Ecological roles of arbuscular mycorrhizal fungi in two wild legume plants

Two wild legume plants,Glycine soja andCassia mimosoides var.nomame, and a cultivated plant, soybean (Glycine max), were employed for a study of triple symbiosis with an inoculum ofScutellispora heterogama harvested from natural soils and an inoculum of their own rhizobial cells. The dry weight, colonization of arbuscular mycorrhizal fungus, nodule formation and N₂-fixation activity were estimated as the parameters of triple symbiosis. The two wild legume plants showed greater growth with colonization of arbuscular mycorrhizae than with nodulation, whereas the cultivated legume showed more nodulation than colonization of arbuscular mycorrhizae. Moreover,S. heterogama appeared to stimulate the triple symbiosis for the wild legume plants. The results suggested that spores ofS. heterogama are important in disturbed soils in Korea.     

Early responses of wild plant seedlings to arbuscular mycorrhizal fungi and pathogens

Many plants form associations with arbuscular mycorrhizal fungi (AMF) because they profit from improved phosphorus nutrition and from protection against pathogens. Whereas mycorrhiza-induced pathogen protection is well understood in agricultural plant species, it is rarely studied in wild plants. As many pathogens infest plants in the first days after germination, mycorrhiza-induced pathogen protection may be especially important in the first few weeks of plant establishment.Here, we investigated interacting effects of AMF and the seedling pathogen Pythium ultimum on the performance of six- to seven-week-old seedlings of six wild plant species of the family Asteraceae in a full factorial experiment.Plant species differed in their response to AMF, the pathogen and their interactions. AMF increased and the pathogen decreased plant biomass in one and three species, respectively. Two plant species were negatively affected by AMF in the absence, but positively or not affected in the presence of the pathogen, indicating protection by AMF. This mycorrhiza-induced pathogen protection is especially surprising as we could not detect mycorrhizal structure in the roots of any of the plants.Our results show that even seedlings without established intraradical hyphal network can profit from AMF, both in terms of growth promotion in the absence of a pathogen and pathogen protection. The function of AMF is highly species-specific, but tends to be similar for more closely related plant species, suggesting a phylogenetic component of mycorrhizal function. Further studies should test a wider range of plant species, as our study was restricted to one plant family, and investigate whether plants profit from early mycorrhizal benefits in the long term.     

Evidence for functional divergence in arbuscular mycorrhizal fungi from contrasting climatic origins

A considerable amount of phenotypic, genetic and symbiotic functional variability has been documented in arbuscular mycorrhizal fungi (AMF). However, little is known about whether distinct AMF ecotypes have evolved within their geographic range. We tested the hypothesis that AMF growing at temperatures closer to those prevalent within their origin would benefit their host and grow more than isolates distant from their native conditions. For each of six AMF species, we chose pairs of isolates that originated from distant areas with contrasting climates. Each isolate was grown in association with two grass species of different thermal optima at two temperature settings. Thus, we also tested whether AMF from different climatic origins were dependent on the thermal adaptation of the host plant species or to temperature per se. Although fungal growth was not directly affected by temperature, we found that AMF isolates originating from contrasting climates consistently and differentially altered plant growth. Our results suggest that AMF from contrasting climates have altered symbiotic function, thus linking an abiotic factor to ecotypic differentiation of putatively important symbionts.     

Indigenous and introduced arbuscular mycorrhizal fungi contribute to plant growth in two agricultural soils from south-western Australia

Arbuscular mycorrhizal (AM) fungi occur in all agricultural soils but it is not easy to assess the contribution they make to plant growth under field conditions. Several approaches have been used to investigate this, including the comparison of plant growth in the presence or absence of naturally occurring AM fungi following soil fumigation or application of fungicides. However, treatments such as these may change soil characteristics other than factors directly involving AM fungi and lead to difficulties in identifying the reason for changes in plant growth. In a glasshouse experiment, we assessed the contribution of indigenous AM fungi to growth of subterranean clover in undisturbed cores of soil from two agricultural field sites (a cropped agricultural field at South Carrabin and a low input pasture at Westdale). We used the approach of estimating the benefit of AM fungi by comparing the curvature coefficients ( C) of the Mitscherlich equation for subterranean clover grown in untreated field soil, in field soil into which inoculum of Glomus invermaium was added and in soil fumigated with methyl bromide. It was only possible to estimate the benefit of mycorrhizas using this approach for one soil (Westdale) because it was the only soil for which a Mitscherlich response to the application of a range of P levels was obtained. The mycorrhizal benefit ( C of mycorrhizal vs. non-mycorrhizal plants or C of inoculated vs. uninoculated plants) of the indigenous fungi corresponded with a requirement for phosphate by plants that were colonised by AM fungi already present in the soil equivalent to half that required by non-mycorrhizal plants. This benefit was independent of the plant-available P in the soil. There was no additional benefit of inoculation on plant growth other than that due to increased P uptake. Indigenous AM fungi were present in both soils and colonised a high proportion of roots in both soils. There was a higher diversity of morphotypes of mycorrhizal fungi in roots of plants grown in the Westdale soil than in the South Carrabin soil that had a history of high phosphate fertilizer use in the field. Inoculation with G. invermaium did not increase the level of colonisation of roots by mycorrhizal fungi in either soil, but it replaced approximately 20% of the root length colonised by the indigenous fungi in Westdale soil at all levels of applied P. The proportion of colonised root length replaced by G. invermaium in South Carrabin soil varied with the level of application of P to the soil; it was higher at intermediate levels of recently added soil P.     

Differential influence of native and introduced arbuscular mycorrhizal fungi on growth of dominant and subordinate plants

The impact of arbuscular mycorrhizal fungi (AMF) on plant ecosystems has been intensively reported. In this research, we explored the difference between native and introduced AMF in promoting the growth of dominant and subordinate plant species. In glasshouse experiments, dominants and subordinates from subtropical grasslands were colonized by native AMF or introduced AMF, Glomus versiforme. The biomass revealed that mycorrhizal dependencies (MD) on the native AMF of the dominants were much higher than those of the subordinates, while MD on the introduced AMF changed following the replacement of native AMF with introduced AMF. A close relationship between biomass promotion and increase in phosphorus uptake was observed, indicating the important role of AMF-enhanced nutrient acquisition by roots. Our results show that plant community structures are partly determined by MD on native AMF, and could be modified by introducing exogenous AMF species.     

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.     

Spatial variation of arbuscular mycorrhizal fungi in two vegetation types in Gurbantonggut Desert

Geostatistical techniques were used to assess the spatial patterns of spores densities and biovolume of arbuscular mycorrhizal fungi (AMF) in soils from two contrasting vegetation communities: an Ephedra distachya-ephemeral plant vegetation community and an Eremurus anisopteris vegetation community. Also evaluated the relationship between the spatial distribution of spore densities and biovolume of AMF and soil properties. Spatial dependence of spore densities and biovolume of AMF were exhibited further by kriged maps. The results showed spore density and biovolume indicated strong spatial autocorrelation and a patchy distribution within both sites. However, the patch size of genera and biovolume of AMF differed between the two communities. The correlation between distribution of spore and biovolume of AMF and distribution of soil parameters was expressed by Spearman rank-correlations coefficients. These results suggest that spore or biovolume distribution of AMF was affected significantly by some soil properties.     

Differential plant response to inoculation with two VA mycorrhizal fungi isolated from a low-pH soil

Sericea lespedaza and ladino clover were inoculated withAcaulospora laevis andGigaspora margarita VA mycorrhizal fungi, both isolated from a local soil having a pH of 4.4. Plants were grown in a greenhouse in fumigated (methyl bromide) soil with four rates of applied P. This soil had a pH of 5.1 for theG. margarita experiment and a 5.8 for theA. laevis experiment. Neither plant species responded to theG. margarita isolate in terms of mycorrhizal infection of roots, top growth, or elemental uptake. TheA. laevis isolate caused increased growth of Ladino clover at the lowest rate of P application and increased growth of sericea lespedeza at the two lowest P application rates. Shoot tissue concentrations of P for both plant species were greater at the two lowest rates of P application when inoculated withA. laevis. Inoculation withA. laevis also resulted in different VAM fungal root colonization patterns between the two plant species as a function of P application rate. Roots of both plants had high infection rates (near 70%) for the two lowest P application rates but sericea lespedeza declined to 40 and 6% at the next two P application rates, respectively, whereas infection in Ladino clover was 74 and 41% at the next two P application rates, respectively. The results of this study support the concept that plant host-VAM fungal endophyte interactions are extremely variable and that characterization of individual combinations must be done if types and magnitudes of responses are to be defined.     

Towards growth of arbuscular mycorrhizal fungi independent of a plant host

When surface-sterilized spores of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices Sy167 were germinated on agar plates in the slightly modified minimum mineral medium described by G. Bécard and J. A. Fortin (New Phytol. 108:211-218, 1988), slime-forming bacteria, identified as Paenibacillus validus, frequently grew up. These bacteria were able to support growth of the fungus on the agar plates. In the presence of P. validus, hyphae branched profusely and formed coiled structures. These were much more densely packed than the so-called arbuscule-like structures which are formed by AMF grown in coculture with carrot roots transformed with T-DNA from Agrobacterium rhizogenes. The presence of P. validus alone also enabled G. intraradices to form new spores, mainly at the densely packed hyphal coils. The new spores were not as abundant as and were smaller than those formed by AMF in the monoxenic culture with carrot root tissues, but they also contained lipid droplets and a large number of nuclei. In these experiments P. validus could not be replaced by bacteria such as Escherichia coli K-12 or Azospirillum brasilense Sp7. Although no conditions under which the daughter spores regerminate and colonize plants have been found yet, and no factor(s) from P. validus which stimulates fungal growth has been identified, the present findings might be a significant step forward toward growth of AMF independent of any plant host.     

Relatedness among arbuscular mycorrhizal fungi drives plant growth and intraspecific fungal coexistence

Arbuscular mycorrhizal fungi (AMF) form symbioses with most plant species. They are ecologically important determinants of plant growth and diversity. Considerable genetic variation occurs in AMF populations. Thus, plants are exposed to AMF of varying relatedness to each other. Very little is known about either the effects of coexisting AMF on plant growth or which factors influence intraspecific AMF coexistence within roots. No studies have addressed whether the genetics of coexisting AMF, and more specifically their relatedness, influences plant growth and AMF coexistence. Relatedness is expected to influence coexistence between individuals, and it has been suggested that decreasing ability of symbionts to coexist can have negative effects on the growth of the host. We tested the effect of a gradient of AMF genetic relatedness on the growth of two plant species. Increasing relatedness between AMFs lead to markedly greater plant growth (27% biomass increase with closely related compared to distantly related AMF). In one plant species, closely related AMF coexisted in fairly equal proportions but decreasing relatedness lead to a very strong disequilibrium between AMF in roots, indicating much stronger competition. Given the strength of the effects with such a shallow relatedness gradient and the fact that in the field plants are exposed to a steeper gradient, we consider that AMF relatedness can have a strong role in plant growth and the ability of AMF to coexist. We conclude that AMF relatedness is a driver of plant growth and that relatedness is also a strong driver of intraspecific coexistence of these ecologically important symbionts.     

Influence of arbuscular mycorrhizal fungi on growth and selenium uptake by garlic plants

The aim of this study is to investigate the effects of arbuscular mycorrhizal fungi (AMF) on garlic plants growth and the uptake of selenium (Se). Garlic plants were grown in the pots inoculated with Glomus fasciculatum and G. mosseae and maintained in a greenhouse. Three weeks after planting, the pots had received different concentrations of Se (5, 10, 15, 20, 25 mg kg^?1 of soil) in the form of selenium dioxide (SeO₂) at 3 weeks intervals up to 12 weeks. For physiological and biochemical analysis, the samples were randomly collected from five plants of each experiment. Maximum AM infection, spore population and plant biomass were observed in the roots of mycorrhizal-mediated plants without Se, and they were gradually declined in both mycorrhizal and non-mycorrhizal (NM) plants with increasing concentrations of Se. Among the two Glomus species tested, G. fasciculatum-mediated plants showed higher AM infection, spore population and plant biomass than G. mosseae. No differences were observed for the uptake of Se in mycorrhizal plants and NM plants. However, NM plants uptake more Se than mycorrhizal plants. Higher contents of total chlorophyll and sugars were observed in plants inoculated with G. fasciculatum without Se and they were decreased in the presence of Se. In contrast, increased amount of glutathione peroxidase was observed at increasing concentrations of Se up to 20 mg kg^?1. High-performance liquid chromatography data revealed that SeO₂ converted to organic form of Se as γ-glutamyl-Se-methylselenocysteine. These results are basis for further investigations on the role of AMF on plant growth and uptake of Se in crop plants.     

Dispersal of arbuscular mycorrhizal fungi and plants during succession

Arbuscular mycorrhizal (AM) fungi are important root symbionts that enhance plant nutrient uptake and tolerance to pathogens and drought. While the role of plant dispersal in shaping successional vegetation is well studied, there is very little information about the dispersal abilities of AM fungi. We conducted a trap-box experiment in a recently abandoned quarry at 10 different distances from the quarry edge (i.e. the potential propagule source) over eleven months to assess the short term, within-year, arrival of plant and AM fungal assemblages and hence their dispersal abilities. Using DNA based techniques we identified AM fungal taxa and analyzed their phylogenetic diversity. Plant diversity was determined by transporting trap soil to a greenhouse and identifying emerging seedlings. We recorded 30 AM fungal taxa. These contained a high proportion of ruderal AM fungi (30% of taxa, 79% of sequences) but the richness and abundance of AM fungi were not related to the distance from the presumed propagule source. The number of sequences of AM fungi decreased over time. Twenty seven plant species (30% of them ruderal) were recorded from the soil seed traps. Plant diversity decreased with distance from the propagule source and increased over time. Our data show that AM fungi with ruderal traits can be fast colonizers of early successional habitats.     

Differential strategies of two species of arbuscular mycorrhizal fungi in the protection of maize plants grown in chromium-contaminated soils

Chromium (Cr) is a nonessential element for plants that is extremely toxic at high concentrations. Zea mays L. is a species of plant that has developed adaptive mechanisms to increase its tolerance and absorption capacity for this metal. One effective mechanism is to form associations with arbuscular mycorrhizal fungi (AMF), which help the plant cope with stress from heavy metals such as Cr. However, it is still not clear which species of AMF are more efficient as bioremediating agents for plants of agricultural importance. Here, we evaluated the effect of Gigaspora gigantea and Rhizophagus irregularis as protective agents of maize plants in soils contaminated with Cr at concentrations of 0, 48.5, 97 and 194 mg kg^?1 under greenhouse conditions. Maize tolerance to Cr was corroborated, as well as increased absorption of this element by plants associated with both species of AMF. G. gigantea caused an increase in nitrogen content and greater translocation of Cr to the aerial part of the plant; R. irregularis registered an increase in the formation of arbuscules and vesicles with increasing metal concentration and greater retention of Cr in the roots of the plants. Based on these results, we can conclude that the analyzed species of fungi use different strategies, with similar effectiveness, to enhance the absorption capacity of Cr by the plant and influence the differential deposition of this metal in various parts of the plant.

     

Validation of plant functional types across two contrasting landscapes

Abstract. The validation of plant functional type models across contrasting landscapes is seen as a step towards the claim that plant functional types should recur regionally or even globally. I sampled the vegetation of an urban landscape on a range of sites representing gradients of resource supply and disturbance intensity. A group of plants with similar attributes was considered a ‘functional type’, if the species significantly co‐occurred in a certain segment of the gradient plane of resource supply and disturbance intensity. Vegetative and regeneration traits were considered. A similar study was performed in a nearby agricultural landscape (Kleyer 1999). The logistic regression models from the urban landscape were applied to the data set of the agricultural landscape and vice versa. Although the overall environment of the two landscapes was very different, recurrent patterns of several functional types were found. At high fertility and high disturbance levels, annual species predominated with a persistent seed bank, high seed output, and short vertical expansion. When disturbances changed from below‐ground to above‐ground, the sexual regeneration mode was replaced by the vegetative mode, while vertical expansion remained low. At medium disturbance intensities, the vertical expansion and vegetative regeneration increased with fertility, while the seed bank remained mostly transient to short‐term persistent and lateral expansion and sexual regeneration was intermediate. At low disturbances and low resource supplies, seed bank longevity, and vertical and lateral expansion tended to be long. Diversity of groups of plants with similar attributes was highest at intermediate disturbance levels and low fertility. These results correspond with Grime's humped‐back model and Connell's intermediate disturbance hypothesis.     

一种改进的丛枝菌根染色方法

研究改进了Vierheilig等描述的AM菌根染色法:将根样于20%KOH溶液中60℃水浴透明40-120 min,5%醋酸酸化5min后,用5%醋酸墨水染色液(派克纯黑书写墨水Quink),于60℃水浴染色30 min,清水浸泡脱色(14h)后即可镜检。根皮层细胞内AM真菌的丛枝结构清晰可见,并且能够明确地分辨AM真菌与其它未知真菌。此外,Quink初染后,再经过SudanⅣ复染(60℃、60 min),70%乙醇脱色5min,暗隔真菌的透明菌丝内所积聚的脂类颗粒被SudanⅣ染上鲜红色,在复式显微镜下能够观察到此类透明菌丝在根皮层组织内的存在状况。采用甘油明胶为封固剂制片,根的染色效果可以保存长久。此项技术可以对同一种植物的多个根样进行同步的透明和染色处理,而且操作简便、低毒性、成本低廉、染色效果极佳,适用于野生和栽培草本植物AM菌根的染色和制片观察。     

Detection and identification of bacterial endosymbionts in arbuscular mycorrhizal fungi belonging to the family Gigasporaceae

Intracellular bacteria have been found previously in one isolate of the arbuscular mycorrhizal (AM) fungus Gigaspora margarita BEG 34. In this study, we extended our investigation to 11 fungal isolates obtained from different geographic areas and belonging to six different species of the family Gigasporaceae. With the exception of Gigaspora rosea, isolates of all of the AM species harbored bacteria, and their DNA could be PCR amplified with universal bacterial primers. Primers specific for the endosymbiotic bacteria of BEG 34 could also amplify spore DNA from four species. These specific primers were successfully used as probes for in situ hybridization of endobacteria in G. margarita spores. Neighbor-joining analysis of the 16S ribosomal DNA sequences obtained from isolates of Scutellospora persica, Scutellospora castanea, and G. margarita revealed a single, strongly supported branch nested in the genus Burkholderia.     

Fresh perspectives on the roles of arbuscular mycorrhizal fungi in plant nutrition and growth

Recent research on arbuscular mycorrhizas has demonstrated that AM fungi play a significant role in plant phosphorus (P) uptake, regardless of whether the plant responds positively to colonization in terms of growth or P content. Here we focus particularly on implications of this finding for consideration of the balance between organic carbon (C) use by the fungi and P delivery (i.e. the C-P trade between the symbionts). Positive growth responses to arbuscular mycorrhizal (AM) colonization are attributed frequently to increased P uptake via the fungus, which results in relief of P deficiency and increased growth. Zero AM responses, compared with non-mycorrhizal (NM) plants, have conventionally been attributed to failure of the fungi to deliver P to the plants. Negative responses, combined with excessive C use, have been attributed to this failure. The fungi were viewed as parasites. Demonstration that the AM pathway of P uptake operates in such plants indicates that direct P uptake by the roots is reduced and that the fungi are not parasites but mutualists because they deliver P as well as using C. We suggest that poor plant growth is the result of P deficiency because AM fungi lower the amount of P taken up directly by roots but the AM uptake of P does compensate for the reduction. The implications of interplay between direct root uptake and AM fungal uptake of P also include increased tolerance of AM plants to toxins such as arsenate and increased success when competing with NM plants. Finally we discuss the new information on C-P trade in the context of control of the symbiosis by the fungus or the plant, including new information (from NM plants) on sugar transport and on the role of sucrose in the signaling network involved in responses of plants to P deprivation.     

Contribution of arbuscular mycorrhizal symbiosis to plant growth under different types of soil stress

The development of symbioses between soil fungi, arbuscular mycorrhizae (AM), and most terrestrial plants can be very beneficial to both partners and hence to the ecosystem. Among such beneficial effects, the alleviation of soil stresses by AM is of especial significance. It has been found that AM fungi can alleviate the unfavourable effects on plant growth of stresses such as heavy metals, soil compaction, salinity and drought. In this article, such mechanisms are reviewed, in the hope that this may result in more efficient use of AM under different stress conditions.     

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.