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1.
MethodsX ; 11: 102477, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38023315

RESUMEN

Fungal mycelium is a major component of the soil microbiome. The soil hyphosphere represents a complex and dynamic niche for specific microorganisms, where multitrophic interactions occur, affecting ecosystem processes. However, extracting fungal mycelium from the soil to enable its taxonomical, chemical, and structural characterisation is challenging in the absence of a fast, efficient, and low-cost procedure. In this study, an old method (Bingle and Paul 1985), based on successive soil wet filtrations and density gradient centrifugation, was improved and tested in three different soil types (silty clay, silty clay loam, and loamy sand). The improved method reduced the number of filtrations by about five times and the centrifugation time from 40 min to 1 min. It avoided using any chemical substance which may impair further chemical analyses or DNA isolation and amplification. The method efficiency was about 50 % in the clay and 23 % in the sandy soils. However, a pre-step consisting of removing the fine-root fragments and other debris under the stereomicroscope may increase the method efficiency to more than 65 %, independent of the soil type.•A simple, efficient, and low-cost method suitable for extracting soil mycelium from a large number of samples.•The protocol includes successive soil wet filtrations and sucrose gradient centrifugation.•The method efficiency increases if the fine-root fragments and other debris are previously removed from the soil.

2.
Mycorrhiza ; 33(5-6): 399-408, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37814097

RESUMEN

Specific biomarker molecules are increasingly being used for detection and quantification in plant and soil samples of arbuscular mycorrhizal (AM) fungi, an important and widespread microbial guild heavily implicated in transfers of nutrients and carbon between plants and soils and in the maintenance of soil physico-chemical properties. Yet, concerns have previously been raised as to the validity of a range of previously used approaches (e.g., microscopy, AM-specific fatty acids, sterols, glomalin-like molecules, ribosomal DNA sequences), justifying further research into novel biomarkers for AM fungal abundance and/or functioning. Here, we focused on complex polar lipids contained in pure biomass of Rhizophagus irregularis and in nonmycorrhizal and mycorrhizal roots of chicory (Cichorium intybus), leek (Allium porrum), and big bluestem (Andropogon gerardii). The lipids were analyzed by shotgun lipidomics using a high-resolution hybrid mass spectrometer. Size range between 1350 and 1550 Da was chosen for the detection of potential biomarkers among cardiolipins (1,3-bis(sn-3'-phosphatidyl)-sn-glycerols), a specific class of phospholipids. The analysis revealed a variety of molecular species, including cardiolipins containing one or two polyunsaturated fatty acids with 20 carbon atoms each, i.e., arachidonic and/or eicosapentaenoic acids, some of them apparently specific for the mycorrhizal samples. Although further verification using a greater variety of AM fungal species and samples from various soils/ecosystems/environmental conditions is needed, current results suggest the possibility to identify novel biochemical signatures specific for AM fungi within mycorrhizal roots. Whether they could be used for quantification of both root and soil colonization by the AM fungi merits further scrutiny.


Asunto(s)
Micorrizas , Cardiolipinas , Ecosistema , Hongos , Plantas , Cebollas , Suelo/química , Carbono , Raíces de Plantas/microbiología
3.
FEMS Microbiol Ecol ; 99(8)2023 07 21.
Artículo en Inglés | MEDLINE | ID: mdl-37481697

RESUMEN

Truffle growers devote great efforts to improve black truffle productivity, developing agronomic practices such as 'truffle nests' (peat amendments that are supplemented with truffle spore inoculum). It has been hypothesized that improved fruiting associated with nests is linked to stimulation of truffle mycelia previously established in soil or to changes generated in soil fungal community. To assess this, we used real-time PCR to quantify black truffle extraradical mycelium during 2 years after nests installation. We also characterized the fungal community via high-throughput amplicon sequencing of the ITS region of rRNA genes. We found that neither the abundance of truffle mycelium in nests nor in the soil-nest interphase was higher than in the bulk soil, which indicates that nests do not improve mycelial growth. The fungal community in nests showed lower richness and Shannon index and was compositionally different from that of soil, which suggests that nests may act as an open niche for fungal colonization that facilitates truffle fruiting. The ectomycorrhizal fungal community showed lower richness in nests. However, no negative relationships between amount of truffle mycelium and reads of other ectomycorrhizal fungi were found, thus countering the hypothesis that ectomycorrhizal competition plays a role in the nest effect.


Asunto(s)
Ascomicetos , Micobioma , Micorrizas , Microbiología del Suelo , Ascomicetos/fisiología , Suelo
4.
Bull Environ Contam Toxicol ; 108(3): 451-456, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33891141

RESUMEN

Sustainable agricultural practices based on the development of native arbuscular mycorrhizal fungi (AMF) can improve crop growth and stress tolerance in acidic soils with manganese toxicity. The beneficial effects are stronger when crops are colonized early in development by an intact extraradical mycelium (ERM), but are dependent on AMF assemblage. In wheat colonized by AMF associated to Lolium rigidum L. (LOL) or Ornithopus compressus (ORN), growth and stress tolerance are differently influenced. In the present study, this functional diversity was studied by evaluating the activity of ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol peroxidase (GPX), superoxide dismutase (SOD) and Mn-SOD. ORN treatment promoted higher wheat shoot and root dry weights, a higher root protein content, decreased root APX, GR and SOD activities but a higher proportion of MnSOD activity. ORN associated microbiota differently manage antioxidant enzyme activity of succeeding wheat to improve growth.


Asunto(s)
Antioxidantes , Micorrizas , Antioxidantes/metabolismo , Ascorbato Peroxidasas/metabolismo , Micelio/metabolismo , Micorrizas/metabolismo , Triticum/metabolismo
5.
Front Plant Sci ; 12: 642094, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33777077

RESUMEN

Fungicides are widely used in conventional agriculture to control fungal diseases, but may also affect non-target microorganisms such as arbuscular mycorrhizal (AM) fungi. These root symbionts develop extended mycelial networks within the soil via mechanisms such as anastomosis that indistinctly concerns intact and damaged hyphae, the latter being named hyphal healing mechanism (HHM). The HHM differs between Glomeraceae and Gigasporaceae. However, the effects of fungicides on this mechanism in unknown. Here, the impact of azoxystrobin, pencycuron, flutolanil, and fenpropimorph at 0.02 and 2 mg L-1 were tested in vitro on the HHM of Gigaspora sp. MUCL 52331 and Rhizophagus irregularis MUCL 41833, and repair events visualized carefully under a dissecting bright-field light microscope. Azoxystrobin was the more detrimental for both AM fungi at 2 mg L-1, while fenpropimorph impacted only R. irregularis (stimulating at low and inhibiting at high concentration). Conversely, flutolanil and pencycuron did not impact any of the two AM fungi. The mechanisms involved remains to be elucidated, but perturbation in the still-to-be firmly demonstrated spitzenkörper or in sterols content as well as a process of hormesis are possible avenues that deserve to be explored in view of a rationale management of chemicals to control fungal pathogens without harming the beneficial AM fungi.

6.
New Phytol ; 230(4): 1609-1622, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33091152

RESUMEN

In forests, ectomycorrhizal mycelium is pivotal for driving soil carbon and nutrient cycles, but how ectomycorrhizal mycelial dynamics vary in ecosystems with drought periods is unknown. We quantified the production and turnover of mycorrhizal mycelium in Mediterranean Pinus pinaster, Pinus sylvestris and Quercus ilex forests and related the estimates to standardised precipitation index (SPI), to study how mycelial dynamics relates to tree species and drought-moisture conditions. Production and turnover of mycelium was estimated between July and February, by quantifying the fungal biomass (ergosterol) in ingrowth mesh bags and using statistical modelling. SPI for time scales of 1-3 months was calculated from precipitation records and precipitation data over the study period. Forests dominated by Pinus trees displayed higher biomass but were seasonally more variable, as opposed to Q. ilex forests where the mycelial biomass remained lower and stable over the season. Production and turnover, respectively, varied between 1.4-5.9 kg ha-1  d-1 and 7.2-9.9 times yr-1 over the different forest types and were positively correlated with 2-month and 3-month SPI over the study period. Our results demonstrated that mycorrhizal mycelial biomass varied with season and tree species and we speculate that production and turnover are related to physiology and plant host performance during drought.


Asunto(s)
Micorrizas , Pinus sylvestris , Pinus , Quercus , Sequías , Ecosistema , Bosques , Micelio , Suelo , Árboles
7.
Methods Mol Biol ; 2146: 19-31, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32415592

RESUMEN

The mycorrhizal donor plant (MDP) in vitro culture system allows the fast and homogeneous colonization of a wide range of photosynthetically active plants. Here we detailed the setup of the system and its potential applications for basic studies as well as mass production and applied purposes.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Micorrizas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Simbiosis/genética
8.
Methods Mol Biol ; 2146: 33-41, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32415593

RESUMEN

An in vivo whole-plant bi-dimensional experimental system has been devised and tested with different host plants, in order to obtain extraradical mycelium (ERM) produced by different arbuscular mycorrhizal fungi (AMF). In this system, a host plant germling is inoculated with AMF to establish mycorrhizal symbiosis, and, after colonization, newly formed extraradical hyphae and spores are removed. Then the mycorrhizal root system is wrapped in a nylon net and placed between membranes in a Petri dish, allowing ERM to grow on the membrane surface. Such extraradical hyphae may be used for in situ morphometric analyses or collected for molecular or biochemical assays: in the latter case, the plant with its root sandwich may be reassembled to renew mycelium production. In this experimental system, which was tested with diverse host plant species and lines, values of explored membrane surface areas and densities of ERM showed wide ranges of variation, and its length ranged from 9.7 ± 2.0 to 48.8 ± 9.9 m per plant, depending on host and AMF identity. Across the different plant-AMF combinations tested, the whole-plant system produced 2.0 ± 0.6 to 5.3 ± 0.3 mg of ERM fresh biomass per plant per harvest. This experimental system can be used for a wide range of AMF and host plants species, either establishing arbuscular mycorrhizas or other mycorrhizal interactions. ERM produced and collected in the whole-plant system is suitable for morphological, physiological, and molecular analyses, facilitating studies on the different aspects of mycorrhizal symbiotic interactions.


Asunto(s)
Técnicas de Cultivo/métodos , Micorrizas/crecimiento & desarrollo , Simbiosis/genética , Hifa/citología , Hifa/crecimiento & desarrollo , Micelio/genética , Micelio/crecimiento & desarrollo , Micorrizas/citología , Raíces de Plantas/microbiología , Plantas/microbiología
9.
Mycorrhiza ; 30(2-3): 229-242, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32300867

RESUMEN

In plant-fungus phenotyping, determining fungal hyphal and plant root lengths by digital image analysis can reduce labour and increase data reproducibility. However, the degree of software sophistication is often prohibitive and manual measuring is still used, despite being very time-consuming. We developed the HyLength tool for measuring the lengths of hyphae and roots in in vivo and in vitro systems. The HyLength was successfully validated against manual measures of roots and fungal hyphae obtained from all systems. Compared with manual methods, the HyLength underestimated Medicago sativa roots in the in vivo system and Rhizophagus irregularis hyphae in the in vitro system by about 12 cm per m and allowed to save about 1 h for a single experimental unit. As regards hyphae of R. irregularis in the in vivo system, the HyLength overestimated the length by about 21 cm per m compared with manual measures, but time saving was up to 20.5 h per single experimental unit. Finally, with hyphae of Aspergillus oryzae, the underestimation was about 8 cm per m with a time saving of about 10 min for a single germinating spore. By benchmarking the HyLength against the AnaMorf plugin of the ImageJ/Fiji, we found that the HyLength performed better for dense fungal hyphae, also strongly reducing the measuring time. The HyLength can allow measuring the length over a whole experimental unit, eliminating the error due to sub-area selection by the user and allowing processing a high number of samples. Therefore, we propose the HyLength as a useful freeware tool for measuring fungal hyphae of dense mycelia.


Asunto(s)
Hifa , Micorrizas , Raíces de Plantas , Reproducibilidad de los Resultados , Esporas Fúngicas
10.
Mycorrhiza ; 30(2-3): 389-396, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32215759

RESUMEN

Arbuscular mycorrhizal fungi (AMF) absorb and translocate nutrients from soil to their host plants by means of a wide network of extraradical mycelium (ERM). Here, we assessed whether nitrogen-fixing rhizobia can be transferred to the host legume Glycine max by ERM produced by Glomus formosanum isolate CNPAB020 colonizing the grass Urochloa decumbens. An H-bridge experimental system was developed to evaluate the migration of ERM and of the GFP-tagged Bradyrhizobium diazoefficiens USDA 110 strain across an air gap compartment. Mycorrhizal colonization, nodule formation in legumes, and occurrence of the GFP-tagged strain in root nodules were assessed by optical and confocal laser scanning microscopy. In the presence of non-mycorrhizal U. decumbens, legume roots were neither AMF-colonized nor nodulated. In contrast, G. formosanum ERM crossing the discontinuous compartment connected mycorrhizal U. decumbens and G. max roots, which showed 30-42% mycorrhizal colonization and 7-11 nodules per plant. Fluorescent B. diazoefficiens cells were detected in 94% of G. max root nodules. Our findings reveal that, besides its main activity in nutrient transfer, ERM produced by AMF may facilitate bacterial translocation and the simultaneous associations of plants with beneficial fungi and bacteria, representing an important structure, functional to the establishment of symbiotic relationships.


Asunto(s)
Fabaceae , Micorrizas , Bacterias , Nitrógeno , Raíces de Plantas , Simbiosis
11.
Biology (Basel) ; 9(3)2020 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-32138156

RESUMEN

The sustainability of agriculture requires the adoption of agricultural soil conservation practices with positive impacts on soil quality, which can promote beneficial soil microbiota like arbuscular mycorrhizal fungi (AMF) and its diversity. This study aims to assess the influence of the presence of intact extraradical mycelium as a preferential source of inoculum of the native AMF in order to guarantee a better colonization as well as its possible bioprotective effect against Magnaporthiopsis maydis. In order to vary the available extraradical mycelium, two experiments, with and without cover crop, were carried out, in which two tillage systems and two maize varieties were studied. The capitalization of the benefits, in terms of grain production and M. maydis presence, associated to the cover crop were only achieved with minimum tillage. Therefore, both cultural practices are necessary to reduce the fungus presence, coupling the effect of mycorrhization together with other benefits associated with the cover crop. Although in the absence of a cover crop and using conventional tillage, yields and lower levels of M. maydis are possibly achieved, this system is more dependent on the variety used, does not benefit from the advantages associated with the cover crop, is more expensive, and environmentally unsustainable.

12.
Fungal Biol ; 124(2): 91-101, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-32008757

RESUMEN

Arbuscular Mycorrhizal fungi (AMF, Glomeromycota) form obligate symbiotic associations with the roots of most terrestrial plants. Our understanding of the molecular mechanisms enabling AMF propagation and AMF-host interaction is currently incomplete. Analysis of AMF proteomes could yield important insights and generate hypotheses on the nature and mechanism of AMF-plant symbiosis. Here, we examined the extraradical mycelium proteomic profile of the arbuscular mycorrhizal fungus Rhizophagus irregularis grown on Ri T-DNA transformed Chicory roots in a root organ culture setting. Our analysis detected 529 different peptides that mapped to 474 translated proteins in the R. irregularis genome. R. irregularis proteome was characterized by a high proportion of proteins (9.9 % of total, 21.4 % of proteins with functional prediction) mediating a wide range of signal transduction processes, e.g. Rho1 and Bmh2, Ca-signaling (calmodulin, and Ca channel protein), mTOR signaling (MAP3K7, and MAPKAP1), and phosphatidate signaling (phospholipase D1/2) proteins, as well as members of the Ras signaling pathway. In addition, the proteome contained an unusually large proportion (53.6 %) of hypothetical proteins, the majority of which (85.8 %) were Glomeromycota-specific. Forty-eight proteins were predicted to be surface/membrane associated, including multiple hypothetical proteins of yet-unrecognized functions. However, no evidence for the overproduction of specific proteins, previously implicated in promoting soil health and aggregation was obtained. Finally, the comparison of R. irregularis proteome to previously published AMF proteomes identified a core set of pathways and processes involved in AMF growth. We conclude that R. irregularis growth on chicory roots requires the activation of a wide range of signal transduction pathways, the secretion of multiple novel hitherto unrecognized Glomeromycota-specific proteins, and the expression of a wide array of surface-membrane associated proteins for cross kingdom cell-to-cell communications.


Asunto(s)
Hongos , Micelio/metabolismo , Proteoma , Comunicación Celular , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hongos/genética , Hongos/metabolismo , Genoma Fúngico , Glomeromycota/genética , Glomeromycota/metabolismo , Glicoproteínas/genética , Glicoproteínas/metabolismo , Micorrizas/metabolismo , Raíces de Plantas/microbiología , Técnicas de Embriogénesis Somática de Plantas/métodos , Plantas/microbiología , Proteoma/genética , Proteoma/metabolismo , Proteómica/métodos , Transducción de Señal , Simbiosis/genética , Simbiosis/fisiología
13.
Mycorrhiza ; 29(4): 341-349, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31190279

RESUMEN

The extraradical mycelium (ERM) produced by arbuscular mycorrhizal fungi is fundamental for the maintenance of biological fertility in agricultural soils, representing an important inoculum source, together with spores and mycorrhizal root fragments. Its viability and structural traits, such as density, extent and interconnectedness, which are positively correlated with the growth and nutrition of host plants, may be affected by different agronomic practices, including the use of pesticides and by different mycorrhizospheric communities. This work, carried out using a whole-plant experimental model system, showed that structural traits of ERM, such as length and density, were strongly decreased by the herbicides dicamba and glufosinolate and the fungicides benomyl and fenhexamid, while anastomosis frequency and hyphal branching were differentially modulated by singly inoculated mycorrhizospheric bacteria, depending on their identity.


Asunto(s)
Fenómenos Fisiológicos Bacterianos , Cichorium intybus/microbiología , Fungicidas Industriales/farmacología , Glomeromycota/efectos de los fármacos , Glomeromycota/crecimiento & desarrollo , Herbicidas/farmacología , Micelio/crecimiento & desarrollo , Micorrizas/efectos de los fármacos , Bacterias/genética , Bacterias/aislamiento & purificación , Benomilo/farmacología , Cichorium intybus/crecimiento & desarrollo , Dicamba/farmacología , Hifa/efectos de los fármacos , Hifa/crecimiento & desarrollo , Micelio/efectos de los fármacos , Micorrizas/crecimiento & desarrollo , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Esporas Bacterianas/genética , Esporas Bacterianas/aislamiento & purificación , Esporas Bacterianas/fisiología
14.
Front Plant Sci ; 10: 1617, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31921260

RESUMEN

In arbuscular mycorrhizal (AM) symbiosis, key components of nutrient uptake and exchange are specialized transporters that facilitate nutrient transport across membranes. As phosphate is a nutrient and a regulator of nutrient exchanges, we investigated the effect of P availability to extraradical mycelium (ERM) on both plant and fungus transcriptomes and metabolomes in a symbiocosm system. By perturbing nutrient exchanges under the control of P, our objectives were to identify new fungal genes involved in nutrient transports, and to characterize in which extent the fungus differentially modulates its metabolism when interacting with two different plant species. We performed transportome analysis on the ERM and intraradical mycelium of the AM fungus Rhizophagus irregularis associated to Populus trichocarpa and Sorghum bicolor under high and low P availability in ERM, using quantitative RT-PCR and Illumina mRNA-sequencing. We observed that mycorrhizal symbiosis induces expression of specific phosphate and ammonium transporters in both plants. Furthermore, we identified new AM-inducible transporters and showed that a subset of phosphate transporters is regulated independently of symbiotic nutrient exchange. mRNA-Sequencing revealed that the fungal transportome was not similarly regulated in the two host plant species according to P availability. Mirroring this effect, many plant carbohydrate transporters were down-regulated in P. trichocarpa mycorrhizal root tissue. Metabolome analysis revealed further that AM root colonization led to a modification of root primary metabolism under low and high P availability and to a decrease of primary metabolite pools in general. Moreover, the down regulation of the sucrose transporters suggests that the plant limits carbohydrate long distance transport (i.e. from shoot to the mycorrhizal roots). By simultaneous uptake/reuptake of nutrients from the apoplast at the biotrophic interface, plant and fungus are both able to control reciprocal nutrient fluxes.

15.
Front Microbiol ; 9: 2920, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30542338

RESUMEN

The symbiosis established between arbuscular mycorrhizal fungi (AMF) and roots of most land plants plays a key role in plant nutrient acquisition and alleviation of environmental stresses. Despite the ubiquity of the symbiosis, AMF and host species display significant specificity in their interactions. To clarify preferential associations between wheat (Triticum aestivum) and AMF, we characterized root AMF communities in the transition from two first host species, ryegrass (Lolium rigidum) and yellow-serradella (Ornithopus compressus), grown separately or together, to a second host (wheat), by sequencing the large subunit ribosomal DNA (LSU rDNA) gene. The response of AMF communities in wheat to prior soil disturbance - and consequently of the mycelial network [intact extraradical mycelium (ERM) vs. disrupted mycelium] established with either of the first hosts - was also investigated. Since the outcome of a specific host-symbiont interaction depends on the molecular responses of the host plant upon microbial colonization, we studied the expression of six key symbiosis-related genes in wheat roots. AMF communities on L. rigidum and O. compressus roots were clearly distinct. Within an undisturbed ERM, wheat AMF communities were similar to that of previous host, and O. compressus-wheat-AMF interactions supported a greater growth of wheat than L. rigidum-wheat-AMF interactions. This effect declined when ERM was disrupted, but generated a greater activation of symbiotic genes in wheat, indicating that plant symbiotic program depends on some extent on the colonizing symbiont propagule type. When a mixture of L. rigidum and O. compressus was planted, the wheat colonization pattern resembled that of O. compressus, although this was not reflected in a greater growth. These results show a lasting effect of previous hosts in shaping wheat AMF communities through an efficient use of the established ERM, although not completely obliterating host-symbiont specificity.

16.
Mycorrhiza ; 27(7): 659-668, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28573458

RESUMEN

Arbuscular mycorrhizal fungi (AMF) establish beneficial mutualistic symbioses with land plants, receiving carbon in exchange for mineral nutrients absorbed by the extraradical mycelium (ERM). With the aim of obtaining in vivo produced ERM for gene expression analyses, a whole-plant bi-dimensional experimental system was devised and tested with three host plants and three fungal symbionts. In such a system, Funneliformis mosseae in symbiosis with Cichorium intybus var. foliosum, Lactuca sativa, and Medicago sativa produced ERM whose lengths ranged from 9.8 ± 0.8 to 20.8 ± 1.2 m per plant. Since ERM produced in symbiosis with C. intybus showed the highest values for the different structural parameters assessed, this host was used to test the whole-plant system with F. mosseae, Rhizoglomus irregulare, and Funneliformis coronatus. The whole-plant system yielded 1-7 mg of ERM fresh biomass per plant per harvest, and continued producing new ERM for 6 months. Variable amounts of high-quality and intact total RNA, ranging from 15 to 65 µg RNA/mg ERM fresh weight, were extracted from the ERM of the three AMF isolates. Ammonium transporter gene expression was successfully determined in the cDNAs obtained from ERM of the three fungal symbionts by RT-qPCR using gene-specific primers designed on available (R. irregulare) and new (F. mosseae and F. coronatus) ammonium transporter gene sequences. The whole-plant experimental system represents a useful research tool for large production and easy collection of ERM for morphological, physiological, and biochemical analyses, suitable for a wide variety of AMF species, for a virtually limitless range of host plants and for studies involving diverse symbiotic interactions.


Asunto(s)
Cichorium intybus/microbiología , Perfilación de la Expresión Génica/métodos , Glomeromycota/genética , Micorrizas/genética , Transcriptoma , Cichorium intybus/fisiología , Micorrizas/fisiología
17.
New Phytol ; 214(1): 424-431, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27997034

RESUMEN

In boreal forest soils, ectomycorrhizal fungi are fundamentally important for carbon (C) dynamics and nutrient cycling. Although their extraradical mycelium (ERM) is pivotal for processes such as soil organic matter build-up and nitrogen cycling, very little is known about its dynamics and regulation. In this study, we quantified ERM production and turnover, and examined how these two processes together regulated standing ERM biomass in seven sites forming a chronosequence of 12- to 100-yr-old managed Pinus sylvestris forests. This was done by determining ERM biomass, using ergosterol as a proxy, in sequentially harvested in-growth mesh bags and by applying mathematical models. Although ERM production declined with increasing forest age from 1.2 to 0.5 kg ha-1  d-1 , the standing biomass increased from 50 to 112 kg ha-1 . This was explained by a drastic decline in mycelial turnover from seven times to one time per year with increasing forest age, corresponding to mean residence times from 25 d up to 1 yr. Our results demonstrate that ERM turnover is the main factor regulating biomass across differently aged forest stands. Explicit inclusion of ERM parameters in forest ecosystem C models may significantly improve their capacity to predict responses of mycorrhiza-mediated processes to management and environmental changes.


Asunto(s)
Biomasa , Micelio/fisiología , Micorrizas/fisiología , Pinus sylvestris/microbiología , Geografía , Suecia , Factores de Tiempo
18.
Mycorrhiza ; 27(3): 201-210, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27838855

RESUMEN

The formation of storage organs, such as spores and vesicles, is a central part of the life cycle of an arbuscular mycorrhizal fungus (AMF), but the conditions under which this occurs in AMF are not well understood. Here, quantity and distribution of storage organs formed by the arbuscular mycorrhizal fungus (AMF) Funneliformis mosseae within dead (excised) roots were characterised. 'Trap roots' (TR), separated from the growth substrate by a 30-µm mesh, supported hyphal growth and formation of storage organs of the AMF. Hyphae developed both inside and on the outside of the TR and also within air gaps of surrounding nylon mesh compartments, but formation of vesicles and spores was confined to the interior and to the surface of the TR. Up to 20 % of the TR length harboured newly formed storage organs, resulting in a number of about 60 per mg TR dry weight. The portion of TR length containing storage organs was greater in coarse (diameter >300 µm) than in thin (<150 µm) TR, irrespective of whether the TR were sourced from an AMF host or non-host plant. We conclude that the AMF's extraradical mycelium produces its storage organs within dead roots in preference to air space in the substrate. Dead roots may indirectly supply nutrients to AMF (once they have been mineralised) or represent a protected space for the fungal structures to develop. The experimental technique described here allows for the preparation of AMF spores and vesicles of F. mosseae free of any mineral substrate.


Asunto(s)
Micorrizas/metabolismo , Esporas Fúngicas/metabolismo , Zea mays/microbiología , Hifa/crecimiento & desarrollo , Micelio/crecimiento & desarrollo , Micorrizas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Microbiología del Suelo , Simbiosis
19.
Front Plant Sci ; 7: 679, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27252708

RESUMEN

Nutrient acquisition and transfer are essential steps in the arbuscular mycorrhizal (AM) symbiosis, which is formed by the majority of land plants. Mineral nutrients are taken up by AM fungi from the soil and transferred to the plant partner. Within the cortical plant root cells the fungal hyphae form tree-like structures (arbuscules) where the nutrients are released to the plant-fungal interface, i.e., to the periarbuscular space, before being taken up by the plant. In exchange, the AM fungi receive carbohydrates from the plant host. Besides the well-studied uptake of phosphorus (P), the uptake and transfer of nitrogen (N) plays a crucial role in this mutualistic interaction. In the AM fungus Rhizophagus irregularis (formerly called Glomus intraradices), two ammonium transporters (AMT) were previously described, namely GintAMT1 and GintAMT2. Here, we report the identification and characterization of a newly identified R. irregularis AMT, GintAMT3. Phylogenetic analyses revealed high sequence similarity to previously identified AM fungal AMTs and a clear separation from other fungal AMTs. Topological analysis indicated GintAMT3 to be a membrane bound pore forming protein, and GFP tagging showed it to be highly expressed in the intraradical mycelium of a fully established AM symbiosis. Expression of GintAMT3 in yeast successfully complemented the yeast AMT triple deletion mutant (MATa ura3 mep1Δ mep2Δ::LEU2 mep3Δ::KanMX2). GintAMT3 is characterized as a low affinity transport system with an apparent Km of 1.8 mM and a V max of 240 nmol(-1) min(-1) 10(8) cells(-1), which is regulated by substrate concentration and carbon supply.

20.
New Phytol ; 208(1): 280-93, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26011828

RESUMEN

A significant fraction of carbon stored in the Earth's soil moves through arbuscular mycorrhiza (AM) and ectomycorrhiza (EM). The impacts of AM and EM on the soil carbon budget are poorly understood. We propose a method to quantify the mycorrhizal contribution to carbon cycling, explicitly accounting for the abundance of plant-associated and extraradical mycorrhizal mycelium. We discuss the need to acquire additional data to use our method, and present our new global database holding information on plant species-by-site intensity of root colonization by mycorrhizas. We demonstrate that the degree of mycorrhizal fungal colonization has globally consistent patterns across plant species. This suggests that the level of plant species-specific root colonization can be used as a plant trait. To exemplify our method, we assessed the differential impacts of AM : EM ratio and EM shrub encroachment on carbon stocks in sub-arctic tundra. AM and EM affect tundra carbon stocks at different magnitudes, and via partly distinct dominant pathways: via extraradical mycelium (both EM and AM) and via mycorrhizal impacts on above- and belowground biomass carbon (mostly AM). Our method provides a powerful tool for the quantitative assessment of mycorrhizal impact on local and global carbon cycling processes, paving the way towards an improved understanding of the role of mycorrhizas in the Earth's carbon cycle.


Asunto(s)
Ciclo del Carbono , Carbono/metabolismo , Micorrizas/metabolismo , Raíces de Plantas , Plantas , Microbiología del Suelo , Suelo/química , Biomasa , Micelio , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Plantas/metabolismo , Plantas/microbiología , Simbiosis
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