Arbuscular Mycorrhizal Fungus Rhizophagus Irregularis Research Articles

Inoculation of plant growth promoting bacteria and arbuscular mycorrhizal fungi improve chickpea performance under water deficit conditions

Chickpea (Cicer arietinum L.) currently ranks second in the world's production of grain legumes and it is considered a cheap source of plant-based protein. In Mediterranean regions, predicted changes in climate are likely to further worsen drought stress and increase the economic vulnerability of chickpea production. Plant growth promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF) have the potential to improve plant growth and ameliorate the adverse effects of drought stress. The aim of this study was to assess the effects of inoculation with PGPB (Mesorhizobium sp., Burkholderia sp. and Pseudomonas sp.) and AMF (Rhizophagus irregularis, Funneliformis geosporum and Claroideoglomus claroideum) on the growth, grain yield and protein content of chickpea in a field experiment under different irrigation regimes throughout the growing season (100% water requirements (WR), 50% WR, 25% WR, 100% WR only during reproductive stages, and rainfed).Based on two years of results, the beneficial effects of co-inoculation (PGPB+AMF) on plant growth parameters of chickpea allow a cumulative grain yield of about 13,838 kg ha−1, resulting in an increase of 6% when as compared to a single inoculation and 24% over the non-inoculated plants. Plants inoculated with PGPB+AMF, and irrigated only during the reproductive stages, had the highest cumulative grain yield (18,157 kg ha−1), resulting in an increase of 16% and 237% over fully irrigated plants inoculated with PGPB+AMF and non-inoculated plants under rainfed conditions, respectively.In water-scarce environments, deficit irrigation only during the reproductive stage allows farmers to achieve higher yields with less water consumption, which, when combined with microbial inoculation, has the potential to benefit agricultural production of chickpea.

First Insights into the Effect of Mycorrhizae on the Expression of Pathogen Effectors during the Infection of Grapevine with Plasmopara viticola

Grapevine (Vitis vinifera L.), widely used for berry and wine production, is highly susceptible to the pathogenic oomycete Plasmopara viticola, the etiological agent of grapevine downy mildew disease. The method commonly used to prevent and control P. viticola infection relies on multiple applications of chemical fungicides. However, with European Union goals to lower the usage of such chemicals in viticulture there is a need to develop new and more sustainable strategies. The use of beneficial microorganisms with biocontrol capabilities, such as the arbuscular mycorrhizal fungi (AMF), has been pointed out as a viable alternative. With this study, we intended to investigate the effect of AMF colonization on the expression of P. viticola effectors during infection of grapevine. Grapevine plants were inoculated with the AMF Rhizophagus irregularis and, after mycorrhizae development, plants were infected with P. viticola. The expression of P. viticola RxLR effectors was analyzed by real-time PCR (qPCR) during the first hours of interaction. Results show that pre-mycorrhizal inoculation of grapevine alters the expression of several P. viticola effectors; namely, PvRxLR28, which presented decreased expression in mycorrhizal plants at the two time points post-infection tested. These results suggest that the pre-inoculation of grapevine with AMF could interfere with the pathogen’s ability to infect grapevine by modulation of pathogenicity effectors expression, supporting the hypothesis that AMF can be used to increase plant resistance to pathogens and promote more sustainable agriculture practices, particularly in viticulture.

Genetically Different Isolates of the Arbuscular Mycorrhizal Fungus Rhizophagus irregularis Induce Differential Responses to Stress in Cassava.

Water scarcity negatively impacts global crop yields and climate change is expected to greatly increase the severity of future droughts. The use of arbuscular mycorrhizal fungi (AMF) can potentially mitigate the effects of water stress in plants. Cassava is a crop that feeds approximately 800 million people daily. Genetically different isolates of the AMF R. irregularis as well as their clonal progeny have both been shown to greatly alter cassava growth in field conditions. Given that cassava experiences seasonal drought in many of the regions in which it is cultivated, we evaluated whether intraspecific variation in R. irregularis differentially alters physiological responses of cassava to water stress. In a first experiment, conducted in field conditions in Western Kenya, cassava was inoculated with two genetically different R. irregularis isolates and their clonal progeny. All cassava plants exhibited physiological signs of stress during the dry period, but the largest differences occurred among plants inoculated with clonal progeny of each of the two parental fungal isolates. Because drought had not been experimentally manipulated in the field, we conducted a second experiment in the greenhouse where cassava was inoculated with two genetically different R. irregularis isolates and subjected to drought, followed by re-watering, to allow recovery. Physiological stress responses of cassava to drought differed significantly between plants inoculated with the two different fungi. However, plants that experienced higher drought stress also recovered at a faster rate following re-watering. We conclude that intraspecific genetic variability in AMF significantly influences cassava physiological responses during water stress. This highlights the potential of using naturally existing variation in AMF to improve cassava tolerance undergoing water stress. However, the fact that clonal progeny of an AMF isolate can differentially affect how cassava copes with natural drought stress in field conditions, highlights the necessity to understand additional factors, beyond genetic variation, which can account for such large differences in cassava responses to drought.

Phytohormone production by the arbuscular mycorrhizal fungus Rhizophagus irregularis

Arbuscular mycorrhizal symbiosis is a mutualistic interaction between most land plants and fungi of the glomeromycotina subphylum. The initiation, development and regulation of this symbiosis involve numerous signalling events between and within the symbiotic partners. Among other signals, phytohormones are known to play important roles at various stages of the interaction. During presymbiotic steps, plant roots exude strigolactones which stimulate fungal spore germination and hyphal branching, and promote the initiation of symbiosis. At later stages, different plant hormone classes can act as positive or negative regulators of the interaction. Although the fungus is known to reciprocally emit regulatory signals, its potential contribution to the phytohormonal pool has received little attention, and has so far only been addressed by indirect assays. In this study, using mass spectrometry, we analyzed phytohormones released into the medium by germinated spores of the arbuscular mycorrhizal fungus Rhizophagus irregularis. We detected the presence of a cytokinin (isopentenyl adenosine) and an auxin (indole-acetic acid). In addition, we identified a gibberellin (gibberellin A4) in spore extracts. We also used gas chromatography to show that R. irregularis produces ethylene from methionine and the α-keto γ-methylthio butyric acid pathway. These results highlight the possibility for AM fungi to use phytohormones to interact with their host plants, or to regulate their own development.

Arbuscular mycorrhizal fungi can ameliorate salt stress in Elaeagnus angustifolia by improving leaf photosynthetic function and ultrastructure

Arbuscular mycorrhizal fungi (AMF) can form symbiosis with Elaeagnus angustifolia, allowing this species to tolerate salt stress. However, the physiological mechanism through which AMF improve E. angustifolia tolerance is still unclear. In this study, we examined E. angustifolia inoculated with AMF Rhizophagus irregularis (M) or inactivated inoculum (NM) under 0 and 300mM NaCl stress for the determination of photosynthetic gas exchange, pigment content, chlorophyll fluorescence, antioxidant capacity and chloroplast ultrastructural in leaves. Photosynthetic gas exchange parameters in the leaves of M and NM decreased significantly under salt stress, while the M treatment significantly reduced the effect of salt stress compared with NM. Various chlorophyll components in the M treatment were two- to three-fold higher than in NM, together with a much more complex chloroplast structure and higher number of plastoglobules. The total flavonoid and proline content in leaves of M increased significantly, while the concentration of malondialdehyde (MDA) decreased significantly under salt stress. Chlorophyll fluorescence data also showed good PSII function in the M treatment, together with salt stress reduction of photochemical reactions and sharp enhancements in non-photosynthetic quenching (NPQ). AMF inoculation ameliorated the inhibition on the actual PSII efficiency (ФPSII) and the photochemical quenching coefficient (qP ) by 10-15%. Our results clearly demonstrate that R. irregularis can improve the salt tolerance of plants by improving leaf photosynthetic performance, PSII function, antioxidant capacity and leaf chloroplast ultrastructure, and that E. angustifolia inoculated with AMF could enhance saline soil rehabilitation.

SeSaMe: Metagenome Sequence Classification of Arbuscular Mycorrhizal Fungi-associated Microorganisms

Arbuscular mycorrhizal fungi (AMF) are plant root symbionts that play key roles in plant growth and soil fertility. They are obligate biotrophic fungi that form coenocytic multinucleated hyphae and spores. Numerous studies have shown that diverse microorganisms live on the surface of and inside their mycelia, resulting in a metagenome when whole-genome sequencing (WGS) data are obtained from sequencing AMF cultivated in vivo. The metagenome contains not only the AMF sequences, but also those from associated microorganisms. In this study, we introduce a novel bioinformatics program, Spore-associated Symbiotic Microbes (SeSaMe), designed for taxonomic classification of short sequences obtained by next-generation DNA sequencing. A genus-specific usage bias database was created based on amino acid usage and codon usage of a three consecutive codon DNA 9-mer encoding an amino acid trimer in a protein secondary structure. The program distinguishes between coding sequence (CDS) and non-CDS, and classifies a query sequence into a genus group out of 54 genera used as reference. The mean percentages of correct predictions of the CDS and the non-CDS test sets at the genus level were 71% and 50% for bacteria, 68% and 73% for fungi (excluding AMF), and 49% and 72% for AMF (Rhizophagus irregularis), respectively. SeSaMe provides not only a means for estimating taxonomic diversity and abundance but also the gene reservoir of the reference taxonomic groups associated with AMF. Therefore, it enables users to study the symbiotic roles of associated microorganisms. It can also be applicable to other microorganisms as well as soil metagenomes. SeSaMe is freely available at www.fungalsesame.org.

Arbuscular Mycorrhizal Symbiosis Enhances Photosynthesis in the Medicinal Herb Salvia fruticosa by Improving Photosystem II Photochemistry.

We investigated the influence of Salvia fruticosa colonization by the arbuscular mycorrhizal fungi (AMF) Rhizophagus irregularis on photosynthetic function by using chlorophyll fluorescence imaging analysis to evaluate the light energy use in photosystem II (PSII) of inoculated and non-inoculated plants. We observed that inoculated plants used significantly higher absorbed energy in photochemistry (ΦPSII) than non-inoculated and exhibited significant lower excess excitation energy (EXC). However, the increased ΦPSII in inoculated plants did not result in a reduced non-regulated energy loss in PSII (ΦNO), suggesting the same singlet oxygen (1O2) formation between inoculated and non-inoculated plants. The increased ΦPSII in inoculated plants was due to an increased efficiency of open PSII centers to utilize the absorbed light (Fv’/Fm’) due to a decreased non-photochemical quenching (NPQ) since there was no difference in the fraction of open reaction centers (qp). The decreased NPQ in inoculated plants resulted in an increased electron-transport rate (ETR) compared to non-inoculated. Yet, inoculated plants exhibited a higher efficiency of the water-splitting complex on the donor side of PSII as revealed by the increased Fv/Fo ratio. A spatial heterogeneity between the leaf tip and the leaf base for the parameters ΦPSII and ΦNPQ was observed in both inoculated and non-inoculated plants, reflecting different developmental zones. Overall, our findings suggest that the increased ETR of inoculated S. fruticosa contributes to increased photosynthetic performance, providing growth advantages to inoculated plants by increasing their aboveground biomass, mainly by increasing leaf biomass.

Functional analysis of the OsNPF4.5 nitrate transporter reveals a conserved mycorrhizal pathway of nitrogen acquisition in plants

Low availability of nitrogen (N) is often a major limiting factor to crop yield in most nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi are beneficial symbionts of most land plants that enhance plant nutrient uptake, particularly of phosphate. A growing number of reports point to the substantially increased N accumulation in many mycorrhizal plants; however, the contribution of AM symbiosis to plant N nutrition and the mechanisms underlying the AM-mediated N acquisition are still in the early stages of being understood. Here, we report that inoculation with AM fungus Rhizophagus irregularis remarkably promoted rice (Oryza sativa) growth and N acquisition, and about 42% of the overall N acquired by rice roots could be delivered via the symbiotic route under N-NO3- supply condition. Mycorrhizal colonization strongly induced expression of the putative nitrate transporter gene OsNPF4.5 in rice roots, and its orthologs ZmNPF4.5 in Zea mays and SbNPF4.5 in Sorghum bicolor OsNPF4.5 is exclusively expressed in the cells containing arbuscules and displayed a low-affinity NO3- transport activity when expressed in Xenopus laevis oocytes. Moreover, knockout of OsNPF4.5 resulted in a 45% decrease in symbiotic N uptake and a significant reduction in arbuscule incidence when NO3- was supplied as an N source. Based on our results, we propose that the NPF4.5 plays a key role in mycorrhizal NO3- acquisition, a symbiotic N uptake route that might be highly conserved in gramineous species.

Ectomycorrhizal Fungal Inoculation of Sphaerosporella brunnea Significantly Increased Stem Biomass of Salix miyabeana and Decreased Lead, Tin, and Zinc, Soil Concentrations during the Phytoremediation of an Industrial Landfill.

Fast growing, high biomass willows (Salix sp.) have been extensively used for the phytoremediation of trace element-contaminated environments, as they have an extensive root system and they tolerate abiotic stressors such as drought and metal toxicity. Being dual mycorrhizal plants, they can engage single or simultaneous symbiotic associations with both arbuscular mycorrhizal (AM) fungi and ectomycorrhizal (EM) fungi, which can improve overall plant health and growth. The aim of this study was to test the effect of these mycorrhizal fungi on the growth and trace element (TE) extraction potential of willows. A field experiment was carried out where we grew Salix miyabeana clone SX67 on the site of a decommissioned industrial landfill, and inoculated the shrubs with an AM fungus Rhizophagus irregularis, an EM fungus Sphaerosporella brunnea, or a mixture of both. After two growing seasons, the willows inoculated with the EM fungus S. brunnea produced significantly higher biomass. Ba, Cd and Zn were found to be phytoextracted to the aerial plant biomass, where Cd presented the highest bioconcentration factor values in all treatments. Additionally, the plots where the willows received the S. brunnea inoculation showed a significant decrease of Cu, Pb, and Sn soil concentrations. AM fungi inoculation and dual inoculation did not significantly influence biomass production and soil TE levels.

Inoculation with Different Nitrogen-Fixing Bacteria and Arbuscular Mycorrhiza Affects Grain Protein Content and Nodule Bacterial Communities of a Fava Bean Crop

The introduction of nitrogen fixing bacteria (NFB) and arbuscular mycorrhizal fungi (AMF) into the soil is an advisable agricultural practice for the crop, since it enhances nutrient and water uptake and tolerance to biotic and abiotic stresses. The aim of this work was to study plant nutrition, biological nitrogen fixation (BNF) and crop yield and quality, after inoculating seeds with NFBs ((Rhizobium leguminosarum, Burkholderia cenocepacia, Burkholderia vietnamiensis)) and/or AMFs (Rhizophagus irregularis, Claroideoglomus etunicatum, Claroideoglomus claroideum and Funneliformis mosseae) in a fava bean crop in two seasons. The composition of the nodule bacterial community was evaluated by the high-throughput sequencing analysis of bacterial 16 S rRNA genes. It was found that microbial inoculation accompanied by a 20% decrease in mineral fertilization had no significant effect on crop yield or the nutritional characteristics compared with a non-inoculated crop, except for an increase in the grain protein content in inoculated plants. None of the inoculation treatments increased biological nitrogen fixation over a non-inoculated level. The bacterial rRNA analysis demonstrated that the genus Rhizobium predominated in all nodules, both in inoculated and non-inoculated treatments, suggesting the previous presence of these bacteria in the soil. In our study, inoculation with Rhizobium leguminosarum was the most effective treatment for increasing protein content in seeds, while Burkholderia sp. was not able to colonise the plant nodules. Inoculation techniques used in fava beans can be considered an environmentally friendly alternative, reducing the input of fertilizers, while maintaining crop yield and quality, with the additional benefit of increasing the grain protein content. However, further research is required on the selection and detection of efficient rhizobial strains under local field conditions, above all those related to pH and soil type, in order to achieve superior nitrogen-fixing bacteria.

Common Mycorrhizal Network Induced JA/ET Genes Expression in Healthy Potato Plants Connected to Potato Plants Infected by Phytophthora infestans.

Most plants are connected belowground via common mycorrhizal networks (CMNs). In their presence, the transmission of warning signals from diseased to uninfected plants has been reported. However, current studies have all been conducted in pots making it difficult to discriminate direct from indirect contribution of hyphae to the transmission of the signals. Here, we conducted an in vitro study with potato plantlets connected by a CMN of the arbuscular mycorrhizal fungus Rhizophagus irregularis. The plantlets were grown in physically separated compartments and their connection ensured only by the CMN. The donor potato plantlets were infected by Phytophthora infestans and defense genes analyzed 24, 48 and 120 h post-infection (hpi) in the uninfected receiver potato plantlets. Twenty-four hpi by the pathogen, PAL, PR-1b, ERF3, and LOX genes were significantly upregulated, whereas no significant transcript variation was noticed 48 and 120 hpi. The exact nature of the warning signals remains unknown but was not associated to microorganisms other than the AMF or to diffusion mechanisms through the growth medium or induced by volatile compounds. The defense response appeared to be transitory and associated with the jasmonic acid or ethylene pathway. These findings demonstrate the direct involvement of hyphae in the transmission of warning signals from diseased to uninfected potato plantlets and their indubitable role in providing a route for activating defense responses in uninfected plants.

Impact of arbuscular mycorrhizal fungus (<i>Rhizophagus irregularis</i>) on disease symptoms caused by the ascomycete fungus (<i>Mycosphaerella fijiensis M.</i>) in Black Sigatoka-resistant banana plantain

Banana (Musa spp.) is grown throughout the tropical and subtropical areas. The Black Sigatoka, however, represents a major threat to world production. This study evaluates the effect of the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis MUCL 41833 on four plantain cultivars with different resistance against Mycosphaerella fijiensis. The mycorrhized banana plantlets of four cultivars were grown under greenhouse and the effect of AMF on micropropagated banana plantlets was evaluated. After 35 days, the height and the diameter of the pseudostem, then the leaf area were measured. The significant effect of the cultivar on the height of the pseudostem and the leaf area and significant effect of M. fijiensis on the diameter of the pseudostem, the height of the pseudostem and the leaf area were obtained. Plants infected with M. fijiensis show more symptoms of the disease than healthy plants. Symptoms were delayed in bananas inoculated with AMF compared to bananas not inoculated with AMF. The disease progressed more rapidly in cultivars Bâtard and C292 than in cultivars CRBP39 and F568. These results suggest that AMF may decrease symptoms of Sigatoka, at the early stage of infection with M. fijiensis. AMF may be a promising tool for the pre-adaptation of micropropagated banana plantlets.
 Keywords: Plantain, Rhizophagus irregularis, arbuscular mycorrhiza fungi, Mycosphaerella fijiensis, disease control method.

Comparative effect of Rhizophagus irregularis strain on cassava root development and Phosphorus uptake under acidic soils conditions of Walungu territory, Eastern DR Congo.

Objective: In the highlands of South-Kivu province of DR Congo, cassava is grown on marginal land not suitable for other crops. Walungu territory for instance is dominated by acidic soils and has the highest nutrient depletion rate in the country. On such types of soil, nutrient depletion is accompanied by a decrease in the availability of phosphorus and many other nutrients. The use of Arbuscular mycorrhizal fungi (AMF) would therefore be an alternative to improve the phosphate nutrition of cassava under different soil conditions. Methodology and results: The experiment reported in the current study was conducted in pots under semi-controlled conditions. Ferrallitic soil from Walungu was used. Soil was sterilized or not and then inoculated with the AMF Rhizophagus irregularis (RI) strain. Results showed that inoculation with an exotic AMF strain (Rhizophagus iregularis) has significantly increased root abundance (number of roots) in both sterilized and unsterilized soil and root dry weight in sterilized soil only. However, in unsterilized soil, root dry weight decreased with AMF inoculation. In Walungu acidic soils, P supply could significantly influence the effect of mycorrhizal inoculation on root development and root abundance. AMF inoculation has significantly increased the shoot P concentration when P was supplied. In sterilized soil, mycorrhizal inoculation had a depressant effect on soil phosphorus concentration, especially when P was not supplied. Conclusion and application of results: Our results suggest that the introduced Rhizophagus iregularis strain increases P uptake in the rhizospheric soil, especially when phosphorus is not applied to the sterilized or unsterilized soil. The use of the Rhizophagus irregularis strain as a biofertilizer could improve phosphorus nutrition and root development in cassava. Keywords: Arbuscular mycorrhizae, Rhizophagus irregularis, P uptake, Cassava, Ferrallitic soil

Rhizophagus irregularis MUCL 41,833 Association with Green Cuttings of Prunus sp. Rootstocks

Sour cherry ‘Latvijas Zemais’ (Prunus cerasus) is a promising dwarfing rootstock for sweet cherries in Latvia, but low growing rate of newly propagated sour cherry plants limits their use. Plum rootstock PU 20651 was selected from hybrids of P. cerasifera and P. salicina ssp. usuriensis recently. The aim of our study was to estimate the effect of arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis MUCL 41833 (Ri) on the growth and P content of rooted green cuttings of sour cherry ‘Latvijas Zemais’ 52, cherry rootstock Gisela 5 (P. cerasus x P. canescens), and plum rootstock PU 20651. The cuttings were planted in peat substrate and grown in plastic tunnel equipped with automatic air moistening system and controllable floor heating. We determined growth parameters and P content in shoots and roots, intensity of mycorrhizal colonization, and frequency of mycorrhiza in root system. In all three Prunus sp. genotypes, the inoculation with Ri resulted in the root colonization. PU 20651 had highest level of colonization at all arbuscular mycorrhiza (AM) estimation parameters. A common effect of colonization was increased diameter of root collar and positive correlation between the leaf number and AM parameters. The effect of Ri on plant biomass and P content depended on the genotype. Beneficial effect of AM on root mass and root P content was detected in ‘Latvijas Zemais’ 52. However, no AM effects on P content were found in Gisela 5, but little negative impact of AM on shoot mass and shoot P content—in PU 20651.

ALLEVIATION OF SOIL SALINITY ON PHYSIOLOGICAL AND AGRONOMIC TRAITS OF RICE CULTIVARS USING Arbuscular mycorrhizal fungi AND Pseudomonas STRAINS UNDER FIELD CONDITIONS

Rice cultivation in Iran has a special place and its cultivation area is about 640,000 ha. Soil salinization is one of the most factor in reducing of grain yields and cultivation of paddy rice. Currently, half of Iran's cultivated lands (9.5 million ha) are affected by salinity, which has a major impact on crop yields. For this purpose, an field experiment conducted with the use of microorganisms (Pseudomonas putida strain S34, Pseudomonas fluorescens strain R167, Arbuscular mycorrhizal fungi (AMF) (Rhizophagus irregularis), co-inoculation of P.putida + P.fluorescens + AMF, and a non-inoculated treatment as control) on two rice cultivars (Hashemi and Gilane) in saline and normal soil in north of Iran during the 2018 and 2019. The results showed interactions of microorganisms and soil condition could alleviate the adverse effects of salinity by decreasing H2O2 (14-71% in normal soil and 44-281% in saline soil) and increasing catalase (CAT) activity (39.5-65% in Gilane and 45-61% in Hashemi), proline (17-46% in Gilane and 13.5-52% in Hashemi) and nitrogen (N) concentrations (12-27% in normal soil and 7-23% in saline soil) of leaves. Also microorganisms significantly increased tiller, panicle and grain number plant-1, and biological and grain yield (23-44.5% and 32.5-56% in normal and saline soil, respectively). Furthermore, the greatest mitigating effects were observed in treated by P.putida + P.fluorescens + AMF plants. This study indicated that P.putida + P.fluorescens + AMF synergistically mitigate harmful impacts of soil salinity in rice cultivars.

DES2 is a fatty acid Δ11 desaturase capable of synthesizing palmitvaccenic acid in the arbuscular mycorrhizal fungus Rhizophagusirregularis.

Arbuscular mycorrhizal (AM) fungi are oleaginous organisms, and the most abundant fatty acyl moiety usually found in their lipids is palmitvaccenic acid (16:1Δ11cis). However, it is not known how this uncommon fatty acid species is made. Here, we have cloned two homologues of lepidopteran fatty acyl‐coenzyme A Δ11 desaturases from the AM fungus Rhizophagus irregularis. Both enzymes, DES1 and DES2, are expressed in intraradical mycelium and can complement the unsaturated fatty acid‐requiring auxotrophic growth phenotype of the Saccharomyces cerevisiae ole1Δ mutant. DES1 expression leads almost exclusively to oleic acid (18:1Δ9cis) production, whereas DES2 expression results in the production of 16:1Δ11cis and vaccenic acid (18:1Δ11cis). DES2 therefore encodes a Δ11 desaturase that is likely to be responsible for the synthesis of 16:1Δ11cis in R. irregularis.

The effect of zinc fertilisation and arbuscular mycorrhizal fungi on grain quality and yield of contrasting barley cultivars.

Zinc is essential for the functioning of many enzymes and plant processes and the malting process. Arbuscular mycorrhizal fungi (AMF) can improve zinc (Zn) uptake in the important cereal crop barley (Hordeum vulgare) on Zn-deficient soils. Here we investigated the impacts of Zn fertilisation and AMF on the yield and grain quality of malting barley cultivars. Five barley genotypes were inoculated or not with the AMF Rhizophagus irregularis, and grown in pots either fertilised with Zn or not. Measurements of Zn nutrition and yield were made for all cultivars. Further analyses of grain biochemical composition, including starch, β-glucan and arabinoxylan contents, and analysis of ATR-MIR spectra were made in two contrasting cultivars. Mycorrhizal colonisation generally resulted in decreased biomass, but increased grain dimensions and mean grain weight. Barley grain yield and biochemical qualities were highly variable between cultivars, and the ATR-MIR spectra revealed grain compositional differences between cultivars and AMF treatments. Mycorrhizal fungi can affect barley grain Zn concentration and starch content, but grain biochemical traits including β-glucan and arabinoxylan contents were more conserved by the cultivar, and unaffected by AMF inoculation. The ATR-MIR spectra revealed that there are other grain characteristics affected by AMF that remain to be elucidated.

Arbuscular Mycorrhizal Fungi Can Compensate for the Loss of Indigenous Microbial Communities to Support the Growth of Liquorice (Glycyrrhiza uralensis Fisch.).

Soil microorganisms play important roles in nutrient mobilization and uptake of mineral nutrition in plants. Agricultural management, such as soil sterilization, can have adverse effects on plant growth because of the elimination of indigenous microorganisms. Arbuscular mycorrhizal (AM) fungi are one of the most important beneficial soil microorganisms for plant growth. However, whether AM fungi can compensate for the loss of indigenous microbial communities to support plant growth and metabolism is largely unknown. In this study, a pot experiment was conducted to investigate the effects of AM fungi on plant growth and secondary metabolism in sterilized and unsterilized soil. We used liquorice (Glycyrrhiza uralensis Fisch.), an important medicinal plant as the host, which was inoculated with the AM fungus Rhizophagus irregularis or not and grown in unsterilized or sterilized soil. Plant photosynthesis traits, plant growth and nutrition level, concentrations of the secondary metabolites, and expression levels of biosynthesis genes were determined. The results showed that soil sterilization decreased plant growth, photosynthesis, and glycyrrhizin and liquiritin accumulation, and moreover, downregulated the expression of related biosynthesis genes. Inoculation with R. irregularis in sterilized soil offset the loss of indigenous microbial communities, resulting in plant growth and glycyrrhizin and liquiritin concentrations similar to those of plants grown in unsterilized soil. Thus, AM fungi could compensate for the loss of indigenous microbial communities by soil sterilization to support plant growth and secondary metabolism.

Imbalanced Regulation of Fungal Nutrient Transports According to Phosphate Availability in a Symbiocosm Formed by Poplar, Sorghum, and Rhizophagus irregularis.

In arbuscular mycorrhizal (AM) symbiosis, key components of nutrient uptake and exchange are specialized transporters that facilitate nutrient transport across membranes. As phosphate is a nutrient and a regulator of nutrient exchanges, we investigated the effect of P availability to extraradical mycelium (ERM) on both plant and fungus transcriptomes and metabolomes in a symbiocosm system. By perturbing nutrient exchanges under the control of P, our objectives were to identify new fungal genes involved in nutrient transports, and to characterize in which extent the fungus differentially modulates its metabolism when interacting with two different plant species. We performed transportome analysis on the ERM and intraradical mycelium of the AM fungus Rhizophagus irregularis associated to Populus trichocarpa and Sorghum bicolor under high and low P availability in ERM, using quantitative RT-PCR and Illumina mRNA-sequencing. We observed that mycorrhizal symbiosis induces expression of specific phosphate and ammonium transporters in both plants. Furthermore, we identified new AM-inducible transporters and showed that a subset of phosphate transporters is regulated independently of symbiotic nutrient exchange. mRNA-Sequencing revealed that the fungal transportome was not similarly regulated in the two host plant species according to P availability. Mirroring this effect, many plant carbohydrate transporters were down-regulated in P. trichocarpa mycorrhizal root tissue. Metabolome analysis revealed further that AM root colonization led to a modification of root primary metabolism under low and high P availability and to a decrease of primary metabolite pools in general. Moreover, the down regulation of the sucrose transporters suggests that the plant limits carbohydrate long distance transport (i.e. from shoot to the mycorrhizal roots). By simultaneous uptake/reuptake of nutrients from the apoplast at the biotrophic interface, plant and fungus are both able to control reciprocal nutrient fluxes.

The extraradical proteins of Rhizophagus irregularis: A shotgun proteomics approach

Arbuscular Mycorrhizal fungi (AMF, Glomeromycota) form obligate symbiotic associations with the roots of most terrestrial plants. Our understanding of the molecular mechanisms enabling AMF propagation and AMF-host interaction is currently incomplete. Analysis of AMF proteomes could yield important insights and generate hypotheses on the nature and mechanism of AMF-plant symbiosis. Here, we examined the extraradical mycelium proteomic profile of the arbuscular mycorrhizal fungus Rhizophagus irregularis grown on Ri T-DNA transformed Chicory roots in a root organ culture setting. Our analysis detected 529 different peptides that mapped to 474 translated proteins in the R. irregularis genome. R. irregularis proteome was characterized by a high proportion of proteins (9.9 % of total, 21.4 % of proteins with functional prediction) mediating a wide range of signal transduction processes, e.g. Rho1 and Bmh2, Ca-signaling (calmodulin, and Ca channel protein), mTOR signaling (MAP3K7, and MAPKAP1), and phosphatidate signaling (phospholipase D1/2) proteins, as well as members of the Ras signaling pathway. In addition, the proteome contained an unusually large proportion (53.6 %) of hypothetical proteins, the majority of which (85.8 %) were Glomeromycota-specific. Forty-eight proteins were predicted to be surface/membrane associated, including multiple hypothetical proteins of yet-unrecognized functions. However, no evidence for the overproduction of specific proteins, previously implicated in promoting soil health and aggregation was obtained. Finally, the comparison of R. irregularis proteome to previously published AMF proteomes identified a core set of pathways and processes involved in AMF growth. We conclude that R. irregularis growth on chicory roots requires the activation of a wide range of signal transduction pathways, the secretion of multiple novel hitherto unrecognized Glomeromycota-specific proteins, and the expression of a wide array of surface-membrane associated proteins for cross kingdom cell-to-cell communications.