.     

Suppression of the Biocontrol Agent Trichoderma harzianum by Mycelium of the Arbuscular Mycorrhizal Fungus Glomus intraradices in Root-Free Soil

Trichoderma harzianum is an effective biocontrol agent against several fungal soilborne plant pathogens. However, possible adverse effects of this fungus on arbuscular mycorrhizal fungi might be a drawback in its use in plant protection. The objective of the present work was to examine the interaction between Glomus intraradices and T. harzianum in soil. The use of a compartmented growth system with root-free soil compartments enabled us to study fungal interactions without the interfering effects of roots. Growth of the fungi was monitored by measuring hyphal length and population densities, while specific fatty acid signatures were used as indicators of living fungal biomass. Hyphal ³³P transport and β-glucuronidase (GUS) activity were used to monitor activity of G. intraradices and a GUS-transformed strain of T. harzianum, respectively. As growth and metabolism of T. harzianum are requirements for antagonism, the impact of wheat bran, added as an organic nutrient source for T. harzianum, was investigated. The presence of T. harzianum in root-free soil reduced root colonization by G. intraradices. The external hyphal length density of G. intraradices was reduced by the presence of T. harzianum in combination with wheat bran, but the living hyphal biomass, measured as the content of a membrane fatty acid, was not reduced. Hyphal ³³P transport by G. intraradices also was not affected by T. harzianum. This suggests that T. harzianum exploited the dead mycelium but not the living biomass of G. intraradices. The presence of external mycelium of G. intraradices suppressed T. harzianum population development and GUS activity. Stimulation of the hyphal biomass of G. intraradices by organic amendment suggests that nutrient competition is a likely means of interaction. In conclusion, it seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum; in contrast, T. harzianum was adversely affected by G. intraradices.     

Response to cadmium of Daucus carota hairy roots dual cultures with Glomus intraradices or Gigaspora margarita

Ri T-DNA-transformed carrot roots were cultivated in two experiments either non-inoculated or inoculated with the arbuscular mycorrhizal (AM) fungi Glomus intraradices or Gigaspora margarita. The influence of two concentrations of cadmium (Cd) in the medium (2 mg l^–1, 4 mg l^–1) on both root and mycelium growth was tested. Both parameters were estimated at 10-day intervals for 70 or 100 days for G. intraradices and Gi. margarita, respectively. In the first experiment, G. intraradices showed a rapid spread of extraradical mycelium (ERM) and reached average densities per treatment of about 90 cm cm^–2 agar medium after 70 days. At the higher Cd level, the growth of ERM was delayed in comparison to the treatment without Cd addition. Root growth was inhibited by both Cd levels; the inhibition was, however, significantly lower in the treatments inoculated with G. intraradices compared to the non-inoculated control. In the second experiment, the ERM of Gi. margarita started to grow after a period of 50 days and reached average densities per treatment of only up to 27 cm cm^–2 by the end of the cultivation. The growth of Gi. margarita mycelium was not inhibited by Cd. No differences in root growth were observed between the Gi. margarita inoculated and non-inoculated treatments. The inhibitory effect of Cd on root growth differed between the non-inoculated treatments in both experiments. The study has shown that the AM fungus Glomus intraradices can alleviate Cd-induced growth inhibition to carrot hairy roots. The potential and limits of the monoxenic system in studying the interaction between AM fungi and heavy metals are discussed.     

Flavonoids exclusively present in mycorrhizal roots of white clover exhibit a different effect on arbuscular mycorrhizal fungi than flavonoids exclusively present in non-mycorrhizal roots of white clover

Abstract

The flavonoids 5,6,7,8,9-hydroxy chalcone, 3,7-hydroxy-4′methoxy flavone, 5,6,7,8-hydroxy-4′-methoxy flavone and 3,5,6,7,4′-hydroxy flavone can be detected only in non-mycorrhizal roots of white clover, but not in mycorrhizal roots, whereas the flavonoids acacetin, quercetin and rhamnetin are only present in mycorrhizal roots. We tested the effect of several concentrations of these compounds on spore germination, hyphal growth, hyphal branching, formation of clusters of auxiliary cells and of secondary spores of the arbuscular mycorrhizal fungi Gigaspora rosea, Gigaspora margarita, Glomus mosseae and Glomus intraradices. Our results indicate that depending on the flavonoid, the tested compounds are involved at different stages in the regulation of mycorrhization. This hypothesis is strengthened by their differing effect on several AM fungal growth parameters. Furthermore, our study provides more data on the AM fungus genus/species specificity of flavonoids.     

Phosphorus and carbon availability regulate structural composition and complexity of AM fungal mycelium

The regulation of the structural composition and complexity of the mycelium of arbuscular mycorrhizal (AM) fungi is not well understood due to their obligate biotrophic nature. The aim of this study was to investigate the structure of extraradical mycelium at high and low availability of carbon (C) to the roots and phosphorus (P) to the fungus. We used monoxenic cultures of the AM fungus Rhizophagus irregularis (formerly Glomus intraradices) with transformed carrot roots as the host in a cultivation system including a root-free compartment into which the extraradical mycelium could grow. We found that high C availability increased hyphal length and spore production and anastomosis formation within individual mycelia. High P availability increased the formation of branched absorbing structures and reduced spore production and the overall length of runner hyphae. The complexity of the mycelium, as indicated by its fractal dimensions, increased with both high C and P availability. The results indicate that low P availability induces a growth pattern that reflects foraging for both P and C. Low C availability to AM roots could still support the explorative development of the mycelium when P availability was low. These findings help us to better understand the development of AM fungi in ecosystems with high P input and/or when plants are subjected to shading, grazing or any management practice that reduces the photosynthetic ability of the plant.     

Dependence of Arbuscular-Mycorrhizal Fungi on Their Plant Host for Palmitic Acid Synthesis

Lipids are the major form of carbon storage in arbuscular-mycorrhizal fungi. We studied fatty acid synthesis by Glomus intraradices and Gigaspora rosea. [¹⁴C]Acetate and [¹⁴C]sucrose were incorporated into a synthetic culture medium to test fatty acid synthetic ability in germinating spores (G. intraradices and G. rosea), mycorrhized carrot roots, and extraradical fungal mycelium (G. intraradices). Germinating spores and extraradical hyphae could not synthesize 16-carbon fatty acids but could elongate and desaturate fatty acids already present. The growth stimulation of germinating spores by root exudates did not stimulate fatty acid synthesis. 16-Carbon fatty acids (16:0 and 16:1) were synthesized only by the fungi in the mycorrhized roots. Our data strongly suggest that the fatty acid synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that fatty acid metabolism may play a major role in the obligate biotrophism of arbuscular-mycorrhizal fungi.     

Real-time RT-PCR profiling of transcription factors including 34 MYBs and signaling components in white lupin reveals their P status dependent and organ-specific expression

Two controlled microcosm experiments aimed at a critical re-assessment of the contributions of divergent arbuscular mycorrhizal (AM) fungi to plant mineral nutrition were established that specifically targeted Plantago lanceolata–Glomus intraradices (B.B/E) and –Gigaspora margarita (BEG 34) symbioses developed in a native, nutrient limited, coastal dune soil. Plant tissue nitrogen (N), phosphorus (P) and potassium (K) status as well as plant growth parameters and levels of mycorrhizal colonization were assessed at harvest. In addition to the general well-established mycorrhizal facilitation of P uptake, the study was able to demonstrate a G. intraradices-specific contribution to improved plant nitrogen and potassium nutrition. In the two respective experiments, G. intraradices-inoculated plants had 27.8% and 40.8% more total N and 55.8% and 23.3% more total K when compared to Gi. margarita inoculated counterparts. Dissimilar overall contribution of the two isolates to plant nutrition was identified in AM-genus specific differences in plant tissue N:P:K ratios. G. intraradices inoculated and non-mycorrhizal plants generally exhibited N:P:K ratios indicative of P limitation whereas for Gi. margarita mycorrhizal plants, corresponding ratios strongly implied either N or K limitation. The study provides further evidence highlighting AM functional biodiversity in respect to plant nutrient limitation experienced by mycorrhizal P. lanceolata in an ecologically relevant soil system.     

Different native arbuscular mycorrhizal fungi influence the coexistence of two plant species in a highly alkaline anthropogenic sediment

Different species of arbuscular mycorrhizal fungi (AMF) can produce different amounts of extraradical mycelium (ERM) with differing architectures. They also have different efficiencies in gathering phosphate from the soil. These differences in phosphate uptake and ERM length or architecture may contribute to differential growth responses of plants and this may be an important contributor to plant species coexistence. The effects of the development of the ERM of AMF on the coexistence of two co-occurring plant species were investigated in root-free hyphal chambers in a rhizobox experimental unit. The dominant shrub (Salix atrocinerea Brot.) and herbaceous (Conyza bilbaoana J. Rémy) plant species found in a highly alkaline anthropogenic sediment were studied in symbiosis with four native AMF species (Glomus intraradices BEG163, Glomus mosseae BEG198, Glomus geosporum BEG199 and Glomus claroideum BEG210) that were the most abundant members of the AMF community found in the sediment. Different AMF species did not influence total plant productivity (sum of the biomass of C. bilbaoana and S. atrocinerea), but had a great impact on the individual biomass of each plant species. The AMF species with greater extracted ERM lengths (G. mosseae BEG198, G. claroideum BEG210 and the four mixed AMF) preferentially benefited the plant species with a high mycorrhizal dependency (C. bilbaoana), while the AMF species with the smallest ERM length (G. geosporum BEG199) benefited the plant species with a low mycorrhizal dependency (S. atrocinerea). Seed production of C. bilbaoana was only observed in plants inoculated with G. mosseae BEG198, G. claroideum BEG210 or the mixture of the four AMF. Our results show that AMF play an important role in the reproduction of C. bilbaoana coexisting with S. atrocinerea in the alkaline sediment and have the potential to stimulate or completely inhibit seed production. The community composition of native AMF and the length of the mycelium they produce spreading from roots into the surrounding soil can be determinant of the coexistence of naturally co-occurring plant species.     

Heavy-Metal Stress and Developmental Patterns of Arbuscular Mycorrhizal Fungi

The rate of global deposition of Cd, Pb, and Zn has decreased over the past few decades, but heavy metals already in the soil may be mobilized by local and global changes in soil conditions and exert toxic effects on soil microorganisms. We examined in vitro effects of Cd, Pb, and Zn on critical life stages in metal-sensitive ecotypes of arbuscular mycorrhizal (AM) fungi, including spore germination, presymbiotic hyphal extension, presymbiotic sporulation, symbiotic extraradical mycelium expansion, and symbiotic sporulation. Despite long-term culturing under the same low-metal conditions, two species, Glomus etunicatum and Glomus intraradices, had different levels of sensitivity to metal stress. G. etunicatum was more sensitive to all three metals than was G. intraradices. A unique response of increased presymbiotic hyphal extension occurred in G. intraradices exposed to Cd and Pb. Presymbiotic hyphae of G. intraradices formed presymbiotic spores, whose initiation was more affected by heavy metals than was presymbiotic hyphal extension. In G. intraradices grown in compartmentalized habitats with only a portion of the extraradical mycelium exposed to metal stress, inhibitory effects of elevated metal concentrations on symbiotic mycelial expansion and symbiotic sporulation were limited to the metal-enriched compartment. Symbiotic sporulation was more sensitive to metal exposure than symbiotic mycelium expansion. Patterns exhibited by G. intraradices spore germination, presymbiotic hyphal extension, symbiotic extraradical mycelium expansion, and sporulation under elevated metal concentrations suggest that AM fungi may be able to survive in heavy metal-contaminated environments by using a metal avoidance strategy.     

Increased Sporulation of Vesicular-Arbuscular Mycorrhizal Fungi by Manipulation of Nutrient Regimens

Adjustment of pot culture nutrient solutions increased root colonization and sporulation of vesicular-arbuscular mycorrhizal (VAM) fungi. Paspalum notatum Flugge and VAM fungi were grown in a sandy soil low in N and available P. Hoagland nutrient solution without P enhanced sporulation in soil and root colonization of Acaulospora longula, Scutellospora heterogama, Gigaspora margarita, and a wide range of other VAM fungi over levels produced by a tap water control or nutrient solutions containing P. However, Glomus intraradices produced significantly more spores in plant roots in the tap water control treatment. The effect of the nutrient solutions was not due solely to N nutrition, because the addition of NH₄NO₃ decreased both colonization and sporulation by G. margarita relative to levels produced by Hoagland solution without P.     

Intracellular pH in Arbuscular Mycorrhizal Fungi : A Symbiotic Physiological Marker

A method was developed to perform real-time analysis of cytosolic pH of arbuscular mycorrhizal fungi in culture using dye and ratiometric measurements (490/450 nm excitations). The study was mainly performed using photometric analysis, although some data were confirmed using image analysis. The use of nigericin allowed an in vivo calibration. Experimental parameters such as loading time and concentration of the dye were determined so that pH measurements could be made for a steady-state period on viable cells. A characteristic pH profile was observed along hyphae. For Gigaspora margarita, the pH of the tip (0–2 μm) was typically 6.7, increased sharply to 7.0 behind this region (9.5 μm), and decreased over the next 250 μm to a constant value of 6.6. A similar pattern was obtained for Glomus intraradices. The pH profile of G. margarita germ tubes was higher when cultured in the presence of carrot (Daucus carota) hairy roots (nonmycorrhizal). Similarly, extraradical hyphae of G. intraradices had a higher apical pH than the germ tubes. The use of a paper layer to prevent the mycorrhizal roots from being in direct contact with the medium selected hyphae with an even higher cytosolic pH. Results suggest that this method could be useful as a bioassay for studying signal perception and/or H⁺ cotransport of nutrients by arbuscular mycorrhizal hyphae.     

Opening the black box: outcomes of interactions between arbuscular mycorrhizal (AM) and non‐host genotypes of Medicago depend on fungal identity,interplay between P uptake pathways and external P supply

We investigated the physiology that underlies the influence of arbuscular mycorrhizal (AM) colonization on outcomes of interactions between plants. We grew Medicago truncatula A17 and its AM‐defective mutant dmi1 in intragenotypic (two plants per pot of the same genotype, x2) or intergenotypic (one plant of each genotype, 1 + 1) combinations, inoculated or not with Rhizophagus irregularis (formerly Glomus intraradices) or Gigaspora margarita. We measured plant growth, colonization, contributions of AM and direct P uptake pathways using ³²P, and expression of plant P_i transporter genes at two levels of P supply. A17 (x2) responded positively to inoculation only at low P. The response was enhanced with 1 + 1 even at high P where colonization in A17 was reduced. With R. irregularis P uptake by the AM pathway was unaffected by P supply, whereas with G. margarita, the AM pathway was lower at high P, and direct uptake higher. Gene expression varied and was unrelated to P uptake through the two pathways. There was no evidence of plant control of P uptake via R. irregularis at high P but there was via G. margarita. Importantly, growth responses of plant genotypes grown alone did not predict outcomes of intergenotypic interactions.     

Tryptophan dimer produced by water-stressed bahia grass is an attractant for <Emphasis Type="Italic">Gigaspora margarita</Emphasis> and <Emphasis Type="Italic">Glomus caledonium</Emphasis>

We isolated and elucidated the structure of several stimulants for arbuscular mycorrhizal fungi (AMF) in water-stressed bahia grass roots. We could isolate some compounds that promoted the growth of Gigaspora margarita Becker and Hall and Glomus caledonium (Nicol. and Gerd.) Trappe and Gerd. In these compounds, tryptophan dimer (Trp–Trp) was elucidated the structure. Trp–Trp was abundantly produced in water-stressed bahia grass roots and exuded to the soil, although it was scarcely detected in non-stressed root exudates. Interestingly, this peptide strongly attracted the hyphae of Gi. margarita and G. caledonium and promoted their hyphal growth in vitro (1.8 × longer than the control). Tryptophan, however, had no effect on hyphal growth and attraction. Thus, Trp–Trp exuded from water-stressed roots would play an important role as a major signal for AMF. An erratum to this article can be found at     

Effects of arbuscular mycorrhizal fungi on zinnia and the different colonization between Gigaspora and Glomus

Zinnia (Zinnia elegans) was inoculated with four arbuscular mycorrhizal fungi (AMF) i.e. Gigaspora margarita, Gigaspora rosea, Glomus intraradices, and Glomus mosseae, either singly or mixture of two species of Gigaspora and Glomus. Results indicated that Glomus significantly enhanced the leaf size and the shoot biomass. G. mosseae was more effective than G. intraradices. Only G. mosseae increased number and size of flowers. Mixed inoculations were not much effective in the growth-promotion than the corresponding singly inoculation with Glomus. Comparison of colonization percent demonstrated that the highest colonization by G. mosseae, and followed by G. intraradices and Gigaspora species. In semi-quantitative PCR amplifications, Glomus was dominant in the roots. Our results suggest that G. mosseae is good for inoculation to zinnia and the interaction between different AMF species should be given full consideration in the application.     

Comparison of <Superscript>233</Superscript>U and <Superscript>33</Superscript>P uptake and translocation by the arbuscular mycorrhizal fungus<Emphasis Type="Italic"> Glomus intraradices</Emphasis> in root organ culture conditions

This study aimed to quantify and compare ²³³U and ³³P uptake and translocation by hyphae of the arbuscular mycorrhizal (AM) fungus Glomus intraradices in root organ culture conditions with transformed carrot (Daucus carota L.) roots as host. Mycorrhizal roots were grown in two-compartment Petri dishes to spatially separate a root compartment (RC) and a hyphal compartment (HC). The HC was labelled with 8.33 Bq ²³³U ml^–1 and 13.33 Bq ³³P ml^–1. After 2 weeks contact between hyphae and the labelled solution, ²³³U and ³³P activities were measured in the RC and in the HC. ²³³U and ³³P were taken up by the extraradical AM mycelium grown in the HC and this uptake represented 4.4% and 16% of the initial isotope supply, respectively. The translocation into roots developing in the RC via hyphae accounted for 5.9% and 72% of the initial isotope supply, respectively. Thus, both uptake and translocation were much higher for ³³P than for ²³³U. This suggests (1) the existence in hyphal tissues of efficient mechanisms limiting the uptake and translocation of non-essential elements such as U, and (2) that the hyphae have a higher sequestration than translocation function for U, and the converse for P.     

Occurrence of arbuscular mycorrhiza in Castanospermum australe A. Cunn. & C. Fraser and effects on growth and production of castanospermine

 Castanospermum australe A. Cunn. & C. Fraser is the only species of the genus Castanospermum (the Moreton Bay chestnut or black bean) native to NE Australia. One constituent of the plant, castanospermine, can inhibit the AIDS virus. The present study investigated possible symbioses between its roots and arbuscular mycorrhizal (AM) fungi. The effects of mycorrhizal fungi on the growth of the plant and yield of alkaloid castanospermine were also studied. The mycorrhizosphere soil and roots of C. australe collected from various sites in and around Sydney, Australia showed AM symbiotic associations with roots, with arbuscules and vesicles in the root cortices. Wet sieving and decanting yielded AM fungal spores, mainly Glomus spp. A positive correlation was found between AM fungal infection and the castanospermine content of seeds of field-grown trees. Field study results were confirmed by growing seedlings under greenhouse conditions and inoculating them with Glomus intraradices Schenck and Smith (INVAM isolate KS906) and Gigaspora margarita Becker & Hall (INVAM isolate BR444–2). The AM fungi increased the growth and P contents of plants and the yield of castanospermine in the leaves, irrespective of the P treatment. No correlation was found between the alkaloid contents of leaves from mycorrhizal seedlings and from non-mycorrhizal plants which received P. No significant difference in the production of castanospermine was found between P treatments when G. margarita was used as inoculum. Accepted: 14 April 1999     

Influence of nitrogen and phosphorus on polyphosphate accumulation in Gigaspora margarita during spore germination

Polyphosphate (polyP) is the form in which phosphorus (P) is transferred from extraradical hyphae into arbuscles in the symbiotic stage of arbuscular mycorrhizal fungi. However, polyP dynamics in the presymbiotic stage are less understood. In this study, we aimed to investigate polyP accumulation in Gigaspora margarita as influenced by nitrogen (N) and/or P supply during germination. Spores of G. margarita were incubated on medium with or without P or N addition. PolyP content in the fungal tissue was monitored using a polyP kinase/luciferase system, and polyP synthetic activity was determined with ³²P labeling. The results showed that both N and P were necessary for polyP accumulation in germ tubes. Nitrate increased the polyP content in germ tubes, but ammonium did not. Along with germination, polyP content decreased in spores, but increased in germ tubes. ³²P labeling indicated that polyP synthetic activity increased in germ tubes along with germination, but was negligible in spores. Our results suggest that, in the presymbiotic stage of G. margarita, uptake of environmental N and P increases polyP content in germ tubes, and that polyP synthesis occurs mainly therein, leading to polyP accumulation. The possible mechanism of transfer of polyP from spores to hyphae remains to be elucidated.     

Extensive In Vitro Hyphal Growth of Vesicular-Arbuscular Mycorrhizal Fungi in the Presence of CO2 and Flavonols

Various flavonoids were tested for their ability to stimulate in vitro growth of germinated spores of vesicular-arbuscular mycorrhizal fungi. Experiments were performed in the presence of 2% CO₂, previously demonstrated to be required for growth of Gigaspora margarita (G. Bécard and Y. Piché, Appl. Environ. Microbiol. 55:2320-2325, 1989). Only the flavonols stimulated fungal growth. The flavones, flavanones, and isoflavones tested were generally inhibitory. Quercetin (10 μM) prolonged hyphal growth from germinated spores of G. margarita from 10 to 42 days. An average of more than 500 mm of hyphal growth and 13 auxiliary cells per spore were obtained. Quercetin also stimulated the growth of Glomus etunicatum. The glycosides of quercetin, rutin, and quercitrin were not stimulatory. The axenic growth of G. margarita achieved here under rigorously defined conditions is the most ever reported for a vesicular-arbuscular mycorrhizal fungus.