Interactions between arbuscular mycorrhizal fungi and soil bacteria

The soil environment is interesting and complicated. There are so many interactions taking place in the soil, which determine the properties of soil as a medium for the growth and activities of plants and soil microorganisms. The soil fungi, arbuscular mycorrhiza (AM), are in mutual and beneficial symbiosis with most of the terrestrial plants. AM fungi are continuously interactive with a wide range of soil microorganisms including nonbacterial soil microorganisms, plant growth promoting rhizobacteria, mycorrhiza helper bacteria and deleterious bacteria. Their interactions can have important implications in agriculture. There are some interesting interactions between the AM fungi and soil bacteria including the binding of soil bacteria to the fungal spore, the injection of molecules by bacteria into the fungal spore, the production of volatiles by bacteria and the degradation of fungal cellular wall. Such mechanisms can affect the expression of genes in AM fungi and hence their performance and ecosystem productivity. Hence, consideration of such interactive behavior is of significance. In this review, some of the most important findings regarding the interactions between AM fungi and soil bacteria with some new insights for future research are presented.     

Community structure of arbuscular mycorrhizal fungi at different soil depths in extensively and intensively managed agroecosystems

The vertical distribution of spores of arbuscular mycorrhizal fungi (AMF) was investigated in soil profiles of extensively and intensively managed agroecosystems, including two permanent grasslands, a vineyard and two continuously mono-cropped maize fields. The number of AMF spores decreased with increasing soil depth--most drastically in the grasslands and the vineyard--but there was a large diversity of AMF species even in the deepest soil layers (50-70 cm). This was particularly striking in the maize fields where the highest species numbers were found below ploughing depth. Some species sporulated mainly, or exclusively, in the deep soil layers, others mainly in the top layers. Soil samples were used to inoculate trap cultures. Up to 18 months after inoculation, there was no conspicuous difference in the species composition among the trap cultures representing different soil depths, and only a weak match to the species composition determined by analysis of field samples. Our results indicate that the AMF communities in deep soil layers are surprisingly diverse and different from the topsoil. Thus, deep soil layers should be included in studies to get a complete picture of AMF diversity.     

Organic amendments increase phylogenetic diversity of arbuscular mycorrhizal fungi in acid soil contaminated by trace elements

In 1998, a toxic mine spill polluted a 55-km² area in a basin southward to Doñana National Park (Spain). Subsequent attempts to restore those trace element-contaminated soils have involved physical, chemical, or biological methodologies. In this study, the restoration approach included application of different types and doses of organic amendments: biosolid compost (BC) and leonardite (LEO). Twelve years after the last addition, molecular analyses of arbuscular mycorrhizal (AM) fungal communities associated with target plants (Lamarckia aurea and Chrysanthemum coronarium) as well as analyses of trace element concentrations both in soil and in plants were performed. The results showed an improved soil quality reflected by an increase in soil pH and a decrease in trace element availability as a result of the amendments and dosages. Additionally, the phylogenetic diversity of the AM fungal community increased, reaching the maximum diversity at the highest dose of BC. Trace element concentration was considered the predominant soil factor determining the AM fungal community composition. Thereby, the studied AM fungal community reflects a community adapted to different levels of contamination as a result of the amendments. The study highlights the long-term effect of the amendments in stabilizing the soil system.     

Uptake and transport of organic and inorganic nitrogen by arbuscular mycorrhizal fungi

New information on N uptake and transport of inorganic and organic N in arbuscular mycorrhizal fungi is reviewed here. Hyphae of the arbuscular mycorrhizal fungus Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe (BEG 107) were shown to transport N supplied as ¹⁵N-Gly to wheat plants after a 48 h labelling period in semi-hydroponic (Perlite), non-sterile, compartmentalised pot cultures. Of the ¹⁵N supplied to hyphae in pot cultures over 48 h, 0.2 and 6% was transported to plants supplied with insufficient N or sufficient N, respectively. The increased ¹⁵N uptake at the higher N supply was related to the higher hyphal length density at the higher N supply. These findings were supported by results from in vitro and monoxenic studies. Excised hyphae from four Glomus isolates (BEG 84, 107, 108 and 110) acquired N from both inorganic (¹⁵NH₄ ¹⁵NO₃, ¹⁵NO₃ ⁻ or ¹⁵NH₄ ⁺) and organic (¹⁵N-Gly and ¹⁵N-Glu, except in BEG 84 where amino acid uptake was not tested) sources in vitro during short-term experiments. Confirming these studies under sterile conditions where no bacterial mineralisation of organic N occurred, monoxenic cultures of Glomus intraradices Schenk and Smith were shown to transport N from organic sources (¹⁵N-Gly and ¹⁵N-Glu) to Ri T-DNA transformed, AM-colonised carrot roots in a long-term experiment. The higher N uptake (also from organic N) by isolates from nutrient poor sites (BEG 108 and 110) compared to that from a conventional agricultural field implied that ecotypic differences occur. Although the arbuscular mycorrhizal isolates used contributed to the acquisition of N from both inorganic and organic sources by the host plants/roots used, this was not enough to increase the N nutritional status of the mycorrhizal compared to non-mycorrhizal hosts. This revised version was published online in June 2006 with corrections to the Cover Date.     

Reduced growth of autumn-sown wheat in a low-P soil is associated with high colonisation by arbuscular mycorrhizal fungi

Autumn-sown wheat (Triticum aestivum) was studied over two seasons in south-eastern Australia, on a low-P soil where indigenous arbuscular mycorrhizal fungi (AMF) were known to provide little nutritional benefit to crops. It was hypothesised that AMF would be parasitic under these circumstances. Shoot dry mass and water soluble carbohydrate (WSC) reserves in roots and shoots were measured for wheat grown with or without P-fertiliser, in plots where crop sequences had produced either high or low colonisation by AMF. Application of P-fertiliser greatly increased crop growth and decreased colonisation by AMF. At tillering, colonisation by AMF ranged from 24 to 66% of root length when no P was applied and from 11 to 32% when P was applied. At each P-level, high colonisation correlated with reductions of around 20% in stem and root WSC concentrations (first season) or shoot WSC content and shoot dry mass (much drier second season). Impacts on yield were not significant (first season) or largely masked by water-stress and frost (second season). While the major fungal root diseases of the region were absent, interactions between crop sequence and other unknown biotic constraints could not be discounted. The results are consistent with the parasitic impacts of colonisation by AMF being induced primarily through the winter conditions experienced by the crops until anthesis. It is concluded that wheat in south-eastern Australia may benefit from reduced colonisation by AMF, which could achieved through selected crop sequences or, perhaps, targeted wheat breeding programs.     

Determination of the nitrogen source for arbuscular mycorrhizal fungi by 15N application to soil and plants

The source of nitrogen in the spores of arbuscular mycorrhizal (AM) fungi was quantified by a ¹⁵N-labeling technique. N was applied as coated urea to the soil and in solution to plant shoots. Soil-applied fertilizer had a significant effect on spore % ¹⁵N (P<0.01), with a 24–75% contribution to spore N. Fertilizer applied to either alfalfa shoots or bahia grass shoots had little effect on spore % ¹⁵N, accounting for 0–14% or 1–9% of spore N, respectively. These results indicate that AM fungi obtain spore N mostly from the soil. The small amount of spore N originating from shoot-applied N may have been obtained via root exudation. Accepted: 6 November 2000     

Genetic variability in arbuscular mycorrhizal fungi compatibility supports the selection of durum wheat genotypes for enhancing soil ecological services and cropping systems in Canada

Crop nutrient- and water-use efficiency could be improved by using crop varieties highly compatible with arbuscular mycorrhizal fungi (AMF). Two greenhouse experiments demonstrated the presence of genetic variability for this trait in modern durum wheat ( Triticum turgidum L. var. durum Desf.) germplasm. Among the five cultivars tested, 'AC Morse' had consistently low levels of AM root colonization and DT710 had consistently high levels of AM root colonization, whereas 'Commander', which had the highest colonization levels under low soil fertility conditions, developed poor colonization levels under medium fertility level. The presence of genetic variability in durum wheat compatibility with AMF was further evidenced by significant genotype × inoculation interaction effects in grain and straw biomass production; grain P, straw P, and straw K concentrations under medium soil fertility level; and straw K and grain Fe concentrations at low soil fertility. Mycorrhizal dependency was an undesirable trait of 'Mongibello', which showed poor growth and nutrient balance in the absence of AMF. An AMF-mediated reduction in grain Cd under low soil fertility indicated that breeding durum wheat for compatibility with AMF could help reduce grain Cd concentration in durum wheat. Durum wheat genotypes should be selected for compatibility with AMF rather than for mycorrhizal dependency.     

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.     

Isolation and identification of soil bacteria growing at the expense of arbuscular mycorrhizal fungi

Soil-microorganism symbioses are of fundamental importance for plant adaptation to the environment. Research in microbial ecology has revealed that some soil bacteria are associated with arbuscular mycorrhizal fungi (AMF). However, these interactions may be much more complex than originally thought. To assess the type of bacteria associated with AMF, we initially isolated spores of Glomus irregulare from an Agrostis stolonifera rhizosphere. The spores were washed with sterile water and plated onto G. irregulare mycelium growing in vitro in a root-free compartment of bicompartmented Petri dishes. We hypothesized that this system should select for bacteria closely associated with the fungus because the only nutrients available to the bacteria were those derived from the hyphae. Twenty-nine bacterial colonies growing on the AMF hyphae were subcultured and identified using 16S rRNA gene sequences. All bacterial isolates showed high sequence identity to Bacillus cereus, Bacillus megaterium, Bacillus simplex, Kocuria rhizophila, Microbacterium ginsengisoli, Sphingomonas sp. and Variovorax paradoxus. We also assessed bacterial diversity on the surface of spores by PCR-denaturating gradient gel electrophoresis. Finally, we used live cellular imaging to show that the bacteria isolated can grow on the surface of hyphae with different growing patterns in contrast to Escherichia coli as a control.