Mycorrhizal Fungal Colonization Research Articles

Effect of Glomus mosseae, cadmium, and elevated air temperature on main flavonoids and phenolic acids contents in alfalfa.

Global warming and heavy metal-contaminated soils co-occur in natural ecosystems. Flavonoids and phenolic acids in plants have significant antioxidant activity and free radical scavenging ability, which can quickly increase under adverse environments. Arbuscular mycorrhizal fungi (AMF) colonization can affect the synthesis of flavonoids and phenolic acids in host plants. This study focused on the main effect of Glomus mosseae, cadmium (Cd, 8mgkg-1 dry soils), and elevated temperature (ET, + 3°C) on main flavonoids and phenolic acids in 120-d Medicago sativa L. (alfalfa). Elevated temperature decreased G. mosseae colonization ratio by 49.5% under Cd exposure. Except for p-hydroxybenzoic acid, flavonoids and phenolic acids content in shoots increased (p < 0.05) under G. mosseae + Cd relative to Cd only. G. mosseae and Cd showed significant effects on rutin, quercetin, apigenin, liquiritigenin, gallic acid, p-hydroxybenzoic acid, p-coumaric acid, and ferulic acid, and G. mosseae colonization led to increases in these compounds by 41.7%, 35.4%, 32.2%, 267.8%, 84.7%, 33.5%, 102.8%, and 89.4%, respectively, under ET + Cd. Carbon, N, and Cd in alfalfa and G. mosseae colonization rate were significant factors on flavonoids and phenolic acids accumulation. Additionally, P content in shoots significantly influenced flavonoids content. G. mosseae inoculation significantly stimulated the synthesis of main flavonoids and phenolic acids in alfalfa shoots under ET + Cd, which was helpful to understand the regulation of AMF on non-enzyme antioxidant system of plants grown in heavy metal-contaminated soils under global change scenarios.

Absorptive root-multidimension strategy links air temperature and species distribution in a montane forest

BackgroundAir temperature affects absorptive root traits, which are closely related to species distribution. However, it is still unclear how air temperature regulates species distribution through changes in absorptive root traits. Seven functional traits of the absorptive roots of 240 individuals of 52 species, soil properties and air temperature were measured along an elevational gradient on Mt. Fanjingshan, Tongren City, Guizhou, and then the direct and indirect effects of these controls on species distribution were detected. ResultsAbsorptive roots adapted to air temperature with two strategies. The first strategy was positively associated with the specific root area (SRA) and specific root length (SRL) and was negatively associated with the root tissue density (RTD), representing the classic root economics spectrum (RES). The second strategy was represented by the trade-off between root diameter, mycorrhizal fungi colonization (MF) and SRL, representing the collaboration gradient with “do it yourself” resource uptake ranging from “outsourcing” to mycorrhizal resource uptake. Air temperature regulated species distribution in six ways: directly reducing species importance value; indirectly increasing the species importance value by reducing soil nitrogen content or increasing soil pH by reducing soil moisture inducing absorptive roots to change from “do it yourself” resource absorption to “outsourcing” resource absorption; indirectly decreasing the species importance value by decreasing soil moisture to change from “outsourcing” resource absorption to “do it yourself” resource absorption; indirectly increasing the species importance value with increasing soil pH by reducing soil moisture resulting in absorptive root traits turning into nutrient foraging traits; and indirectly decreasing the species importance value by promoting absorptive root traits to nutrient conservation traits. ConclusionsAbsorptive root traits play a crucial role in the regulation of species distribution through multi-approaches of air temperature.

Role of plant relatedness in plant-soil feedback dynamics of sympatric Asclepias species.

Plants affect associated biotic and abiotic edaphic factors, with reciprocal feedbacks from soil characteristics affecting plants. These two-way interactions between plants and soils are collectively known as plant-soil feedbacks (PSFs). The role of phylogenetic relatedness and evolutionary histories have recently emerged as a potential driver of PSFs, although the strength and direction of feedbacks among sympatric congeners are not well-understood. We examined plant-soil feedback responses of Asclepias syriaca, a common clonal milkweed species, with several sympatric congeners across a gradient of increasing phylogenetic distances (A.tuberosa, A.viridis, A.sullivantii, and A.verticillata, respectively). Plant-soil feedbacks were measured through productivity and colonization by arbuscular mycorrhizal (AM) fungi. Asclepias syriaca produced less biomass in soils conditioned by the most phylogenetically distant species (A.verticillata), relative to conspecific-conditioned soils. Similarly, arbuscular mycorrhizal (AM) fungal colonization of A.syriaca roots was reduced when grown in soils conditioned by A.verticillata, compared with colonization in plants grown in soil conditioned by any of the other three Asclepias species, indicating mycorrhizal associations are a potential mechanism of observed positive PSFs. This display of differences between the most phylogenetically distant, but not close or intermediate, paring(s) suggests a potential phylogenetic threshold, although other exogenous factors cannot be ruled out. Overall, these results highlight the potential role of phylogenetic distance in influencing positive PSFs through mutualists. The role of phylogenetic relatedness and evolutionary histories have recently emerged as a potential driver of plant-soil feedbacks (PSFs), although the strength and direction of feedbacks among sympatric congeners are not well-understood. Congeneric, sympatric milkweeds typically generated positive PSFs in terms of productivity and AM fungal colonization, suggesting the low likelihood of coexistence among tested pairs, with a strength of feedback increasing as the phylogenetic distance increases.

Leaf manganese and phenolics as proxies of soil acidification and phosphorus acquisition mechanisms in lentil cultivars on alkaline soil.

Leaf manganese (Mn) concentration has been used as a proxy for root exudation and phosphorus (P) uptake under controlled conditions, but there are limited field studies that confirm its validity. On an alkaline, P-poor soil, four lentil cultivars ('Samos', 'Thessaly', 'Flip', 'Algeria') received two P rates (0 and 26.2kgPha-1 ), for two growing seasons, to study whether aboveground assessments [leaf P, Mn, phenolic concentration (TPhe)] can approximate rhizosphere physiological traits related to P acquisition [soil acidification (ΔpH), arbuscular mycorrhizal fungi (AMF) colonisation, acid phosphatase activity (APase)]. Phosphorus addition had no effect on the determined traits. Cultivars differed in leaf P, Mn, TPhe and AMF, but there was no clear pattern relating aboveground traits to rhizosphere traits related to P acquisition, thus not confirming that leaf Mn can be a proxy of root exudation. Of three growth stages [V 7-8, R1 (first bloom), R4 (flat pod)], R1 seemed to be critical, showing the highest leaf P, ΔpH, AMF and TPhe. Precipitation and temperatures over the growing season were determinants of lentil responses affecting rhizosphere activity, soil P availability and finally leaf traits. In conclusion, in lentil on alkaline and P-limiting soils, high leaf Mn and phenolic concentration are not reliable indicators of rhizosphere P-acquiring mechanisms.

Predicting arbuscular mycorrhizal fungal colonization of soybean in farmers’ fields by using infection unit density

ABSTRACT Estimating arbuscular mycorrhizal (AM) fungal activity to colonize crop root before cultivation is prerequisite for effective utilization of their functions which enhance growth and yield of the plant especially under low fertilizer input. We have hypothesized that the infection unit (IU) density formed on test plant roots grown for short period (12 days) with soil sampled from soybean production fields would be an effective indicator to predict AM fungal colonization intensity to the plant. In order to test this hypothesis, three-year farmland survey was conducted, in which soil samples before sowing soybean and the plant root samples at third trifoliate (V3) and full bloom (R2) stage were collected from farmers’ fields in two regions in Hokkaido, Iwamizawa and Tokachi. For each sampling spot, IU density was determined by using test plants, and intensity of AM fungal colonization of soybean root was measured. Before pursuing field survey, laboratory experiments were conducted to find out proper soil storage condition that keeps IU density unchanged while handling many soil samples. Our results indicated that IU density was almost comparable to the original value after six-month storage if soil samples were kept in a refrigerator, although storing at ambient temperature significantly decreased the measurement. Air drying also had negative impact on IU density. According to the field survey, IU densities determined using field soil were positively and significantly correlated with AM fungal colonization of soybean roots at both V3 and R2 stages. Differences in climate, soil type, and style of agriculture between Iwamizawa and Tokachi seemed to have little effect on IU density-AM fungal colonization relationship. Other than IU density, soil pH and soil penetration resistance at 10 cm depth were selected as significant explanatory variables for predicting AM fungal colonization by multiple regression analysis. However, IU density was the most influential factor among three. Therefore, IU density is recognized as an effective measure to evaluate AM fungal colonizing activity in field soil.

Mycorrhizal Fungi Synergistically Promote the Growth and Secondary Metabolism of Cyclocarya paliurus

Cyclocarya paliurus has traditionally been used as medicine or a nutraceutical food. This study aims at investigating whether the growth and secondary metabolism of C. paliurus could be simultaneously promoted by inoculating with mycorrhizal fungi, and if so, to uncover the underlying regulatory mechanism. A mycorrhizal microbial inoculum, consisting of the superficial layer fine roots and rhizosphere soil collected from the natural forest of C. paliurus, was used to infect aseptic seedlings of C. paliurus. Roots of aseptic seedlings were successfully infected by mycorrhizal fungi with a 59.7% colonization rate. For mycorrhizal seedlings of C. paliurus, the induced endogenous auxin, net photosynthetic rate, nitrogen absorption, and growth-related genes resulted in a significantly higher growth and biomass accumulation. In addition, a systemic defense response was observed in response to mycorrhizal fungal colonization, such that jasmonic acid biosynthesis and signaling were induced and the biosynthesis of secondary metabolites and antioxidant systems were up-regulated. The improved growth and accumulation of secondary metabolites ultimately facilitated the yield of health-promoting substrates per plant. Overall, mycorrhizal fungal colonization had a significant positive effect both on growth and production of secondary metabolites in C. paliurus. The results can provide the basis for overcoming the limitation of soil nutrient regulation in cultivation practice and offering a simpler alternative to improve the quality of medicinal plants.

Interplay between rhizobial nodulation and arbuscular mycorrhizal fungal colonization inLotus japonicusroots

This study aims to identify main factors that influence the tripartite association of legumes with arbuscular mycorrhiza fungi (AMF) and nitrogen-fixing rhizobia. Concurrent inoculations with Mesorhizobium loti and four AMF strains were performed on the model legume Lotus japonicus. Nodulation was significantly enhanced by all AMF strains, under normal conditions, and by specific AMF strains under heat-stress conditions. The impact of rhizobia on mycorrhizal colonization was AMF strain dependent. Co-inoculation trials, where either AMF or rhizobia were restricted outside the root, showed that the symbiotic phenotypes are not influenced by microbial interactions at the pre-symbiotic stage. External application of nutrients showed that P enhances nodulation, while N application does not enhance mycorrhizal colonization. Nodulation and mycorhization affect one another during advanced stages of the symbiosis. AMF strains may enhance nodulation under both normal and high environmental temperatures. Rhizobium-AMF compatibility is critical, as rhizobium may positively affect specific AMF strains, an effect that does not derive from increased N uptake.

Distribution Characteristics and Influence Factors of Rhizosphere Glomalin-Related Soil Protein in Three Vegetation Types of Helan Mountain, China

To reveal distribution characteristics of glomalin-related soil protein (GRSP) and it’s influencing factors under different vegetation types in the drought-tolerant shrubland of Helan Mountain, we chose three vegetation types as study subjects: Stipa breviflora (Grassland, G), Amygdalus mongolica (Shrub, S), and Stipa breviflora-Amygdalus mongolica (Grassland-Shrub, G×S) and bare soil was used as the control (CK). The contents of easily extractable glomalin-related soil protein (EE-GRSP) and total glomalin-related soil protein (T-GRSP), soil physicochemical properties, colonization rate, spore density, and species abundance in the rhizosphere soil were determined. The results indicated that EE-GRSP and T-GRSP showed significant difference (p &lt; 0.05) among vegetation types, with GRSP content highest under G×S (5.68 and 6.27 mg·g−1, respectively) and lowest under CK (3.84 and 4.48 mg·g−1, respectively). EE-GRSP/soil organic carbon (SOC) and T-GRSP/SOC showed no significant difference (p &gt; 0.05). The trends of colonization rate, spore density, and species abundance were the same and were significantly different from those of GRSP content (p &lt; 0.05), with maximum values of 75.6%, 20.7 × 10 g−1, and 29.7, and minimum values of 55.6%, 13.0 × 10 g−1, and 12.7, respectively. Pearson correlation analysis showed that EE-GRSP was significantly positively correlated with SOC, total phosphorus, available phosphorus, and colonization rate (p &lt; 0.05), and it showed an extremely significant positive correlation with available potassium, spore density, and species abundance (p &lt; 0.01). T-GRSP was significantly positively correlated with total phosphorus and available phosphorus (p &lt; 0.05), as well as with soil organic carbon, available potassium, spore density, colonization rate, and species abundance (p &lt; 0.01). The redundancy analysis (RDA) showed similar results. Therefore, the distribution characteristics of GRSP and its influencing factors under different vegetation types in the low elevation area of Helan Mountain were influenced by vegetation types, physicochemical properties of rhizosphere soil, and arbuscular mycorrhizal fungi (AMF) colonization, thus providing a scientific basis for soil quality improvement and vegetation restoration.

Exploration and Assessment on the Agronomic Requirement of Taverniera abyssinica A. Rich: A Critically Endangered Medicinal Plant of Ethiopia

We determined the distribution and abundance of Taverniera abyssinica A. Rich in the Shewa floristic region, Ethiopia. We also carried out a mesh-house experiment to know whether T. abyssinica is able to survive and grow in any soil. From the nine potential locations we made exploration, T. abyssinca populations were found only in the two, Lemen and Mojo. The abundance of mature individuals was estimated to be more than 600/hectare. The one-way ANOVA results indicated that soil does not have a significant (p&gt;0.05) effect on seedlings survival rate. However, it was found to have significant (p&lt;0.05) effects on seedlings growth, root nodulation, and root arbuscular mycorrhizal fungi colonization. Seedlings grown on the soil collected from Lemen and Mojo produced significantly (p&lt;0.05) more number of leaves and grew better than those grown on Addis Ababa (where the species was never reported to grow) soil. The root fresh weight of seedlings grown on Addis Ababa soil was found to be significantly (p&lt;0.05) and 38.89% and 54.17% lower than the root fresh weight of seedlings grown on Lemen and Mojo soils respectively. We report that T. abyssinica is N-fixer and arbuscular mycorrhizal. Seedlings grown on the Addis Ababa soil were not colonized by arbuscular mycorrhizal fungi while those grown on Lemen and Mojo soils were. Although the estimated abundance of mature T. abyssinica individuals was high, there is continued exploitation of the species and habitat loss is imminent. Therefore, integrated conservation program by way of ex situ conservation, in situ conservation, and cultivation should be implemented. Taverniera abyssinica could be cultivated in areas with leptosol and degraded vertisol soils with slightly acidic to basic pH. Arbuscular mycorrhizal fungi could play key role in future conservation and cultivation efforts of the species.

Soil Application of Sewage Sludge Treated with Clay Minerals or Biochar and Its Effect on Soil Properties and White Clover’s (Trifolium repens L.) Growth and Arbuscular Mycorrhizal Fungal Root Colonization

From the perspective of finding new, more environmental-friendly methods for the stabilization of sewage sludge to be used in agriculture, sludge treated with 15% bentonite, vermiculite or biochar was investigated as a soil amendment for white clover (Trifolium repens L.) growth, by means of a pot experiment. The sludge treatments, which were applied to two soils (an acid and an alkaline soil), in three replications, were the addition of 2% (≈80 Mg ha−1) treated sludge with the clay minerals or biochar, as well as limed or untreated (air-dried) sludge (for comparison reasons). Additional treatments with inorganic fertilization or neither organic nor inorganic fertilization (control) were also included. The application of 2% sludge to both soils significantly increased salinity compared to the control, which remained below harmful levels for sensitive crops, except for the case of untreated sludge. Furthermore, it significantly increased the soil-available macronutrients N, P and K, and micronutrients Cu, Zn (several times, especially in the case of untreated sludge) and B (up to three times) compared with the control. Moreover, the pH of the acid soil was improved, except for the case of limed sludge, where an undesirable pH increase close to 8.5 was observed. Addition of the treated sludge with the clay minerals or biochar and untreated sludge to the acid and alkaline soil significantly increased the aboveground biomass yield of white clover by 117–233% and 114–153%, respectively, compared to the control, whereas limed sludge had no effect. Plant nutrient uptake increased as well. In general, the effect of sludge on soil microbiological properties and arbuscular mycorrhizal fungal root colonization was ambiguous. It was concluded that 2% soil addition of sewage sludge treated with 15% bentonite, vermiculite or biochar could improve soil fertility and enhance plant growth; however, caution is needed with respect to potential risks of soil salinization or Zn and B phytotoxicities.

Soil microbial legacies influence plant survival and growth in mine reclamation.

Plants alter soil biological communities, generating ecosystem legacies that affect the performance of successive plants, influencing plant community assembly and successional trajectories. Yet, our understanding of how microbe-mediated soil legacies influence plant establishment is limited for primary successional systems and forest ecosystems, particularly for ectomycorrhizal plants. In a two-phase greenhouse experiment using primary successional mine reclamation materials with or without forest soil additions, we conditioned soil with an early successional shrub with low mycorrhizal dependence (willow, Salix scouleriana) and a later-successional ectomycorrhizal conifer (spruce, Picea engelmannii × glauca). The same plant species and later-successional plants (spruce and/or redcedar, Thuja plicata, a mid- to late-successional arbuscular mycorrhizal conifer) were grown as legacy-phase seedlings in conditioned soils and unconditioned control soils. Legacy effects were evaluated based on seedling survival and biomass, and the abundance and diversity of root fungal symbionts and pathogens. We found negative intraspecific (same-species) soil legacies for willow associated with pathogen accumulation, but neutral to positive intraspecific legacies in spruce associated with increased mycorrhizal fungal colonization and diversity. Our findings support research showing that soil legacy effects vary with plant nutrient acquisition strategy, with plants with low mycorrhizal dependence experiencing negative feedbacks and ectomycorrhizal plants experiencing positive feedbacks. Soil legacy effects of willow on next-stage successional species (spruce and redcedar) were negative, potentially due to allelopathy, while ectomycorrhizal spruce had neutral to negative legacy effects on arbuscular mycorrhizal redcedar, likely due to the trees not associating with compatible mycorrhizae. Thus, positive biological legacies may be limited to scenarios where mycorrhizal-dependent plants grow in soil containing legacies of compatible mycorrhizae. We found that soil legacies influenced plant performance in mine reclamation materials with and without forest soil additions, indicating that initial restoration actions may potentially exert long-term effects on plant community composition, even in primary successional soils with low microbial activity.

Suitability of selected legume (Vachellia spp.) tree species for forest restoration in the Central Ethiopian highlands

Abstract. Asmelash F, Getachew E. 2022. Suitability of selected legume (Vachellia spp.) tree species for forest restoration in the Central Ethiopian highlands. Nusantara Bioscience 14: 195-202. This study aimed to evaluate the comparative suitability of the legume tree species viz., Vachellia abyssinica (Hochst. ex. Benth.) Kyal. &amp; Boatwr., Vachellia etbaica (Schweinf.) Kyal. &amp; Boatwr., Vachellia lahai (Steud. &amp; Hochst. ex Benth.) Kyal. &amp; Boatwr., and Vachellia seyal (Delile) P.J.H.Hurter for restoring forests in Central Ethiopian highlands. The suitability of three accessions of V. seyal was also compared. The correlation between root nodule number and root Arbuscular Mycorrhizal Fungi (AMF) colonization (RC) and seedlings’ growth variables were computed, and the effect of seedling age on nodule number and RC was determined. Seedlings were grown for nine months on degraded local soil in a mesh house in central Ethiopia. We measured shoot height, shoot fresh weight, rooting depth, root nodule number, and RC in the third and ninth months. The one-way ANOVA results indicated that tree species and/or accession (for V. seyal) had a significant (p&lt;0.05) effect on all the measured variables except shoot height and rooting depth at the ninth month. Generally, nodule number and RC increased with seedling age. However, according to the independent t-test results, significant (p&lt;0.05) differences were recorded for V. abyssinica, with a 57.16% reduction in nodule number, and V. seyal accession-1, with a 418.52% increase in RC. The Spearman’s rank correlation results indicated that the correlation between nodule number and RC was weak and non-significant (p&gt;0.05) both in the third and ninth months. Based on the measured growth variables, nodule number (N-fixation potential), and RC, V. etbaica was the least suitable species for forest restoration in central Ethiopian highlands. The remaining species/accessions are comparably suitable. The V. abyssinica lost its comparative fitness with seedling age, maybe because it is a provenance far away from central Ethiopia. However, V. seyal accession-3, the furthest provenance, has performed much better. The legume trees of Ethiopia are less studied. Their role as environmental engineers could be better understood by knowing more about their root traits. Therefore, this study could motivate future research in this regard. Long-term experiments are required to consider more legume tree species and provenances in the future.

Anthocyanin pigmentation as a quantitative visual marker for arbuscular mycorrhizal fungal colonization of Medicago truncatula roots.

Visualization of root colonization by arbuscular mycorrhizal fungi (AMF) is the most elementary experiment in the field of mycorrhizal symbiosis. The most widely used approach for evaluating levels of AMF colonization is staining with trypan blue or ink, which is scored using the time-consuming grid intersection method. Here we demonstrate the use of an anthocyanin-based visual marker system for visualizing AMF colonization of Medicago truncatula roots. Expression of MtLAP1, a transcription factor which regulates the production of anthocyanins, from the AMF-induced Kunitz Protease Inhibitor 106 promoter, allowed the visualization of arbuscules in live plant tissues without microscopy or staining. This marker system allowed straightforward qualitative evaluation of the ram1, vpy and dmi3 AMF phenotypes using Agrobacterium rhizogenes hairy-root transformation. For the strigolactone biosynthesis mutant carotenoid cleavage dioxygenase 8a and a novel mutant scooby, which show quantitative AMF symbiotic phenotypes, the amount of anthocyanins in the roots estimated by spectrophotometry correlated very well with colonization levels estimated by staining and scoring using the grid intersection method. The LAP1-based marker system therefore provides a highly efficient approach for mutant screening and monitoring of AMF colonization in live tissues by eye, or for quantitative assessment using a simple and quick photometric assay.

Influence of Aged Wood Biochar on Mycorrhizal Colonisation, Growth and Leaf Gas Exchange of Wheat and Clover with Different Water Regimes

Water scarcity is one of the most limiting environmental factors for crop productivity globally, where biochar application can play a role. To test this, a glasshouse experiment was conducted with aged biochar application with water regimes on arbuscular mycorrhizal fungi colonisation, plant growth, phosphorus nutrition and leaf gas exchange in wheat and subterranean clover. Six treatment combinations (two biochar levels of 0 and 6 t ha−1; and three water regimes of well-watered, water-stressed and watering intermittently) were arranged in factorial completely randomized block design with three replications. The wheat and subterranean clover were grown and harvested 14, 24, 34 and 44 days after sowing. In this study, aged biochar had no significant effect on plant growth for both wheat and clover, regardless of water regimes. Shoot and root dry weights increased in well-watered conditions compared to water-stressed conditions. Root length and colonised root length increased with biochar addition for wheat, mostly in well-watered treatment. Phosphorus uptake increased in biochar treatment, and the effect was higher in well-watered conditions. Leaf photosynthetic rate (PN), stomatal conductance (gs), intercellular CO2 concentration (Ci) and transpiration rate (E) increased with biochar addition. For clover, the effect was higher in water-stressed than well-watered conditions. These results suggest that biochar may enhance water availability to clover plants in water-stressed conditions. However, the positive effect of biochar on plant growth under water-stressed conditions, varied with plant species, needs to be explored further in a range of crop species and biochar sources in addition to the material used in the current investigation.

Mixed conifer-broadleaf trees on arbuscular mycorrhizal and ectomycorrhizal communities in rhizosphere soil of different plantation stands in the temperate zone, Northeast China.

In comparison with ectomycorrhizal (EM) tree species, arbuscular mycorrhizal (AM) trees have different litter quality and nitrogen cycle modes, which may affect mycorrhizal colonization and the community composition and diversity. However, available studies addressing the mycorrhizal fungal colonization rate, diversity and community composition in mixed forest stands composed of AM and EM trees are rare. In the present study, we assessed litter quality, soil physicochemical properties and correlated them with mycorrhizal community characteristics in rhizosphere soils of monoculture and mixture plantation stands of AM tree species (Fraxinus mandschurica Rupr.) and EM tree species (Larix gmelinii Rupr., Picea koraiensis Nakai) in Northeast China. We hypothesized that (1) the effect of mixture pattern on mycorrhizal colonization rate and diversity would change with tree species, (2) the effect of mixture pattern on mycorrhizal community composition would be less pronounced in comparison with that of tree species. We found that mixture did not change AMF colonization rate regardless of mixture identity, whereas mixture and tree species exerted significant effects on EMF colonization rate. For AMF community, both M-AS (Fraxinus mandschurica Rupr. and Picea koraiensis Nakai) and M-AL (Fraxinus mandschurica Rupr. and Larix gmelinii Rupr.) mixtures significantly increased Pielou index and Simpson index, whereas only M-AS significantly increased Sobs. For EMF community, mixture significantly affected examined diversity indices except for Chao1. Mixture significantly shifted AMF and EMF community, and the magnitude was tree species dependent. The dominant genera in AMF and EMF communities in plantation stands were Glomus and Tomentella, respectively. The EnvFit analysis showed that the determinant factors of EMF community are soil moisture, pH, nitrate nitrogen content, dissolved organic nitrogen content, soil organic matter content, soil organic carbon/total nitrogen and litter carbon/total nitrogen. In conclusion, mixed conifer-broadleaf trees significantly changed soil physicochemical properties, litter quality as well as mycorrhizal fungi community diversity and composition.

Nutrient management and bioaugmentation interactively shape plant–microbe interactions inMiscanthus × giganteus

AbstractMarginal lands are an untapped source of agricultural potential, particularly regarding high‐yielding, low‐input bioenergy crops likeMiscanthus × giganteus(Miscanthus).Miscanthusis of specific interest because it can be productive and sequester carbon in soil even under the stressful conditions present on some degraded lands. A key component of these abilities is the interaction ofMiscanthuswith its soil and root microbiome. Microbial functions depend on the nutrient status of soil, and hence are sensitive to fertilization regimes. Nevertheless, little is known about how fertilization strategies affect the partnership ofMiscanthuswith its microbial associates. Here, we tested the individual and interactive effects of nutrient addition and bioaugmentation (i.e., the addition of microbial communities) onMiscanthusperformance and microbiome function in marginal soil. We found that the effect of nutrient addition onMiscanthusbiomass yield depended on nutrient addition type (i.e., organic or inorganic nutrients) and whether bioaugmentation was also applied. Some microbial functions, like free‐living nitrogen fixation and carbon use efficiency, were sensitive to nutrient addition depending on whether bioaugmentation was also applied. On the other hand, arbuscular mycorrhizal fungus colonization of roots decreased with fertilization regardless of bioaugmentation. These results imply that managing microbial communities may regulate the effect of nutrient addition on plant–microbe interactions that in part determine system productivity and environmental impact.

Aphids and Mycorrhizal Fungi Shape Maternal Effects in Senecio vulgaris.

Plant performance in any one generation is affected not only by the prevailing environmental conditions, but also by the conditions experienced by the parental generation of those plants. These maternal effects have been recorded in a many plant species, but the influence of external biotic (as opposed to abiotic) factors on shaping maternal effects have been rarely examined. Furthermore, almost all previous studies have taken place over one plant generation, rather than across multiple generations. Here, we studied the influence of insect herbivory and arbuscular mycorrhizal (AM) fungal colonisation on the shaping of maternal effects in the annual forb Senecio vulgaris. We grew plants with and without aphids (Myzus persicae) and AM fungi (hereafter termed ‘induction events’) over four successive generations, wherein seeds from plants in any one treatment were used to grow plants of the same treatment in the next generation, all in identical environmental conditions. We found strong evidence of maternal effects in the second plant generation, i.e., after one induction event. These plants took longer to germinate, flowered in a shorter time, produced lighter seeds and were shorter and of lower biomass than their parents. Aphid attack tended to enhance these effects, whereas AM fungi had little influence. However, thereafter there was a gradual recovery in these parameters, so that plants experiencing three inductions showed similar life history parameters to those in the original generation. We conclude that experiments investigating maternal effects need to be performed over multiple plant generations and that biotic factors such as insects and mycorrhizas must also be taken into account, along with abiotic factors, such as nutrient and water availability.

Rhizobium leguminosarum symbiovar viciae strains are natural wheat endophytes that can stimulate root development.

Although rhizobia that establish a nitrogen-fixing symbiosis with legumes are also known to promote growth in non-legumes, studies on rhizobial associations with wheat roots are scarce. We searched for Rhizobium leguminosarum symbiovar viciae (Rlv) strains naturally competent to endophytically colonize wheat roots. We isolated 20 strains from surface-sterilized wheat roots and found a low diversity of Rlv compared to that observed in the Rlv species complex. We tested the ability of a subset of these Rlv for wheat root colonization when co-inoculated with other Rlv. Only a few strains, including those isolated from wheat roots, and one strain isolated from pea nodules, were efficient in colonizing roots in co-inoculation conditions, while all the strains tested in single strain inoculation conditions were found to colonize the surface and interior of roots. Furthermore, Rlv strains isolated from wheat roots were able to stimulate root development and early arbuscular mycorrhizal fungi colonization. These responses were strain and host genotype dependent. Our results suggest that wheat can be an alternative host for Rlv; nevertheless, there is a strong competition between Rlv strains for wheat root colonization. In addition, we showed that Rlv are endophytic wheat root bacteria with potential ability to modify wheat development.

Biostimulatory Response of Easily Extractable Glomalin-Related Soil Protein on Soil Fertility Mediated Changes in Fruit Quality of Citrus

Arbuscular mycorrhizal fungi secrete glomalin deposited into the soil as glomalin-related soil protein (GRSP), which possess multiple utility to benefit both soil as well as plant health. The present study aimed to assess the effects of the foliar application of an easily extractable GRSP (EE-GRSP) on the fruit quality, soil nutrients, and soil structural changes in three important citrus varieties (Satsuma mandarin Oita 4, Newhall navel orange, and Cocktail grapefruit). The exogenous EE-GRSP significantly elevated root mycorrhizal fungal colonization and soil hyphal length in Newhall and Oita 4 varieties, but without any such response in Cocktail grapefruit variety. The foliar spray of the EE-GRSP improved different external (e.g., pericarp, sarcocarp, and single fruit weight) and internal (e.g., soluble solids, titratable acids, and sugar contents) qualities of fruits to varying magnitudes, depending on citrus variety, with a more prominent effect on Cocktail grapefruit. EE-GRSP-treated fruits of Newhall and Oita 4 were more suitable for processing than non-treated control because of a low fruit hardness. However, no significant effect of the EE-GRSP was observed on the internal quality parameters of Newhall. EE-GRSP-treated citrus trees represented higher soil available nutrients over control, to some extent, especially on Oita 4. The foliar application of the EE-GRSP also increased various GRSP fractions to varying proportions and improved the distribution of water-stable aggregates in the size fraction of 0.25–2 mm, thereby increasing the mean weight diameter, particularly in Newhall and Cocktail grapefruit varieties. These observations provided clues about the stimulatory role of the EE-GRSP through soil structure and nutrient pool-mediated improvements in fruit quality.

Orchid mycorrhizal fungi and ascomycetous fungi in epiphytic Vanda falcata roots occupy different niches during growth and development.

Epiphytic orchids are commonly found in exposed environments, which plausibly lead to different root fungal community structures from terrestrial orchids. Until recently, few studies have been conducted to show the fungal community structure during the growth of a photosynthetic and epiphytic orchid in its natural growing site. In this study, the Vanda falcata (commonly known as Neofinetia falcata), one of Japan's ornamental orchids, was used to characterize the fungal community structure at different developmental stages. Amplicon sequencing analysis showed that all development stages contain a similar fungal community: Ascomycota dominate half of the community while one-third of the community belongs to Basidiomycota. Rhizoctonia-like fungi, a polyphyletic basidiomycetous fungal group forming mycorrhizas in many orchids, exist even in a smaller portion (around one-quarter) compared to other Basidiomycota members. While ascomycetous fungi exhibit pathogenicity, two Ceratobasidium strains isolated from young and adult plants could initiate seed germination in vitro. It was also found that the colonization of mycorrhizal fungi was concentrated in a part of the root where it directly attaches to the phorophyte bark, while ascomycetous fungi were distributed in the velamen but never colonized cortical cells. Additionally, the root parts attached to the bark have denser exodermal passage cells, and these cells were only colonized by mycorrhizal fungi that further penetrated into the cortical area. Therefore, we confirmed a process that physical regulation of fungal entry to partition the ascomycetes and mycorrhizal fungi results in the balanced mycorrhizal symbiosis in this orchid.