Search

We assessed, in a field experiment, the effects of arbuscular mycorrhizal fungi (Rhizophagus intraradices) and plant growth-promoting bacteria (Azospirillum brasilense) on the soil biological activity and the growth of key pioneer species used in the revegetation of coal-mining areas undergoing recovery. We applied four inoculation treatments to the pioneer plant species (Lablab purpureus, Paspalum notatum, Crotalaria juncea, Neonotonia wightii, Stylosanthes guianensis, Andropogon gayanus and Trifolium repens) used in the recovery process: NI (Control - Non-inoculated), AZO (A. brasilense), AMF (R. intraradices), and co-inoculation of AZO and AMF. On the 75th and 180th days, we measured plant dry mass, mycorrhizal colonization, N and P concentration, and accumulation in plant tissue. We collected soil to quantify glomalin content and soil enzyme activity. After 180 days, we did a phytosociological characterization of the remaining spontaneous plants.The both microorganisms, singly or co-inoculated, promoted increases in different fractions of soil glomalin, acid phosphatase activity, and fluorescein diacetate activity at 75 and 180 days. The inoculation was linked to higher plant biomass production (62-89%) and increased plant P and N accumulation by 34-75% and 70-85% at 180 days, compared with the non-inoculated treatment. Among the pioneer species sown Crotalaria juncea produced the highest biomass at the 75th and 180th days (67% and 76% of all biomass), followed by Lablab purpureus (3% and 0.5%), while the other species failed to establish. At 180 days, we observed twenty spontaneous plant species growing in the area, primarily from the Poaceae family (74%). That suggests that the pioneer species present in the area do not hinder the ecological succession process. Inoculation of R. intraradices and A. brasilense, isolated or combined, increases soil biological activity, growth, and nutrient accumulation in key pioneer plant species, indicating the potential of that technique for the recovery of lands degraded by coal mining., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Physio-biochemical regulations governing crop growth period are pivotal for drought adaptation. Yet, the extent to which functionality of arbuscular mycorrhizal fungi (AM fungi) varies across different stages of maize growth under drought conditions remains uncertain. Therefore, periodic functionality of two different AM fungi i.e., Rhizophagus irregularis SUN16 and Glomus monosporum WUM11 were assessed at jointing, silking, and pre-harvest stages of maize subjected to different soil moisture gradients i.e., well-watered (80% SMC (soil moisture contents)), moderate drought (60% SMC), and severe drought (40% SMC). The study found that AM fungi significantly (p < 0.05) affected various morpho-physiological and biochemical parameters at different growth stages of maize under drought. As the plants matured, AM fungi enhanced root colonization, glomalin contents, and microbial biomass, leading to increased nutrient uptake and antioxidant activity. This boosted AM fungal activity ultimately improved photosynthetic efficiency, evident in increased photosynthetic pigments and photosynthesis. Notably, R. irregularis and G. monosporum improved water use efficiency and mycorrhizal dependency at critical growth stages like silking and pre-harvest, indicating their potential for drought resilience to stabilize yield. The principal component analysis highlighted distinct plant responses to drought across growth stages and AM fungi, emphasizing the importance of early-stage sensitivity. These findings underscore the potential of incorporating AM fungi into agricultural management practices to enhance physiological and biochemical responses, ultimately improving drought tolerance and yield in dryland maize cultivation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Rhizosphere microbes are essential partners for plant stress tolerance. Recent studies indicate that arbuscular mycorrhizal fungi (AMF) can facilitate the revegetation of soils contaminated by heavy metals though interacting with rhizosphere microbiome. However, it is unclear how AMF affect rhizosphere microbiome to improve the growth of plant under rare earth elements (REEs) stress. AMF (Claroideoglomus etunicatum) was inoculated to maize grown in soils spiked with Lanthanum (0 mg kg -1 , La0; 10 mg kg -1 , La10; 100 mg kg -1 , La100; 500 mg kg -1 , La500). Plant biomass, nutrient uptake, REE uptake and rhizosphere bacterial and fungal community were evaluated. The results indicated that La100 and La500 decreased significantly root colonization rates and nutrition uptake (K, P, Ca and Mg content). La500 decreased significantly α-diversity indexes of bacterial and fungal community. AMF enhanced significantly the shoot and root fresh and dry weight of maize in all La treatments (except for the root fresh and dry weight of La0 and La10 treatment). For La100 and La500 treatments, AMF increased significantly nutrition uptake (K, P, Ca and Mg content) in shoot of maize by 27.40-441.77%. For La500 treatment, AMF decreased significantly shoot La concentration by 51.53% in maize, but increased significantly root La concentration by 30.45%. In addition, AMF decreased bacterial and fungal Shannon index in La0 treatment, but increased bacterial Shannon index in La500 treatment. Both AMF and La500 affected significantly the bacterial and fungal community composition, and AMF led to more influence than La. AMF promoted the enrichment of bacteria, including Planomicrobium, Lysobacter, Saccharothrix, Agrococcus, Microbacterium, Streptomyces, Penicillium and other unclassified genus, and fungi (Penicillium) in La500, which showed the function for promoting plant growth and tolerance of heavy metal. The study revealed that AMF can regulate the rhizosphere bacterial and fungal composition and foster certain beneficial microbes to enhance the tolerance of maize under La stress. Phytoremediation assisted by AMF is an attractive approach to ameliorate REEs-contaminated soils., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Accumulation of As (metalloid) degrades soil by negatively affecting the activities of soil enzymes, which in turn reduce growth and yield of the inhabiting plant. Arbuscular mycorrhizal (AM) symbiosis can impart metalloid tolerance in plants by secreting glomalin-related soil protein (GRSP) which binds with As or inertly adsorb in the extraradical mycelial surface. However, profitable use of AM requires selection of the most efficient combination of host plant and fungal species. The current study, therefore designed to study the efficacy of 3 a.m. fungal species: Rhizoglomus intraradices (Ri), Funneliformis mosseae (Fm) and Claroideoglomus claroideum (Cc) in imparting arsenate As(V) and arsenite As(III) stress tolerance in Cicer arietinum (chickpea) genotypes (G) - relatively metalloid tolerant- HC 3 and sensitive- C 235. Roots were found to be more severly affected as compared to shoots which resulted into a major decline in uptake of nutrients, chlorophyll concentrations and yield with As(III) inducing more toxic effects than As(V). HC 3 established more effective mycorrhizal symbiosis and was able to extract higher nutrients from the soil than C 235. Ri was most beneficial in improving plant biomass, carbohydrate utilization and productivity followed by Fm and Cc which could be due to its capability to initiate highest percent colonization and least metalloid uptake in roots through higher glomalin production in the soil. Moreover, Ri was highly efficient in improving soil enzymes activities-phosphatases (PHAs), β-glucosidase (BGA) and invertase (INV), thereby, imparting metalloid tolerance in chickpea genotypes. The results suggested use of Ri-chickpea symbiosis as a promising strategy for ameliorating As stress in chickpea., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Arbuscular mycorrhizal (AM) symbiosis is a mutually beneficial association of plants and fungi of the subphylum Glomeromycotina. Endosymbiotic AM fungi colonize the inner cortical cells of the roots, where they form branched hyphae called arbuscules that function in nutrient exchange with the plant. To support arbuscule development and subsequent bidirectional nutrient exchange, the root cortical cells undergo substantial transcriptional reprogramming. REDUCED ARBUSCULAR MYCORRHIZA1 ( RAM1 ), previously studied in several dicot plant species, is a major regulator of this cortical cell transcriptional program. Here, we generated ram1 mutants and RAM1 overexpressors in a monocot, Brachypodium distachyon. The AM phenotypes of two ram1 line s revealed that RAM1 is only partly required to enable arbuscule development in B. distachyon Transgenic lines constitutively overexpressing BdRAM1 showed constitutive expression of AM-inducible genes even in the shoots. Following inoculation with AM fungi, BdRAM1 -overexpressing plants showed higher arbuscule densities relative to controls, indicating the potential to manipulate the relative proportion of symbiotic interfaces via modulation of RAM1 However, the overexpressors also show altered expression of hormone biosynthesis genes and aberrant growth patterns, including stunted bushy shoots and poor seed set. While these phenotypes possibly provide additional clues about the scope of influence of BdRAM1 , they also indicate that directed approaches to increase the density of symbiotic interfaces will require a more focused, potentially cell type specific manipulation of transcription factor gene expression., (© 2020 American Society of Plant Biologists. All Rights Reserved.)

The synergistic interaction between arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing bacteria (PSB) can enhance growth and phosphorous uptake in plants. Since PSBs are well known hyphal colonizers we sought to understand this physical interaction and exploit it in order to design strategies for the application of a combined microbial inoculum. Phosphate-solubilizing bacteria strongly attached to the hyphae of Rhizoglomus irregulare were isolated using a two compartment system (root and hyphal compartments), which were separated by a nylon mesh through which AMF hyphae could pass but not plant roots. Allium ampeloprasum (Leek) was used as the host plant inoculated with R. irregulare. A total of 128 bacteria were isolated, of which 12 showed stable phosphate solubilizing activity. Finally, three bacteria belonging to the genus Pseudomonas showed the potential for inorganic and organic phosphate mobilization along with other plant growth promoting traits. These PSBs were further evaluated for their functional characteristics and their interaction with AMF. The impact of single or co-inoculations of the selected bacteria and AMF on Solanum lycopersicum was tested and we found that plants inoculated with the combination of fungus and bacteria had significantly higher plant biomass compared to single inoculations, indicating synergistic activities of the bacterial-fungal consortium., (Copyright © 2020 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Arbuscular mycorrhizal (AM) fungi, forming symbiotic associations with land plants, are obligate symbionts that cannot complete their natural life cycle without a host. The fatty acid auxotrophy of AM fungi is supported by recent studies showing that lipids synthesized by the host plants are transferred to the fungi, and that the latter lack genes encoding cytosolic fatty acid synthases. Therefore, to establish an asymbiotic cultivation system for AM fungi, we tried to identify the fatty acids that could promote biomass production. To determine whether AM fungi can grow on medium supplied with fatty acids or lipids under asymbiotic conditions, we tested eight saturated or unsaturated fatty acids (C12 to C18) and two β-monoacylglycerols. Only myristate (C14:0) led to an increase in the biomass of Rhizophagus irregularis , inducing extensive hyphal growth and formation of infection-competent secondary spores. However, such spores were smaller than those generated symbiotically. Furthermore, we demonstrated that R. irregularis can take up fatty acids in its branched hyphae and use myristate as a carbon and energy source. Myristate also promoted the growth of Rhizophagus clarus and Gigaspora margarita Finally, mixtures of myristate and palmitate accelerated fungal growth and induced a substantial change in fatty acid composition of triacylglycerol compared with single myristate application, although palmitate was not used as a carbon source for cell wall biosynthesis in this culture system. Our findings demonstrate that myristate boosts the asymbiotic growth of AM fungi and can also serve as a carbon and energy source., Competing Interests: The authors declare no competing interest.

Arbuscular mycorrhizas (AMs) are divided into two types according to morphology: Arum- and Paris-type AMs. Gibberellins (GAs) mainly inhibit the establishment of Arum-type AM symbiosis in most model plants, whereas the effects of GAs on Paris-type AM symbiosis are unclear. To provide insight into the mechanism underlying this type of symbiosis, the roles of GAs were investigated in Eustoma grandiflorum when used as the host plant for Paris-type AM establishment. Eustoma grandiflorum seedlings were inoculated with the model AM fungus, Rhizophagus irregularis, and the effects of GA and the GA biosynthesis inhibitor uniconazole-P on the symbiosis were quantitatively evaluated. Exogenous GA significantly increased hyphopodium formation at the epidermis, thus leading to the promotion of fungal colonization and arbuscule formation in the root cortex. By contrast, the suppression of GA biosynthesis and signaling attenuated fungal entry to E. grandiflorum roots. Moreover, the exudates from GA-treated roots strongly induced the hyphal branching of R. irregularis. Our results show that GA has an contrasting effect on Paris-type AM symbiosis in E. grandiflorum compared with Arum-type AM symbiosis. This finding could be explained by the differential regulation of the early colonization stage, where fungal hyphae make contact with and penetrate the epidermis., (� The Author(s) 2019. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Biomass of arbuscular mycorrhizal fungi (AMF, Glomeromycota) is often only available in small quantities as these fungi are obligate biotrophs and many species are difficult to cultivate under controlled conditions. Here, I describe a simple, efficient approach to produce crude extracts from single or a small number of spores that can be used for genotyping AMF.

Septoglomus mexicanum is here described as a new species of arbuscular mycorrhizal fungi (AMF; Glomeromycota) based on morphological and phylogenetic analyses. It was isolated from rhizospheric soil of two endemic Mexican legumes: Prosopis laevigata and Mimosa luisana , which grow in semiarid regions of central Mexico. Septoglomus mexicanum is characterized by forming globose spores of (154.5-)202.8(-228.9) µm diam and a spore wall consisting of four layers (SWL1-SWL4): outer wall layer (SWL1) hyaline, evanescent, (1.7-)3.2(-4.3) µm thick; SWL2 laminate and smooth, orange to reddish orange, (3.1-)4.5(-6.1) µm thick; SWL3 laminate, smooth, reddish orange to reddish brown, (4.1-)5.1(-5.7) µm thick; and SWL4 hyaline, semiflexible, (0.93-)1.2(-1.4) µm thick. None of the spore wall layers stain with Melzer's reagent. The subtending hypha has a color from yellowish to golden and presents a septum on spore base. Septoglomus mexicanum can be distinguished from all other Septoglomus species by spore size and color, by spore wall structure (four layers), and by color change of the subtending hypha. Phylogenetic analysis based on the AMF extended DNA barcode covering a 1.5-kb fragment of the small subunit (SSU), internal transcribed spacer region (ITS1-5.8S-ITS2), and the large subunit (LSU) of rRNA genes places S. mexicanum in the genus Septoglomus , separated from other described Septoglomus species, especially S. turnauae , with whom it could be confused morphologically. All available sequences in public databases suggest that this new fungal species has not yet been previously detected. Thus, there are currently 149 Glomeromycota species registered in Mexico, representing 47.4% of the known species worldwide.

Arbuscular mycorrhizal fungi (AMF), particularly the Glomerales group, play a paramount role in plant nutrient uptake, and abiotic and biotic stress management in rice, but recent evidence revealed that elevated CO 2 concentration considerably reduces the Glomerales group in soil. In view of this, the present study was initiated to understand the interaction effect of native Glomerales species application in rice plants (cv. Naveen) under elevated CO 2 concentrations (400 ± 10, 550 ± 20, and 700 ± 20 ppm) in open-top chambers. Three different modes of application of the AMF inoculum were evaluated, of which, combined application of AMF at the seedling production and transplanting stages showed increased AMF colonization, which significantly improved grain yield by 25.08% and also increased uptake of phosphorus by 18.2% and nitrogen by 49.5%, as observed at 700-ppm CO 2 concentration. Organic acids secretion in rice root increased in AMF-inoculated plants exposed to 700-ppm CO 2 concentration. To understand the overall effect of CO 2 elevation on AMF interaction with the rice plant, principal component and partial least square regression analysis were performed, which found both positive and negative responses under elevated CO 2 concentration., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Phytoremediation is an alternative for remediating soil contamination by copper, and its efficiency has been shown to increase when arbuscular mycorrhizal fungi (AMF) and earthworms are separately inoculated into the soil. This study evaluated the isolated and combined effects of inoculating earthworms and arbuscular mycorrhizal fungi into a sandy soil on copper phytoremediation by Canavalia ensiformis. The plants were grown in a greenhouse in soil contaminated with 100 mg Cu kg -1 with and without being inoculated with the arbuscular mycorrhizal fungus Rhizoglomus clarum and the earthworm Eisenia andrei. The availabilities of solid-phase Cu and other nutrients in the soil solution and plant growth were evaluated along with Cu phytotoxicity based on photochemical efficiency and oxidative stress enzyme activity. Accumulation of Cu and other nutrients in the shoots and roots; mycorrhizal colonization, nodulation, and reproduction; and Cu accumulation in the earthworm tissues were also evaluated. The copper caused photosynthetic and biochemical damage that reduced the shoot dry weight by 44% and the root dry weight by 29%. However, the arbuscular mycorrhizal fungus alleviated the Cu toxicity to the plant and increased the shoot dry weight by 81% in the contaminated soil. The earthworms increased the Cu uptake and translocation to the shoot by 31%. The combined presence of the arbuscular mycorrhizal fungus and earthworms in the contaminated soil increased the growth and Cu content of the aerial plant tissues, yielding a 200% increase in Cu accumulation (metal content × biomass) in the C. ensiformis shoots. Combined inoculation with earthworms and arbuscular mycorrhizal fungi increased copper phytoextraction by Canavalia ensiformis in a sandy soil., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This research is aimed to investigate the effect of arbuscular mycorrhiza (AM) fungi on soil microbial activity and carbon mineralization in the maize rhizosphere under potted condition. Glomus etunicatum was used for our experiment. Results showed that AM symbiosis increased the levels of microorganism in the maize rhizosphere soil, and enhanced activity of soil microbial enzymes. After inoculating AM fungi, the contents of dissolved organic carbon (DOC), microbial biomass carbon (MBC) and readily oxidizable carbon (ROC) in the rhizosphere soil of maize increased with varying degrees. We obtained strong evidence that higher contents of MBC, DOC, ROC, superior number of microbes and stronger soil enzyme activities could be responsible for the higher rate of carbon mineralization in AM fungi treatment. AM fungi inoculation was confirmed to be effective to improve the soil quality for larger-scale ecoengineering., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Arbuscular mycorrhizal (AM) fungi are obligate symbionts that depend on living host plants to complete their life cycle 1,2 . This feature, which leads to their unculturability in the absence of plants, strongly hinders basic research and agricultural application of AM fungi. However, at least one AM fungus can grow and develop fertile spores independently of a host plant in co-culture with the bacterium Paenibacillus validus 3 . The bacteria-derived substances are thought to act as stimulants or nutrients for fungal sporulation, but these molecules have not been identified. Here, we show that (S)-12-methyltetradecanoic acid 4,5 , a methyl branched-chain fatty acid isolated from bacterial cultures, stimulates the branching of hyphae germinated from mother spores and the formation of secondary spores in axenic culture of the AM fungus Rhizophagus irregularis. Extensive testing of fatty acids revealed that palmitoleic acid induces more secondary spores than the bacterial fatty acid in R. irregularis. These induced spores have the ability to infect host plant roots and to generate daughter spores. Our work shows that, in addition to a major source of organic carbon 6-9 , fatty acids act as stimulants to induce infection-competent secondary spores in the asymbiotic stage and could provide the key to developing the axenic production of AM inoculum.

In vitro propagation of AM fungi using transformed root cultures (TRC) is commonly used to obtain pure AM fungal propagules for use in research and industry. Early observations indicate that such an artificial environment can alter traits and function of AM fungi over time. We hypothesized that increased in vitro cultivation may promote ruderal strategies in fungi by enhancing propagule production and reducing mutualistic quality. To examine the effect of in vitro cultivation on the trait and function of AM fungi, we inoculated plants with 11 Rhizoglomus irregulare isolates which fell along a cultivation gradient spanning 80 generations. We harvested plants at 10, 20 and 30 d post inoculation to observe differences in fungal and plant traits post infection. In vitro cultivation led to increased spore production but reduced plant shoot phosphorus. Our results indicate that in vitro propagation may indirectly select for traits that affect symbiotic quality., (Copyright © 2019 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Detailed information on structural changes that occur during ontogenesis of Rhizophagus irregularis in axenically developed coculture is limited. Our study aims to investigate the series of events that occur during mycorrhizal ontogenesis under axenic condition through basic and advanced microscopic techniques followed by comparison among these to identify the suitable technique for rapid and detailed analysis of mycorrhizal structures. Three stages were identified in mycorrhizal ontogenesis from initiation (preinfection stage of hyphae; its branching, infection and appressoria formation; epidermal opening; and hyphal entry), progression (arbuscular development; hyphal coils and vesicles) to maturity (extraradical spores). Scanning electron microscopy was found to be an efficient tool for studying spatial three-dimensional progression. Adding to the advantages of advanced microscopy, potential of autofluorescence to explore the stages of symbiosis nondestructively was also established. We also report imaging of ultrathin sections by bright field microscopy to provide finer details at subcellular interface. Owing to the merits of nondestructive sampling, ease of sample preparation, autofluorescence (no dye required), no use of toxic chemicals, rapid analysis and in depth characterization confocal laser scanning microscopy was identified as the most preferred technique. The method thus developed can be used for detailed structural inquisition of mycorrhizal symbiosis both in in planta and in an in vitro system., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

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

Interactions between bacteria and arbuscular mycorrhizal (AM) fungi play a significant role in mediating organic phosphorus (P) transformations and turnover in soil. The bacterial community in soil is largely responsible for mobilization of the soil organic P pool, and the released P is taken up by extraradical AM hyphae, which mediate its use for plant growth. However, the functional microbiome involved in organic P mineralization in the hyphosphere remains poorly understood. The aim of this study was to determine how AM hyphae-associated bacterial communities related to P turnover in the hyphosphere of leek (Allium porrum) respond to different forms of soil P. Using a compartmented microcosm, leek was grown with the AM fungus Funneliformis mosseae, and the extraradical mycelium of F. mosseae was allowed to grow into a separate hyphal compartment containing either no added P, or P as KH 2 PO 4 or phytin. High-throughput sequencing showed that the alkaline phosphatase (ALP)-harboring bacterial community associated with the AM hyphae was dominated by Sinorhizobium, Bradyrhizobium, Pseudomonas, and Ralstonia and was significantly changed in response to different P treatments, with Pseudomonas showing higher relative abundance in organic P treatments than in control and inorganic P treatments. Pseudomonas was also the major genus harboring the β-propeller phytase (BPP) gene in the hyphosphere, but the BPP-harboring community structure was not affected by the presence of different P forms. These results demonstrate the profound differences in ALP- and BPP-harboring bacterial communities in the hyphosphere at bacterial genus level, providing new insights to link bacteria and biogeochemical P cycling driven in association with mycorrhizal hyphae.

The herbicide Roundup (and glyphosate, its active ingredient) is extensively used for weed control on a worldwide scale. It is absorbed after foliar application and quickly translocated inside the plant. In this study, we investigated the effects of Roundup speed, a commercial glyphosate formulation, on the structural composition (dominance of microbial groups, phospholipid fatty acid analysis - PLFA) and functional diversity (use of carbon sources, Multiple Substrate Induced Respiration - MSIR) of soil microorganisms. We specifically aimed at understanding the potential impact of biotic interactions on herbicide effects and included plants, earthworms, and endomycorrhizal fungi in the experimental setup. For this, we grew clover (Trifolium repens) in the greenhouse and added mycorrhizal inoculum (Glomus mosseae) and earthworms (Lumbricus terrestris) to the pots. Two weeks after foliar Roundup application and subsequent plant death, the pots were destructively sampled. The application resulted in a significant increase of microbial respiration (SIR) by approximately 30%. A multivariate analysis of the MSIR data exhibited small but significant differences between the microbial communities of treated and untreated pots, while no significant difference was apparent for the PLFA data. Bacterial PLFAs generally decreased following herbicide application, while mycorrhizal and fungal PLFAs were not affected. We did not find a consistent difference between the fatty acid markers of gram negative and gram positive bacteria. For all investigated parameters, there were highly significant differences between the upper (0-5 cm depth) and lower (5-10 cm) soil layers. The fact that rooting density differed by a factor of 3.5 between the two layers indicated that herbicide effects were especially pronounced in the clover rhizosphere and were likely due to changes in root exudate composition. We found significant, though very small, interactions between Roundup and other experimental factors (especially mycorrhizal inoculum)., (Copyright © 2019. Published by Elsevier Inc.)

Arbuscular mycorrhizal (AM) fungi can protect plants against cadmium (Cd) stress, and are the most prominent symbiotic fungi for contribution to phytoremediation. However, the tolerance mechanism for AM symbiosis on Cd toxicity still remains unclear, especially the related molecular mechanisms. In this study, different Cd treatments were applied to two soybean genotypes with different Cd tolerance in the presence or absence of AM fungal inoculation. The results showed that Cd addition obviously decreased AM colonization. AM symbiosis significantly increased plant dry weight, root growth, and P acquisition in Cd-tolerant HX3 genotype at Cd addition treatments. The effectiveness was associated with a concomitant increased expression of the AM inducible phosphate (Pi) transporter genes GmPT8, GmPT9, GmPT10, and upregulated expression of P-type heavy metal ATPase gene GmHMA19. Additionally, AM fungal inoculation effectively impacted the partitioning of Mg, Cu and Zn, including increased Mg, and decreased Cu and Zn relative concentrations in shoots of Cd tolerant HX3. Taken together, these results suggest that AM symbiosis can alleviate Cd toxicity in soybean through enhanced P nutrition, up-regulated expression of AM inducible GmPTs and GmHMA19, as well as, the alteration of the partitioning of essential nutrient elements., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Soil contaminated with heavy metals (HMs) is a serious problem throughout the world that threatens all living organisms in the soil. Therefore, large-scale remediation is necessary. This study investigated a new combination of remediation techniques on heavy metal contaminated soil, phytoremediation, and soil amendment with nano-sized zero-valent iron (nZVI) and rhizosphere microorganisms. White willow (Salix alba L.) was grown for 160 days in pots containing Pb, Cu, and Cd and amended with 0, 150, and 300 (mg kg -1 ) of nZVI and rhizosphere microorganisms, including the arbuscular mycorrhizal fungus (AMF), Rhizophagus irregularis, and the plant growth promoting rhizobacteria (PGPR), Pseudomonas fluorescens. The results showed that inoculation with PGPR and AMF, particularly dual inoculation, improved plant growth as well as the physiological and biochemical parameters of white willow, and increased the bioconcentration factor (BCF) of Pb, Cu, and Cd. The low dose of nZVI significantly increased the root length and the leaf area of the seedlings and increased the BCF of Cd. In contrast, the high dose of nZVI had negative effects on the seedlings growth and the BCF of Pb and Cu, about - 32% and - 63%, respectively. Our results demonstrate that nZVI at low doses can improve plant performance in a phytoremediation context and that the use of beneficial rhizosphere microorganisms can minimize nZVI stress in plants and make them less susceptible to stress even under high dose conditions.

Arbuscular mycorrhizal fungi (AMF) are obligate plant root symbionts delivering a range of benefits to the host plant such as improved nutrient acquisition and resistance to pathogens and abiotic stress. However, whether they can enhance the function of plant root systems damaged due to subsidence caused by excessive coal mining has not been well explored. In the present study, we investigated the effects of AMF using Funneliformis mosseae (FM) as the test fungus on maize (Zea mays L.) growth and hormone levels under different levels of root damage stress by simulating mining subsidence. The results show that plants treated with FM had more shoots, roots, mycorrhizal colonization and higher hyphal density than those without FM under the same simulated mining-induced subsidence conditions. In addition, plants treated with FM also possessed higher N, P, K, Ca, and Mg contents in the shoots and the roots and higher indole-3-acetic acid, gibberellin (GA), and cytokinin (CTK) contents in the roots, indicating that the mycorrhizal association promoted plant biomass and nutrient uptake. FM treatment was no longer beneficial when root damage due to mining-induced subsidence affected more than half of the roots. Soil SOC, AK, and TG were identified as key factors affecting GA, CTK, IAA, and ABA, and AMF can alter plant hormones directly via the hyphae and indirectly by altering soil physicochemical properties under root damage stress. Overall, our results provide baseline data for assessing the biological reclamation effects of AMF on coal mining-induced subsidence.

Continuous cropping in soybean is increasingly practiced in Heilongjiang Province, leading to substantial yield reductions and quality degradation. Arbuscular mycorrhizal fungi (AMF) are soil microorganisms that form mutualistic interactions with plant roots and can restore the plant rhizosphere microenvironment. In this study, two soybean lines (HN48 and HN66) were chosen as experimental materials, which were planted in different years of continuous cropping soybean soils and were inoculated or not with Funneliformis mosseae in potted-experiments. Ultimately, analysis of root tissue metabolome and root exudates, soil physicochemical properties, plant biomass, as well as rhizosphere soil properties in different experimental treatments, inoculated or not with F. mosseae , was performed. Experimental results showed that: (a) The disease index of soybean root rot was significantly lower in the treatment group than in the control group, and there were differences in disease index and the resistance effect of F. mosseae between the two cultivars; (b) compared with the control, the root tissue metabolome and root exudates remained unchanged, but there were changes in the relative amounts in the treatment group, and the abundant metabolites differed by soybean cultivar; (c) soybean biomass was significantly higher in the treatment group than in the control group, and the effect of F. mosseae on biomass differed with respect to the soybean cultivar; and (d) there were differences in the physiochemical indexes of soybean rhizosphere soil between the treatment and control groups, and the repairing effect of F. mosseae differed between the two cultivars. Therefore, F. mosseae can increase the biomass of continuously cropped soybean, improve the physicochemical properties of the rhizosphere soil, regulate the root metabolite profiles, and alleviate barriers to continuous cropping in potted-experiments of soybean.

Background: Mycorrhizal symbiosis is one of the most fundamental types of mutualistic plant-microbe interaction. Among the many classes of mycorrhizae, the arbuscular mycorrhizae have the most general symbiotic style and the longest history. However, the genomes of arbuscular mycorrhizal (AM) fungi are not well characterized due to difficulties in cultivation and genetic analysis. In this study, we sequenced the genome of the AM fungus Rhizophagus clarus HR1, compared the sequence with the genome sequence of the model species R. irregularis, and checked for missing genes that encode enzymes in metabolic pathways related to their obligate biotrophy., Results: In the genome of R. clarus, we confirmed the absence of cytosolic fatty acid synthase (FAS), whereas all mitochondrial FAS components were present. A KEGG pathway map identified the absence of genes encoding enzymes for several other metabolic pathways in the two AM fungi, including thiamine biosynthesis and the conversion of vitamin B6 derivatives. We also found that a large proportion of the genes encoding glucose-producing polysaccharide hydrolases, that are present even in ectomycorrhizal fungi, also appear to be absent in AM fungi., Conclusions: In this study, we found several new genes that are absent from the genomes of AM fungi in addition to the genes previously identified as missing. Missing genes for enzymes in primary metabolic pathways imply that AM fungi may have a higher dependency on host plants than other biotrophic fungi. These missing metabolic pathways provide a genetic basis to explore the physiological characteristics and auxotrophy of AM fungi.

Over the last three decades, the presence of arbuscular mycorrhizal fungi (AMF) in wetland habitats had been proven, and their roles played in wetland ecosystems and potential functions in wastewater bioremediation technical installations are interesting issues. To increase knowledge on the functions of AMF in the plant-based bioremediation of wastewater, we constructed two vertical-flow wetlands planting with Phragmites australis and investigated AMF distribution in plant roots and their roles played in purification of wastewater polluted by heavy metals (HMs), utilizing the Illumina sequencing technique. A total of 17 operational taxonomic units (OTUs) from 33,031 AMF sequences were obtained, with Glomus being the most dominant. P. australis living in the two vertical-flow constructed wetlands (CWs) harbored diverse AMF comparable with the AM fungal communities in upland habitats. The AMF composition profiles of CW1 (vegetated with non-inoculated plants) and CW2 (vegetated with mycorrhizal plants inoculated with Rhizophagus intraradices) were significantly different. CW1 (15 OTUs) harbored more diverse AMF than CW2 (7 OTUs); however, CW2 harbored much more OTU13 than CW1. In addition, a zipf species abundance distribution (SAD), which might due to the heavy overdominance of OTU13, was observed across AM fugal taxa in P. australis roots of the two CWs. CW1 and CW2 showed high (> 70%) removal capacity of HMs. CW2 exhibited significant higher Cd and Zn removal efficiencies than CW1 (CK) (p = 0.005 and p = 0.008, respectively). It was considered that AMF might play a role in HM removal in CWs.

The phytoremediation of copper (Cu)-contaminated sandy soils can be influenced by the addition of vermicompost to the soil and the mycorrhization of plants. The objective of this study was to evaluate the effects of inoculation with the mycorrhizal fungus Rhizophagus clarus and the addition of different doses of bovine manure vermicompost on the phytoremediation of a sandy soil with a high Cu content using Canavalia ensiformis. Soil contaminated with 100 mg kg -1 Cu received five doses of vermicompost and was cultivated with C. ensiformis, with and without inoculation with mycorrhizal fungus, and the Cu and nutrients in the soil and soil solution were evaluated. The concentrations of Cu and other nutrients and the biomass and Cu phytotoxicity in the plants were quantified by gauging the photochemical efficiency, concentration of photosynthetic pigments and activity of oxidative stress enzymes. The vermicompost increased the soil pH and nutrient concentrations and reduced the Cu content of the solution. When the vermicompost was applied at a dose equivalent to 80 mg phosphorus (P) kg -1 , the phytoextraction efficiency was higher, but the phytostabilization efficiency was higher for vermicompost doses of 10 and 20 mg P kg -1 . The presence of mycorrhizal fungi increased Cu phytostabilization, especially at vermicompost doses of 10 and 20 mg P kg -1 . The use of vermicompost at low doses and inoculation with mycorrhizal fungi increase the phytostabilization potential of C. ensiformis in sandy soil contaminated by Cu.

Arbuscular mycorrhizal fungi (AMF) are widespread, important plant symbionts. They absorb and translocate mineral nutrients from the soil to host plants through an extensive extraradical mycelium, consisting of indefinitely large networks of nonseptate, multinucleated hyphae which may be interconnected by hyphal fusions (anastomoses). This work investigated whether different lineages of the same isolate may lose the ability to establish successful anastomoses, becoming vegetatively incompatible, when grown separately. The occurrence of hyphal incompatibility among five lineages of Funneliformis mosseae, originated from the same ancestor isolate and grown in vivo for more than 20 years in different European locations, was assessed by systematic detection of anastomosis frequency and cytological studies. Anastomosis frequencies ranged from 60 to 80% within the same lineage and from 17 to 44% among different lineages. The consistent detection of protoplasm continuity and nuclei in perfect fusions showed active protoplasm flow both within and between lineages. In pairings between different lineages, post-fusion incompatible reactions occurred in 6-48% of hyphal contacts and pre-fusion incompatibility in 2-17%. Molecular fingerprinting profiles showed genetic divergence among lineages, with overall Jaccard similarity indices ranging from 0.85 to 0.95. Here, phenotypic divergence among the five F. mosseae lineages was demonstrated by the reduction of their ability to form anastomosis and the detection of high levels of vegetative incompatibility. Our data suggest that potential genetic divergence may occur in AMF over only 20 years and represent the basis for detailed studies on the relationship between genes regulating anastomosis formation and hyphal compatibility in AMF.

Phragmites australis (Cav.) Trin. ex Steud. has been reported to form a symbiosis with arbuscular mycorrhizal fungus (AMF). However, the tolerance mechanism for AMF symbiosis on cadmium (Cd) phytotoxicity still remains unclear. In this study, we investigated the effects of Rhizophagus irregularis inoculation on Cd-stressed (0, 1, and 20mgL -1 ) roots, stems, and leaves of P. australis with regard to subcellular Cd distribution and chemical forms of Cd. In addition, transmission electron microscopy and Fourier transform infrared spectroscopy were used to investigate variations in subcellular structures and functional groups in plant organs. The results showed that AMF inoculation could induce selective Cd distribution at subcellular levels, depending on different Cd treatments. The investigation of the chemical forms illustrated that AMF inoculation could alleviate Cd toxicity in all organs. Increases were observed in the ratios of undissolved Cd (F HAc ) and oxalate Cd (F HCl ), while decreases were observed in pectates and protein-integrated Cd (F NaCl ) as well as water soluble Cd (F W ). Hydroxyl (-OH), amide (-NH), carboxyl (C=O), and phosphate (P=O) groups as well as C-O and C-N stretching played predominant roles for the enhancement of Cd tolerance in response to AMF inoculation. These results provide instructive evidence for the mechanisms by which AMF inoculation enhances the Cd tolerance of P. australis via Cd uptake and distribution., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Root-hair growth and development regulated by soil microbes is associated with auxin. In this background, we hypothesized that mycorrhizal fungal inoculation induces greater root-hair growth through stimulated auxin synthesis and transport under water stress conditions. Trifoliate orange (Poncirus trifoliata) was inoculated with an arbuscular mycorrhizal (AM) fungus (Funneliformis mosseae) under well-watered (WW) and drought stress (DS) for 9 weeks. Compared with non-AM seedlings, AM seedlings displayed significantly higher density, length, and diameter of root hairs and root indoleacetic acid (IAA) level, whereas lower total root IAA efflux, regardless of soil moisture status. Root PtYUC3 and PtYUC8 involved in IAA biosynthesis were up-regulated by mycorrhization under WW and DS, whereas AM-modulated expression in PtTAA1, PtTAR2, PtYUC4, and PtYUC6 depended on status of soil moisture. Mycorrhizal inoculation down-regulated the transcript level of root auxin efflux carriers like PtPIN1 and PtPIN3, whereas significantly up-regulated the expression of root auxin-species influx carriers like PtABCB19 and PtLAX2 under DS. These results indicated that AMF-stimulated greater root-hair growth of trifoliate orange under DS that is independent on AMF species is related with mycorrhiza-modulated auxin synthesis and transport, which benefits the host plant to enhance drought tolerance.

Arbuscular mycorrhizal fungi (AMF) improve plant ability to uptake P and tolerate heavy metals. This study aimed to evaluate the effect of available P and the inoculation of Rhizophagus clarus in a Cu-contaminated soil (i) on the activity of acid phosphatases (soil and plant), the presence of glomalin, and (ii) in the biochemical and physiological status of Mucuna cinereum. A Typic Hapludalf soil artificially contaminated by adding 60 mg kg -1 Cu was used in a 3 × 2 factorial design with three replicates. Treatments consisted of three P levels: 0, 40, and 100 mg kg -1 P. Each P treatment level was inoculated (+AMF)/non-inoculated (-AMF) with 200 spores of R. clarus per pot, and plants grown for 45 days. The addition of at least 40 mg kg -1 P and the inoculation of plants with R. clarus proved to be efficient to reduce Cu phytotoxicity and increase dry matter yield. Mycorrhization and phosphate fertilization reduced the activity of enzymes regulating oxidative stress (SOD and POD), and altered the chlorophyll a fluorescence parameters, due to the lower stress caused by available Cu. These results suggest a synergism between the application of P and the inoculation with R. clarus, favoring the growth of M. cinereum in a Cu-contaminated soil. This study shows that AMF inoculation represents an interesting alternative to P fertilization to improve plant development when exposed to excess Cu., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF) are widespread root symbionts that perform important ecological services, such as improving plant nutrient and water acquisition. Some AMF from the Gigasporaceae family host a population of endobacteria, Candidatus Glomeribacter gigasporarum (Cagg). The analysis of the Cagg genome identified six putative toxin-antitoxin modules (TAs), consisting of pairs of stable toxins and unstable antitoxins that affect diverse physiological functions. Sequence analysis suggested that these TA modules were acquired by horizontal transfer. Gene expression patterns of two TAs (yoeB/yefM and chpB/chpS) changed during the fungal life cycle, with the expression during the pre-symbiotic phase higher than during the symbiosis with the plant host. The heterologous expression in Escherichia coli demonstrated the functionality only for the YoeB-YefM pair. On the basis of these observations, we speculate that TA modules might help Cagg adapt to its intracellular habitat, coordinating its proliferation with the physiological state of the AMF host.

Arsenic (As) gets accumulated in plants via phosphorous transporters and water channels and interferes with nutrient and water uptake, adversely affecting growth and productivity. Although, Si and AM have been reported to combat arsenic stress, their comparative and interactive roles in ameliorating As V and As III toxicities have not been reported. Study evaluated effects of Si and Rhizophagus irregularis on growth, As uptake and yield under arsenate and arsenite stress in two pigeonpea genotypes (metal tolerant-Pusa 2002 and metal sensitive-Pusa 991). Higher As accumulation and translocation was observed in As III treated roots of Pusa 991 than those of Pusa 2002 when compared with As V. Roots were more negatively affected than shoots which led to a significant decline in nutrient uptake, leaf chlorophylls, and yield, with As III inducing more negative effects. Pusa 2002 established more effective mycorrhizal symbiosis and had higher biomass than Pusa 991. Si was more effective in inducing shoot biomass while AM inoculation significantly improved root biomass. AM enhanced Si uptake in roots and leaves in a genotype dependent manner. Combined application of Si and AM were highly beneficial in improving leaf water status, chlorophyll pigments, biomass, and productivity. Complete amelioration of negative impacts of both concentrations of As V and lower concentration of As III were recorded under +Si +AM in Pusa 2002. Results highlighted great potential of Si in improving growth and productivity of pigeonpea through R. irregularis under As V and As III stresses.

One of the bottlenecks in mycorrhiza research is that arbuscular mycorrhizal fungi (AMF) have to be cultivated with host plant roots. Some AMF species, such as Rhizophagus irregularis, can be grown in vitro on dual-compartment plates, where fungal material can be harvested from a fungus-only compartment. Plant roots often grow into this fungus compartment, and regular root trimming is required if the fungal material needs to be free of traces of plant material. Trimming also increases unwanted contamination by other microorganisms. We compared 22 different culture types and conditions to a widely used dual-compartment culture system that we refer to as the "standard system." We found two modified culture systems that allowed high spore production and low rates of contamination. We then compared the two modified culture systems with the standard system in more detail. In the two modified culture systems versus the standard system, a comparable number of spores were produced per plate, the necessity for root trimming was reduced, and there was significantly diminished contamination in the fungal compartment. A cost analysis showed that both modified culture systems were more economic than the standard culture system for the production of the same number of non-contaminated spores. The two modified culture systems provide an economic alternative for the production of contaminant-free fungal material which is ideal for studies requiring AMF DNA or RNA for genetics, genomics, and transcriptomic studies or for studies requiring relatively large amounts of fungal material for greenhouse experiments.

Arbuscular mycorrhizal (AM) fungi facilitate plant uptake of mineral nutrients and draw organic nutrients from the plant. Organic nutrients are thought to be supplied primarily in the form of sugars. Here we show that the AM fungus Rhizophagus irregularis is a fatty acid auxotroph and that fatty acids synthesized in the host plants are transferred to the fungus to sustain mycorrhizal colonization. The transfer is dependent on RAM2 (REQUIRED FOR ARBUSCULAR MYCORRHIZATION 2) and the ATP binding cassette transporter-mediated plant lipid export pathway. We further show that plant fatty acids can be transferred to the pathogenic fungus Golovinomyces cichoracerum and are required for colonization by pathogens. We suggest that the mutualistic mycorrhizal and pathogenic fungi similarly recruit the fatty acid biosynthesis program to facilitate host invasion., (Copyright © 2017, American Association for the Advancement of Science.)

Although there is a growing amount of evidence that arbuscular mycorrhizal fungi (AMF) influence the decomposition process, the extent of their involvement remains unclear. Therefore, given this knowledge gap, our aim was to test how AMF influence the soil decomposer communities. Dual compartment microcosms, where AMF (Glomus mosseae) were either allowed access (AM+) to or excluded (AM-) from forest soil compartments containing litterbags (leaf litter from Calophyllum polyanthum) were used. The experiment ran for six months, with destructive harvests at 0, 90, 120, 150, and 180 days. For each harvest we measured AMF colonization, soil nutrients, litter mass loss, and microbial biomass (using phospholipid fatty acid analysis (PLFA)). AMF significantly enhanced litter decomposition in the first 5 months, whilst delaying the development of total microbial biomass (represented by total PLFA) from T 150 to T 180 . A significant decline in soil available N was observed through the course of the experiment for both treatments. This study shows that AMF have the capacity to interact with soil microbial communities and inhibit the development of fungal and bacterial groups in the soil at the later stage of the litter decomposition (180 days), whilst enhancing the rates of decomposition., Competing Interests: The authors declare no competing financial interests.

Aims: This study aimed at evaluating the impact of seven plant growth-promoting rhizobacteria (PGPR) on root colonization and life cycle of Rhizophagus irregularis MUCL 41833 when co-entrapped in alginate beads., Methods and Results: Two in vitro experiments were conducted. The first consisted of the immobilization of R. irregularis and seven PGPR isolates into alginate beads to assess the effect of the bacteria on the pre-symbiotic growth of the fungus. In the second experiment, the best performing PGPR from experiment 1 was tested for its ability to promote the symbiotic development of the AMF in potato plantlets from three cultivars. Results showed that only one isolate identified as Pseudomonas plecoglossicida (R-67094) promoted germ tube elongation and hyphal branching of germinated spores during the pre-symbiotic phase of the fungus. This PGPR further promoted the symbiotic development of the AMF in potato plants., Conclusions: The co-entrapment of Ps. plecoglossicida R-67094 and R. irregularis MUCL 41833 in alginate beads improved root colonization by the AMF and its further life cycle under the experimental conditions., Significance and Impact of the Study: Co-entrapment of suitable AMF-PGPR combinations within alginate beads may represent an innovative technology that can be fine-tuned for the development of efficient consortia-based bioformulations., (© 2016 The Society for Applied Microbiology.)

Pesticide contamination of the environment can result from agricultural practices. Persistence of pesticide residues is a threat to the soil biota including plant roots and beneficial microorganisms, which support an important number of soil ecosystem services. Arbuscular mycorrhizal fungi (AMF) are key symbiotic microorganisms contributing to plant nutrition. In the present study, we assessed whether AMF could indicate eventual side effects of pesticides when directly applied to field soils. We evaluated the ecotoxicological impact of a cocktail of three commonly used agricultural pesticides (fenhexamid, folpel, deltamethrin) on the abundance and composition of the AMF community in vineyard (Montagne de Saint-Emilion) and arable (Martincourt) soils subjected to different agricultural practices. The dissipation of applied pesticides was monitored by multiresidual analyses to determine the scenario of exposure of the AMF community. Diversity analysis before application of the pesticide cocktail showed that the AMF communities of vineyard soils, subjected to mechanical weeding or grass cover, and of the arable soil subjected to intensive agriculture, were dominated by Glomerales. Ribotypes specific to each soil and to each agricultural practice in the same soil were found, with the highest abundance and diversity of AMF being observed in the vineyard soil with a grass-cover. The abundance of the global AMF community (Glomeromycota) and of three taxa of AMF (Funneliformis mosseae, Claroideoglomus etunicatum/C. claroideum) was evaluated after pesticide application. The abundance of Glomeromycota decreased in both soils after pesticide application while the abundance of Claroideoglomus and F. mosseae decreased only in the arable soil. These results show that higher doses of pesticide exposure did not affect the global abundance, but altered the composition, of the AMF community. Resilience of the AMF community composition was observed only in the vineyard soil, where F. mosseae was the most tolerant taxon to pesticide exposure., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Arbuscular mycorrhizal (AM) symbiosis associates most plants with fungi of the phylum Glomeromycota. The fungus penetrates into roots and forms within cortical cell branched structures called arbuscules for nutrient exchange. We discovered that miR171b has a mismatched cleavage site and is unable to downregulate the miR171 family target gene, LOM1 (LOST MERISTEMS 1). This mismatched cleavage site is conserved among plants that establish AM symbiosis, but not in non-mycotrophic plants. Unlike other members of the miR171 family, miR171b stimulates AM symbiosis and is expressed specifically in root cells that contain arbuscules. MiR171b protects LOM1 from negative regulation by other miR171 family members. These findings uncover a unique mechanism of positive post-transcriptional regulation of gene expression by miRNAs and demonstrate its relevance for the establishment of AM symbiosis., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Breeding selection of germplasm under fertilized conditions may reduce the frequency of genes that promote mycorrhizal associations. This study was developed to compare variability in mycorrhizal colonization and its effect on mycorrhizal dependency (MD) in improved soybean genotypes (I-1 and I-2) with differential tolerance to drought stress, and in unimproved soybean genotypes (UI-3 and UI-4). As inoculum, a mixed native arbuscular mycorrhizal fungi (AMF) was isolated from soybean roots, showing spores mostly of the species Funneliformis mosseae. At 20 days, unimproved genotypes followed by I-2, showed an increase in arbuscule formation, but not in I-1. At 40 days, mycorrhizal plants showed an increase in nodulation, this effect being more evident in unimproved genotypes. Mycorrhizal dependency, evaluated as growth and biochemical parameters from oxidative stress was increased in unimproved and I-2 since 20 days, whereas in I-1, MD increased at 40 days. We cannot distinguish significant differences in AMF colonization and MD between unimproved and I-2. However, variability among improved genotypes was observed. Our results suggest that selection for improved soybean genotypes with good and rapid AMF colonization, particularly high arbuscule/hyphae ratio could be a useful strategy for the development of genotypes that optimize AMF contribution to cropping systems.

Background and Aims: Arbuscular mycorrhizal (AM) fungi play a key role in the phosphate (P) uptake of many important crop species, but the mechanisms that control their efficiency and their contribution to the P nutrition of the host plant are only poorly understood., Methods: The P uptake and growth potential of two soybean genotypes that differ in their root architectural traits and P acquisition efficiency were studied after colonization with different AM fungi and the transcript levels of plant P transporters involved in the plant or mycorrhizal P uptake pathway were examined., Key Results: The mycorrhizal growth responses of both soybean genotypes ranged from highly beneficial to detrimental, and were dependent on the P supply conditions, and the fungal species involved. Only the colonization with Rhizophagus irregularis increased the growth and P uptake of both soybean genotypes. The expression of GmPT4 was downregulated, while the mycorrhiza-inducible P transporter GmPT10 was upregulated by colonization with R. irregularis Colonization with both fungi also led to higher transcript levels of the mycorrhiza-inducible P transporter GmPT9, but only in plants colonized with R. irregularis were the higher transcript levels correlated to a better P supply., Conclusions: The results suggest that AM fungi can also significantly contribute to the P uptake and growth potential of genotypes with a higher P acquisition efficiency, but that mycorrhizal P benefits depend strongly on the P supply conditions and the fungal species involved., (© The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The positive effects of arbuscular mycorrhizal fungi (AM) on the survival, growth and physiology of plants under various stress conditions have been widely recognized. However, whether sex-dependent susceptibility to AM colonization exists, which can induce a differential tolerance between the sexes to stress conditions, is still unclear. In this study, we investigated the effects of Glomus intraradices on Cd-stressed males and females of Populus deltoides (spiked with 10 mg Cd per kg dry substrate) in terms of morphology, physiology, biochemistry, ultrastructure, and toxin storage and translocation. Exposure to Cd promoted the colonization by G. intraradices in males, but not in females. Generally, females suffered more impairments than males in response to Cd stress, reflected by leaf symptoms, the extent of lipid peroxidation, and integrity of the cellular ultrastructure, whether they were inoculated or not. Inoculation with G. intraradices alleviated the phytotoxic effects of Cd in females by stimulating antioxidant enzymes, decreasing levels of reactive oxygen species (ROS) and restricting Cd transfer to the shoots. In contrast, these beneficial effects induced by AM were not detected in mycorrhizal males compared to non-mycorrhizal males, based on thiobarbituric acid-reactive substances (TBARS) and cellular ultrastructure. Inoculation with AM promoted Cd accumulation in males but not in females, and caused the sequestration of more toxic Cd in the root systems in both sexes. Therefore, our results suggest that inoculated males of P. deltoides are suitable candidates for phytostabilization in Cd-polluted soils, due to their higher accumulation ability and greater tolerance relative to inoculated females., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In a greenhouse pot experiment, lettuce plants (Lactuca sativa L.) were grown in a Hg-contaminated sandy soil with and without inoculation with arbuscular mycorrhizal fungi (AMF) (a commercial inoculum containing infective propagules of Rhizophagus irregularis and Funneliformis mosseae) amended with different rates of a humic acid (0, 1, and 2 g kg(-1) of soil), with the objective of verifying the synergistic effects of the two soil treatments on the Hg tolerance of lettuce plants. Our results indicated that the plant biomass was significantly increased by the combined effect of AMF and humic acid treatments. Addition of humic matter to soil boosted the AMF effect on improving the nutritional plant status, enhancing the pigment content in plant leaves, and inhibiting both Hg uptake and Hg translocation from the roots to the shoots. This was attributed not only to the Hg immobilization by stable complexes with HA and with extraradical mycorrhizal mycelium in soil and root surfaces but also to an improved mineral nutrition promoted by AMF. This work indicates that the combined use of AMF and humic acids may become a useful practice in Hg-contaminated soils to reduce Hg toxicity to crops.

A pot experiment was carried out to investigate the effects of water management and mycorrhizal inoculation on arsenic (As) uptake by two rice varieties, the As-resistant BRRI dhan 47 (B47) and As-sensitive BRRI dhan 29 (B29). Grain As concentration of B47 plants was significantly lower than that of B29, and grain As concentration of B47 was higher under flooding conditions than that under aerobic conditions. In general, mycorrhizal inoculation (Rhizophagus irregularis) had no significant effect on grain As concentrations, but decreased the proportion of inorganic arsenic (iAs) in grains of B47. The proportion of dimethylarsinic acid (DMA) in the total grain As was dramatically higher under flooding conditions. Results demonstrate that rice variety selection and appropriate water management along with mycorrhizal inoculation could be practical countermeasures to As accumulation and toxicity in rice grains, thus reducing health risks of As exposure in rice diets.

Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21% of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9%. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2-7.7, while it ranged from 25.8-48 to 35.6-53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2%). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots.

Long-lived radionuclides such as (90)Sr and (137)Cs can be naturally or accidentally deposited in the upper soil layers where they emit β/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a (137)Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.

In many deforested regions of the tropics, afforestation with native tree species could valorize a growing reservoir of degraded, previously overused and abandoned land. The inoculation of tropical tree seedlings with arbuscular mycorrhizal fungi (AM fungi) can improve tree growth and viability, but efficiency may depend on plant and AM fungal genotype. To study such effects, seven phylogenetically diverse AM fungi, native to Ecuador, from seven genera and a non-native AM fungus (Rhizophagus irregularis DAOM197198) were used to inoculate the tropical potential crop tree (PCT) species Handroanthus chrysanthus (synonym Tabebuia chrysantha), Cedrela montana, and Heliocarpus americanus. Twenty-four plant-fungus combinations were studied in five different fertilization and AMF inoculation treatments. Numerous plant growth parameters and mycorrhizal root colonization were assessed. The inoculation with any of the tested AM fungi improved seedling growth significantly and in most cases reduced plant mortality. Plants produced up to threefold higher biomass, when compared to the standard nursery practice. AM fungal inoculation alone or in combination with low fertilization both outperformed full fertilization in terms of plant growth promotion. Interestingly, root colonization levels for individual fungi strongly depended on the host tree species, but surprisingly the colonization strength did not correlate with plant growth promotion. The combination of AM fungal inoculation with a low dosage of slow release fertilizer improved PCT seedling performance strongest, but also AM fungal treatments without any fertilization were highly efficient. The AM fungi tested are promising candidates to improve management practices in tropical tree seedling production.

To exploit the potential benefits of mycorrhizas, we need to investigate the processes that occur in these symbiotic interactions, particularly in the arbuscular compartment where nutrients are exchanged between the plant and the fungus. Progress in this area is restricted due to the intricacy and complexity of this plant-fungus interface and many techniques that have been employed successfully in other plants and animal systems cannot be used. An effective approach to study processes in arbuscules is to examine transcript composition and dynamics. We applied laser capture microdissection (LCM) to isolate approximately 3000 arbuscules from Glomus intraradices colonised Me- dicago truncatula roots. Total RNA was extracted from microdissected arbuscules and subjected to T7 RNA polymerase-based linear amplification. Amplified RNA was then usedfor construction of a cDNA library. The presence and level of enrichment of mycorrhiza-specific transcripts was determined by quantitative Real-time and conventional PCR. To improve enrichment a cDNA library subtraction was performed. Complementation of yeast mutants deficient in the uptake of.potassium, phosphate, sulphate, amino acids, ammonium and of a Mn²⁺sensitive strain, demonstrates the functionality of our cDNA library.

Arbuscular mycorrhizal (AM) fungi provide benefits to host plants and show functional diversity, with evidence of functional trait conservation at the family level. Diverse communities of AM fungi ought therefore to provide increased benefits to the host, with implications for the management of sustainable agroecosystems. However, this is often not evident in the literature, with diversity saturation at low species number. Growth and nutrient uptake were measured in onions in the glasshouse on AM-free phosphorus (P)-poor soil, inoculated with between one and seven species of AM fungi in all possible combinations. Inoculation with AM fungi increased shoot dry weight as well as P and copper concentrations in shoots but reduced the concentration of potassium and sulphur. There was little evidence of increased benefit from high AM fungal diversity, and increasing diversity beyond three species did not result in significantly higher shoot weight or P or Cu concentrations. Species of Glomeraceae had the greatest impact on growth and nutrient uptake, while species of Acaulospora and Racocetra did not have a significant impact. Failure to show a benefit from high AM fungal diversity in this and other studies may be the result of experimental conditions, with the benefits of AM fungal diversity only becoming apparent when the host plant is faced with multiple stress factors. Replicating the complex interactions between AM fungi, the host plant and their environment in the laboratory in order to fully understand these interactions is a major challenge to AM research.

Arbuscular mycorrhizal fungi (AMF) are promoted as biofertilizers for sustainable agriculture. So far, most researchers have investigated the effects of AMF on plant growth under highly controlled conditions with sterilized soil, soil substrates or soils with low available P or low inoculum potential. However, it is still poorly documented whether inoculated AMF can successfully establish in field soils with native AMF communities and enhance plant growth. We inoculated grassland microcosms planted with a grass-clover mixture (Lolium multiflorum and Trifolium pratense) with the arbuscular mycorrhizal fungus Rhizoglomus irregulare. The microcosms were filled with eight different unsterilized field soils that varied greatly in soil type and chemical characteristics and indigenous AMF communities. We tested whether inoculation with AMF enhanced plant biomass and R. irregulare abundance using a species specific qPCR. Inoculation increased the abundance of R. irregulare in all soils, irrespective of soil P availability, the initial abundance of R. irregulare or the abundance of native AM fungal communities. AMF inoculation had no effect on the grass but significantly enhanced clover yield in five out of eight field soils. The results demonstrate that AMF inoculation can be successful, even when soil P availability is high and native AMF communities are abundant., (© 2015 John Wiley & Sons Ltd.)

The characterization of mycorrhizal status in hosts can be a good indicator of symbiotic associations in inoculation experiments or in ecological research. The most common microscopic-based observation methods, such as (i) the gridline intersect method, (ii) the magnified intersections method and (iii) the five-class system of Trouvelot were tested to find the most simple, easily executable, effective and objective ones and their appropriate parameters for characterization of mycorrhizal status. In a pot experiment, white clover (Trifolium repens L.) host plant was inoculated with 6 (BEG144; syn. Rhizophagus intradices) in pumice substrate to monitor the AMF colonization properties during host growth. Eleven (seven classical and four new) colonization parameters were estimated by three researchers in twelve sampling times during plant growth. Variations among methods, observers, parallels, or individual plants were determined and analysed to select the most appropriate parameters and sampling times for monitoring. The comparability of the parameters of the three methods was also tested. As a result of the experiment classical parameters were selected for hyphal colonization: colonization frequency in the first stage or colonization density in the later period, and arbuscular richness of roots. A new parameter was recommended to determine vesicule and spore content of colonized roots at later stages of symbiosis.