Fungal Root Research Articles

Synergy of biochar and biofertilizers to improve bell pepper fruit biochemical quality with increased soil carbon, Azospirillum population and mycorrhization

Sustainable crop production depends more on maintaining good soil health. Biochar significantly improves the soil's physical, chemical, biological and nutritional properties while aiding in carbon sequestration. Biofertilizers, which promote nutrient cycling, heavy metal chelation and the production of plant growth substances are crucial to sustainable agriculture. While the individual benefits of biochar and biofertilizers are well-documented, their combined effect in vegetable farming is less studied. The synergistic application of biochar and biofertilizers can improve soil health and nutrient dynamics, enhancing crops' biochemical properties. In a pot culture experiment, California Wonder bell peppers were treated with biochar and biofertilizers, including Azospirillum, Vesicular Arbuscular Mycorrhiza (VAM), and Serendipita indica. These were applied directly to the soil during transplanting. Results showed that plants treated with biochar + Azospirillum had higher flavonoid, total soluble protein and ascorbic acid content. Biochar + VAM treatment resulted in higher polyphenol content, total dry matter and root volume. S. indica reduced the vegetative period and induced earlier flowering. Biochar also increased soil organic carbon (2.89 %), improved the rhizosphere Azospirillum population and enhanced fungal endophytes' root colonization. Combining biochar and biofertilizer improves bell pepper plant performance, enhances biochemical parameters and improves soil health.

Fire-associated microbial shifts in soils of western conifer forests with Armillaria root disease.

With its influence on ecosystem processes and functions, the soil microbiome can interact with soilborne pathogens to facilitate or suppress plant disease development. These forest ecosystems are experiencing increased wildfire frequency and burn severity that could influence the fungal root pathogen, Armillaria solidipes, and interactions with the soil microbiome. We examined changes to the soil microbiome following three levels of burn severity, and examined how these changes correspond with A. solidipes, and a putatively beneficial species, A. altimontana. Following severe burn, there was a decreased relative abundance of ectomycorrhizal fungi associated A. altimontana. A. solidipes and associated microbial taxa dominated following high-severity burns, suggesting that severe fires provide suitable environmental conditions for these species. Our results suggest that shifts in the soil microbiome and an associated increase in the activity of A. solidipes following high-severity burns in conifer forests may result in priority areas for monitoring and proactive management of Armillaria root disease.

Anthoxanthum odoratum (Poaceae) as a bioindicator of arbuscular mycorrhizal fungi abundance prior to native revegetation on agricultural soils

ABSTRACT Mycorrhizal fungi, which form symbiotic associations with plant roots, can be critical to the successful establishment of plants. Up to 90% of land plant species form associations with arbuscular mycorrhizal fungi (AMF), which develop fungal structures inside host roots. Common site preparation practices prior to ecological restoration, such as soil disturbance and herbicide application, may lessen the availability of AMF, potentially jeopardising plant establishment. However, the state of the soil microbiome can be difficult to assess directly. This study examines the potential of the widespread pasture grass Anthoxanthum odoratum (Sweet Vernal, Poaceae) as a bioassay of AMF abundance in soils. We present a rapid clearing and staining protocol for visualising AMF structures in the roots of A. odoratum. We then compare % colonisation on roots of naturally established A. odoratum plants collected from undisturbed pastures vs. sprayed and tilled pastures. Roots of A. odoratum collected from sprayed and tilled pastures showed a significantly lower average AMF colonisation at 43.8% compared with 57.6% on roots from undisturbed pastures. However, all plants sourced from the sprayed and tilled pastures still showed some AMF structures. Our results strongly suggest that A. odoratum can be a useful bioindicator of AMF presence and abundance in soils, offering a promising tool for seed-based restoration on retired agricultural land.

TML1 AND TML2 SYNERGISTICALLY REGULATE NODULATION AND AFFECT ARBUSCULAR MYCORRHIZA IN MEDICAGO TRUNCATULA.

Two symbiotic processes, nodulation and arbuscular mycorrhiza, are primarily controlled by the plant's need for nitrogen (N) and phosphorus (P), respectively. Autoregulation of Nodulation (AON) and Autoregulation of Mycorrhization (AOM) both negatively regulate their respective processes and share multiple components - plants that make too many nodules usually have higher AM fungal root colonization. The protein TML (TOO MUCH LOVE) was shown to function in roots to maintain susceptibly to rhizobial infection under low N conditions and control nodule number through AON in Lotus japonicus . M. truncatula has two sequence homologs: Mt TML1 and Mt TML2. We report the generation of stable single and double mutants harboring multiple allelic variations in MtTML1 and MtTML2 using CRISPR-Cas9 targeted mutagenesis and screening of a transposon mutagenesis library. Plants containing single mutations in Mt TML1 or Mt TML2 produced 2-3 times the nodules of wild-type plants whereas plants containing mutations in both genes displayed a synergistic effect, forming 20x more nodules compared to wild type plants. Examination of expression and heterozygote effects suggest genetic compensation may play a role in the observed synergy. Plants with mutations in both TMLs only showed mild increases in AM fungal root colonization at later timepoints in our experiments, suggesting these genes may also play a minor role in AM symbiosis regulation. The mutants created will be useful tools to dissect the mechanism of synergistic action of Mt TML1 and Mt TML2 in M. truncatula symbiosis with beneficial microbes.

Conserved perception of host and non-host signals via the a-pheromone receptor Ste3 in Colletotrichum graminicola.

Understanding the interactions between fungal plant pathogens and host roots is crucial for developing effective disease management strategies. This study investigates the molecular mechanisms underpinning the chemotropic responses of the maize anthracnose fungus Colletotrichum graminicola to maize root exudates. Combining the generation of a deletion mutant with monitoring of disease symptom development and detailed analysis of chemotropic growth using a 3D-printed device, we identify the 7-transmembrane G-protein coupled receptor (GPCR) CgSte3 as a key player in sensing both plant-derived class III peroxidases and diterpenoids. Activation of CgSte3 initiates signaling through CgSo, a homolog to the Cell Wall Integrity Mitogen-Activated Protein Kinase (CWI MAPK) pathway scaffold protein identified in other filamentous fungi, facilitating the pathogen's growth towards plant defense molecules. The NADPH oxidase CgNox2 is crucial for peroxidase sensing but not for diterpenoid detection. These findings reveal that CgSte3 and CWI MAPK pathways are central to C. graminicola's ability to hijack plant defense signals, highlighting potential targets for controlling maize anthracnose.

Hongos micorrícicos arbusculares y endófitos septados oscuros en raíces de helechos nativos de uso medicinal de las Sierras Centrales de San Luis, centro-oeste de Argentina

Background and aims: The plant´s roots and fungi establish symbiotic endophytic associations that can coexist: arbuscular mycorrhizae and “dark septate” endophytes. Ferns and lycophytes are vascular plants that can form both fungal symbioses. The objective of this work is to detect the presence of fungal root endosymbiosis, know the mycorrhizal status, and analyze the type of colonizationin in native and ornamental ferns from the “Sierras Centrales de San Luis”. M&M: In three sites in the “Sierras Centrales de San Luis”, 9-10 individuals of each studied species were collected, their roots were processed to study these fungal. Characterization and quantification of their root colonization was evaluated. Results: In the three sites studied, Anemia tomentosa var. anthriscifolia formed mycorrhizal associations, the colonization presented abundant hyphal and arbusculated intracellular coils. In addition, glomoid spores of mycorrhizal fungi were found in their rhizospheres. Dark septate endophytes were absent in all species analyzed, none hyphae neither microsclerotia were recorded in roots of these native medicinal ferns. Conclusions: The mycorrhizal status of Anemia tomentosa var. anthriscifolia, one of the five medicinal fern species from the Sierras Centrales de San Luis studied, is presented. The arbuscular mycorrhizal colonization is described and quantified, and its type is determined as Paris. None of the fern species studied was associated with dark septate endophytes.

Mycorrhizal fungi in Annona muricata L. rhizosphere in two agricultural production systems in Nayarit, Mexico

Mycorrhizal symbiosis is the association between the roots of several plant species and soil fungi of the Glomeromycotina subphylum. This symbiosis plays a crucial role in plant growth, development, and defense; therefore, understanding the abundance and diversity of arbuscular mycorrhizal fungi (AMF) species associated with plants of economic importance is of utmost concern. The objective of this study was to compare the abundance and composition of AMF communities in two Annona muricata cultivation sites, and in propagated soil, under different agricultural practices. The first cultivation site is under technified management with periodic fertilization and irrigation (TS), while the second cultivation site is under agroecological management without fertilization, irrigation, and with the presence of livestock (non-technified site; NTS). The extracted spores from the collected samples and the trap cultures were taxonomically identified based on their morphology. 13 species associated with A. muricata belonging to seven genera were identified in soil samples. The most abundant species were Funneliformis geosporum, Acaulospora kentinensis and Rhizophagus intraradices with a relative abundance of 45.9, 19.0 and 15.8%, respectively. In the propagation substrates, only 69% of the AMF species found in the field were identified. 11 species of AMF were found in the site with non-technified agronomic management, while only five species were found in the technified site.

A fungal endophyte increases plant resilience to low nutrient availabilities: a case of Fe acquisition in legumes.

Microbial inocula are considered a promising and effective alternative solution to the use of chemical fertilizers to support plant growth and productivity since they play a key role in the availability and uptake of nutrients. Here, the effect of a beneficial of a fungal root endophyte, Fusarium solani strain K (FsK), on nutrient acquisition efficiency of the legume Lotus japonicus was studied, and putative mode-of-action of the endophyte at a molecular level was determined. Plant colonization with the endophyte resulted in increased shoot and root fresh weight under Fe deficiency compared to control nutrient conditions. Plant inoculation with FsK was associated with a significant increase in macro- and micronutrient concentration in leaves at an early stage of endophyte inoculation and a replenishment of Fe content under prolonged iron starvation. The mechanistic basis of the plant growth promotion capabilities of the endophyte is exerted at the transcriptional level since we recorded changes in the expression levels of genes related to iron uptake in FsK-colonized plants under stress conditions compared to uninoculated plants. In addition, the observed changes in the ethylene biosynthesis-related genes suggest a possible implication of ethylene in the mode of action used by FsK to enhance plant response to nutrient stress conditions. Finally, we demonstrated that the endophyte possesses a reductive high-affinity Fe uptake system and identified a ferric reductase that was induced in planta under Fe deficiency conditions, indicating that this fungal Fe homeostasis mechanism may result in a benefit in nutrient acquisition for the plant as well.

Mycorrhizal inoculation alters rhizosphere fungal community and root morphology to improve drought tolerance of resource-acquisitive but not resource-conservative Quercus species

The mutualistic association between mycorrhizal fungi and plants is well known to improve plant drought resistance. However, the influence of external inoculants on the interactions between soil microbial communities and plant functional traits and how these interactions improve plant drought tolerance under drought stress remain unclear. We conducted a pot inoculation experiment to assess the effect of two inoculated fungal strains (Clitopolius hobsonii NL-19 and C. sp. HSL-YX-7-A) on morphological traits and rhizosphere fungal communities of eight Quercus species seedlings. Plant and soil fungal traits were measured 2 months after inoculation and then underwent a short-term (33 days) drought rewetting treatment. The biomass of all tree species reduced significantly under drought stress followed by rewetting (drought treatment) compared with that in the control treatment (well-watered); however, inoculated mycorrhizal fungi (drought + mycorrhizal inoculation treatment) increased the biomass of five Quercus species (growth-promoting species (GPS) exhibiting a significant increase in biomass under mycorrhizal inoculation treatment than under the uninoculated treatment during drought stress, characterized by a resource-use acquisitive strategy) and had no effect on the other three Quercus species (non-growth-promoting species (NGPS) exhibiting no significant difference in biomass between mycorrhizal inoculation and uninoculated treatments during drought stress, characterized by a resource-use conservative strategy). The leaf traits of the two species groups (GPS and NGPS) varied little among the three treatments. The values of most root traits (e.g., specific root length and root length) of GPS increased significantly under drought and drought + mycorrhizal inoculation treatments compared with those in the control, whereas relatively minor variations were observed in NGPS among the three treatments. The fungal community composition and functional groups (primary trophic modes) in the drought treatment were altered for NGPS but not for GPS; additionally, they changed in the drought + mycorrhizal inoculation treatment for GPS but not for NGPS when compared with those in the drought treatment. Furthermore, the drought tolerance of GPS was improved directly by fungal functional groups (i.e., Pathotroph-Saprotroph- Symbiotrophs proliferation) and indirectly by root traits (e.g., increased specific root length), whereas NGPS may adapt to drought stress through other pathways (e.g., physiological and biochemical regulation). Overall, drought and mycorrhizal inoculation affected the root traits and fungal community structure of Quercus seedlings, which rely on plant resource-use strategies. Our results provide new insights into the relationship between plant resource-use strategies and soil microbes for improving plant performance under drought stress.

Intraspecific diploidization of a halophyte root fungus drives heterosis

How organisms respond to environmental stress is a key topic in evolutionary biology. This study focused on the genomic evolution of Laburnicola rhizohalophila, a dark-septate endophytic fungus from roots of a halophyte. Chromosome-level assemblies were generated from five representative isolates from structured subpopulations. The data revealed significant genomic plasticity resulting from chromosomal polymorphisms created by fusion and fission events, known as dysploidy. Analyses of genomic features, phylogenomics, and macrosynteny have provided clear evidence for the origin of intraspecific diploid-like hybrids. Notably, one diploid phenotype stood out as an outlier and exhibited a conditional fitness advantage when exposed to a range of abiotic stresses compared with its parents. By comparing the gene expression patterns in each hybrid parent triad under the four growth conditions, the mechanisms underlying growth vigor were corroborated through an analysis of transgressively upregulated genes enriched in membrane glycerolipid biosynthesis and transmembrane transporter activity. In vitro assays suggested increased membrane integrity and lipid accumulation, as well as decreased malondialdehyde production under optimal salt conditions (0.3 M NaCl) in the hybrid. These attributes have been implicated in salinity tolerance. This study supports the notion that hybridization-induced genome doubling leads to the emergence of phenotypic innovations in an extremophilic endophyte.

Composition and Assembly of the Endophytic Fungal Community of Alpine Rhododendron Hosts Along Elevation Gradients

Alpine mountain ecosystems are highly sensitive to global warming and threatened by climate change. Elevation gradients are frequently used as space-for-time surrogates for climate change in mountain ecosystems due to their correspondence with climate variables over short geographic distances. However, the distribution patterns of fungal communities in perennial woody plants along elevations in this ecosystem and how elevation and host identity affect them remain unclear. In this work, we used Illumina MiSeq DNA sequencing technology to study the rhizosphere soil fungal community and root and leaf endophytic fungal community assembly of alpine Rhododendron communities across a 3,100- to 4,100-m elevation gradient on Jiaozi Mountain, Southwest China. The abundance and diversity of fungi in different compartments varied among elevation gradients and hosts. The fungal community assembly in each compartment was mainly influenced by stochastic processes. However, the contribution of homogeneous selection, dispersal limitation, and drift varied among host identities and elevations in different compartments. The diversity and composition of rhizosphere soil, root, and leaf guilds and the overall abundance of fungal functional groups tracked the Rhododendron host identity more closely than elevation did. Most indicator taxa of fungi were putative saprotrophs. These findings will improve our understanding of the maintenance of alpine Rhododendron communities along elevation gradients. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Arbuscular mycorrhizal fungi attenuate negative impact of drought on soil functions.

Although positive effects of arbuscular mycorrhizal (AM) fungi on plant performance under drought have been well documented, how AM fungi regulate soil functions and multifunctionality requires further investigation. In this study, we first performed a meta-analysis to test the potential role of AM fungi in maintaining soil functions under drought. Then, we conducted a greenhouse experiment, using a pair of hyphal ingrowth cores to spatially separate the growth of AM fungal hyphae and plant roots, to further investigate the effects of AM fungi on soil multifunctionality and its resistance against drought. Our meta-analysis showed that AM fungi promote multiple soil functions, including soil aggregation, microbial biomass and activities of soil enzymes related to nutrient cycling. The greenhouse experiment further demonstrated that AM fungi attenuate the negative impact of drought on these soil functions and thus multifunctionality, therefore, increasing their resistance against drought. Moreover, this buffering effect of AM fungi persists across different frequencies of water supply and plant species. These findings highlight the unique role of AM fungi in maintaining multiple soil functions by mitigating the negative impact of drought. Our study highlights the importance of AM fungi as a nature-based solution to sustaining multiple soil functions in a world where drought events are intensifying.

Wheat varieties show consistent differences in root colonization by mycorrhiza across a European pedoclimatic gradient

AbstractArbuscular mycorrhizal (AM) fungi form mutualistic relationships with the majority of land plants and are an important part of the soil microbial community in natural and agricultural ecosystems. These fungi promote water and nutrient acquisition by their host plant and regulate the allocation of photosynthetic carbon to soil. Both crop variety and environment affect naturally occurring mycorrhizal abundance in roots, but the relative importance of those factors for mycorrhization is largely unknown. In a field study covering a large pedoclimatic gradient across four European sites, we (i) compared the abundance of AM fungi in the roots of 10 modern winter wheat (Triticum aestivum L.) varieties, (ii) evaluated the relative importance of variety and site for the variability in root colonization by AM fungi and (iii) tested the relationship between mycorrhizal abundance and grain yield. Root colonization by arbuscules and hyphae ranged from 10% to 59% and 20% to 91%, respectively, across all samples and varied by 8% and 18%, respectively, among varieties when averaged across sites. Variance decomposition analysis revealed a 10 times higher importance of site than variety for AM fungal root colonization. Specifically, we found the highest mycorrhizal abundance on the site with the most arid conditions and the lowest on the sites with low soil pH and high nutrient availability. Despite the low variability in mycorrhizal abundance among varieties, there were significant differences in both arbuscular and hyphal root colonization. However, this did not translate into an increase in yield as no significant relationships between mycorrhizal abundance at flowering and grain yield were detected. The consistent differences between wheat varieties in root colonization by AM fungi across European field sites underline that genetic drivers of mycorrhization are to some extent independent of the site. This highlights the relevance of breeding practices to shape a wheat variety's capacity for mycorrhizal symbiosis across a range of environmental conditions.

In vitro potential control of white root fungus in nutmeg (Myristica fragrans) based on botanical fungicide of coconut shell liquid smoke

Nutmeg (Myristica fragrans) is spice plant which has an important position as a source of essential oils. An important problem in nutmeg cultivation is white root disease which can reduce nutmeg production. Until now, white root disease is still a problem in nutmeg plantations in South Aceh district. One of the solution that can be explored as an environmentally friendly control measure is to use the botanical fungicide coconut shell liquid smoke. This research aims to determine the potential of coconut shell liquid smoke in controlling white root fungus in vitro. The research was carried out in the Plant Protection Laboratory, Faculty of Agriculture, Teuku Umar University. The research design used was a non-factorial Completely Randomized Design (CRD) consisting of nine levels of liquid smoke concentration treatment; control 0%, 0.5%, 1%, 2%, 3%, 4%, 6%, 8%, 10%. Each treatment involved three replications. Data analysis was carried out using analysis of variance and Tukey's least significant difference (HSD) test.The research stages carried out consisted of producing coconut shell liquid smoke, isolating white root fungus from the roots of infected nutmeg plants and potential assay of liquid smoke in inhibiting the growth of white root fungus. In vitro bioassay result showed that liquid smoke treatment 3% was the best treatment because it was the lowest concentration causing total inhibition of the growth of white root fungus.

First report of root rot of strawberry caused by Fusarium cugenangense, a member of the F. oxysporum species complex, in Tennessee, USA.

Strawberry (Fragaria × ananassa Duch) in Tennessee is cultivated on plastic mulched beds annually, and production is limited primarily by multiple oomycete and fungal root rot pathogens that result in reduced vigor and black root rot disease symptoms. In early June 2018, plants (cv. Chandler) with reduced shoot vigor and size, and black, necrotic stunted roots were collected from Rhea County, TN. Roots and crowns of 10 plants were cut into 1-3 cm pieces and surface sterilized with 0.6% NaOCl, followed by 70% ethanol for 1 min each, and plated on water agar. White mycelia produced after 3 days were transferred to potato dextrose agar amended with 10 mg/liter rifampicin. After 10 days, fungal colonies were light purple on the surface and dark purple on the colony underside, later developing blue-black pigmentation on the underside. Microconidia on carnation leaf agar were ovoid to ellipsoid, aseptate or septate and 8.0 to 24.2 (13.7) × 3.0 to 4.5 (3.8) μm in size, macroconidia were 3 to 5 septate and falcate to almost straight and 33.7 to 52.8 (44.4) × 4.0 to 5.5 (4.9) μm in size (n=80); both conidia were produced on monophialides. Chlamydospores were globose and subglobose, formed terminally and intercalary on aerial, submerged, and surface mycelium, singly or in pairs and were abundantly produced in sucrose broth and on synthetic nutrient-poor agar (SNA) (diam. 7.6 μm). Morphology was consistent with Fusarium oxysporum (Leslie and Summerell, 2006) and F. cugenangense, a member of the F. oxysporum species complex, as described by Maryani et al. (2019). Fungal mycelia were used for PCR (Phire Plant Direct PCR Master Mix, Thermo Scientific, CA) and the translational elongation factor 1-α (EF1α) region was amplified with primers EF-1/EF-2 (O'Donnell et al., 1998), internal transcribed spacer (ITS) regions amplified with primers ITS1/ITS2 (White et al. 1990), and the RNA polymerase second largest subunit region (RPB2) with primer pairs 5f2/7cr and 7cf/11ar (O'Donnell et al., 2022). PCR products of isolate SC5 were sequenced, and sequences compared to all sequences in the FUSARIOID-ID database using polyphasic identification (Crous et al., 2021) with EF1α (GenBank Accession No. ON703236) and RPB2 (OR472390) sequences. The highest similarity (100%) was with isolates of F. cugenangense, including ex-type isolate InaCC F984 (99.94% similarity) (Maryani et al., 2019). F. cugenangense is closely related to F. callistephi and F. elaeidis, but both species lack chlamydospores, and F. elaeidis has polyphialides (Lombard et al, 2019). To satisfy Koch's postulates, healthy rooted strawberry plants produced in soilless media were transplanted into 4 plastic pots (1.2-liter) containing 5% (w/v) fungal inoculum (grown on barley grain) and mixed into the top 5-cm of peat-based soilless medium. Pots were incubated at 25°C and 50% RH in a growth chamber. Four pots without inoculum served as controls. The trial was repeated. Within 8 weeks, all inoculated plants had low vigor, with necrotic and stunted roots. Root sections of control and inoculated plants were plated, and the pathogen was re-isolated from diseased roots of all inoculated plants only and confirmed as F. cugenangense based on morphology and sequence analysis. To our knowledge, this is the first report of F. cugenangense, or any member of the F. oxysporum species complex, causing root rot of strawberry in Tennessee and could be an important component of the production-limiting black root rot disease complex of strawberry.

Assessing the biodiversity of rhizosphere and endophytic fungi in Knoxia valerianoides under continuous cropping conditions

BackgroundRhizosphere and endophytic fungi play important roles in plant health and crop productivity. However, their community dynamics during the continuous cropping of Knoxia valerianoides have rarely been reported. K. valerianoides is a perennial herb of the family Rubiaceae and has been used in herbal medicines for ages. Here, we used high-throughput sequencing technology Illumina MiSeq to study the structural and functional dynamics of the rhizosphere and endophytic fungi of K. valerianoides.ResultsThe findings indicate that continuous planting has led to an increase in the richness and diversity of rhizosphere fungi, while concomitantly resulting in a decrease in the richness and diversity of root fungi. The diversity of endophytic fungal communities in roots was lower than that of the rhizosphere fungi. Ascomycota and Basidiomycota were the dominant phyla detected during the continuous cropping of K. valerianoides. In addition, we found that root rot directly affected the structure and diversity of fungal communities in the rhizosphere and the roots of K. valerianoides. Consequently, both the rhizosphere and endophyte fungal communities of root rot-infected plants showed higher richness than the healthy plants. The relative abundance of Fusarium in two and three years old root rot-infected plants was significantly higher than the control, indicating that continuous planting negatively affected the health of K. valerianoides plants. Decision Curve Analysis showed that soil pH, organic matter (OM), available K, total K, soil sucrase (S_SC), soil catalase (S_CAT), and soil cellulase (S_CL) were significantly related (p < 0.05) to the fungal community dynamics.ConclusionsThe diversity of fungal species in the rhizosphere and root of K. valerianoides was reported for the first time. The fungal diversity of rhizosphere soil was higher than that of root endophytic fungi. The fungal diversity of root rot plants was higher than that of healthy plants. Soil pH, OM, available K, total K, S_CAT, S_SC, and S_CL were significantly related to the fungal diversity. The occurrence of root rot had an effect on the community structure and diversity of rhizosphere and root endophytic fungi.

Fast, Easy, and Comprehensive Techniques for Microscopic Observations of Fungal and Oomycete Organisms Inside the Roots of Herbaceous and Woody Plants.

The roots of herbaceous and woody plants growing in soil are complex structures that are affected by both natural and artificial fungal colonization to various extents. To obtain comprehensive information about the overall distribution of fungi or oomycetes inside a plant root system, rapid, effective, and reliable screening methods are required. To observe both fine roots, i.e., a common site for penetration of fungi and oomycetes, and mature roots, different techniques are required to overcome visual barriers, such as root browning or tissue thickening. In our protocol, we propose using fast, cost-effective, and non-harmful methods to localize fungal or oomycete structures inside plant roots. Root staining with a fluorescent dye provides a quick initial indication of the presence of fungal structures on the root surfaces. The protocol is followed by clearing and staining steps, resulting in a deeper insight into the root tissue positioning, abundance, and characteristic morphological/reproductive features of fungal or oomycete organisms. If required, the stained samples can be prepared by using freeze-drying for further observations, including advanced microscopic techniques. Key features • The protocol enhances tissue-clearing techniques employing KOH or NaOH and is applicable to a broad range of roots from different plant species. • Hydroxides are mixed with hydrogen peroxide to obtain an efficient bleaching solution, which effectively clears roots without causing significant tissue damage. • The protocol could also be used for staining of fungi or oomycetes localized both on the root surface or inside the root tissues. • Simple combination of non-fluorescent methyl blue and fluorescent solophenyl flavine dyes allows the observation of fungal organisms in both brightfield and fluorescence microscopy.

The role of the plant microbiome for forestry, agriculture and urban greenspace in times of environmental change.

This Lilliput article provides a literature overview on ecological effects of the plant microbiome with a focus on practical application in forestry, agriculture and urban greenspace under the spectre of climate change. After an overview of the mostly bacterial microbiome of the model plant Arabidopsis thaliana, worldwide data from forests reveal ecological differentiation with respect to major guilds of predominantly fungal plant root symbionts. The plant-microbiome association forms a new holobiont, an integrated unit for ecological adaptation and evolutionary selection. Researchers explored the impact of the microbiome on the capacity of plants to adapt to changing climate conditions. They investigated the impact of the microbiome in reforestation programs, after wildfire, drought, salination and pollution events in forestry, grasslands and agriculture. With increasing temperatures plant populations migrate to higher latitudes and higher altitudes. Ecological studies compared the dispersal capacity of plant seeds with that of soil microbes and the response of soil and root microbes to experimental heating of soils. These studies described a succession of microbiome associations and the kinetics of a release of stored soil carbon into the atmosphere enhancing global warming. Scientists explored the impact of synthetic microbial communities (SynComs) on rice productivity or tea quality; of whole soil addition in grassland restoration; or single fungal inoculation in maize fields. Meta-analyses of fungal inoculation showed overall a positive effect, but also a wide variation in effect sizes. Climate change will be particularly prominent in urban areas ("urban heat islands") where more than half of the world population is living. Urban landscape architecture will thus have an important impact on human health and studies started to explore the contribution of the microbiome from urban greenspace to ecosystem services.

Two novel Archaeorhizomyces species isolated from ericoid mycorrhizal roots and their association with ericaceous plants in vitro

Archaeorhizomyces is a diverse and ubiquitous genus of the subphylum Taphrinomycotina, which contains soil-inhabiting/root-associated fungi. Although ecological importance and root-associating lifestyles of Archaeorhizomyces can be postulated, morphological aspects of fungal body and root colonization are largely unknown due to the scarcity of cultures. We obtained three unidentified Archaeorhizomyces isolates from ericoid mycorrhizal (ErM) roots of Rhododendron scabrum and Rhododendron × obtusum collected in Japan. To advance our understanding of lifestyle of the genus, we investigated their general morphology, phylogeny, and in vitro root-colonizing ability in ericoid mycorrhizal hosts, Vaccinium virgatum and Rhododendron kaempferi. Some morphological characteristics, such as slow glowing white-to-creamy-colored colonies and formation of yeast-like or chlamydospore-like cells, were shared between our strains and two described species, Archaeorhizomycesfinlayi and Archaeorhizomyces borealis, but they were phylogenetically distant. Our strains were clearly distinguished as two undescribed species based on morphology and phylogenetic relationship. As seen in typical ErM fungi, both species frequently formed hyphal coils within vital rhizodermal cells of ErM plants in vitro. The morphology of hyphal coils was also different between species. Consequently, two novel species, Archaeorhizomyces notokirishimae sp. nov. and Archaeorhizomyces ryukyuensis sp. nov., were described.

The community of root fungi is associated with the growth rate of Norway spruce (Picea abies).

Our study delved into the relationship between root-associated fungi, gene expression and plant morphology in Norway spruce cuttings derived from both slow-and fast-growing trees. We found no clear link between the gene expression patterns of adventitious roots and the growth phenotype, suggesting no fundamental differences in the receptiveness to fungal symbionts between the phenotypes. Interestingly, saplings from slow-growing parental trees exhibited a higher richness of ectomycorrhizal species and larger roots. Some ectomycorrhizal species, typically found on mature spruces, were more prevalent on saplings from slow-growing spruces. The ericoid mycorrhizal fungus, Hyaloscypha hepaticola, showed a stronger association with saplings from fast-growing spruces. Moreover, saplings from slow-growing spruces had a greater number of Ascomycete taxa and free-living saprotrophic fungi. Aboveground sapling stems displayed some phenotypic variation; saplings from fast-growing phenotypes had longer branches but fewer whorls in their stems compared to those from the slow-growing group. In conclusion, the observed root-associated fungi and phenotypic characteristics in young Norway spruces may play a role in their long-term growth rate. This suggests that the early interactions between spruces and fungi could potentially influence their growth trajectory.