Spin angular momentum-encoded single-photon emitters in a chiral nanoparticle-coupled WSe2 monolayer.

Spin angular momentum (SAM)-encoded single-photon emitters, also known as circularly polarized single photons, are basic building blocks for the advancement of chiral quantum optics and cryptography. Despite substantial efforts such as coupling quantum emitters to grating-like optical metasurfaces and applying intense magnetic fields, it remains challenging to generate circularly polarized single photons from a subwavelength-scale nanostructure in the absence of a magnetic field. Here, we demonstrate single-photon emitters encoded with SAM in a strained WSe2 monolayer coupled with chiral plasmonic gold nanoparticles. Single-photon emissions were observed at the nanoparticle position, exhibiting photon antibunching behavior with a g(2)(0) value of ~0.3 and circular polarization properties with a slight preference for left-circular polarization. Specifically, the measured Stokes parameters confirmed strong circular polarization characteristics, in contrast to emitters coupled with achiral gold nanocubes. Therefore, this work provides potential insights to make SAM-encoded single-photon emitters and understand the interaction of plasmonic dipoles and single photons, facilitating the development of chiral quantum optics.

Laboratory Evaluation of Storage Stability for Asphalt Binder Modified with Crumb Rubber and Styrene-Isoprene-Styrene Depending on Evaluation Factors and Blending Condition.

Modified asphalt binders are still considered important in asphalt pavement. However, the comprehensive use of various modifiers is limited due to storage stability issues. Moreover, there is a scarcity of detailed analyses regarding the degree of separation for asphalt binders among each method despite the utilization of various methods to assess the storage stability of binders. Therefore, a comprehensive analysis was conducted to assess the storage stability of asphalt binder modified with a crumb rubber modifier (CRM) and styrene-isoprene-styrene (SIS), utilizing five evaluation factors following the ASTM D7173 guidelines based on four mixing methods (A: high-shear mixing method, B: low-speed agitating method, C: high-shear mixing method + low mixing method, D: low-speed agitating method + low mixing method). To produce the modified asphalt binder, the proportions of the CRM were 5% and 10% for each binder, and 10% SIS was added to all binders. The results in this study convey that (1) the addition of the modifier led to an increase in G*/sin δ with different mixing methods, but using mixing methods (C and D) for a relatively long time resulted in a lower G*/sin δ, indicating suboptimal performance; (2) through the multiple stress creep recovery (MSCR), rheological properties of Jnr and % rec exhibited trends similar to G*/sin δ evaluation, highlighting an improved elastic recovery with a higher modifier content; (3) storage stability assessment revealed consistent trends in high-shear mixing groups (A and C), while low-speed mixing groups (B and D) exhibited an elevated separation index (SI), suggesting a sensitivity to modification conditions; (4) evaluation using the MSCR method indicated that % rec with a 3.2 kPa load is effective for the sensitive assessment of binder storage stability and Jnr showed a limited sensitivity across varying loads, advocating for % rec for precise evaluation; and (5) despite permitting various tests, achieving consistent results remains challenging. Future research should explore diverse modifiers and optimal evaluation methods to enhance knowledge of binder behavior and separation dynamics.

Facultative mycorrhization in a fern (Struthiopteris spicant L. Weiss) is bound to light intensity.

BACKGROUND: The establishment of mycorrhizal relationships between a fungus and a plant typically enhances nutrient and water uptake for the latter while securing a carbon source for the fungus. However, under a particular set of environmental conditions, such as low availability of light and abundant nutrients in the soil, the resources invested in the maintenance of the fungi surpass the benefits obtained by the host. In those cases, facultative mycorrhizal plants are capable of surviving without symbiosis. Facultative mycorrhization in ferns has been overlooked until now. The present study measured the response of Struthiopteris spicant L. Weiss, and its root-associated fungi to different levels of light and nutrient availability in terms of growth, mycorrhizal presence, and leaf nutrient content. This fern species exhibits a great tolerance to variable light, nutrient, and pH conditions, and it has been found with and without mycorrhizae. We conducted a greenhouse experiment with 80 specimens of S. spicant and three factors (Light, Phosphorus, and Nitrogen) resulting in eight treatments. RESULTS: We found a significant influence of the factor light on fungal community composition, plant biomass, and nutrient accumulation. Departing from a lack of colonization at the initial stage, plants showed a remarkable increment of more than 80% in the arbuscular mycorrhizal fungi (AMF) richness and abundance in their roots when grown under high light conditions, compared with the ones in low light. We also observed an upward trend of C:P and C:N ratios and the above- and belowground biomass production when AMF abundance increased. Furthermore, the compositional analysis of the whole fungal communities associated with S. spicant roots revealed clear differences among low-light and high-light treatments. CONCLUSIONS: This study is the first to investigate the importance of light and nutrient availability in determining fern-AMF relationships. We confirmed that Struthiopteris spicant is a facultative mycorrhizal plant. The composition and diversity of AMF found in the roots of this fern are strongly influenced by light and less by nutrient conditions. Our study shows that ferns respond very sensitively to changes in environmental factors, leading to shifts in the associated mycorrhizal communities.

Draft Genome of Nocardia canadensis sp. nov. Isolated from Petroleum-Hydrocarbon-Contaminated Soil.

The bacterial strain WB46 was isolated from the rhizosphere of willow plants (Salix purpurea L.) growing in soil contaminated with petroleum hydrocarbons. The strain was subjected to whole-genome shotgun sequencing using Illumina HiSeq. Its draft genome is 7.15 Mb, with a 69.55% GC content, containing 6387 protein-coding genes and 51 tRNA and 15 rRNA sequences. The quality and reliability of the genome were assessed using CheckM, attaining an estimated genome completeness of 98.75% and an estimated contamination of 1.68%. These results indicate a high-quality genome (>95%) and low contamination (<5%). Many of these genes are responsible for petroleum hydrocarbon degradation, such as alkane 1-monooxygenase (alkB) and naphthalene dioxygenase (ndo). 16S rRNA gene analysis, including in silico DNA-DNA hybridization (DDH) and average nucleotide identity (ANI), showed that strain WB46 belongs to the genus Nocardia, and the most closely related species is Nocardia asteroides. The strain WB46 showed a distance of 63.4% and sequence identity of 88.63%, respectively. These values fall below the threshold levels of 70% and 95%, respectively, suggesting that the strain WB46 is a new species. We propose the name of Nocardia canadensis sp. nov. for this new species. Interestingly, the sequence divergence of the 16S rRNA gene showed that the divergence only occurred in the V2 region. Therefore, the conventional V3-V4, V5-V7, or V8-V9 targeting metabarcoding, among others, would not be able to assess the diversity related to this new species.

From microbiome composition to functional engineering, one step at a time.

SUMMARYCommunities of microorganisms (microbiota) are present in all habitats on Earth and are relevant for agriculture, health, and climate. Deciphering the mechanisms that determine microbiota dynamics and functioning within the context of their respective environments or hosts (the microbiomes) is crucially important. However, the sheer taxonomic, metabolic, functional, and spatial complexity of most microbiomes poses substantial challenges to advancing our knowledge of these mechanisms. While nucleic acid sequencing technologies can chart microbiota composition with high precision, we mostly lack information about the functional roles and interactions of each strain present in a given microbiome. This limits our ability to predict microbiome function in natural habitats and, in the case of dysfunction or dysbiosis, to redirect microbiomes onto stable paths. Here, we will discuss a systematic approach (dubbed the N+1/N-1 concept) to enable step-by-step dissection of microbiome assembly and functioning, as well as intervention procedures to introduce or eliminate one particular microbial strain at a time. The N+1/N-1 concept is informed by natural invasion events and selects culturable, genetically accessible microbes with well-annotated genomes to chart their proliferation or decline within defined synthetic and/or complex natural microbiota. This approach enables harnessing classical microbiological and diversity approaches, as well as omics tools and mathematical modeling to decipher the mechanisms underlying N+1/N-1 microbiota outcomes. Application of this concept further provides stepping stones and benchmarks for microbiome structure and function analyses and more complex microbiome intervention strategies.

Using enzyme activities as an indicator of soil fertility in grassland - an academic dilemma.

Grasslands play an important role in conserving natural biodiversity and providing ecosystem functions and services for societies. Soil fertility is an important property in grassland, and the monitoring of soil fertility can provide crucial information to optimize ecosystem productivity and sustainability. Testing various soil physiochemical properties related to fertility usually relies on traditional measures, such as destructive sampling, pre-test treatments, labor-intensive procedures, and costly laboratory measurements, which are often difficult to perform. However, soil enzyme activity reflecting the intensity of soil biochemical reactions is a reliable indicator of soil properties and thus enzyme assays could be an efficient alternative to evaluate soil fertility. Here, we review the latest research on the features and functions of enzymes catalyzing the biochemical processes that convert organic materials to available plant nutrients, increase soil carbon and nutrient cycling, and enhance microbial activities to improve soil fertility. We focus on the complex relationships among soil enzyme activities and functions, microbial biomass, physiochemical properties, and soil/crop management practices. We highlight the biochemistry of enzymes and the rationale for using enzyme activities to indicate soil fertility. Finally, we discuss the limits and disadvantages of the potential new molecular tool and provide suggestions to improve the reliability and feasibility of the proposed alternative.

Effect of Kaolinite and Cloisite Na+ on Storage Stability of Rubberized Binders.

This study aimed to evaluate the impact of a two-step modification process involving kaolinite and cloisite Na+ on the storage stability of rubberized binders. The process involved the manual combination of virgin binder PG 64-22 with crumb rubber modifier (CRM), which was heated to condition it. The preconditioned rubberized binder was then modified for two hours at a high speed of 8000 rpm using wet mixing. The second stage modification was performed in two parts, with part 1 using only crumb rubber as the modifier and part 2 involving the use of kaolinite and montmorillonite nano clays at a replacement percentage of 3% to the original weight of the binder along with the crumb rubber modifier. The Superpave and multiple shear creep recovery (MSCR) test methods were used to calculate the performance characteristics and separation index % of each modified binder. The results showed that the viscosity properties of kaolinite and montmorillonite improved the performance class of the binder, with montmorillonite demonstrating greater viscosity values than kaolinite even at high temperatures. Additionally, kaolinite with rubberized binders showed higher resistance to rutting, and the % recovery value from multiple shear creep recovery testing indicated that kaolinite with rubberized binders was more effective than montmorillonite with rubberized binders, even at higher load cycles. The use of kaolinite and montmorillonite reduced phase separation between the asphaltene phase and rubber-rich phase at higher temperatures, but the performance of the rubber binder was affected by higher temperatures. Overall, kaolinite with the rubber binder generally demonstrated greater binder performance.

A metabarcoding protocol targeting two DNA regions to analyze root-associated fungal communities in ferns and lycophytes.

Premise: Detailed studies of the fungi associated with lycophytes and ferns provide crucial insights into the early evolution of land plants. However, most investigations to date have assessed fern-fungus interactions based only on visual root inspection. In the present research, we establish and evaluate a metabarcoding protocol to analyze the fungal communities associated with fern and lycophyte roots. Methods: We used two primer pairs focused on the ITS rRNA region to screen the general fungal communities, and the 18S rRNA to target Glomeromycota fungi (i.e., arbuscular mycorrhizal fungi). To test these approaches, we collected and processed roots from 12 phylogenetically distant fern and lycophyte species. Results: We found marked compositional differences between the ITS and 18S data sets. While the ITS data set demonstrated the dominance of orders Glomerales (phylum Glomeromycota), Pleosporales, and Helotiales (both in phylum Ascomycota), the 18S data set revealed the greatest diversity of Glomeromycota. Non-metric multidimensional scaling (NMDS) ordination suggested an important geographical effect in sample similarities. Discussion: The ITS-based approach is a reliable and effective method to analyze the fungal communities associated with fern and lycophyte roots. The 18S approach is more appropriate for studies focused on the detailed screening of arbuscular mycorrhizal fungi.

Premisa: El estudio de los hongos asociados a licofitas y helechos proporciona información crucial sobre la evolución temprana de las plantas terrestres. Sin embargo, hasta el momento, la mayoría de las investigaciones ha evaluado las interacciones helecho­hongo basándose solamente en la observación directa de las raíces. En la presente investigación, establecemos y evaluamos un protocolo de metabarcoding enfocado en dos regiones de ADN para analizar las comunidades fúngicas asociadas a las raíces de helechos y licofitas. Métodos: Utilizamos dos pares de primer orientados hacia la región ITS ARNr, para la detección de las comunidades fúngicas generales, y la región 18S ARNr, para captar hongos pertenecientes al phylum Glomeromycota (i.e., hongos micorrícicos arbusculares). Para evaluar estos procedimientos, nosotros recolectamos y procesamos raíces de 12 especies de helechos y licofitas distantes desde el punto de vista filogenético. Resultados: Se observaron claras diferencias de composición entre los sets de datos ITS y 18S: mientras el primero demostró un predominio de los órdenes Glomerales (phylum Glomeromycota), Pleosporales y Helotiales (ambos en phylum Ascomycota), el set 18S reveló la mayor diversidad de hongos micorrizógenos arbusculares. Ninguno de los marcadores moleculares utilizados detectó miembros del phylum Mucoromycota en las muestras. El escalamiento multidimensional no métrico (NMDS) sugirió un papel importante de la región geográfica de origen en la determinación de las similitudes entre muestras. Discusión: El método basado en la región ITS es consistente, replicable y eficaz para analizar las comunidades fúngicas asociadas con raíces de helechos y licofitos. El enfoque 18S es más apropiado para estudios centrados en la detección de los hongos micorrizógenos arbusculares.

Effect of Blending and Curing Conditions on the Storage Stability of Rubberized Asphalt Binders.

Crumb rubber modifier (CRM) binders easily suffer from instability at high temperatures, with many suggestions being developed to evaluate their storage stability. However, much uncertainty around CRM binders still exists regarding the relationship between mixing methods and experiments in order to calculate the separation index. In this study, a laboratory investigation into how CRM binders behave regarding storage stability using different mixing methods and experiments was conducted. The CRM asphalt binder in this study was prepared through a wet mixing process with the addition of 5% and 10% crumb rubber modifier (by weight) at 200 °C. The three main modification methods were method A: high-shear mixing (8000 rpm) for 2 h, method B: low mixing (300 rpm) for 8 h, and method C: high-shear mixing (8000 rpm) for 2 h + low mixing (300 rpm) for 6 h. In addition, the effect of separation index (SI) on storage stability was assessed, measuring viscosity, G*/sin δ, and percentage recovery. In general, the results of this study revealed that method C appeared to have the most prominent effect on decreasing the difference between the values of the top and bottom parts; the results for 5% CRM using method C showed that there were no differences among the values for the top, middle, and bottom parts; from the MSCR test, method C was effective in decreasing the difference between the values of the top and bottom parts. It was observed that method C improved storage stability. However, the results for percentage recovery were relatively higher than the separation index when using rotational viscosity and G*/sin δ.

Laboratory Evaluation of Asphalt Binders Containing Styrene-Butadiene-Styrene (SBS) and Processed Oil.

The study presents an experimental evaluation to improve the resistivity of binders with "Styrene-Butadiene-Styrene" (SBS) and "Processed oil" by studying the physical properties, rheology, and cracking. For this experiment, PG 64-22 was mixed with SBS at different percentages of 5%, 10%, and 15% by weight of the original binder with two processed oil contents of 6% and 12% by weight of the binder. Laboratory tests have been conducted at various high, medium, and low temperature ranges to evaluate their properties. The processed oil polymer modified asphalt (PMA) binder is artificially aged in both the short and long-term using a Rolling Thin Film Oven (RTFO) and a Pressure Aging Vessel (PAV). The Superpave testing method was performed on modified binders using a Rotational Viscometer (RV), Dynamic Shear Rheometer (DSR), and Bending Beam Rheometer (BBR). The results of this study illustrate (1) The addition of SBS leads to higher viscosity, but the co-modification of asphalt binder with the processed oil shows a significant modulation of the viscosity value. (2) In addition, processed oil reduced the resistance to rutting, but the addition of SBS significantly improved the rutting resistance of the asphalt binder. (3) The addition of SBS and processed oil improved the value of G sin δ, notably. (4) According to BBR, it has been shown that the addition of SBS in addition to the processed oil improves the stiffness values of modified asphalt binders.

Reciprocal recombination genomic signatures in the symbiotic arbuscular mycorrhizal fungi Rhizophagus irregularis.

Arbuscular mycorrhizal fungi (AMF) are part of the most widespread fungal-plant symbiosis. They colonize at least 80% of plant species, promote plant growth and plant diversity. These fungi are multinucleated and contain either one or two haploid nuclear genotypes (monokaryon and dikaryon) identified by the alleles at a putative mating-type locus. This taxon has been considered as an ancient asexual scandal because of the lack of observable sexual structures. Despite identification of a putative mating-type locus and functional activation of genes related to mating when two isolates co-exist, it remains unknown if the AMF life cycle involves a sexual or parasexual stage. We used publicly available genome sequences to test if Rhizophagus irregularis dikaryon genomes display signatures of sexual reproduction in the form of reciprocal recombination patterns, or if they display exclusively signatures of parasexual reproduction involving gene conversion. We used short-read and long-read sequence data to identify nucleus-specific alleles within dikaryons and then compared them to orthologous gene sequences from related monokaryon isolates displaying the same putative MAT-types as the dikaryon. We observed that the two nucleus-specific alleles of the dikaryon A5 are more related to the homolog sequences of monokaryon isolates displaying the same putative MAT-type than between each other. We also observed that these nucleus-specific alleles displayed reciprocal recombination signatures. These results confirm that dikaryon and monokaryon isolates displaying the same putative MAT-type are related in their life-cycle. These results suggest that a genetic exchange mechanism, involving reciprocal recombination in dikaryon genomes, allows AMF to generate genetic diversity.

Effect of Processed Oil on Asphalt Binder Properties.

This study investigates the effectiveness of processed oil in the modification of PG 64-22 and PG 76-22 by assessing their physical and rheological properties, and multiple comparison was conducted between the two binders. The base binders PG 64-22 and PG 76-22 were blended with processed oil at four different percentages of contents (3%, 6%, 9% and 12% by the weight of the binder) and compared with the control binder in each test. The base and modified binders were artificially short-term and long-term aged using a rolling thin film oven (RTFO) and pressure aging vessel (PAV) procedures. Superpave binder tests were performed on the modified binders by applying a rotational viscometer (RV), dynamic shear rheometer (DSR), and bending beam rheometer (BBR). The comparisons and results presented in this study indicate that (1) the processed oil has a significant effect on the binders' viscosity, which changes with respect to the increment of processed oil content. The viscosity of both modified binders decreased with the addition of 3, 6, 9 and 12% processed oil; (2) the performed DSR test showed that the addition of processed oil had a negative effect on the rutting resistance for both binders, since in PG 64-22, G*/Sin δ values decreased by 55, 65, 75 and 83% with the addition of 3, 6, 9 and 12% processed oil, respectively, while a decrement of G*/Sin δ of 24, 45, 58 and 65% with the addition of 3, 6, 9 and 12% processed oil was observed in PG 76-22; meanwhile, the fatigue cracking performance was improved and was found to be effective, while G* Sin δ in PG76-22 decreased by 9, 30, 36, and 52% and in PG 64-22 by 27, 44, 53, and 67% with the addition of 3, 6, 9 and 12% processed oil; (3) the results from the BBR test indicate significant improvement in the thermal cracking properties of the binders. The addition of 3, 6, 9 and 12% processed oil resulted in a decrease in the stiffness of both the PG 64-22 and PG 76-22 binders, with a positive effect consequently being observed on the m-values of the binders.

Laboratory Characterization of Asphalt Binders Modified with Styrene Butadiene Rubber (SBR).

The study describes the laboratory assessment (physical and rheological properties) of the binders (PG 64-22 and PG 76-22) modified with Styrene Butadiene Rubber (SBR), and a comprehensive comparison between these two modified binder types. PG 64-22 and PG 76-22 were used as base binders. Both of the base binders were blended with SBR at four different percentages of content (0%, 4%, 6%, and 8% by the weight of the binder). The base and modified binders were artificially short-term and long-term aged using a rolling thin film oven (RTFO) and pressure aging vessel (PAV) procedures. Superpave binder tests were conducted on the SBR modified binder using rotational viscometer (RV), dynamic shear rheometer (DSR), and bending beam rheometer (BBR). In depth rutting performance was investigated using Multiple Stress Creep Recovery (MSCR). The results of this study indicated that (1) the addition of SBR into both binders increased the viscosity and polymer modified asphalt (PMA) binders observed to have more significant effect on its viscosity property; (2) the higher the SBR content, the better the rutting resistance of the binder and it is observed that the effect is prominent on the control binder; (3) MSCR test results showed that the SBR modified binders improved the binder percentage recovery and found to have a more significant effect on the PG 76-22 binder compared to PG 64-22; and (4) both the control PG 64-22 and PMA PG 76-22 binders resulted in similar trends on the cracking properties and were found to have insignificant effects due to the addition of an SBR modifier.

Salix purpurea and Eleocharis obtusa Rhizospheres Harbor a Diverse Rhizospheric Bacterial Community Characterized by Hydrocarbons Degradation Potentials and Plant Growth-Promoting Properties.

Phytoremediation, a method of phytomanagement using the plant holobiont to clean up polluted soils, is particularly effective for degrading organic pollutants. However, the respective contributions of host plants and their associated microbiota within the holobiont to the efficiency of phytoremediation is poorly understood. The identification of plant-associated bacteria capable of efficiently utilizing these compounds as a carbon source while stimulating plant-growth is a keystone for phytomanagement engineering. In this study, we sampled the rhizosphere and the surrounding bulk soil of Salixpurpurea and Eleocharis obusta from the site of a former petrochemical plant in Varennes, QC, Canada. Our objectives were to: (i) isolate and identify indigenous bacteria inhabiting these biotopes; (ii) assess the ability of isolated bacteria to utilize alkanes and polycyclic aromatic hydrocarbons (PAHS) as the sole carbon source, and (iii) determine the plant growth-promoting (PGP) potential of the isolates using five key traits. A total of 438 morphologically different bacterial isolates were obtained, purified, preserved and identified through PCR and 16S rRNA gene sequencing. Identified isolates represent 62 genera. Approximately, 32% of bacterial isolates were able to utilize all five different hydrocarbons compounds. Additionally, 5% of tested isolates belonging to genera Pseudomonas, Acinetobacter, Serratia, Klebsiella, Microbacterium, Bacillus and Stenotrophomonas possessed all five of the tested PGP functional traits. This culture collection of diverse, petroleum-hydrocarbon degrading bacteria, with multiple PGP traits, represents a valuable resource for future use in environmental bio- and phyto-technology applications.

The effects of mycorrhizal colonization on phytophagous insects and their natural enemies in soybean fields.

The use of belowground microorganisms in agriculture, with the aim to stimulate plant growth and improve crop yields, has recently gained interest. However, few studies have examined the effects of microorganism inoculation on higher trophic levels in natural conditions. We examined how the diversity of phytophagous insects and their natural enemies responded to the field-inoculation of soybean with a model arbuscular mycorrhizal fungus (AMF), Rhizophagus irregularis, combined with a nitrogen-fixing bacterium, Bradyrhizobium japonicum, and a plant growth-promoting bacterium, Bacillus pumilus. We also investigate if the absence or presence of potassium fertilizer can affect this interaction. We found an increase in the abundance of piercing-sucking insects with the triple inoculant irrespective of potassium treatment, whereas there were no differences among treatments for other insect groups. A decrease in the abundance of the soybean aphid, Aphis glycines, with the double inoculant Rhizophagus + Bradyrhizobium was observed in potassium enriched plots and in the abundance of Empoasca spp. with potassium treatment independent of inoculation type. Although it was not possible to discriminate the mycorrhization realized by inoculum from that of the indigenous AMF in the field, we confirmed global negative effects of overall mycorrhizal colonization on the abundance of phytophagous piercing-sucking insects, phytophagous chewing insects, and the alpha diversity of phytophagous insects. In perspective, the use of AMF/Rhizobacteria inoculants in the field should focus on the identity and performance of strains to better understand their impact on insects.

Lasing Action from Anapole Metasurfaces.

We study active dielectric metasurfaces composed of two-dimensional arrays of split-nanodisk resonators fabricated in InGaAsP membranes with embedded quantum wells. Depending on the geometric parameters, such split-nanodisk resonators can operate in the optical anapole regime originating from an overlap of the electric dipole and toroidal dipole Mie-resonant optical modes, thus supporting strongly localized fields and high-Q resonances. We demonstrate room-temperature lasing from the anapole lattices of engineered active metasurfaces with low threshold and high coherence.

The Phosphate Inhibition Paradigm: Host and Fungal Genotypes Determine Arbuscular Mycorrhizal Fungal Colonization and Responsiveness to Inoculation in Cassava With Increasing Phosphorus Supply.

A vast majority of terrestrial plants are dependent on arbuscular mycorrhizal fungi (AMF) for their nutrient acquisition. AMF act as an extension of the root system helping phosphate uptake. In agriculture, harnessing the symbiosis can potentially increase plant growth. Application of the AMF Rhizophagus irregularis has been demonstrated to increase the yields of various crops. However, there is a paradigm that AMF colonization of roots, as well as the plant benefits afforded by inoculation with AMF, decreases with increasing phosphorus (P) supply in the soil. The paradigm suggests that when fertilized with sufficient P, inoculation of crops would not be beneficial. However, the majority of experiments demonstrating the paradigm were conducted in sterile conditions without a background AMF or soil microbial community. Interestingly, intraspecific variation in R. irregularis can greatly alter the yield of cassava even at a full application of the recommended P dose. Cassava is a globally important crop, feeding 800 million people worldwide, and a crop that is highly dependent on AMF for P uptake. In this study, field trials were conducted at three locations in Kenya and Tanzania using different AMF and cassava varieties under different P fertilization levels to test if the paradigm occurs in tropical field conditions. We found that AMF colonization and inoculation responsiveness of cassava does not always decrease with an increased P supply as expected by the paradigm. The obtained results demonstrate that maximizing the inoculation responsiveness of cassava is not necessarily only in conditions of low P availability, but that this is dependent on cassava and fungal genotypes. Thus, the modeling of plant symbiosis with AMF under different P levels in nature should be considered with caution.

The methylome of the model arbuscular mycorrhizal fungus, Rhizophagus irregularis, shares characteristics with early diverging fungi and Dikarya.

Early-diverging fungi (EDF) are distinct from Dikarya and other eukaryotes, exhibiting high N6-methyldeoxyadenine (6mA) contents, rather than 5-methylcytosine (5mC). As plants transitioned to land the EDF sub-phylum, arbuscular mycorrhizal fungi (AMF; Glomeromycotina) evolved a symbiotic lifestyle with 80% of plant species worldwide. Here we show that these fungi exhibit 5mC and 6mA methylation characteristics that jointly set them apart from other fungi. The model AMF, R. irregularis, evolved very high levels of 5mC and greatly reduced levels of 6mA. However, unlike the Dikarya, 6mA in AMF occurs at symmetrical ApT motifs in genes and is associated with their transcription. 6mA is heterogeneously distributed among nuclei in these coenocytic fungi suggesting functional differences among nuclei. While far fewer genes are regulated by 6mA in the AMF genome than in EDF, most strikingly, 6mA methylation has been specifically retained in genes implicated in components of phosphate regulation; the quintessential hallmark defining this globally important symbiosis.

Generation of unequal nuclear genotype proportions in Rhizophagus irregularis progeny causes allelic imbalance in gene transcription.

Arbuscular mycorrhizal fungi (AMF) form mutualisms with most plant species. The model AMF Rhizophagus irregularis is common in many ecosystems and naturally forms homokaryons and dikaryons. Quantitative variation in allele frequencies in clonally dikaryon offspring suggests they disproportionately inherit two distinct nuclear genotypes from their parent. This is interesting, because such progeny strongly and differentially affect plant growth. Neither the frequency and magnitude of this occurrence nor its effect on gene transcription are known. Using reduced representation genome sequencing, transcriptomics, and quantitative analysis tools, we show that progeny of homokaryons and dikaryons are qualitatively genetically identical to the parent. However, dikaryon progeny differ quantitatively due to unequal inheritance of nuclear genotypes. Allele frequencies of actively transcribed biallelic genes resembled the frequencies of the two nuclear genotypes. More biallelic genes showed transcription of both alleles than monoallelic transcription, but biallelic transcription was less likely with greater allelic divergence. Monoallelic transcription levels of biallelic genes were reduced compared with biallelic gene transcription, a finding consistent with genomic conflict. Given that genetic variation in R. irregularis is associated with plant growth, our results establish quantitative genetic variation as a future consideration when selecting AMF lines to improve plant production.