In this paper we review the membrane transport processes that are involved in the transfer of mineral nutrients and organic carbon between the symbiotic partners in mycorrhizas. In particular, we reassess the prevailing hypothesis that transfer in vesicular-arbuscular (VA) mycorrhizas occurs simultaneously and bidirectionally across the same interface and that arbuscules are the main sites of transfer. Using cytochemical techniques, we and our collaborators have reexamined the distribution of ATPases in the arbuscular and intercellular hyphal interfaces in VA mycorrhizas formed between roots ofAllium cepa (onion) and the fungus Glomus intraradices. The results showed that H +-ATPases have different localisation on plant and fungal membranes in arbuscular and hyphal interfaces (Gianinazzi-Pearson et al. 1991). While some arbuscular interfaces had H+-ATPase activity on both fungal and plant membranes, in most cases the fungal membrane lacked this activity. In contrast, the plasma membranes of intercellular hyphae always had H + -ATPase and the adjacent root cells did not. This suggests that the different interfaces in a VA mycorrhiza may have different functions. We propose that passive loss of P from the arbuscules is associated with active uptake by the energised (ATPase-bearing) plant membrane and that passive loss of carbohydrate from the root cells is followed by active uptake by the intercellular hyphae. If this model is correct, then variations in "mycorrhizal efficiency" (i.e. the extent to which mycorrhizal plants grow better than non-mycorrhizal controls) might be determined by differences in the numbers of active arbuscules as a proportion of the total fungal biomass within the root. As a first step towards investigating this possibility, we have developed methods for measuring the surface areas of arbuscular and hyphal interfaces in different fungus-host combinations, Glomus spp./ Allium porrum (leek). We have also measured fluxes of P from fungus to plant and have been able to partition these between the arbuscular and total (arbuscular plus hyphal) interfaces. The implications of this work, and suggestions for future investigations of the molecular mechanisms involved in nutrient transfer in mycorrhizas, are discussed

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 (32)P, and expression of plant Pi 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.

Here, we used phosphorus-32 (32P) labelling in compartmented pots combined with quantitative real-time polymerase chain reaction (PCR) analysis of phosphate(Pi) transporter gene expression to investigate regulation of Pi uptake pathways in barley (Hordeum vulgare), an arbuscular mycorrhizal (AM) plant that does not show strong positive growth responses to colonization.Barley was colonized well by Glomus intraradices and poorly by Glomus geosporum,but both fungi induced significant and similar growth depressions compared with non mycorrhizal controls. The lack of correlation between per cent colonization and extent of growth depression suggests that the latter is not related to carbon drain to the fungus. The contribution of the AM Pi uptake pathway for the two fungi was, in general,related to per cent colonization and expression of the AM-inducible Pi transporter gene, HvPT8, but not to plant responsiveness. Glomus intraradices contributed 48%of total plant P whereas G. geosporum contributed very little.The growth depression in plants where the AM uptake pathway was functional suggests that the contribution of the direct Pi uptake pathway via root hairs and epidermis was decreased. This decrease was not correlated with downregulation of the epidermal-expressed Pi transporter genes, HvPT1 and HvPT2. We hypothesize post-transcriptional or post-translational control of this transport process by AM colonization.

*Here, we used barley (Hordeum vulgare) grown in normal and compartmented pots to investigate sensitivity to arsenic (As) in the absence of a positive growth response to arbuscular mycorrhizas (AM). *We tested the hypothesis that upon inoculation with AM fungi downregulation of HvPht1;1 and HvPht1;2 genes (encoding high-affinity inorganic orthophosphate (P(i))-uptake systems in a direct pathway via root epidermis and root hairs) and upregulation of the AM-induced HvPht1;8 (encoding the P(i)-uptake system responsible for transfer of P(i) from the symbiotic interface to cortical cells) play a role in decreased As uptake and hence reduced As sensitivity in AM plants. *Barley did not respond, or responded negatively to colonization by Glomus intraradices in terms of growth. In terms of specific phosphorus (P) uptake (P uptake per g of root) barley was nonresponsive. There was a significant interaction between As treatment and colonization, resulting in a lower As concentration and uptake in AM compared with nonmycorrhizal (NM) plants. *The decreased uptake of As and higher P : As molar ratios in the AM barley can be explained by the operation of the AM pathway as indicated by induction of HvPht1;8 and by down-regulation of HvPht1;1 and HvPht1;2.

This study investigated the potential of the arbuscular mycorrhizal (AM) fungus Glomus mosseae to protect basil (Ocimum basilicum) against Fusarium oxysporum f.sp. basilici (Fob). It was hypothesised that G. mosseae could confer a bioprotective effect against Fob as a result of increases in leaf rosmarinic (RA) and caffeic acids (CA) or essential oil concentrations. Glomus mosseae conferred a bioprotective effect against Fob by reducing plant mortality to 20% compared to 33% in non‐mycorrhizal (NM) plants. This bioprotective effect was not related to improved phosphorus (P) nutrition, as AM and NM plants treated with Fob had similar shoot P concentrations (6 and 8 mg g−1 dry weight (DW), respectively). Both AM and NM plants treated with Fob had similar leaf and root RA and CA concentrations. Furthermore, phenolic (40–70 mg CA g−1 DW) or essential oil concentrations (0·1–0·6 mg g−1 DW) were not increased in plants treated with the AM fungus and Fob. Therefore, the bioprotective effect conferred by G. mosseae was not a result of increases in the phytochemicals tested in this study. However, under the AM symbiosis, basil plants treated with Fob had lower methyleugenol concentrations in their leaves (0·1 mg g−1 DW) than NM plants treated with the pathogen (0·6 mg g−1 DW).

We investigated the effects of phosphorus (P) supply on the uptake and toxicity of arsenate (As(V)) in two plant species (Medicago truncatula and Hordeum vulgare) grown in soil/sand mixes. Our initial hypothesis was that competition between phosphate (Pi) and As(V) for uptake would be observed, and that this would be the basis for the 'protective' effect of P with respect to As toxicity, as shown in solution culture. Addition of P to the soil/sand mixes did not have major effects on water extractable As, or vice versa. We observed that toxic effects of As(V) on plant growth were ameliorated by increased P in both plant species. However, we found no evidence that increased P supply reduced specific uptake of As(V) on a molar basis, so that competition with Pi could not be the basis for the effect. A more complex mechanism of protection is indicated which might relate to different Pi transport systems being expressed at different P levels.

The potential of three arbuscular mycorrhizal fungi (AMF) to enhance the production of antioxidants (rosmarinic and caffeic acids, RA and CA) was investigated in sweet basil (Ocimum basilicum). After adjusting phosphorus (P) nutrition so that P concentrations and yield were matched in AM and non-mycorrhizal (NM) plants we demonstrated that Glomus caledonium increased RA and CA production in the shoots. Glomus mosseae also increased shoot CA concentration in basil under similar conditions. Although higher P amendments to NM plants increased RA and CA concentrations, there was higher production of RA and CA in the shoots of AM plants, which was not solely due to better P nutrition. Therefore, AMF potentially represent an alternative way of promoting growth of this important medicinal herb, as natural ways of growing such crops are currently highly sought after in the herbal industry.

This review commemorates and examines the significance of the work of Isobel Gallaud more than 100 years ago that first established the existence of distinct structural classes ( Arum -type and Paris -type) within arbuscular mycorrhizal (AM) symbioses. We add new information from recent publications to the previous data last collated 10 years ago to consider whether any patterns have emerged on the basis of different fungal morphology within plant species or families. We discuss: (1) possible control exerted by the fungus over AM morphology; (2) apparent lack of plant phylogenetic relationships between the classes; (3) functions of the interfaces in different structural classes in relation to nutrient transfer in particular; and (4) the occurrence of plants with both of the major classes, and with intermediate AM structures, in different plant habitats. We also give suggestions for future research to help remove uncertainties about the functional and ecological significance of differences in AM morphology. Lastly, we urge retention of the terms Arum - and Paris -type, which are now well recognised by those who study AM symbioses.

A greenhouse experiment was conducted to evaluate the potential role of arbuscular mycorrhizal fungi (AMF) in encouraging revegetation of copper (Cu) mine tailings. Two native plant species, Coreopsis drummondii and Pteris vittata, together with a turf grass, Lolium perenne and a leguminous plant Trifolium repens associated with and without AMF Glomus mosseae were grown in Cu mine tailings to assess mycorrhizal effects on plant growth, mineral nutrition and metal uptake. Results indicated that symbiotic associations were successfully established between G. mosseae and all plants tested, and mycorrhizal colonization markedly increased plant dry matter yield except for L. perenne. The beneficial impacts of mycorrhizal colonization on plant growth could be largely explained by both improved P nutrition and decreased shoot Cu, As and Cd concentrations. The experiment provided evidence for the potential use of local plant species in combination with AMF for ecological restoration of metalliferous mine tailings.

Two cultivars of winter wheat (Triticum aestivum L.) (Jing 411 and Lovrin 10) were used to investigate arsenate (As) uptake and distribution in plants grown in hydroponic culture and in the soil. Results showed that without As addition, Lovrin 10 had higher biomass than Jing 411 in the soil pot experiment; in the hydroponic experiment Lovrin 10 had similar root biomass to and lower shoot biomass than Jing 411. Increasing P supply from 32 to 161 microM resulted in lower tissue As concentrations, and increasing As supply from 0 to 2,000 microM resulted in lower tissue P concentrations. Increasing P supply tended to increase shoot-to-root ratios of As concentrations, and increasing As supply tended to decrease shoot-to-root ratios of As concentrations. Both cultivars invested more in root production under P deficient conditions than under P sufficient conditions. Lovrin 10 invested more biomass production to roots than Jing 411, which might be partly responsible for higher shoot P and As concentrations and higher shoot-to-root ratios of As concentrations. Moreover, Lovrin 10 allocated less As to roots than Jing 411 and the difference disappeared with decreasing P supply.

A tomato mutant with reduced mycorrhizal colonisation, rmc, confers resistance to almost all arbuscular mycorrhizal (AM) fungal species tested, although there is variation in colonisation of different root cell layers by different fungi and one species of AM fungus can colonise this mutant relatively normally. These variations indicate a high degree of specificity in relation to AM colonisation. We explored the possibility of specificity or otherwise in interactions between rmc and three non-AM root-infecting fungi, Rhizoctonia solani anastomosis groups (AG) 4 and AG8, and binucleate Rhizoctonia (BNR). There were no differences between the wild type tomato 76R and rmc in the speed or extent to which these fungi infected roots or caused disease. Infection by R. solani induced high levels of defence-related gene expression in both tomato genotypes relative to non-infected plants. In contrast, with BNR the expression of these genes was not induced or induced to a much lower extent than with R. solani. The expression of defence-related genes with these two non-AM fungi was very similar in the two plant genotypes. It was different from effects observed during colonisation by AM fungi, which enhanced expression of defence-related genes in rmc compared with the wild type tomato. The specificity and molecular mechanisms of rmc in control of AM colonisation are discussed.

The aim of this paper was to investigate the effects of soil conditions and distance from a host plant on the ability of hyphae of arbuscular mycorrhizal (AM) fungi to grow and colonise a new host. Two glasshouse experiments were conducted using compartmented pots. The first investigated the effects of distance between a colonised and uncolonised host plant (Trifolium subterraneum L.) and average pore size of the growth substrate (100μm, 38μm) on the ability of two AM fungi, G. intraradices and G. mosseae, to colonise a new host plant. The second experiment determined if the pore size of the substrate (100μm, 38μm) affected the growth of AM fungi in the absence of a new host. In Experiment 1, both G. mosseae and G. intraradices grew successfully through the two sand substrates and colonised new host plants. Both fungi reached and colonised new hosts fastest when hosts were separated by the shortest distance (2.5cm), with largest pore size substrate (100μm). G. mosseae produced more external hyphae per unit of colonised root and colonised new host plants more rapidly than G. intraradices. However, receiver plants colonised by G. mosseae exhibited a negative mycorrhizal growth response following colonisation. Experiment 2 showed that G. mosseae grew further from its host than G. intraradices. The results support the theory that some AM fungal species may produce large amounts of external hyphae primarily to increase the probability of locating and colonising a new host plant.

Two hydroponic experiments were conducted to investigate the antioxidant response of winter wheat (Triticum aestivum L.) to cadmium (Cd)-zinc (Zn) interactions, Seedlings of winter wheat (cv. Yuandong 977), were grown in modified Hoagland nutrient solution with the addition of increasing concentrations of Cd (0, 10, 25, 50 AM). In experiment 2, the seedlings of the same cultivar were treated with constant concentration of Cd (25 mu M) and varying levels of Zn (0, 1, 10, 50 mu M). Hydrogen peroxide (H2O2) and malondialdehyde (MDA) as well as the activities of three antioxidant enzymes, catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX) were monitored to estimate the amount of oxidative stress and the antioxidant ability of seedlings treated with Cd and Zn for 10 days. The results showed that levels of H2O2 and MDA in experiment I were significantly increased with increasing Cd concentrations. The data indicated that Cd could induce oxidative stress and lipid peroxidation in the plants. While H2O2 and MDA levels were significantly reduced by addition of Zn in experiment 2, the activities antioxidant enzymes were enhanced. A concentration of 10 mu M Zn appeared to be the optimal level in this experiment for seedlings' growth, chlorophyll synthesis and antioxidant status, indicating that Zn alleviated the oxidative stress induced by Cd.

Colonization of two plant species by Glomus intraradices was studied to investigate the two morphological types (Arum and Paris), their symbiotic interfaces and metabolic activities. Root pieces and sections were stained to observe the colonization and metabolic activity of all mycorrhizal structures. There were no growth responses observed in the plants caused by mycorrhizal symbiosis. The two morphological types had a similar percentage of root colonized, but the Arum-type had higher metabolic activity. Most of the mycorrhizal structures (88%) showed succinate dehydrogenase activity; about half showed acid phosphatase activity; and a small percentage showed alkaline phosphatase activity. Phosphatase activity was highest in arbuscules and low in intercellular hyphae in the Arum-type colonization. In the Paris-type, hyphal coils and arbusculate coils showed a similar intermediate percentage of phosphatase activity. We conclude that acid phosphatase is more important than alkaline phosphatase in both colonization types. We discuss the possibility that, whereas arbuscules in Arum-type are the main site for phosphorus release to the host plant, both the hyphal and arbusculate coils may be involved in the Paris-type.

We evaluated the roles of arbuscular mycorrhizal (AM) fungi in growth and phosphorus (P) nutrition of wheat ( Triticum aestivum L.) in a highly calcareous soil and compared the responses of wheat with those of clover ( Trifolium subterraneum L). In the first experiment wheat (cv. Brookton) was harvested at 6 wk. Colonisation by four AM fungi was low (<20%). Clover was harvested at 8 wk. Colonisation varied with different fungi, with the highest value (52%) obtained with Glomus intraradices . Although suffering from P deficiency, non-mycorrhizal (NM) wheat grew relatively well with no added P (P0) and application of P at 100 mg kg −1 (P100) increased the dry weight (DW). Shoot P concentrations increased with P application and there were positive effects of all AM fungi at P100. In contrast, NM clover grew very poorly at P0 and did not respond to P application. Clover responded positively to all AM fungi at both P levels, associated with increases in P uptake. In the second experiment colonisation by a single AM fungus ( G. intraradices ) of two wheat cultivars (Brookton and Krichauff) was well established at 6 wk (~50% in P0 plants) and continued to increase up to maturity (~70%), but decreased greatly at both harvests as P supply was increased (up to 150 mg P kg −1 : P150). Addition of P significantly increased plant growth, grain yield and P uptake irrespective of cultivar and harvest time, and the optimum soil P for grain yield was P100. In both cultivars, a growth depression in AM plants occurred at 6 wk at all P levels, but disappeared at 19 wk with added P. At P0, AM plants also produced lower grain yield (weight) per plant, but with higher P, AM plants produced higher grain yields than NM plants. There was a significant positive effect of AM on grain P concentration at P0, but not at other P levels. Brookton was somewhat more P efficient than Krichauff, and the latter responded more to AM fungi. This study showed that responses of wheat to AM inoculation and P supply were quite different from those of clover, and changed during development. Results are discussed in relation to the underlying soil properties.

Effects of application of zinc (Zn) (0, 1, 5, 10 mg kg(-1) soil) and phosphorus (P) (0, 10, 50, 100 mg kg(-1) soil) on growth and cadmium (Cd) accumulations in shoots and roots of winter wheat (Triticum aestivum L.) seedlings were investigated in a pot experiment. All soils were supplied with a constant concentration of Cd (6 mg kg(-1) soil). Phosphorus application resulted in a pronounced increase in shoot and root biomass. Effects of Zn on plant growth were not as marked as those of P. High Zn (10 mg kg(-1)) decreased the biomass of both shoots and roots; this result may be ascribed to Zn toxicity. Phosphor-us and Zn showed complicated interactions in uptake by plants within the ranges of P and Zn levels used. Cadmium in shoots decreased significantly with increasing Zn (P < 0.001) except at P addition of 10 mg kg(-1). In contrast, root Cd concentrations increased significantly except at Zn addition of 5 mg kg(-1) (P < 0.001). These results indicated that Zn might inhibit Cd translocation from roots to shoots. Cadmium concentrations increased in shoots (P < 0.001) but decreased in roots (P < 0.001) with increasing P supply. The interactions between Zn and P had a significant effect on Cd accumulation in both shoots (p = 0.002) and roots (P < 0.001).

A Lycopersicon esculentum mutant (rmc) is resistant to colonization by most arbuscular mycorrhizal fungi (AMF), but one Glomus intraradices isolate (WFVAM 23) develops arbuscules and vesicles in the rmc cortex. It is unknown whether the symbiotic phosphate (Pi)-uptake pathway is operational in this interaction. Hyphal uptake of (32)Pi and expression of plant Pi transporter genes were investigated in the rmc mutant and its wild-type progenitor (76R) associated with three AMF. Hyphae transferred (32)Pi in all symbioses with 76R and in the rmc-G. intraradices WFVAM 23 symbiosis. The other AMF did not colonize rmc. The Pi transporter-encoding LePT1 and LePT2 were expressed constitutively or in P-starved roots, respectively. The mycorrhiza-inducible Pi transporters LePT3 and LePT4 were expressed only in plants with AMF colonization and symbiotic (32)Pi transfer. LePT3 and LePT4 transcripts were reliable markers for a functional mycorrhizal uptake pathway in rmc. Our novel approach to the physiology and molecular biology of P transport can be applied to other arbuscular-mycorrhizal symbioses, irrespective of the size of plant responses.

ABSTRACTMost 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.

A greenhouse pot experiment was conducted to investigate heavy metal [copper (Cu), zinc (Zn), lead (Pb), and cadmium (Cd)] uptake by two upland rice cultivars, '91B3 and '277, grown in a sterilized field soil contaminated by a mixture of Cu, Zn, Pb, and Cd. Rice plants were inoculated with each of three arbuscular mycorrhizal fungi (AMF), Glomus versiforme (GV), Glomus mosseae (GM), and Glomus diaphanum (GD), or remained noninoculated (NM). Both rice cultivars could be colonized by the three AMF used in this experiment. The percentage of mycorrhizal colonization by the three AMFs on the two rice cultivars ranged from 30% to 70%. Mycorrhizal colonization of both upland rice cultivars had a large influence on plant growth by increasing the shoot and root biomass compared with non-inoculated (NM) plants. The results indicate that mycorrhiza exert some protective effects against the combined toxicity of Cu, Zn, Pb, and Cd in the contaminated soil. This conclusion is supported by the partitioning of heavy metals (HMs) in the two cultivars. In the two cultivars, colonization by AMF reduced the translocation of HMs from root to shoot (except that the colonization of AMF increased the Cu translocation of HMs in cultivar '277). Immobilization of the HMs in roots can alleviate the potential toxicity to shoots induced by the mixture of Cu, Zn, Pb, and Cd. The two rice cultivars showed significant differences in uptake of Cu, Zn, Pb, and Cd when uninoculated. GM inoculation gave the most protective effects on the two cultivars under the combined soil contamination.