Direct application of carbendazim and propiconazole at field rates to the external mycelium of three arbuscular mycorrhizal fungi species: effect on 32P transport and succinate dehydrogenase activity

 The influence of the systemic fungicides propiconazole (Tilt 250E) and carbendazim (Bavistin) at field application rates on the functioning of three arbuscular mycorrhizal fungi was studied. Short-term fungal ³²P transport and succinate dehydrogenase (SDH) activity in external hyphae of Glomus intraradices Schenck and Smith, G. claroideum Schenck and Smith and G. invermaium Hall in symbiosis with pea (Pisum sativum L.) were measured. In the experimental system used, the hyphae grew into two root-free hyphal compartments (HCs). The fungicides were applied to each HC 24 days after sowing and ³²P was added to one HC of each pot. Four days later, the fungicide effect on fungal P transport was measured as the difference in ³²P content of treated and untreated plants. SDH activity in fungal hyphae was determined in the HCs given no ³²P. Carbendazim severely inhibited ³²P transport and SDH activity in external hyphae at an application rate of 0.5 μg g^–1 soil. The ergosterol inhibitor propiconazole affected none of these parameters. The fungicides had similar effects on all three fungal species, although P transport efficiency and SDH activity differed markedly between the fungi. Accepted: 12 December 1996     

Influence of two legume species on hyphal production and activity of two arbuscular mycorrhizal fungi

 Two arbuscular mycorrhizal (AM) fungi (Glomus mosseae and G. intraradices) were compared for abundance of intraradical and soil-borne hyphae in association with Astragalus sinicum, a small-seeded, and Glycine max, a large-seeded legume. A. sinicum was more responsive than G. max to mycorrhizal formation, especially at early growth stages. Biomass allocation was greater in roots than shoots for mycorrhizal A. sinicum, while the opposite was true for G. max. Hyphal development in root and soil compartments was estimated by trypan blue staining and after staining for succinate dehydrogenase (SDH) or alkaline phosphatase (ALP) activity. Total fungal abundance increased steadily in roots and soil with time to a maximum 8 weeks after planting. SDH- and ALP-active AM hyphae increased in roots during plant growth but decreased in soil at later harvests. Mycorrhizal root mass in A. sinicum and G. max increased about 14-fold and 2.5-fold, respectively, but total length of soil hyphae produced per plant differed little, so that the pattern of AM soil to root abundance of the two fungi varied considerably with the host plant. Accepted: 23 July 1997     

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

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

The BEG Expert System – a multimedia identification system for arbuscular mycorrhizal fungi

 Knowledge of the biodiversity of arbuscular mycorrhizal fungi is limited and keys for their identification are not in a user-friendly format for the non-expert. An Expert System, linked to the BEG (La Banque Européenne des Glomales), has been developed on a CD-ROM to produce a multimedia identification system for these fungi. Accepted: 12 March 1996     

Effects of Pseudomonas fluorescens DF57 on growth and P uptake of two arbuscular mycorrhizal fungi in symbiosis with cucumber

 The effect of Pseudomonas fluorescens DF57 on growth and P uptake of two arbuscular mycorrhizal (AM) fungi in symbiosis with cucumber plants was studied in compartmentalised growth systems. Hyphae of Glomus intraradices Schenck & Smith (BEG87) or G. caledonium (Nicol. & Gerd.) Trappe & Gerdeman (BEG15) grew into lateral root-free compartments. Non-mycorrhizal plants served as control. The soil in half of the growth units of each mycorrhizal treatment was inoculated with P. fluorescens DF57. P. fluorescens DF57 enhanced hyphal length density of one of the AM fungi, G. caledonium, but this was not reflected in a higher hyphal transport of P from the root-free soil to the plant. The total P content was higher in plants grown in symbiosis with G. intraradices than in plants in the other treatments. G. caledonium and P. fluorescens DF57 had a synergistic effect in that total P content in plants inoculated with G. caledonium was higher in the presence than in the absence of P. fluorescens DF57. Accepted: 7 January 1999     

Different growth response of five co-existing stoloniferous plant species to inoculation with native arbuscular mycorrhizal fungi

Five species of stoloniferous plants originating from the same field site (Galeobdolon montanum, Glechoma hederacea, Potentilla anserina, Ranunculus repens and Trifolium repens) were studied with respect to their interaction with arbuscular mycorrhizal (AM) fungi. More specifically, the question was addressed whether mycorrhizal growth response of host plant species could be related to their vegetative mobility. The roots of all the species examined were colonised with AM fungi in the field, with the percentage of colonisation varying among species from approximately 40% to 90%. In a subsequent pot experiment, plants of all the species were either left non-inoculated or were inoculated with a mixture of three native AM fungi isolated from the site of plant origin (Glomus mosseae, G. intraradices and G. microaggregatum). AM fungi increased phosphorus uptake in all the plant species; however, plant growth response to inoculation varied widely from negative to positive. In addition to the biomass response, AM inoculation led to a change in clonal growth traits such as stolon number and length or ramet number in some species. Possible causes of the observed differences in mycorrhizal growth response of various stoloniferous plants are discussed.     

Response of endangered plant species to inoculation with arbuscular mycorrhizal fungi and soil bacteria

Three endangered plant species, Plantago atrata and Pulsatilla slavica, which are on the IUCN red list of plants, and Senecio umbrosus, which is extinct in the wild in Poland, were inoculated with soil microorganisms to evaluate their responsiveness to inoculation and to select the most effective microbial consortium for application in conservation projects. Individuals of these taxa were cultivated with (1) native arbuscular mycorrhizal fungi (AMF) isolated from natural habitats of the investigated species, (2) a mixture of AMF strains available in the laboratory, and (3) a combination of AMF lab strains with rhizobacteria. The plants were found to be dependent on AMF for their growth; the mycorrhizal dependency for P. atrata was 91%, S. umbrosus-95%, and P. slavica-65%. The applied inocula did not significantly differ in the stimulation of the growth of P. atrata and S. umbrosus, while in P. slavica, native AMF proved to be the less efficient. We therefore conclude that AMF application can improve the ex situ propagation of these three threatened taxa and may contribute to the success of S. umbrosus reintroduction. A multilevel analysis of chlorophyll a fluorescence transients by the JIP test permitted an in vivo evaluation of plant vitality in terms of biophysical parameters quantifying photosynthetic energy conservation, which was found to be in good agreement with the results concerning physiological parameters. Therefore, the JIP test can be used to evaluate the influence of AMF on endangered plants, with the additional advantage of being applicable in monitoring in a noninvasive way the acclimatization of reintroduced species in nature.     

Response of two tomato cultivars differing in salt tolerance to inoculation with mycorrhizal fungi under salt stress

Effects of arbuscular mycorrhizal fungi (AMF) and salt stress on nutrient acquisition and growth of two tomato cultivars exhibiting differences in salt tolerance were investigated. Plants were grown in a sterilized, low-P (silty clay) soil-sand mix. Salt was applied at saturation extract (EC_e) values of 1.4 (control), 4.9 (medium) and 7.1 dS m^–1 (high salt stress). Mycorrhizal colonization occurred irrespective of salt stress in both cultivars, but AMF colonization was higher under control than under saline soil conditions. The salt-tolerant cultivar Pello showed higher mycorrhizal colonization than the salt-sensitive cultivar Marriha. Shoot dry matter (DM) yield and leaf area were higher in mycorrhizal than nonmycorrhizal plants of both cultivars. Shoot DM and leaf area but not root DM were higher in Pello than Marriha. The enhancement in shoot DM due to AMF inoculation was 22% and 21% under control, 31% and 58% under medium, and 18% and 59% under high salinity for Pello and Marriha, respectively. For both cultivars, the contents of P, K, Zn, Cu, and Fe were higher in mycorrhizal than nonmycorrhizal plants under control and medium saline soil conditions. The enhancement in P, K, Zn, Cu, and Fe acquisition due to AMF inoculation was more pronounced in Marriha than in the Pello cultivar under saline conditions. The results suggest that Marriha benefited more from AMF colonization than Pello under saline soil conditions, despite the fact that Pello roots were highly infected with the AMF. Thus, it appears that Marriha is more dependent on AMF symbiosis than Pello. Accepted: 22 January 2001     

Growth and survival of seedlings of native plants in an impoverished and highly disturbed soil following inoculation with arbuscular mycorrhizal fungi

We investigated whether arbuscular mycorrhizas influenced growth and survival of seedlings in an extremely impoverished and highly disturbed soil. Seedlings of four plants species native to the site were either inoculated with native sporocarpic arbuscular mycorrhizal (AM) fungi or fertilised prior to transplanting, and followed over 86 weeks at the site. One treatment was also irrigated with N-rich leachate from the site. In a laboratory experiment, seedlings were fertilised with excess P for 6 weeks, and location of the P store determined. Growth and survival of AM and fertilised seedlings were similar at the site. Inoculated mycorrhizal fungi and roots appeared to extend into the surrounding soil together. P concentration in leaves of all plants was extremely low. Irrigation with leachate increased growth of seedlings. In the laboratory experiment, significantly more P was stored in roots than shoots. We suggest that successful revegetation of extremely disturbed and impoverished sites requires selection of mycorrhizal fungi and plants to suit the edaphic conditions and methods of out-planting.     

Effect of root exudate fractions from P-deficient and P-sufficient onion plants on root colonisation by the arbuscular mycorrhizal fungus Gigaspora margarita

 The effect of root exudates from P-deficient onion on root colonisation by an arbuscular mycorrhizal fungus was examined. Onions (Allium cepa L.) were grown in solution culture at phosphorus concentrations of 0 (P0) and 2 (P2) mg P l^–1. Root exudates were collected and fractionated with Amberlite XAD-4 resin to give EtOH and water soluble fractions. Onions inoculated with the arbuscular mycorrhizal fungus Gigaspora margarita Becker & Hall were grown with or without (control) root exudates and exudate fractions in a growth chamber. After 24 days, arbuscular mycorrhiza levels and appressoria formation had increased in plants treated with P0-root exudate or the P0-EtOH fraction when compared to corresponding P2 treatments or control plants. P0 and P2 water-soluble fractions did not significantly affect either aspect of fungal development. These results suggest that hydrophobic compounds found in root exudates from P-deficient onion increase appressorium formation and, therefore, enhance mycorrhiza development. Accepted: 2 June 1998     

Accumulation of secondary compounds in barley and wheat roots in response to inoculation with an arbuscular mycorrhizal fungus and co-inoculation with rhizosphere bacteria

 Colonization of Hordeum vulgare L. cv. Salome (barley)and Triticum aestivum L. cv. Caprimus (wheat) roots by the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith leads to de novo synthesis of isoprenoid cyclohexenone derivatives with blumenin [9-O-(2′-O-β-glucuronosyl)-β-glucopyranoside of 6-(3-hydroxybutyl)-1,1,5-trimethyl-4-cyclohexen-3-one] as the major constituent and to transient accumulation of hydroxycinnamate amides (4-coumaroylagmatine and -putrescine). Accumulation of these compounds in mycorrhizal wheat roots started 2 weeks after sowing together with the onset of arbuscule formation and proceeded with mycorrhizal progression. Highest levels were reached in 3- to 4-week-old secondary roots (root branches of first and higher order) characterized by the formation of vesicles. In the final developmental stages, the fungus produced massive amounts of spores, enclosing the stele of older root parts (older than 5 weeks) characterized by cortical death. In these root parts, the secondary compounds were detected in trace amounts only, indicating that they were located in the cortical tissues. Some rhizosphere bacteria tested, i.e. Agrobacterium rhizogenes, Pseudomonas fluorescens, and Rhizobium leguminosarum, markedly stimulated both fungal root colonization and blumenin accumulation, thus, acting as mycorrhiza-helper bacteria (MHB). Application of blumenin itself strongly inhibited fungal colonization and arbuscule formation at early stages of mycorrhiza development. This was associated with a markedly reduced accumulation of the hydroxycinnamate amides 4-coumaroylputrescine and -agmatine. The results suggest that both the isoprenoid and the phenylpropanoid metabolism are closely linked to the developmental stage and the extent of fungal colonization. Their possible involvement in the regulation of mycorrhiza development is discussed. Accepted: 18 September 1998     

Comparison of two test systems for measuring plant phosphorus uptake via arbuscular mycorrhizal fungi

 Plant phosphorus uptake via external hyphae of arbuscular mycorrhizal fungi has been measured using compartmented systems where a hyphal compartment is separated from a rooting compartment by a fine mesh. By labelling the soil within the hyphal compartment with a radioactive phosphorus (P) isotope, hyphal uptake of P into the plant can be traced. The objective of this growth chamber study was to test two hyphal compartments of different design with respect to their suitabilities for measurement of hyphal P uptake. One hyphal compartment was simply a nylon mesh bag filled with ³²P-labelled soil. The labelled soil in the other hyphal compartment was completely surrounded by an 8–10 mm layer of unlabelled soil that served as a buffer zone. Mycorrhizal and non-mycorrhizal subterranean clover plants were grown in pots with a centrally positioned hyphal compartment. Uptake of radioactive P by non-mycorrhizal control plants was 25% of that by mycorrhizal plants with the mesh bag but only 3% when including the buffer zone. Based on this good control of non-mycorrhizal P uptake from within the hyphal compartment and its greater ease of handling once produced, we judged the hyphal compartment including a buffer zone to be superior to the mesh bag. Accepted: 15 September 1998     

Response of strawberry to inoculation with arbuscular mycorrhizal fungi under very high soil phosphorus conditions

A field study was done to assess the potential benefit of arbuscular mycorrhizal (AM) inoculation of elite strawberry plants on plant multiplication, under typical strawberry nursery conditions and, in particular, high soil P fertility (Mehlich-3 extractible P=498 mg kg⁻¹). Commercially in vitro propagated elite plants of five cultivars (‘Chambly,’ ‘Glooscap,’ ‘Joliette,’ ‘Kent,’ and ‘Sweet Charlie’) were transplanted in noninoculated growth substrate or in substrate inoculated with Glomus intraradices or with a mixture of species (G. intraradices, Glomus mosseae, and Glomus etunicatum) at the acclimation stage and were grown for 6 weeks before transplantation in the field. We found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P. Inoculated mother plants produced about 25% fewer daughter plants than the control in Chambly (P=0.03), and Glooscap produced about 50% more (P=0.008) daughter plants when inoculated with G. intraradices, while the productivity of other cultivars was not significantly decreased. Daughter plant shoot mass was not affected by treatments, but their roots had lower, higher, or similar mass, depending on the cultivar–inoculum combination. Root mass was unrelated to plant number. The average level of AM colonization of daughter plants produced by noninoculated mother plants did not exceed 2%, whereas plants produced from inoculated mothers had over 10% of their root length colonized 7 weeks after transplantation of mother plants and ∼6% after 14 weeks (harvest), suggesting that the AM fungi brought into the field by inoculated mother plants had established and spread up to the daughter plants. The host or nonhost nature of the crop species preceding strawberry plant production (barley or buckwheat) had no effect on soil mycorrhizal potential, on mother plant productivity, or on daughter plant mycorrhizal development. Thus, in soil excessively rich in P, inoculation may be the only option for management of the symbiosis.     

Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland

 Analysis of the community of arbuscular mycorrhizal (AM) fungi in roots of Fragaria vesca growing in a heavy metal contaminated site was carried out on a Zn waste site near Chrzanow (southern Poland). The waste substratum was characterized by high contents of Pb, Zn, Cd, Cu and As, and by low levels of N, P and organic matter. Spores of Glomales were isolated by wet sieving and DNA was isolated from individual spores. Nested polymerase chain reaction (PCR) with taxon-specific primers was used to identify the species Glomus mosseae, Glomus intraradices and Glomus claroideum. Spores of other fungi were morphologically characterized and new taxon-discriminating molecular probes were developed for two of them (Glomus sp. HM-CL4 and HM-CL5) based on variations in the large ribosomal subunit (25S rDNA). High sequence similarities were found between Glomus sp. HM-CL4 and Glomus gerdemanii, and between Glomus sp. HM-CL5 and Glomus occultum. The designed primers were used to characterize the population of AM fungi colonizing the roots of F. vesca collected from the Zn waste site. The analysis, carried out on roots stained with trypan blue, showed that the most effective colonizer was closely related to G. gerdemannii. G. claroideum and the G. occultum-like fungus were slightly less common whilst frequencies of G. intraradices and G. mosseae in roots were much lower. The analysis of mycorrhiza stained with rhodizoniate to localize heavy metal accumulation showed that the stain does not influence the PCR reaction. Seventy percent of the root samples containing positively stained fungal hyphae were found to be colonized by G. mosseae. The data obtained demonstrate the usefulness of nested PCR for studies carried out in polluted areas. It will enable selection of AM fungi which are able to colonize plant roots under heavy metal stress conditions, as well as the identification of fungi showing high in situ accumulation of potentially toxic elements. Accepted: 7 July 2000     

Effect of host shoot clipping on carbon and nitrogen sources for arbuscular mycorrhizal fungi

The effect of clipping of the host-plant shoot on the sources of carbon and nitrogen for the arbuscular mycorrhizal (AM) fungus Gigaspora margarita was determined by measuring ¹³C in spores and hyphae in cocultures of C₃ and C₄ plants and by differential ¹⁵N labeling. C₃ and C₄ plants, which have different δ¹³C values, were grown in the same container separated by a series of hyphal compartments. The C₃ and C₄ plants were applied with ¹⁴N- and ¹⁵N-urea, respectively. After clipping of the C₃ shoots, spore δ ¹³C gradually approached that of the C₄ roots. Hyphal δ ¹³C paralleled that of spores. Spore % ¹⁵N was similar to that of mineral N in the C₄ plant compartment. Thus C in G. margarita coming from the clipped plants decreased with time. This demonstrates that C in AM fungi comes from living plants, whilst the N in spores comes mostly from the soil. Accepted: 28 November 2000     

Building a mycorrhizal cell: How to reach compatibility between plants and arbuscular mycorrhizal fungi

Abstract

Arbuscular mycorrhizal fungi occur throughout the majority of ecosystems supporting host plant nutrition. Recent findings describe the accommodation of the fungus by the root cell as a crucial step for compatibility between the partners. We discuss here the novel aspects of cellular plant-fungus interactions, with a particular attention to the interface compartment, the unique apoplastic space hosting intracellular fungal structures. The main features of arbuscular mycorrhizal colonization are examined and recent information in the field of plant and fungal cell responses during the establishment of the symbiosis is discussed. Differences between the colonization of root epidermal and cortical tissues are discussed, highlighting the growing interest in the role of epidermal cells during the first and decisive steps of the symbiosis. New approaches such as root organ cultures, in vivo observations, GFP tagging and mutant plant analysis are commented on and information from these is compared with that gained from more traditional methods. In particular, the use of plant mutants is depicted as a powerful tool for dissecting and understanding the genetic and cellular aspects of plant/fungus compatibility. Finally, perspectives in this field are outlined through the application of these approaches to the currently unanswered questions.     

Effects of inoculation of phosphate-solubilizing microorganisms and an arbuscular mycorrhizal fungus on mungbean grown under natural soil conditions

 The effect of inoculation of the phosphate-solubilizing microorganisms (PSM) Bacillus circulans and Cladosporium herbarum and the arbuscular mycorrhizal (AM) fungus Glomus fasciculatum with or without Mussoorie rockphosphate (MRP) was studied in a P-deficient natural non-disinfected sandy soil on mungbean (Vigna radiata). The AM levels increased following the addition of MRP or inoculation with PSM or G. fasciculatum. Both grain and straw yield of mungbean increased following inoculation with PSM or the AM fungus. In general, the increase in yield was higher in the presence of MRP and inoculation with a combination of PSM and AM fungus. Highest N and P uptake by mungbean was recorded after treatment with a combination of B. circulans, C. herbarum and G. fasciculatum in the presence of MRP. Generally the PSM population increased after AM fungus inoculation. Accepted: 13 October 1997     

Interactions between a mycophagous Collembola,dry yeast and the external mycelium of an arbuscular mycorrhizal fungus

 A plant growth system with root-free hyphal compartments was used to examine the interactions between a mycophagous Collembola (Folsomia candida Willem), dry yeast and an arbuscular mycorrhizal (AM) fungus [Glomus caledonium (Nicol. & Gerd.) Trappe and Gerdemann] in terms of Collembola reproduction, AM-hyphal length and AM-hyphal P transport. Collembola reproduction was unaffected by AM mycelium, but a supplement of dry yeast increased the Collembola population size. The addition of dry yeast increased AM-hyphal P transport by increasing hyphal length. Collembola without yeast affected neither AM-hyphal growth nor AM-hyphal P transport, whereas Collembola with yeast decreased AM-hyphal P transport by 75% after 8 weeks. The hyphal density of G. caledonium remained unaffected by Collembola except after 4 weeks in combination with yeast, when a 33% reduction was observed. The results of this experiment show that the interaction between F. candida and the external mycelium of G. caledonium is limited under the conditions imposed. Accepted: 27 February 1996