Arbuscular mycorrhizal fungi in a mountain grassland II: Seasonal variation of colonization studied, along with its relation to grazing and metabolic host type

The relationships among seasons, host metabolic type, grazing and arbuscular mycorrhizal colonization were analyzed in a high South American native grassland. This study investigated seasonal changes and grazing effects on the symbiotic endomycorrhizal interaction in 5 Poaceae [C(3) metabolic pathway: Briza subaristata Lam., Deyeuxia hieronymi (Hack.) Türpe and Poa stuckertii (Hack.) Parodi; with C(4) metabolic pathway: Eragrostis lugens Nees and Sorghastrum pellitum (Hack.) Parodi; and a Rosaceae (Alchemilla pinnata Ruíz & Pav.)]. All hosts were dominant species in the mountain grassland in central Argentina. It was found that the seasons markedly influenced endomycorrhizal colonization, whereas grazing did not affect this interaction. C(4) grasses presented the highest root colonization. Hosts Briza subaristata (C(3) metabolic pathway) and Sorghastrum pellitum (C(4) metabolic pathway) showed Arum- and Paris-type colonization and intermediate forms.     

Identification of root-colonizing AM fungal communities and their responses to short-term climate change and grazing on Tibetan plateau

The molecular diversity of arbuscular mycorrhizal (AM) fungi remains largely unexplored in sedge-dominated ecosystems, much less their responses to global climate change and grazing. To determine the molecular diversity of AM fungi and how they are affected by climate change and grazing, we examined AM fungal communities inside roots in a sedge meadow ecosystem (4875 m a.s.l.) undergoing 4 yr. of fully factorial manipulations of experimental warming, snow addition and grazing on the Tibetan Plateau. A total of 21 AM fungal virtual taxa (VTs) were identified, including one new genus-like clade and one new VT unrecorded in the MaarjAM database. However, we did not observe significant effects of climate change and/or grazing on AM fungal abundance, diversity and community composition. These results indicate that diverse AM fungal taxa inhabit in the sedge-dominated meadows on the Tibetan Plateau, but the AM fungal communities were not sensitive to short-term climate change and grazing.     

Degradation of 4-fluorobiphenyl by mycorrhizal fungi as determined by (19)F nuclear magnetic resonance spectroscopy and (14)C radiolabelling analysis.

The pathways of biotransformation of 4-fluorobiphenyl (4FBP) by the ectomycorrhizal fungus Tylospora fibrilosa and several other mycorrhizal fungi were investigated by using (19)F nuclear magnetic resonance (NMR) spectroscopy in combination with (14)C radioisotope-detected high-performance liquid chromatography ((14)C-HPLC). Under the conditions used in this study T. fibrillosa and some other species degraded 4FBP. (14)C-HPLC profiles indicated that there were four major biotransformation products, whereas (19)F NMR showed that there were six major fluorine-containing products. We confirmed that 4-fluorobiphen-4'-ol and 4-fluorobiphen-3'-ol were two of the major products formed, but no other products were conclusively identified. There was no evidence for the expected biotransformation pathway (namely, meta cleavage of the less halogenated ring), as none of the expected products of this route were found. To the best of our knowledge, this is the first report describing intermediates formed during mycorrhizal degradation of halogenated biphenyls.     

丛枝菌根真菌对植物次生代谢的影响

丛枝菌根(AM)是自然界中分布最为广泛、最为重要的一类菌根,许多研究已经观察到丛枝菌根真菌与植物次生代谢的相关性,丛枝菌根真菌能够直接或间接地影响植物的次生代谢过程。植物的次生代谢产物主要分为萜类物质、酚类物质和含氮化合物(主要是生物碱)三大类群,该文简要介绍了丛枝菌根真菌对这3类植物次生代谢产物的影响。丛枝菌根真菌与萜类物质代谢关系的研究比较细致和深入,有些工作已经从细胞及分子水平探讨其间的作用机制,如Blumenin、类胡萝卜素等。丛枝菌根真菌与酚类物质代谢关系的研究也比较深入,其中具有特殊功能的酚类物质——植保素、细胞壁酚酸、类黄酮/异类黄酮等倍受关注。目前有关丛枝菌根真菌与生物碱关系的研究相对较少,不过现有的研究表明,菌根的形成有助于生物碱积累。     

菌根真菌影响森林生态系统碳循环研究进展

森林生态系统在全球碳(C)储量中占据极为重要的地位。菌根真菌广泛存在于森林生态系统中,在森林生态系统C循环过程中发挥重要的作用。阐述了不同菌根类型真菌在森林生态系统C循环过程中的功能,对比了温带/北方森林与热带/亚热带森林中菌根真菌介导的C循环研究方面新近取得的研究结果。发现温带和北方森林的外生菌根(EcM)植物对地上生物量C的贡献相对较小,然而是地下C储量的主要贡献者;以丛枝菌根(AM)共生为主的热带/亚热带森林地表生物量占比较高,表明AM植被对热带/亚热带森林地上生物量C的贡献相对较大。我们还就全球变化背景下,菌根真菌及其介导的森林生态系统C汇功能,以及不同菌根类型树种影响C循环的机制等进行了总结。菌根真菌通过影响凋落物分解、土壤有机质形成及地下根系生物量,进而影响整个森林生态系统的C循环功能。菌根介导的森林C循环过程很大程度上取决于(优势)树木的菌根类型和森林土壤中菌根真菌的群落结构。最后指出了当前研究存在的主要问题以及未来研究展望。本文旨在明确菌根真菌在森林生态系统C循环转化过程中的重要生态功能,有助于准确地评估森林生态系统C汇现状,为应对全球变化等提供重要的依据。     

甘肃盐碱土植物VA菌根真菌侵染研究

对甘肃盐碱土中植物的VA菌根真菌共生状况进行研究,结果表明：在10科17种植物中,除碱蓬（Suaeda glauca Beg.）外均被菌根真菌侵染,占94.1%;盐碱土中孢子密度较高,表明甘肃盐碱土生态系统中植物对菌根真菌具有较高的依赖性,菌根真菌在盐碱土中产孢能力较强;所调查植物的VA菌根结构类型Arum型占68.75%,Pris型占31.25%;菌根结构类型与宿主植物类型有关,禾本科（Poaceae）和鸢尾科（Iridaceae）植物为P型菌根,百合科（Liliaceae）、胡颓子科（Elaeagnaceae）等其它科植物均为A型菌根;土壤类型影响了宿主植物的菌根侵染率和根际土的孢子密度,相同宿主植物在不同类型土壤中的菌根侵染率和孢子密度具有很大的差异.     

Shifts in mycorrhizal fungi during the evolution of autotrophy to mycoheterotrophy in Cymbidium (Orchidaceae)

? Premise of the study: Mycoheterotrophic plants, which completely depend upon mycorrhizal fungi for their nutrient supply, have unusual associations with fungal partners. The processes involved in shifts in fungal associations during cladogenesis of plant partners from autotrophy to mycoheterotrophy have not been demonstrated using a robust phylogenetic framework. ? Methods: Consequences of a mycorrhizal shift were examined in Cymbidium (Orchidaceae) using achlorophyllous and sister chlorophyllous species. Fungal associates of the two achlorophyllous mycoheterotrophs (C. macrorhizon and C. aberrans), their close relatives, the chlorophyllous mixotrophs (C. goeringii and C. lancifolium) and an outgroup, the chlorophyllous autotroph C. dayanum, were identified by internal transcribed spacers of the nuclear ribosomal DNA sequences. ? Key results: Molecular identification of mycorrhizal fungi revealed: (1) the outgroup autotroph is predominantly dependent on saprobic Tulasnellaceae, (2) the mixotrophs associate with the Tulasnellaceae and ectomycorrhizal groups including the Sebacinales, Russulaceae, Thelephoraceae and Clavulinaceae, and (3) the two mycoheterotrophs are mostly specialized with ectomycorrhizal Sebacinales. ? Conclusion: Fungal partners in Cymbidium have shifted from saprobic to ectomycorrhizal fungi via a phase of coexistence of both nutritional types of fungi. These three phases correspond to the evolution from autotrophy to mycoheterotrophy via mixotrophy in Cymbidium. We demonstrate that shifts in mycorrhizal fungi correlate with the evolution of nutritional modes in plants. Furthermore, gradual shifts in fungal partners through a phase of coexistence of different types of mycobionts may play a crucial role in the evolution of mycoheterotrophic plants.     

Arbuscular mycorrhizal fungi alter phosphorus relations of broomsedge (Andropogon virginicus L.) plants

Broomsedge (Andropogon virginicus L.) is a dominant grass revegetating many abandoned coal-mined lands in West Virginia, USA. Residual soils on such sites are often characterized by low pH, low nutrients, and high aluminium. Experiments were conducted to assess the resistance of broomsedge to limited phosphorus (Pi) availability and to investigate the role that arbuscular mycorrhizal (AM) fungi play in aiding plant growth under low Pi conditions. Pregerminated mycorrhizal and non-mycorrhizal seedlings were grown in a sand-culture system with nutrient solutions containing Pi concentrations ranging from 10 to 100 microM for 8 weeks. Non-mycorrhizal plants exhibited severe inhibition of growth under Pi limitation (<60 microM). Colonization by AM fungi (combined Glomus clarum Nicolson & Schenck and Gigaspora gigantea (Nicol. & Gerd.) Gerd. & Trappe) greatly enhanced host plant growth at low Pi concentrations, but did not benefit growth when Pi was readily available (100 microM). In comparison to non-mycorrhizal plants, mycorrhizal plants had higher phosphorus use efficiency at low Pi concentrations and maintained nearly constant tissue nutrient concentrations across the gradient of Pi concentrations investigated. Manganese (Mn) and sodium (Na) accumulated in shoots of non-mycorrhizal plants under Pi limitation. Mycorrhizal plants exhibited lower instantaneous Pi uptake rates and significantly lower C(min) values compared to non-mycorrhizal plants. These patterns suggest that the symbiotic association between broomsedge roots and AM fungi effectively maintains nutrient homeostasis through changes in physiological properties, including nutrient uptake, allocation and use. The mycorrhizal association is thus a major adaptation that allows broomsedge to become established on infertile mined lands.     

Effects of livestock grazing intensity on soil arbuscular mycorrhizal fungi and glomalin-related soil protein in a mountain forest steppe and a desert steppe of Mongolia

Arbuscular mycorrhizal fungi (AMF) are important components of the grassland ecosystems in terms of plant phosphorus uptake and accumulation of glomalin-related soil protein (GRSP). Though Mongolian grasslands are seriously degraded by livestock grazing, the effects of grazing on soil AMF and GRSP remain unclear. Here, we examined community composition and diversity of AMF as well as amount of GRSP at three different grazing intensities: lightly grazed (LG), moderately grazed (MG), and heavily grazed (HG) under two different types of grassland, mountain forest steppe at Hustai and desert steppe at Mandalgobi. The diversity and biomass of AMF-host and non-AMF plants strongly affected the overall AMF community composition and its diversity. When we separately analyzed the factors affecting soil AMF diversity at Hustai and Mandalgobi, decrease in the shoot biomass of Poaceae plants at Hustai and decreases in the species number and shoot biomass of AMF-host plants at Mandalgobi were significantly correlated with AMF diversity. GRSP decreased with increasing grazing intensity, which was significantly correlated with soil pH and total root biomass at Hustai. The decrease in plant biomass caused by grazing thus led to GRSP reduction. Our results showed that change in soil AMF community caused by livestock grazing were associated with change in the biomass and diversity of functional vegetation groups such as Poaeceae, AMF-host and non-AMF plants, indicating the importance to focus on such functional vegetation groups to evaluate the effect of grazing on AMF.

     

水分胁迫下丛枝菌根真菌对枳实生苗生长和渗透调节物质含量的影响

采用盆栽试验研究了水分胁迫下接种丛枝菌根真菌摩西球囊霉(Glomaus mosseae)对枳[Poncirustrifoliat(L.)Raf.]实生苗的生长和渗透调节物质含量的影响.结果表明,在土壤含水量为20%、16%和12%条件下,接种G.mosseae能够增加植株的生长(株高、茎粗、叶面积、地上部干重、地下部干重和植株干重),促进植株根系活跃吸收面积和根际土壤有效磷的吸收,提高叶片和根系可溶性糖含量的积累,降低叶片脯氨酸含量,增强植株的水分利用效率(达20%～40%),使枳实生苗的抗旱能力得到增强.土壤含水量为20%和16%条件下接种G.mosseae对植株的效果较土壤含水量为12%条件下更显著.12%的土壤含水量严重抑制Gmosseae的侵染,说明丛枝菌根侵染程度轻,其对植物的效果也差.     

Effects of genetic modifications to flax (Linum usitatissimum) on arbuscular mycorrhiza and plant performance

Although arbuscular mycorrhizal fungi (AMF) are known for their positive effect on flax growth, the impact of genetic manipulation in this crop on arbuscular mycorrhiza and plant performance was assessed for the first time. Five types of transgenic flax that were generated to improve fiber quality and resistance to pathogens, through increased levels of either phenylpropanoids (W92.40), glycosyltransferase (GT4, GT5), or PR2 beta-1,3-glucanase (B14) or produce polyhydroxybutyrate (M50), were used. Introduced genetic modifications did not change the degree of mycorrhizal colonization as compared to parent cultivars Linola and Nike. Arbuscules were well developed in each tested transgenic type (except M50). In two lines (W92.40 and B14), a higher abundance of arbuscules was observed when compared to control, untransformed flax plants. However, in some cases (W92.40, GT4, GT5, and B14 Md), the mycorrhizal dependency for biomass production of transgenic plants was slightly lower when compared to the original cultivars. No significant influence of mycorrhiza on the photosynthetic activity of transformed lines was found, but in most cases P concentration in mycorrhizal plants remained higher than in nonmycorrhizal ones. The transformed flax lines meet the demands for better quality of fiber and higher resistance to pathogens, without significantly influencing the interaction with AMF.     

Frost hardiness of mycorrhizal (Hebeloma sp.) and non-mycorrhizal Scots pine roots

The frost hardiness (FH) of mycorrhizal [ectomycorrhizal (ECM)] and non-mycorrhizal (NM) Scots pine (Pinus sylvestris) seedlings was studied to assess whether mycorrhizal symbiosis affected the roots’ tolerance of below-zero temperatures. ECM (Hebeloma sp.) and NM seedlings were cultivated in a growth chamber for 18 weeks. After 13 weeks’ growth in long-day and high-temperature (LDHT) conditions, a half of the ECM and NM seedlings were moved into a chamber with short-day and low-temperature (SDLT) conditions to cold acclimate. After exposures to a range of below-zero temperatures, the FH of the roots was assessed by means of the relative electrolyte leakage test. The FH was determined as the inflection point of the temperature-response curve. No significant difference was found between the FH of mycorrhizal and non-mycorrhizal roots in LDHT (?8.9 and ?9.8 °C) or SDLT (?7.5 and ?6.8 °C). The mycorrhizal treatment had no significant effect on the total dry mass, the allocation of dry mass among the roots and needles or nutrient accumulation. The mycorrhizal treatment with Hebeloma sp. did not affect the FH of Scots pine in this experimental setup. More information is needed on the extent to which mycorrhizas tolerate low temperatures, especially with different nutrient contents and different mycorrhiza fungi.     

Divergence in mycorrhizal specialization within Hexalectris spicata (Orchidaceae), a nonphotosynthetic desert orchid

Evidence is accumulating for specialized yet evolutionarily dynamic associations between orchids and their mycorrhizal fungi. However, the frequency of tight mycorrhizal specificity and the phylogenetic scale of changes in specificity within the Orchidaceae are presently unknown. We used microscopic observations and PCR-based methods to address these questions in three taxa of nonphotosynthetic orchids within the Hexalectris spicata complex. Fungal ITS RFLP analysis and sequences of the ITS and nuclear LSU ribosomal gene fragments allowed us to identify the fungi colonizing 25 individuals and 50 roots. Thanatephorus ochraceus (Ceratobasidiaceae) was an occasional colonizer of mycorrhizal roots and nonmycorrhizal rhizomes. Members of the Sebacinaceae were the primary mycorrhizal fungi in every Hexalectris root and were phylogenetically intermixed with ectomycorrhizal taxa. These associates fell into six ITS RFLP types labeled B through G. Types B, C, D, and G were found in samples of H. spicata var. spicata, while only type E was found in H. spicata var. arizonica and only type F was found in H. revoluta. These results provide preliminary evidence for divergence in mycorrhizal specificity between these two closely related orchid taxa. We hypothesize that mycorrhizal interactions have contributed to the evolutionary diversification of the Orchidaceae.     

Nitrogen decreases and precipitation increases ectomycorrhizal extramatrical mycelia production in a longleaf pine forest

The rates and controls of ectomycorrhizal fungal production were assessed in a 22-year-old longleaf pine (Pinus palustris Mill.) plantation using a complete factorial design that included two foliar scorching (control and 95% plus needle scorch) and two nitrogen (N) fertilization (control and 5 g N m⁻² year⁻¹) treatments during an annual assessment. Ectomycorrhizal fungi production comprised of extramatrical mycelia, Hartig nets and mantles on fine root tips, and sporocarps was estimated to be 49 g m⁻² year⁻¹ in the control treatment plots. Extramatrical mycelia accounted for approximately 95% of the total mycorrhizal production estimate. Mycorrhizal production rates did not vary significantly among sample periods throughout the annual assessment (p = 0.1366). In addition, reduction in foliar leaf area via experimental scorching treatments did not influence mycorrhizal production (p = 0.9374), suggesting that stored carbon (C) may decouple the linkage between current photosynthate production and ectomycorrhizal fungi dynamics in this forest type. Nitrogen fertilization had a negative effect, whereas precipitation had a positive effect on mycorrhizal fungi production (p = 0.0292; r ² = 0.42). These results support the widely speculated but poorly documented supposition that mycorrhizal fungi are a large and dynamic component of C flow and nutrient cycling dynamics in forest ecosystems.     

13C and 15N in microarthropods reveal little response of Douglas-fir ecosystems to climate change

Understanding ecosystem carbon (C) and nitrogen (N) cycling under global change requires experiments maintaining natural interactions among soil structure, soil communities, nutrient availability, and plant growth. In model Douglas-fir ecosystems maintained for five growing seasons, elevated temperature and carbon dioxide (CO₂) increased photosynthesis and increased C storage belowground but not aboveground. We hypothesized that interactions between N cycling and C fluxes through two main groups of microbes, mycorrhizal fungi (symbiotic with plants) and saprotrophic fungi (free-living), mediated ecosystem C storage. To quantify proportions of mycorrhizal and saprotrophic fungi, we measured stable isotopes in fungivorous microarthropods that efficiently censused the fungal community. Fungivorous microarthropods consumed on average 35% mycorrhizal fungi and 65% saprotrophic fungi. Elevated temperature decreased C flux through mycorrhizal fungi by 7%, whereas elevated CO₂ increased it by 4%. The dietary proportion of mycorrhizal fungi correlated across treatments with total plant biomass (n= 4, r²= 0.96, P= 0.021), but not with root biomass. This suggests that belowground allocation increased with increasing plant biomass, but that mycorrhizal fungi were stronger sinks for recent photosynthate than roots. Low N content of needles (0.8–1.1%) and A horizon soil (0.11%) coupled with high C : N ratios of A horizon soil (25–26) and litter (36–48) indicated severe N limitation. Elevated temperature treatments increased the saprotrophic decomposition of litter and lowered litter C : N ratios. Because of low N availability of this litter, its decomposition presumably increased N immobilization belowground, thereby restricting soil N availability for both mycorrhizal fungi and plant growth. Although increased photosynthesis with elevated CO₂ increased allocation of C to ectomycorrhizal fungi, it did not benefit plant N status. Most N for plants and soil storage was derived from litter decomposition. N sequestration by mycorrhizal fungi and limited N release during litter decomposition by saprotrophic fungi restricted N supply to plants, thereby constraining plant growth response to the different treatments.     

Arbuscular Mycorrhizal Fungi Promote the Growth of Ceratocarpus arenarius (Chenopodiaceae) with No Enhancement of Phosphorus Nutrition

The mycorrhizal status of plants in the Chenopodiaceae is not well studied with a few controversial reports. This study examined arbuscular mycorrhizal (AM) colonization and growth response of Ceratocarpus arenarius in the field and a greenhouse inoculation trial. The colonization rate of AM fungi in C. arenarius in in-growth field cores was low (around 15%). Vesicles and intraradical hyphae were present during all growth stages, but no arbuscules were observed. Sequencing analysis of the large ribosomal rDNA subunit detected four culturable Glomus species, G. intraradices, G. mosseae, G. etunicatum and G. microaggregatum together with eight unculturable species belong to the Glomeromycota in the root system of C. arenarius collected from the field. These results establish the mycotrophic status of C. arenarius. Both in the field and in the greenhouse inoculation trial, the growth of C. arenarius was stimulated by the indigenous AM fungal community and the inoculated AM fungal isolates, respectively, but the P uptake and concentration of the mycorrhizal plants did not increase significantly over the controls in both experiments. Furthermore, the AM fungi significantly increased seed production. Our results suggest that an alternative reciprocal benefit to carbon-phosphorus trade-off between AM fungi and the chenopod plant might exist in the extremely arid environment.     

Symbiotic germination capability of four Epipactis species (Orchidaceae) is broader than expected from adult ecology

? Premise of the study: Both abiotic and biotic factors shape species distributions. Orchids produce minute seeds with few nutrient reserves, thus requiring mycorrhizal fungi for germination. Therefore, both environmental conditions and mycorrhizal fungi distribution affect their germination success, but these ecological requirements and their congruence with habitat preferences of adults remain poorly understood. We investigated the importance of these factors during germination in four forest orchid species of the genus Epipactis. ? Methods: We sowed seeds of three habitat specialists and one generalist in different forest types at sites harboring adults of at least one of these ecologically diverging species. We analyzed germination pattern and identified mycorrhizal fungi of both seedlings and adults. ? Key results: Habitat conditions had little influence on germination pattern as seedlings grew in more habitats than expected from the adults' ecology. Ectomycorrhizal fungi availability did not limit germination. Suitable mycorrhizal fungi, mostly pezizalean ascomycetes, were recruited in various forest types, though the fungal communities differed according to habitat type. Finally, orchids with divergent ecological preferences shared similar mycorrhizal fungi. ? Conclusions: Limited adult distribution contrasted with successful seed germination at diverse sites and indicates existence of niche differentiation between adults and seedlings. Ecological specialization may thus be determined by factors other than mycorrhizal fungi that act later in the ontogeny, perhaps during the transition to above-ground development.     

Mycorrhizal diversity and specificity in Lecanorchis (Orchidaceae)

Lecanorchis is a nonphotosynthetic plant genus in Vanilloideae, Orchidaceae. Because of the distribution of many Lecanorchis taxa in various climate conditions, we hypothesized that mycorrhizal diversity and specificity are different among the different taxa of Lecanorchis. In the present study, identities of mycorrhizal fungi were examined for 90 individuals of 10 Lecanorchis taxa at 26 sites from Niigata to Okinawa Prefectures in Japan. Phylogenetic analyses of Lecanorchis taxa based on the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene (rDNA) divided the examined Lecanorchis taxa into three groups, groups A, B, and C. ITS rDNA sequences suggested that fungi associating with Lecanorchis were ectomycorrhiza-forming fungi in Lactarius, Russula, Atheliaceae, and Sebacina, with Lactarius and Russula dominant. Our results suggested some degree of mycorrhizal specialization among Lecanorchis taxa. Interestingly, the Lecanorchis group C had some specific relationships with Lactarius, whereas less specificity was found in the relationships with Russula. However, observed specificity results may be biased by geographic opportunity, and we suggest further research to assess whether Lecanorchis species are limited to the associations we observed.     

Influence of vesicular-arbuscular mycorrhizal infection and P addition on growth and P nutrition of Anthyllis cytisoides L. and Brachypodium retusum (Pers.) Beauv.

The effect of P applications and mycorrhizal inoculation on the growth and P nutrition of Anthyllis cytisoides L. (Fabaceae) and Brachypodium retusum (Pers.) Beauv. (Poaceae) was studied. Both plants are widely distributed and well adapted to semi-arid habitats in southern Spain. In all treatments, even with high P doses, mycorrhizal plants showed a higher concentration of phosphorus in their tissues than non-mycorrhizal plants. Mycorrhizal inoculation enhanced the growth of the plants when no P was applied. At high P addition, non-mycorrhizal plants showed higher growth than mycorrhizal plants. The response of each plant type to P application was somewhat different.     

Comparison of phosphatase localization in the intraradical hyphae of arbuscular mycorrhizal fungi,Glomus spp. and Gigaspora spp.

The localization of acid and alkaline phosphatases in the intraradical hyphae of the arbuscular mycorrhizal fungi, Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe (Gm), Gl. etunicatum Becker and Gerd. (Ge) and Gigaspora rosea Nicol. and Schenck (Gir) were compared. Marigold (Tagetes patula L.) and leek (Allium porrum L.) were inoculated with each of the three fungi. The mycorrhizal roots were harvested at 3, 4, 5 and 6 weeks after sowing (WAS), treated with a digestion solution containing cellulase and pectinase, and then stained for phosphatase activities at pH 5.0 and pH 8.5. The development of fungal structures in the host root was also examined. Gm formed fine-branched (mature) arbuscules only at the early phase of infection (3 to 4 WAS). Mature arbuscules of Ge and Gir were observed from the early phase (4 WAS) up to the end of experiment. At pH 5.0, the localization of the phosphatase activities of the three fungi were similar irrespective to host plant species. The activity appeared in mature arbuscules and intercellular hyphae, whereas the collapsed arbuscules were inactive. Ten millimolar NaF, an acid phosphatase inhibitor, inhibited the phosphatase activities of Gm and Ge but did not affect that of Gir. At pH 8.5, a difference among the fungal species was found in the localization of phosphatase activity while that between host species was not. The mature arbuscules were also the active sites in all three species. Only Gir showed the activity in the intercellular hyphae while the two Glomus spp. did not. Five millimolar EDTA inhibited the activity of Gir at pH 8.5 while the activities of Ge and Gm were not affected by either 5 mM EDTA or 10 mM KCN (both are alkaline phosphatase inhibitors).