Intact Extraradical Mycelium Research Articles

Mycorrhizal Colonization of Wheat by Intact Extraradical Mycelium of Mn-Tolerant Native Plants Induces Different Biochemical Mechanisms of Protection.

Soil with excess Mn induces toxicity and impairs crop growth. However, with the development in the soil of an intact extraradical mycelia (ERM) from arbuscular mycorrhizal fungi (AMF) symbiotic to native Mn-tolerant plants, wheat growth is promoted due to a stronger AMF colonization and subsequent increased protection against Mn toxicity. To determine the biochemical mechanisms of protection induced by this native ERM under Mn toxicity, wheat grown in soil from previously developed Lolium rigidum (LOL) or Ornithopus compressus (ORN), both strongly mycotrophic plants, was compared to wheat grown in soil from previously developed Silene gallica (SIL), a non-mycotrophic plant. Wheat grown after LOL or ORN had 60% higher dry weight, ca. two-fold lower Mn levels and almost double P contents. Mn in the shoots was preferentially translocated to the apoplast along with Mg and P. The activity of catalase increased; however, guaiacol peroxidase (GPX) and superoxide dismutase (SOD) showed lower activities. Wheat grown after ORN differed from that grown after LOL by displaying slightly higher Mn levels, higher root Mg and Ca levels and higher GPX and Mn-SOD activities. The AMF consortia established from these native plants can promote distinct biochemical mechanisms for protecting wheat against Mn toxicity.

Subcellular Element Distribution in Shoots of Wheat Grown in an Acidic Soil with Native AMF Extraradical Mycelium

Soil acidity can reduce crop growth by increasing bioavailable soil Al, Fe, and/or Mn to toxic levels. The presence of an intact extraradical mycelium (ERM) of arbuscular mycorrhizal fungi (AMF), developed by the native Ornithopus compressus in the acidic soil, can increase wheat growth and prevent symptoms of Mn toxicity. To understand the protective effect of the intact ERM of this native plant on wheat element balance and distribution, in the present study, shoot Al, Fe, K, Zn, Na, and Si levels and their subcellular partitioning were determined by inductively coupled plasma mass spectrometry (ICP-MS), for the first time, for this system. In undisturbed soil, where an intact ERM structure is maintained, wheat shoot growth was promoted, probably due to faster root mycorrhizal colonization. The levels of potentially toxic Al and Fe were reduced, the proportions of the macronutrient K and micronutrient Zn were higher in the symplast, and the Na proportion increased in the vacuole, while Si increased in the apoplast. Overall, the undisturbed soil from O. compressus treatment appeared to influence the uptake and distribution of essential and beneficial elements, as a strategy to reduce the negative effect of soil acidity on wheat growth. Understanding the dynamics of element distribution influenced by stress-adapted AMF on wheat growth can provide more sustainable approaches to intensive agriculture.

Manganese Uptake to Wheat Shoot Meristems Is Differentially Influenced by Arbuscular Mycorrhiza Fungal Communities Adapted to Acidic Soil

Soil acidity is a strong promoter of the bioavailability of Al, Fe, and Mn, whose concentrations can sometimes reach toxic levels for plants. In agricultural soils, the use of arbuscular mycorrhizal fungi (AMF) has shown a protective influence on wheat growth under Mn toxicity. The intact extraradical mycelium (ERM) promotes faster AMF colonization, leading to a higher wheat shoot weight, lower Mn uptake, and changes in antioxidant enzyme activity. Its effect on the uptake and distribution of plant nutrients according to the developmental stage of shoot organs has seldomly been analyzed. In the present study, Mn, Mg, Ca, and K were quantified by ICP–MS in leaves and apical meristems of wheat grown in soil with two different ERM consortia, developed from the native plants Lolium rigidum (LOL) and Ornithopus compressus (ORN). The ORN treatment induced the highest wheat shoot weights and the lowest Mn levels. In the leaves, no significant differences were detected for Mg, Ca, or K, but in the apical meristems, the ORN treatment slightly lowered the Ca concentration. The AMF associated with ORN was seen to enhance wheat weight partly by protecting the zones of active growth against high Mn levels in Mn toxic soils. The use of ORN in acidic soils with Mn toxicity provides a sustainable alternative and an efficient complement to current farming practices to lower the negative impacts of farming on the environment.

Diversity of Native Arbuscular Mycorrhiza Extraradical Mycelium Influences Antioxidant Enzyme Activity in Wheat Grown Under Mn Toxicity.

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.

The Protective Biochemical Properties of Arbuscular Mycorrhiza Extraradical Mycelium in Acidic Soils Are Maintained throughout the Mediterranean Summer Conditions

In acidic soils with manganese (Mn) toxicity, arbuscular mycorrhizal fungi (AMF) can improve plant host growth by enhancing nutrition and protecting against environmental stress. The intact extraradical mycelium (ERM) of AMF is able to survive Mediterranean summer conditions and provide an earlier colonization of winter crops. This study evaluated if summer season conditions hindered the beneficial effects of wheat colonization by the intact ERM associated with a native plant, in acidic soil. Wheat was grown in soil with intact or disrupted ERM associated with Ornithopus compressus (ORN), developed for 7 or 24 weeks, to simulate ERM summer survival. The activity of antioxidant enzymes was determined, and the quantitative analysis of Mn and macronutrients was performed by inductively coupled plasma mass spectrometry (ICP-MS), in wheat shoots and respective subcellular fractions. Wheat colonization by intact ERM decreased shoot Mn concentration but increased the proportion of Mn in the apoplast. Overall, antioxidant enzymatic activity decreased but the proportion of Mn-superoxide dismutase activity over the remaining isoforms increased, suggesting its important role in the AMF-mediated mitigation of Mn toxicity. Summer conditions did not substantially reduce the benefits provided by ORN ERM. A no-till strategy allied to the development of native microbiota can contribute to the sustainable optimization of acidic soil use.

Effect of Tillage System and Cover Crop on Maize Mycorrhization and Presence of Magnaporthiopsis maydis.

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.

Symbiosis Specificity of the Preceding Host Plant Can Dominate but Not Obliterate the Association Between Wheat and Its Arbuscular Mycorrhizal Fungal Partners.

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.

Agronomic management of AMF functional diversity to overcome biotic and abiotic stresses - The role of plant sequence and intact extraradical mycelium

Intentional use of arbuscular mycorrhizal fungi (AMF) in cropping systems has been marginal, owing to the high cost and limited biodiversity of commercial inocula, together with the timeliness of colonization to achieve benefits. Additionally, mycorrhiza are considered incompatible with high input cropping systems. Combining results from 4 different experiments resulted in a strategy for the earlier and faster colonization by AMF, through an extensive extraradical mycelium (ERM) acting as a preferential source of inoculum if kept intact by the adoption of appropriate tillage techniques. Selection of host plants on which the ERM develops, provides the tool to manage AMF functional diversity. This strategy resulted in protection of sensitive crop species against biotic and abiotic stresses and can be implemented in low- and high-input cropping systems. Under Mn toxicity arbuscular colonization increased 2.6-fold and shoot dry weight 2.3-fold. In presence of Fusarium, arbuscular colonization increased 2.1-fold and shoot dry weight 1.5-fold.

The effect of arbuscular mycorrhiza fungal propagules on the growth of subterranean clover (Trifolium subterraneum L.) under Mn toxicity in ex situ experiments

AbstractThe role of intact extraradical mycelium (ERM) as the most effective fungal propagule in the formation of the tripartite symbiosis between indigenous arbuscular mycorrhizal fungi (AMF), rhizobia and subterranean clover was investigated under conditions of Mn toxicity. ERM was previously developed in 8 L pots under greenhouse conditions by growing plants, which exhibited various levels of mycotrophicity and were tolerant to the levels of Mn in the soil used in the experiment (Silene gallica L, Lolium rigidum L, Ornithopus compressus L. and Rumex bucephalophorus L). Contrasting conditions of the integrity of the ERM at the planting of subterranean clover were created by soil disturbance (ERM fragmented – soil disturbed; ERM intact – soil undisturbed). Where an intact ERM was present at the time of planting, growth of subterranean clover was 2.5 times greater after 21 days and 3.9 times after 42 days relative to other forms of AMF propagule. This enhanced growth was associated with a reduction in the Mn concentration of roots due to a greater AMF colonization at 21 days after planting. The protection granted by an enhanced AMF root colonization allowed a greater root nodule development, leading to more N acquisition and plant growth. The ERM can be developed in the soil by mycotrophic plants tolerant to the stressing agent and kept intact at the seeding of the crop to be protected by adopting appropriate tillage techniques.

Managing arbuscular mycorrhizal fungi for bioprotection: Mn toxicity

We investigated whether an intact extraradical mycelium (ERM) is more effective than other forms of propagule from indigenous arbuscular mycorrhizal fungi (AMF) in providing protection against stress to a host plant. The response of wheat (Triticum aestivum L.) to Mn toxicity was studied in a two-phase greenhouse experiment. In Phase 1, four Mn tolerant species from the natural vegetation, ranging from strongly mycotrophic to non- or weakly mycotrophic, were grown to develop different amounts of ERM. Wheat was then planted (Phase 2) with the ERM fragmented by sieving (Disturbed Treatment) or kept intact with no prior soil disturbance (Undisturbed Treatment). The growth of wheat was doubled by earlier and faster mycorrhizal colonization (AC) in the presence of an intact ERM at planting. There was a positive correlation between plant growth and the reduction of Mn and enhancement of P and S uptake into shoots. However, the growth of plants in undisturbed soil was significantly affected by the ERM developer species, which was not explained by differences in AC. Colonization starting from an intact ERM greatly enhanced the potential of AMF for protection against Mn toxicity. However, the degree of protection depended on the plant previously grown to develop the ERM, suggesting that there may be functional diversity within the ERM developed by mycotrophic plants of the natural vegetation.

Fungal biomass production in response to elevated atmospheric CO2 in a Glomus mosseae–Prunus cerasifera model system

Biomass and length of intraradical and extraradical mycorrhizal mycelium under ambient (aCO2) and elevated (eCO2 ) atmospheric CO2 was investigated using a non-destructive in vivo experimental model system. Time-course experiments allowed measurements of intact extraradical mycelium spreading from mycorrhizal roots of Prunus cerasifera micropropagated plants inoculated with the arbuscular mycorrhizal fungus Glomus mosseae, in controlled environmental chambers. The length of extraradical mycelium was significantly increased at the highest CO2 concentration, ranging from 10.7 to 20.3 m at aCO2 and eCO2, respectively. The biochemical determination of mycelial glucosamine content allowed the evaluation of intraradical and extraradical fungal biomass, which were 2 and 3 times larger at eCO2 than at aCO2. Present data show that Glomus mosseae responds to increases of CO2 concentrations producing larger mycorrhizal networks which may potentially represent carbon sink agents in soil ecosystems.

Functional diversity of arbuscular mycorrhizal fungal isolates in relation to extraradical mycelial networks

We investigated the functional significance of extraradical mycorrhizal networks produced by geographically different isolates of the arbuscular mycorrhizal fungal (AMF) species Glomus mosseae and Glomus intraradices. A two-dimensional experimental system was used to visualize and quantify intact extraradical mycelium (ERM) spreading from Medicago sativa roots. Growth, phosphorus (P) and nitrogen (N) nutrition were assessed in M. sativa plants grown in microcosms. The AMF isolates were characterized by differences in extent and interconnectedness of ERM. Phenotypic fungal variables, such as total hyphal length, hyphal density, hyphal length per mm of total or colonized root length, were positively correlated with M. sativa growth response variables, such as total shoot biomass and plant P content. The utilization of an experimental system in which size, growth rate, viability and interconnectedness of ERM extending from mycorrhizal roots are easily quantified under realistic conditions allows the simultaneous evaluation of different isolates and provides data with a predictive value for selection of efficient AMF.

Kinetics of phosphate absorption by mycorrhizal and non‐mycorrhizal Scots pine seedlings

Kinetics of net phosphate (Pi) uptake was measured on intact ectomycorrhizal and non‐mycorrhizal Pinus sylvestris seedlings using a semihydroponic cultivation method. The depletion of Pi in a nutrient solution was assessed over a 160–0.2 μM Pi gradient. Growth of the pine seedlings was P limited and measurements were performed 7 and 9 weeks after inoculation. Three ectomycorrhizal fungi were studied: Paxillus involutus, Suillus bovinus and Thelephoraterrestris. Pi uptake was extremely fast in plants colonised by P. involutus. The Pi concentration dropped below 0.2 μM within 4–5 h. In plants colonised with S. bovinus this occurred in 5–6 h and in plants associated with T. terrestris 8 h were needed to run through the whole concentration range. Non‐mycorrhizal plants of similar size and nutrient status decreased Pi to a concentration between 1 and 2 μM in 18 h. Data were curve fitted to a two‐phase Michaelis‐Menten equation. The apparent kinetic constants, Km and Vmax, for the high affinity Pi uptake system of the pine roots could be estimated accurately. Vmax of this system was up to 7 times higher in pines associated with P. involutus than in non‐mycorrhizal seedlings. The intact extraradical mycelium greatly increased the absorption surface area of the roots (Vmax). Non‐mycorrhizal plants had a Km between 7.8 and 16.4 μM Pi. Plants mycorrhizal with P. involutus had Km values between 2.4 and 7.2, plants colonised with S. bovinus had a Km between 5.1 and 12.3, and seedlings associated with T. terrestris had a Km from 4.6 to 10.1 μM Pi. All 3 ectomycorrhizal fungi had a strong impact on the Pi absorption capacity of the pine seedlings. The results also demonstrated that there is substantial heterogeneity in kinetic parameters among the different mycorrhizal root systems.