Further root colonization by arbuscular mycorrhizal fungi in already mycorrhizal plants is suppressed after a critical level of root colonization

An established arbuscular mycorrhizal symbiosis suppresses further mycorrhization. It is not clear whether the observed suppressional effect is linked with the level of root colonization or not. In the present work we studied the effect of the degree of root colonization by the arbuscular mycorrhizal fungus Glomus mosseae on further root colonization by G. mosseae. At different time points barley plants grown in split-root compartments were pre-inoculated on one half of the split-root system with G. mosseae. Sequential inoculation resulted in different colonization levels. Thereafter, the second half of the split root system was inoculated. The results indicate an enhanced suppression of root colonization on the second side of the split-root system when colonization levels increased on the first side.     

Nutrients and environment influence arbuscular mycorrhizal colonization both independently and interactively in Schizachyrium scoparium

Plant and Soil - Arbuscular mycorrhizal fungi (AMF) are important for plant nutrient and water acquisition. Much is known about how nutrient addition and environment affect AMF, but little is known...     

Polyphosphate dynamics in mycorrhizal roots during colonization of an arbuscular mycorrhizal fungus

Inorganic polyphosphate (poly P) has been considered to be a translocatable form of phosphate (Pi) in arbuscular mycorrhizal fungi (AMF). Here we examined time-course changes in poly P content during the AMF colonization process. Onion (Allium cepa) plants were cultured with or without inoculation with Gigaspora margarita for 2-8 wk with periodic sampling. Poly P in the extracts, purified through gel filtration, was quantified by the reverse reaction of polyphosphate kinase. The length of poly P in mycorrhizal roots appeared to be shorter than in extraradical hyphae or in spores of the AMF, indicating that AMF depolymerize poly P before providing Pi to the host. The poly P content increased as colonization proceeded, and was highly correlated with the weight of the colonized roots. These results support the model that AMF supply Pi to the host through the poly P pool, and that the poly P content of a mycorrhizal root can be a good indicator of the Pi-supplying activity of AMF.     

Azospirillum and arbuscular mycorrhizal colonization enhance rice growth and physiological traits under well-watered and drought conditions

The response of rice plants to inoculation with an arbuscular mycorrhizal (AM) fungus, Azospirillum brasilense, or combination of both microorganisms, was assayed under well-watered or drought stress conditions. Water deficit treatment was imposed by reducing the amount of water added, but AM plants, with a significantly higher biomass, received the same amount of water as non-AM plants, with a poor biomass. Thus, the water stress treatment was more severe for AM plants than for non-AM plants. The results showed that AM colonization significantly enhanced rice growth under both water conditions, although the greatest rice development was reached in plants dually inoculated under well-watered conditions. Water level did not affect the efficiency of photosystem II, but both AM and A. brasilense inoculations increased this value. AM colonization increased stomatal conductance, particularly when associated with A. brasilense, which enhanced this parameter by 80% under drought conditions and by 35% under well-watered conditions as compared to single AM plants. Exposure of AM rice to drought stress decreased the high levels of glutathione that AM plants exhibited under well-watered conditions, while drought had no effect on the ascorbate content. The decrease of glutathione content in AM plants under drought stress conditions led to enhance lipid peroxidation. On the other hand, inoculation with the AM fungus itself increased ascorbate and proline as protective compounds to cope with the harmful effects of water limitation. Inoculation with A. brasilense also enhanced ascorbate accumulation, reaching a similar level as in AM plants. These results showed that, in spite of the fact that drought stress imposed by AM treatments was considerably more severe than non-AM treatments, rice plants benefited not only from the AM symbiosis but also from A. brasilense root colonization, regardless of the watering level. However, the beneficial effects of A. brasilense on most of the physiological and biochemical traits of rice plants were only clearly visible when the plants were mycorrhized. This microbial consortium was effective for rice plants as an acceptable and ecofriendly technology to improve plant performance and development.     

Polyploidy in tomato roots as affected by arbuscular mycorrhizal colonization

Nuclear changes in roots of tomato (Lycopersicon esculentum), a plant with a small genome, during the establishment of arbuscular mycorrhizal (AM) colonization were studied using light and electron microscopy, as well as flow and static cytometry. Nuclei of mycorrhizal root cortex cells were larger and had more decondensed chromatin than those of controls. Significant ploidy distribution differences were observed between nuclei of AM colonized and control roots, and a strong correlation between nuclear polyploidization and AM colonization was found. Polyploidization and decondensation are usually associated with high metabolic activity. The metabolic activity of mycorrhizal root cells, evaluated in this work as respiratory activity by using a cytochemical assay for succinate dehydrogenase combined with image analysis, increased in comparison to controls. The meaning of polyploidization is discussed in relation to the structural and metabolic modifications induced by mycorrhization.     

Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization : Hyphal density and mycorrhizal root colonization

The influence of tillage practices on native arbuscular mycorrhizal fungi (AMF) was studied in two, consecutive years in eastern Canada, in two 11 year-old long-term tillage-fertilizer experimental field soils, a sandy loam and a clay, growing corn in monoculture. The three tillage practices were: 1) conventional tillage (CT; fall plowing plus spring disking), reduced tillage (RT; spring disking) and no-till (NT). The corn crop received either inorganic (N and K) or organic (liquid dairy manure) fertilizers. Mycorrhizal hyphal density was estimated from soil samples obtained in early spring (before disking), at the 12–14 leaf stage, at silking, and at harvest. The percentage of corn root colonization by AMF at the 12–14 leaf stage, at silking and at harvest was also determined. The sandy loam was sampled over two consecutive seasons and the clay soil over one season.Densities of total and metabolically active soil hyphae, and mycorrhizal root colonization were significantly lower in CT soil than in RT and NT soil. Lowest soil hyphal densities were observed in early spring. The levels of intra- and extraradical fungal colonization always increased from spring to silking and decreased thereafter. Spring disking had only a small and transient negative effect on hyphal abundance in soil. Fertilization did not influence mycorrhizal colonization of corn or abundance of soil hyphae in the sandy loam soil, but in the clay soil metabolically active hyphae were more abundant with manure application than with mineral fertilization. In 1992, in both soils different tillage systems had same grain yield, however, in 1993, corn yield was higher in NT compared to CT system.     

A comparison of visualization techniques for recording arbuscular mycorrhizal colonization

The extent of arbuscular mycorrhizal colonization was assessed in 10 field-collected plant species, representing three annual forbs, three perennial forbs, three perennial grasses and one annual grass. Each root system of each plant was split into four portions, and for each portion, mycorrhizal structures were revealed with epifluorescence microscopy (under which only arbuscules are generally visible) and three commonly used stains (Chlorazol Black E, Acid Fuchsin and Trypan Blue). The aim of the study was not to evaluate the efficacy of each method, but to compare results obtained by each under standard laboratory conditions. The recorded colonization levels of arbuscules, total arbuscular mycorrhizal fungal material and total fungal (arbuscular mycorrhizal+non-arbuscular mycorrhizal) material differed significantly between visualization methods in a number of species. However, there were also interactions between stain and plant species, indicating that the performance of a stain is dependent on the plant species being examined. In some cases (e.g. Plantago lanceolata ), each visualization method produced the same colonization level, while in others (e.g. Dactylis glomerata ), each method gave a different result. These data therefore suggest that the level of mycorrhizal colonization recorded in any particular plant species at a particular time is dependent on the technique employed.     

Changes in arbuscular mycorrhizal associations and fine root traits in sites under different plant successional phases in southern Brazil

Fine root morphological traits and distribution, arbuscular mycorrhizal (AM) fungi, soil fertility, and nutrient concentration in fine root tissue were compared in sites under different successional phases: grass plants, secondary forest, and mature forest in Londrina county, Paraná state, southern Brazil. Soil cores were collected randomly at the 0-10- and 10-20-cm depths in three quadrants (50 m2) in each site. Plants from the different successional stages displayed high differences in fine root distribution, fine root traits, and mycorrhizal root colonization. There were increases in the concentration of nutrients both in soil and fine roots and decrease of bulk soil density along the succession. The fine root biomass and diameter increased with the succession progress. The total fine root length, specific root length, root hair length, and root hair incidence decreased with the succession advance. Similarly, the mycorrhizal root colonization and the density of AM fungi spores in the soil decreased along the succession. Mycorrhizal root colonization and spore density were positively correlated with fine root length, specific root length, root hair length, root hair incidence, and bulk density and negatively correlated with fine root diameter and concentration of some nutrients both in soil and root tissues. Nutrient concentration in root tissue and in soil was positively correlated with fine root diameter and negatively correlated with specific root length, root hair length, and root hair incidence. These results suggest different adaptation strategies of plant roots for soil exploration and mineral acquisition among the different successional stages. Early successional stages displayed plants with fine root morphology and AM fungi colonization to improve the root functional efficiencies for uptake of nutrients and faster soil resource exploration. Late successional stages displayed plants with fine root morphology and mycorrhizal symbiosis for both a lower rate of soil proliferation and soil exploration capacity to acquire nutrients.     

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.     

Influence of soil organic matter decomposition on arbuscular mycorrhizal fungi in terms of asymbiotic hyphal growth and root colonization

Soil organic matter is known to influence arbuscular mycorrhizal (AM) fungi, but limited information is available on the chemical components in the organic matter causing these effects. We studied the influence of decomposing organic matter (pure cellulose and alfalfa shoot and root material) on AM fungi after 30, 100, and 300 days of decomposition in nonsterile soil with and without addition of mineral N and P. Decomposing organic matter affected maize root length colonized by the AM fungus Glomus claroideum in a similar manner as other plant growth parameters. Colonized root length was slightly increased by both nitrogen and phosphorus application and plant materials, but not by application of cellulose. In vitro hyphal growth of Glomus intraradices was increased by soil extracts from the treatments with all types of organic materials independently of mineral N and P application. Pyrolysis of soil samples from the different decomposition treatments revealed in total 266 recognizable organic compounds and in vitro hyphal growth of G. intraradices in soil extract positively correlated with 33 of these compounds. The strongest correlation was found with 3,4,5-trimethoxybenzoic acid methyl ester. This compound is a typical product of pyrolysis of phenolic compounds produced by angiosperm woody plants, but in our experiment, it was produced mainly from cellulose by some components of the soil microflora. In conclusion, our results indicate that mycelia of AM fungi are influenced by organic matter decomposition both via compounds released during the decomposition process and also by secondary metabolites produced by microorganisms involved in organic matter decomposition.     

Metabolic activity of Glomus intraradices in Arum- and Paris-type arbuscular mycorrhizal colonization

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

Characterization of root colonization profiles by a microcosm community of arbuscular mycorrhizal fungi using 25S rDNA-targeted nested PCR

The aim of the present work was to study colonization patterns in roots by different arbuscular mycorrhizal fungi developing from a mixed community in soil. As different fungi cannot be distinguished with certainty in planta on the basis of fungal structures, taxon-discriminating molecular probes were developed. The 5' end of the large ribosomal subunit containing the variable domains D1 and D2 was amplified by PCR from Glomus mosseae (BEG12), G. intraradices (LPA8), Gigaspora rosea (BEG9) and Scutellospora castanea (BEG1) using newly designed eukaryote-specific primers. Sequences of the amplification products showed high interspecies variability and PCR taxon-discriminating primers were designed to distinguish between each of these four fungi. A nested PCR, using universal eukaryotic primers for the first amplification and taxon-discriminating primers for the second, was performed on individual trypan blue-stained mycorrhizal root fragments of onion and leek, and root colonization by four fungi inoculated together in a microcosm experiment was estimated. More than one fungus was detected in the majority of root fragments and all four fungi frequently co-existed within the same root fragment. Root colonization by G. mosseae and G. intraradices was similar from individual and mixed inoculum, whilst the frequency of S. castanea and Gig. rosea increased in the presence of the two Glomus species, suggesting that synergistic interactions may exist between some arbuscular mycorrhizal fungi.     

Suppression of arbuscular mycorrhizal colonization and nodulation in split-root systems of alfalfa after pre-inoculation and treatment with Nod factors

Roots of legumes establish symbiosis with arbuscular mycorrhizal fungi (AMF) and nodule-inducing rhizobia. The existing nodules systemically suppress subsequent nodule formation in other parts of the root, a phenomenon termed autoregulation. Similarly, mycorrhizal roots reduce further AMF colonization on other parts of the root system. In this work, split- root systems of alfalfa (Medicago sativa) were used to study the autoregulation of symbiosis with Sinorhizobium meliloti and the mycorrhizal fungus Glomus mosseae. It is shown that nodulation systemically influences AMF root colonization and vice versa. Nodules on one half of the split-root system suppressed subsequent AMF colonization on the other half. Conversely, root systems pre-colonized on one side by AMF exhibited reduced nodule formation on the other side. An inhibition effect was also observed with Nod factors (lipo-chito-oligosaccharides). NodSm-IV(C16:2, S) purified from S. meliloti systemically suppressed both nodule formation and AMF colonization. The application of Nod factors, however, did not influence the allocation of (14)C within the split-root system, excluding competition for carbohydrates as the regulatory mechanism. These results indicate a systemic regulatory mechanism in the rhizobial and the arbuscular mycorrhizal association, which is similar in both symbioses.     

Role of the modification in root exudation induced by arbuscular mycorrhizal colonization on the intraradical growth of Phytophthora nicotianae in tomato

We studied the role of modification in root exudation induced by colonization with Glomus intraradices and Glomus mosseae in the growth of Phytophthora nicotianae in tomato roots. Plants were grown in a compartmentalized plant growth system and were either inoculated with the AM fungi or received exudates from mycorrhizal plants, with the corresponding controls. Three weeks after planting, the plants were inoculated or not with P. nicotianae growing from an adjacent compartment. At harvest, P. nicotianae biomass was significantly reduced in roots colonized with G. intraradices or G. mosseae in comparison to non-colonized roots. Conversely, pathogen biomass was similar in non-colonized roots supplied with exudates collected from mycorrhizal or non-mycorrhizal roots, or with water. We cannot rule out that a mycorrhiza-mediated modification in root exudation may take place, but our results did not support that a change in pathogen chemotactic responses to host root exudates may be involved in the inhibition of P. nicotianae.