Functionality of arbuscular mycorrhizal fungi varies across different growth stages of maize under drought conditions.

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.

Rhizophagus intraradices and Azospirillum brasilense improve growth of herbaceous plants and soil biological activity in revegetation of a recovering coal-mining area.

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.

Arbuscular mycorrhizal fungi alter microbiome structure of rhizosphere soil to enhance maize tolerance to La.

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.

Relative roles of Arbuscular Mycorrhizae in establishing a correlation between soil properties, carbohydrate utilization and yield in Cicer arietinum L. under As stress.

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), ß-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.

Myristate can be used as a carbon and energy source for the asymbiotic growth of arbuscular mycorrhizal fungi.

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 ß-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.

Constitutive Overexpression of RAM1 Leads to an Increase in Arbuscule Density in Brachypodium distachyon.

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 lines 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.

The interaction between Rhizoglomus irregulare and hyphae attached phosphate solubilizing bacteria increases plant biomass of Solanum lycopersicum.

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.

Single-Spore Extraction for Genetic Analyses of Arbuscular Mycorrhizal Fungi.

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, a new species of arbuscular mycorrhizal fungi from semiarid regions in Mexico.

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.

Gibberellin Promotes Fungal Entry and Colonization during Paris-Type Arbuscular Mycorrhizal Symbiosis in Eustoma grandiflorum.

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.

Understanding interaction effect of arbuscular mycorrhizal fungi in rice under elevated carbon dioxide conditions.

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 CO2 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 CO2 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 CO2 concentration. Organic acids secretion in rice root increased in AMF-inoculated plants exposed to 700-ppm CO2 concentration. To understand the overall effect of CO2 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 CO2 concentration.

The role of in vitro cultivation on symbiotic trait and function variation in a single species of arbuscular mycorrhizal fungus.

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.

Arbuscular mycorrhiza fungi and related soil microbial activity drive carbon mineralization in the maize rhizosphere.

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.

Earthworms and mycorrhization increase copper phytoextraction by Canavalia ensiformis in sandy soil.

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.

Stimulation of asymbiotic sporulation in arbuscular mycorrhizal fungi by fatty acids.

Arbuscular mycorrhizal (AM) fungi are obligate symbionts that depend on living host plants to complete their life cycle1,2. 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 validus3. 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 acid4,5, 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 carbon6-9, 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.

Two herbicides, two fungicides and spore-associated bacteria affect Funneliformis mosseae extraradical mycelium structural traits and viability.

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.

Understanding dynamics of Rhizophagus irregularis ontogenesis in axenically developed coculture through basic and advanced microscopic techniques.

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.

Phosphorus forms affect the hyphosphere bacterial community involved in soil organic phosphorus turnover.

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 KH2PO4 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 ß-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.

Foliar Roundup application has minor effects on the compositional and functional diversity of soil microorganisms in a short-term greenhouse experiment.

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).

The impact of nanoparticles zero-valent iron (nZVI) and rhizosphere microorganisms on the phytoremediation ability of white willow and its response.

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.