Differential host plant performance as a function of soil arbuscular mycorrhizal fungal communities: experimentally manipulating co-occurring Glomus species

In order to evaluate host plant performance relative to different soil arbuscular mycorrhizal fungal (AMF) communities, Andropogon gerardii seedlings were grown with nine different AMF communities. The communities consisted of 0, 10, or 20 spores of Glomus etunicatum and 0, 10, or 20 spores of Glomus intraradices in all possible combinations. Spores were produced by fungal cultures originating on A. gerardii in a serpentine plant community; seeds of A. gerardii were collected at the same site. The experiment was performed in the greenhouse using a mixture of sterilized serpentine soil and sand to which naturally occurring non-mycorrhizal microbes were added. There was no difference in root AMF colonization rates between single species communities of either G. etunicatum or G. intraradices, but G. intraradices enhanced plant growth and G. etunicatum did not. However, plants grew larger with some combinations of G.␣intraradices plus G. etunicatum than with the same quantity of G. intraradices alone. These results suggest the potential for niche complementarity in the mycorrhizal fungi. That G. etunicatum only increased plant growth in the presence of G. intraradices could be illustrative of why AMF that appear to be parasitic or benign when examined in isolation are maintained within multi-species mycorrhizal communities in nature.     

Interactions between<Emphasis Type="Italic"> Trichoderma pseudokoningii</Emphasis> strains and the arbuscular mycorrhizal fungi<Emphasis Type="Italic"> Glomus mosseae</Emphasis> and<Emphasis Type="Italic"> Gigaspora rosea</Emphasis>

The interaction between Trichoderma pseudokoningii (Rifai) 511, 2212, 741A, 741B and 453 and the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG12 and Gigaspora rosea Nicolson & Schenck BEG9 were studied in vitro and in greenhouse experiments. All T. pseudokoningii strains inhibited the germination of G. mosseae and Gi. rosea except the strain 453, which did not affect the germination of Gi. rosea. Soluble exudates and volatile substances produced by all T. pseudokoningii strains inhibited the spore germination of G. mosseae. The germination of Gi. rosea spores was inhibited by the soluble exudates produced by T. pseudokoningii 2212 and 511, whereas T. pseudokoningii 714A and 714B inhibited the germination of Gi. rosea spores by the production of volatile substances. The strains of T. pseudokoningii did not affect dry matter and percentage of root length colonization of soybean inoculated with G. mosseae, except T. pseudokoningii 2212, which inhibited both parameters. However, all T. pseudokoningii strains decreased the shoot dry matter and the percentage of AM root length colonization of soybean inoculated with Gi. rosea. The saprotrophic fungi tested seem to affect AM colonization of root by effects on the presymbiotic phase of the AM fungi. No influence of AM fungi on the number of CFUs of T. pseudokoningii was found. The effect of saprotrophic fungi on AM fungal development and function varied with the strain of the saprotrophic species tested.     

Field response of wheat to arbuscular mycorrhizal fungi and drought stress

Mycorrhizal plants often have greater tolerance to drought than nonmycorrhizal plants. This study was conducted to determine the effects of arbuscular mycorrhizal (AM) fungi inoculation on growth, grain yield and mineral acquisition of two winter wheat (Triticum aestivum L.) cultivars grown in the field under well-watered and water-stressed conditions. Wheat seeds were planted in furrows after treatment with or without the AM fungi Glomus mosseae or G. etunicatum. Roots were sampled at four growth stages (leaf, tillering, heading and grain-filling) to quantify AM fungi. There was negligible AM fungi colonization during winter months following seeding (leaf sampling in February), when soil temperature was low. During the spring, AM fungi colonization increased gradually. Mycorrhizal colonization was higher in well-watered plants colonized with AM fungi isolates than water-stressed plants. Plants inoculated with G. etunicatum generally had higher colonization than plants colonized with G. mosseae under both soil moisture conditions. Biomass and grain yields were higher in mycorrhizal than nonmycorrhizal plots irrespective of soil moisture, and G. etunicatum inoculated plants generally had higher biomass and grain yields than those colonized by G. mosseae under either soil moisture condition. The mycorrhizal plants had higher shoot P and Fe concentrations than nonmycorrhizal plants at all samplings regardless of soil moisture conditions. The improved growth, yield and nutrient uptake in wheat plants reported here demonstrate the potential of mycorrhizal inoculation to reduce the effects of drought stress on wheat grown under field conditions in semiarid areas of the world.     

Diversity and infectivity of arbuscular mycorrhizal fungi in agricultural soils of the Sichuan Province of mainland China

Knowledge about the presence and diversity of arbuscular mycorrhizal fungi (AMF) in a specific area is an essential first step for utilizing these fungi in any application. The community composition of AMF in intensively managed agricultural soil in the Sichuan Province of southwest China currently is unknown. In one set of samples, AMF were trapped in pot cultures from 40 fields growing legumes in the Panxi region, southeast Sichuan. In a second set of samples, the MPN method with four-fold dilutions and maize as host was used to estimate infective propagules in soil from another 50 agricultural sites throughout the province. Soil types were heterogeneous and were classified as purple, yellow, paddy and red. Crops at each site were either maize, wheat or sweet orange. From this set of soil, AMF spores were also extracted and identified. Including all ninety soils, thirty glomeromycotan species in Glomus (20 species), Acaulospora (four species), Scutellospora (three species), Ambispora (one species), Archaeospora (one species) and Paraglomus (one species) were identified. Yellow, red and purple soils yielded similar numbers of AMF species, while AMF species diversity was clearly lower in paddy soil. In trap culture soils, the most frequent species were Glomus aggregatum or Glomus intraradices, Glomus claroideum and Glomus etunicatum. The species Acaulospora capsicula, Acaulospora delicata, G. aggregatum (or intraradices), G. claroideum, Glomus epigaeum, G. etunicatum, Glomus luteum, Glomus monosporum, Glomus mosseae and Glomus proliferum were successfully cultured as single-species pot cultures in Plantago lanceolata. The three most frequent species in field soils were G. mosseae, Glomus caledonium and Glomus constrictum. MPN values varied between 17 and 3334 propagules 100 g soil⁻¹ among the fifty field sites sampled. Regression analysis, including host&soil, log(P) and pH as explanatory variables explained 59% of the variation in log(MPN). The highest MPN estimates were found in purple soil cropped with maize and citrus, 324 and 278 propagules 100 g soil⁻¹, respectively. The lowest MPN value, 54 propagules 100 g soil⁻¹, was measured in wheat in purple and yellow soil. Despite intensive agricultural management that can include often repeated tillage, our examination of 90 agricultural sites revealed that soils of the Sichuan region have moderate to high numbers of infective AMF propagules as well as a high AMF species diversity. This opens possibilities for further studies and utilization of AMF in agriculture and horticulture in the Sichuan province, People’s Republic of China.     

Influence of temperature on colonization of spring barleys by vesicular arbuscular mycorrhizal fungi

The effects of three soil temperatures on growth of spring barleys (Hordeum vulgare L.) and on their root colonization by vesicular arbuscular mycorrhizal (VAM) fungi from agricultural soils in Montana (USA) or Syria at different inoculum concentrations were tested in soil incubators in the greenhouse. The number of mycorrhizal plants as well as the proportion and intensity of roots colonized increased with higher soil temperatures. VAM fungi from Montana, primarily Glomus macrocarpum, were cold tolerant at 11°C while those from Syria, primarily G. hoi, were heat tolerant at 26°C. Inoculum potential of Montana VAM fungi was higher than Syrian VAM fungi in cool soils. Harmal, selected from Syrian barley land races, had the highest colonization by mycorrhizal fungi of the cultivars tested.Journal Series Paper: J-2532 Montana Agricultural Experiment Station.     

Interaction between supernodulating or non-nodulating mutants of soybean and two arbuscular mycorrhizal fungi

 Twelve nodulation mutants (seven non-nodulating and five supernodulating) of soybean [Glycine max (L.) Mirr.] were screened for arbuscular mycorrhizal colonization in the presence of either Glomus etunicatum Becker and Gerdemann or Gigaspora margarita Becker and Hall. The cultivars showed variation in colonization parameters. The two supernodulating mutants En6500 and NOD1–3 had higher frequencies of colonization with 2.5–4.5 times higher arbuscular abundance than the respective wild types. The enhanced mycorrhization resulted in significant enhancement of P uptake by En6500. The non-nodulating mutants showed decreases in mycorrhizal parameters. Mutants En1282 and Harosoy^–exhibited aborted infection after formation of typical appressorium-like structures at some sites. However, none of these had the non-mycorrhizal phenotype. Growth and nutrient-uptake parameters should be considered while studying plant mutants for mycorrhization. Accepted: 7 July 2000     

Effects of NaCl and Mycorrhizal Fungi on Antioxidative Enzymes in Soybean

The effects of different concentrations of NaCl on the activities of antioxidative enzymes in the shoots and roots of soybean (Glycine max [L.] Merr cv. Pershing) inoculated or not with an arbuscular mycorrhizal fungus, Glomus etunicatum Becker & Gerdemann, were studied. Furthermore, the effect of salt acclimated mycorrhizal fungi on the antioxidative enzymes in soybean plants grown under salt stress (100 mM NaCl) was investigated. Activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were increased in the shoots of both mycorrhizal (M) and nonmycorrhizal (NM) plants grown under NaCl salinity. Salinity increased SOD activity in the roots of M and NM plants, but had no effect on CAT and polyphenol oxidase activities in the roots. M plants had greater SOD, POD and ascorbate peroxidase activity under salinity. Under salt stress, soybean plants inoculated with salt pre-treated mycorrhizal fungi showed increased SOD and POD activity in shoots, relative to those inoculated with the non pre-treated fungi.     

Studies on the diversity of the distinct phylogenetic lineage encompassing Glomus claroideum and Glomus etunicatum

Morphological and molecular characters were analysed to investigate diversity within isolates of the Glomus claroideum/Glomus etunicatum species group in the genus Glomus. The inter- and intra-isolate sequence diversity of the large subunit (LSU) rRNA gene D2 region of eight isolates of G. claroideum and G. etunicatum was studied using PCR-single strand conformational polymorphism (SSCP)-sequencing. In addition, two isolates recently obtained from Southern China were included in the analysis to allow for a wider geographic screening. Single spore DNA isolation confirmed the magnitude of gene diversity found in multispore DNA extractions. An apparent overlap of spore morphological characters was found between G. claroideum and G. etunicatum in some isolates. Analysis of the sequence frequencies in all G. etunicatum and G. claroideum isolates (ten) showed that four LSU D2 sequences, representing 32.1% of the clones analysed for multispore extraction (564) were found to be common to both species, and those sequences were the most abundant in four of the ten isolates analysed. The frequency of these sequences ranged between 23.2% and 87.5% of the clones analysed in each isolate. The implications for the use of phenotypic characters to define species in arbuscular mycorrhizal fungi are discussed. The current position of G. claroideum/G.etunicatum in the taxonomy of the Glomeromycota is also discussed.     

Arbuscular mycorrhizal fungi and plant symbiosis in a saline-sodic soil

The seasonality of arbuscular mycorrhizal (AM) fungi–plant symbiosis in Lotus glaber Mill. and Stenotaphrum secundatum (Walt.) O.K. and the association with phosphorus (P) plant nutrition were studied in a saline-sodic soil at the four seasons during a year. Plant roots of both species were densely colonized by AM fungi (90 and 73%, respectively in L. glaber and S. secundatum) at high values of soil pH (9.2) and exchangeable sodium percentage (65%). The percentage of colonized root length differed between species and showed seasonality. The morphology of root colonization had a similar pattern in both species. The arbuscular colonization fraction increased at the beginning of the growing season and was positively associated with increased P concentration in both shoot and root tissue. The vesicular colonization fraction was high in summer when plants suffer from stress imposed by high temperatures and drought periods, and negatively associated with P in plant tissue. Spore and hyphal densities in soil were not associated with AM root colonization and did not show seasonality. Our results suggest that AM fungi can survive and colonize L. glaber and S. secundatum roots adapted to extreme saline-sodic soil condition. The symbiosis responds to seasonality and P uptake by the host altering the morphology of root colonization.     

Impact of Bacillus subtilis JA, a biocontrol strain of fungal plant pathogens, on arbuscular mycorrhiza formation in Zea mays

Summary  In the present study, the influence of Bacillus subtilis JA on arbuscular mycorrhizal fungi (AMF) was evaluated by either pot culture or in vitro conditions, respectively. Under the pot culture conditions, the inoculation of B. subtilis JA decreased the frequency (% F) of the root colonization by indigenous arbuscular mycorrhizal fungi and the shoot dry weight of maize (Zea mays L.), but had no apparent effect on the intensity (% I) of AM fungal root colonization. The unknown volatile emitted from the B. subtilis JA in vitro significantly inhibited spore germination and the hyphal growth in the dual-compartment experiments. Moreover, the data from the direct interaction between B. subtilis JA and Glomus etunicatum showed that soluble antifungal lipopeptides influenced the development of AMF. Therefore, the application of antifungal Bacillus strains should take the compatibility with the indigenous beneficial fungi into consideration.     

Preparation of gel-entrapped mycorrhizal inoculum in the presence or absence of Yarowia lipolytica

Two arbuscular mycorrhizal fungi (Glomus deserticola and Glomus fasciculatum) were entrapped in calcium alginate, alone or in combination with a phosphate-solubilizing yeast (Yarowia lipolytica) and, after storage for 60 days, were inoculated into soil microcosms with tomato as the test plant. The average extent of root colonization by gel-entrapped G. deserticola and G. fasciculatum were 32 ± 5.6 and 24 ± 12.1%, respectively. Improved infective potential and colonization efficiency were observed when Y. lipolytica was co-entrapped with the mycorrhizal fungi. The best value, 49%, of mycorrhizal colonization was in roots of plants inoculated with G. deserticola co-entrapped with Y. lipolytica.     

Ectomycorrhizal and arbuscular mycorrhizal colonization of two species of floodplain willows

To understand the relationships between the distribution of Chosenia arbutifolia and Salix sachalinensis and their mycorrhizal colonization, changes in the quality and types of ectomycorrhizas and arbuscular mycorrhizas of the seedlings of two species were studied at five different sites with different soil conditions in the floodplain of the Satsunai River, Hokkaido. High ectomycorrhizal and low arbuscular mycorrhizal colonization were found in roots of both plants. Ectomycorrhizal colonization of S. sachalinensis in wet sandy or muddy soil conditions was at the same level as that in dry gravelly sites. In contrast, ectomycorrhizal colonization of C. arbutifolia seedlings was lower from wet sandy sites than that from dry gravelly sites. In all study sites, the same three morphological types of ectomycorrhizas were dominant.     

Arbuscular mycorrhizae respond to plants exposed to elevated atmospheric CO2 as a function of soil depth

The importance of arbuscular mycorrhizae (AM) in plant and ecosystem responses to global changes, e.g. elevated atmospheric CO₂, is widely acknowledged. Frequently, increases in AM root colonization occur in response to increased CO₂, but also the lack of significant changes has been reported. The goal of this study was to test whether arbuscular mycorrhizae (root colonization and composition of root colonization) respond to plants grown in elevated CO₂ as a function of soil depth. We grew Bromus hordeaceus L. and Lotus wrangelianus Fischer & C. Meyer monocultures in large pots with a synthetic serpentine soil profile for 4 yr in an experiment, in which CO₂ concentration was crossed factorially with NPK fertilization. When analyzing root infection separately for topsoil (0–15 cm) and subsoil (15–45 cm), we found large (e.g., about 5-fold) increases of AM fungal root colonization in the subsoil in response to CO₂, but no significant changes in the corresponding topsoil of Bromus. Only the coarse endophyte AM fungi, not the fine endophyte AM fungi, were responsible for the observed increase in the bottom soil layer, indicating a depth-dependent shift in the AM community colonizing the roots, even at this coarse morphological level. Other response variables also had significant soil layer ^* CO₂ interaction terms. The subsoil response would have been hidden in an unstratified assessment of the total root system, since most of the root length was concentrated in the top soil layer. The increased presence of mycorrhizae in roots deeper in the soil should be considered in sampling protocols, as it may be indicative of changed patterns of nutrient acquisition and carbon sequestration.     

Attenuation of cadmium toxicity in mycorrhizal celery (<Emphasis Type="Italic">Apium graveolens</Emphasis> L.)

A pot-culture experiment was carried out to investigate the effect of arbuscular mycorrhizal (AM) fungus (Glomus macrocarpum Tul. and Tul.) on plant growth and Cd²⁺uptake by Apium graveolens L. in soil with different levels of Cd²⁺. Mycorrhizal (M) and non-mycorrhizal (NM) plants were grown in soil with 0, 5, 10, 40 and 80 Cd²⁺ mg kg⁻¹soil. The infectivity of the fungus was not affected by the presence of Cd²⁺ in the soil. M plants showed better growth and less Cd²⁺ toxicity symptoms. Cd²⁺ root : shoot ratio was higher in M plants than in NM plants. These differences were more evident at highest Cd²⁺ level (80 mg kg⁻¹ soil). Chlorophyll a and chlorophyll b concentrations were significantly higher in AM-inoculated celery leaves. The dilution effect due to increased biomass, immobilization of Cd²⁺ in root and enhanced P-uptake in M plants may be related to attenuation of Cd²⁺toxicity in celery.     

Host genotype dependency and growth enhancing ability of VA-mycorrhizal fungi for Eleusine coracana (finger millet)

Inoculation of finger millet (Eleusine coracana Gaertn.) plants with one of six different vesicular, arbuscular, mycorrhizal (VAM) fungi increased plant biomass, height, leaf area and absolute growth rate; however, effectiveness of the various VAM fungi varied significantly. Maximum root colonization and mycorrhizal efficacy was observed with plants inoculated with Glomus caledonicum. Among five host genotypes tested for mycorrhizal dependency against G. caledonicum, genotype HR-374 gave the highest plant biomass, mycorrhizal efficacy and root colonization, the inoculation resulting in increased mineral (phosphate, nitrogen, Zn²⁺ and Cu²⁺) content and uptake in shoots.     

Synthesis and establishment of <Emphasis Type="Italic">Tuber melanosporum</Emphasis> Vitt. ectomycorrhizae on two <Emphasis Type="Italic">Nothofagus</Emphasis> species in Chile

Axenically germinated seedlings of two species of Southern beech (Nothofagus obliqua, N. glauca) from Chile were inoculated with spores of the Périgord black truffle (Tuber melanosporum). Ectomycorrhizal development was monitored for 6 months in the greenhouse and compared to the performance of the natural host species Quercus ilex and Quercus robur. Seedling survival and mycorrhization showed major differences in both Nothofagus species: T. melanosporum readily formed ectomycorrhizae with seedlings of N. obliqua, although at a lower rate than with Q. ilex but at a proportion very similar to Q. robur; survival and colonization rates were high, and seedling growth was not visibly affected by the high soil pH required by T. melanosporum. In contrast, more than 50% of N. glauca seedlings died after inoculation, and mycorrhiza formation was very sparse. In both species, no colonization by adventive ectomycorrhizal fungi could be observed, whereas both species of Quercus showed minor colonization by another fungus, probably Inocybe or Hebeloma. Our results show that it is possible to infect N. obliqua with the Périgord black truffle under greenhouse conditions, which opens up the possibility of cultivating this truffle as a secondary crop during reforestation with N. obliqua in Chile.     

Occurrence of some Glomales in Finland

By using trap plants, 17 species of arbuscular mycorrhizal fungi (AMF) belonging to the order Glomales were identified in 266 soil samples collected in the period 1987–1989. Of the identified isolates, 87.1% belonged to the genus Glomus Tulasne & Tulasne, 8.5% to Acaulospora Gerdemann & Trappe, 4.1% to Scutellospora Walker & Sanders and 0.3% to Entrophospora. Of the individual species identified, Glomus hoi Berch & Trappe was the most frequently identified, followed by G. fistulosum Skou & Jakobsen and G. mosseae (Nicol. & Gerd.) Gerdemann & Trappe. Only small differences in AMF trapping ability were observed between the four trap plants used, Trifolium pratense L., Zea mays L., Allium cepa L. var. cepa and Fragaria x ananassa Duch. T. pratense was chosen for further study because it had the highest AMF sporulation index in trap cultures and it also performed better than the other plants when grown in soils with different physical and chemical properties. The proportion of soil samples where AMF were identified decreased from close to 100% in the southern and central parts of Finland to about 50% in northern Finland.     

Weed control and cover crop management affect mycorrhizal colonization of grapevine roots and arbuscular mycorrhizal fungal spore populations in a California vineyard

Arbuscular mycorrhizal (AM) fungi naturally colonize grapevines in California vineyards. Weed control and cover cropping may affect AM fungi directly, through destruction of extraradical hyphae by soil disruption, or indirectly, through effects on populations of mycorrhizal weeds and cover crops. We examined the effects of weed control (cultivation, post-emergence herbicides, pre-emergence herbicides) and cover crops (Secale cereale cv. Merced rye, × Triticosecale cv.Trios 102) on AM fungi in a Central Coast vineyard. Seasonal changes in grapevine mycorrhizal colonization differed among weed control treatments, but did not correspond with seasonal changes in total weed frequency. Differences in grapevine colonization among weed control treatments may be due to differences in mycorrhizal status and/or AM fungal species composition among dominant weed species. Cover crops had no effect on grapevine mycorrhizal colonization, despite higher spring spore populations in cover cropped middles compared to bare middles. Cover crops were mycorrhizal and shared four AM fungal species (Glomus aggregatum, G. etunicatum, G. mosseae, G. scintillans) in common with grapevines. Lack of contact between grapevine roots and cover crop roots may have prevented grapevines from accessing higher spore populations in the middles.     

Trehalase activity in mycorrhizal and nonmycorrhizal roots of leek and soybean

Root extracts of leek (Allium porrum L.) and soybean (Glycine max L. Merr.) showed trehalase activity which was inhibited by phloridzin and was several times higher than the activity of general -glucosidase. The activity had an acidic optimum. Trehalase activity in extracts of sporocarps and extraradical mycelium of the arbuscular mycorrhizal fungus Glomus mosseae Nicol. & Gerd. (Trappe & Gerd.) was higher than in root extracts and had an optimum at pH 7. Following inoculation with G. mosseae, trehalase activity increased in mycorrhizal roots above the levels observed in nonmycorrhizal roots. Irrespective of fungal colonization, root trehalase activity increased in the presence of Mg²⁺, decreased in the presence of Mn²⁺ and Zn²⁺, and was unaffected by Na₂EDTA.     

General microflora,arbuscular mycorrhizal colonization and occurrence of endophytes in the rhizosphere of two age groups of <Emphasis Type="Italic">Ginkgo biloba</Emphasis> L.of Indian Central Himalaya

The populations of the general microflora (bacteria, actinomycetes and fungi) in the rhizosphere and their corresponding non-rhizosphere soil samples of Ginkgo biloba L. of two age groups (Group A, <25 years-young trees; Group B, >60 years-old trees) growing under a temperate location of Indian Himalayan Region (IHR) have been determined. Observations were also made for the diversity, distribution and colonization of arbuscular mycorrhizal (AM) fungi and occurrence of endophytes in roots of G. biloba. The population of general microflora was found to be higher in the rhizosphere of Group B trees, more clearly reflected in terms of rhizosphere: soil (R:S) ratios. Contrary to this, per cent colonization and spore densities of AM fungi were higher in the rhizosphere of Group A trees as compared to the rhizosphere of Group B. AM fungal colonization was observed mostly in form of loose coils. All the spores detected, belonged to the genus Glomus with five different types. Presence of endophytes (both bacteria and fungi) was observed in the cortical cells of G. biloba roots, more profound in case of Group B trees. Data suggest that, while the species of Glomus dominated the rhizosphere of G. biloba, an inverse correlation exist between the colonization of general microflora and the colonization of AM fungi including endophytes.