Modulation of cherry tomato performances in response to molybdenum biofortification and arbuscular mycorrhizal fungi in a soilless system.

Molybdenum (Mo) is a crucial microelement for both, humans and plants. The use of agronomic biofortification techniques can be an alternative method to enhance Mo content in vegetables. Concomitantly, arbuscular mycorrhizal fungi (AMF) application is a valuable strategy to enhance plant performances and overcome plant abiotic distresses such as microelement overdose. The aim of this research was to estimate the direct and/or indirect effects of Mo supply at four doses [0.0, 0.5 (standard dose), 2.0 or 4.0 µmol L-1], alone or combined with AMF inoculation, on plant performances. In particular, plant height and first flower truss emission, productive features (total yield, marketable yield and average marketable fruit weight) and fruit qualitative characteristics (fruit dry matter, soluble solids content, titratable acidity, ascorbic acid, lycopene, polyphenol, nitrogen, copper, iron and molybdenum) of an established cherry tomato genotype cultivated in soilless conditions were investigated. Moreover, proline and malondialdehyde concentrations, as well as Mo hazard quotient (HQ) in response to experimental treatments were determined. A split-plot randomized experimental block design with Mo dosages as plots and +AMF or -AMF as sub-plots was adopted. Data revealed that AMF inoculation enhanced marketable yield (+50.0 %), as well as some qualitative traits, such as fruit soluble solids content (SSC) (+9.9 %), ascorbic acid (+7.3 %), polyphenols (+2.3 %), and lycopene (+2.5 %). Molybdenum application significantly increased SSC, polyphenols, fruit Mo concentration (+29.0 % and +100.0 % in plants biofortified with 2.0 and 4.0 µmol Mo L-1 compared to those fertigated with the standard dose, respectively) and proline, whereas it decreased N (-25.0 % and -41.6 % in plants biofortified with 2.0 and 4.0 µmol Mo L-1 compared to those fertigated with the standard dose, respectively). Interestingly, the application of AMF mitigated the detrimental effect of high Mo dosages (2.0 or 4.0 µmol L-1). A pronounced advance in terms of plant height 45 DAT, fruit lycopene concentration and fruit Fe, Cu and Mo concentrations was observed when AMF treatment and Mo dosages (2.0 or 4.0 µmol Mo L-1) were combined. Plants inoculated or not with AMF showed an improvement in the hazard quotient (HQ) in reaction to Mo application. However, the HQ - for a consumption of 200 g day-1 of biofortified cherry tomato - remained within the safety level for human consumption. This study suggests that Mo-implementation (at 2.0 or 4.0 µmol L-1) combined with AMF inoculation could represent a viable cultivation protocol to enhance yield, produce premium quality tomato fruits and, concomitantly, improve Mo dose in human diet. In the light of our findings, further studies on the interaction between AMF and microelements in other vegetable crops are recommended.

Variability in Chemical Composition and Biochemical Activities of Mentha x piperita L. Essential Oil, in Response to Mycorrhizal Symbiosis and Heavy Metal Stress.

The present paper highlights the effect of Pb/Cd-stress and/or mycorrhizal colonization by Glomus Intraradices on yield, chemical composition, cytotoxicity and antimicrobial activity of Mentha x piperita L. essential oil. Our findings showed that mycorrhizal colonization could be used to improve the essential oil yield of M. x piperita, either in non-stressed or Pb/Cd-stressed plants. GC-MS analysis revealed three chemotypes: linalool/pulegone (32.6/30.8 %) chemotype in essential oils of non-mycorrhizal Pb-stressed plants, menthone/menthyl acetate (30.3/25.1 %) chemotype in essential oils of non-mycorrhizal Cd-stressed plants and menthol (44.6 %) chemotype in essential oils of non-mycorrhizal non-stressed plants, mycorrhizal non-stressed plants and mycorrhizal Pb/Cd-stressed plants. The cytotoxicity of M. x piperita essential oil, evaluated by brine shrimp lethality bioassay, was increased in presence of Pb/Cd-stress (from 379.58 to 72.84 µm/mL) and decreased in mycorrhizal plants (from 379.58 to 482.32 µm/mL). The antimicrobial activity of M. x piperita essential oil, evaluated by disc diffusion method and determination of Minimum Inhibitory Concentration against ten microorganisms, was enhanced by the mycorrhizal colonization and deceased by the Pb/Cd-stress. In conclusion, the inoculation of medicinal plants with mycorrhizal fungi is a real avenue for alleviating abiotic stress and/or increasing the quantity and quality of secondary metabolites in terms of biological activities.

Diversity of arbuscular mycorrhizal fungi in seleniferous soils and their role in plant growth promotion.

The present study aimed to investigate the molecular diversity of arbuscular mycorrhizal fungi (AMF) in natural seleniferous soils and their role in protecting plants from Se toxicity. The genomic DNA extracted from maize roots grown in seleniferous and non-seleniferous regions was amplified using AMF-specific primers by nested PCR. The 1.5 kb amplicon spanning pSSU-ITS-pLSU of 18S rRNA of AMF was deciphered using the Illumina Miseq Next Generation Sequencing (NGS) technique. A total of 17 AMF species from the seleniferous region and 18 AMF species from the non-seleniferous region were identified. The number of reads of Glomus irregularis, G. custos, and G. intraradices was higher in seleniferous soil than in non-seleniferous soil, indicating their tolerance to Se. A consortium of Se-tolerant AMF inoculum was prepared and inoculated to maize plants, grown in natural seleniferous soils. AMF-inoculated plants had healthy growth with higher root, shoot, and grain biomass than non-AMF-inoculated plants. AMF inoculation leads to higher Se accumulation in roots but lesser Se accumulation in shoots and seeds of inoculated maize plants as compared to control plants. Present study results suggest that AMF species from seleniferous soils have the potential to be used as biofertilizers to improve plant growth and tolerate Se toxicity in seleniferous soils.

Effects of Arbuscular Mycorrhizal Fungi on Robinia pseudoacacia L. Growing on Soils Contaminated with Heavy Metals.

Arbuscular mycorrhizal fungi (AMF) have been shown to assist plants in increasing metal tolerance and accumulation in heavy metal (HM)-contaminated soils. Herein, a greenhouse pot experiment was conducted to assess the interactions of growth substrates (S1, S2, and S3, respectively) with various HM contamination and nutrient status sampling from a typical contaminated soil and tailings in Shuikoushan lead/zinc mining in Hunan province, China, and AMF inoculation obtained from plants in uncontaminated areas (Glomus mosseae, Glomus intraradices, and uninoculated, respectively) on the biomass and uptake of HMs and phosphorus (P) by the black locust plant (Robinia pseudoacacia L.). The results indicated that the inoculation with AMF significantly enhanced the mycorrhizal colonization of plant roots compared with the uninoculated treatments, and the colonization rates were found to be higher in S1 and S2 compared with S3, which were characterized with a higher nutrient availability and lead concentration. The biomass and heights of R. pseudoacacia were significantly increased by AMF inoculation in S1 and S2. Furthermore, AMF significantly increased the HM concentrations of the roots in S1 and S2 but decreased the HM concentrations in S3. Shoot HM concentrations varied in response to different AMF species and substrate types. Mycorrhizal colonization was found to be highly correlated with plant P concentrations and biomass in S1 and S2, but not in S3. Moreover, plant biomass was also significantly correlated with plant P concentrations in S1 and S2. Overall, these findings demonstrate the interactions of AMF inoculation and growth substrates on the phytoremediation potential of R. pseudoacacia and highlights the need to select optimal AMF isolates for their use in specific substrates for the remediation of HM-contaminated soil.

Arbuscular Mycorrhizal Symbiosis Improves Ex Vitro Acclimatization of Sugarcane Plantlets (Saccharum spp.) under Drought Stress Conditions.

The symbiotic associations between arbuscular mycorrhizal fungi (AMF) and plants can induce drought stress tolerance. In this study, we evaluated the effect of Glomus intraradices, a mycorrhizal fungus, on the ex vitro development and survival of sugarcane plantlets subjected to drought stress during the acclimatization stage of micropropagation. In vitro obtained sugarcane plantlets (Saccharum spp. cv Mex 69-290) were inoculated with different doses of G. intraradices (0, 100, and 200 spores per plantlet) during greenhouse acclimatization. Sixty days after inoculation, plantlets were temporarily subjected to drought stress. We evaluated the survival rate, total chlorophyll, total protein, carotenoids, proline, betaine glycine, soluble phenolic content, and antioxidant capacity every 3 days for 12 days. Symbiotic interaction was characterized by microscopy. Our results showed that the survival rate of inoculated plants was higher in 45% than the treatment without mycorrhizae. Total chlorophyll, protein, proline, betaine glycine content, and antioxidant capacity were increased in AMF inoculated plants. The soluble phenolic content was higher in non-inoculated plants than the treatment with mycorrhizae during the drought stress period. Microscopy showed the symbiotic relationship between plant and AMF. The early inoculation of 100 spores of G. intraradices per sugarcane plantlet during the acclimatization stage could represent a preconditioning advantage before transplanting into the field and establishing basic seedbeds.

Uptake of different pharmaceuticals in soil and mycorrhizal artichokes from wastewater.

The irrigation with treated wastewater is among the main anthropogenic sources for the release of pharmaceuticals (PhACs) into the soils and their translocation into crops, with possible toxic and adverse effects on humans. The arbuscular mycorrhizal fungi (AMF) can be employed for the reduction of organic soil pollutants, even if their efficiency depends on the mycorrhizal fungi, the plant colonized, and the type and concentration of the contaminant. This study aimed to evaluate the uptake of PhACs from wastewaters of different qualities used for the irrigation of mycorrhizal artichoke plants, the presence in their edible parts and the role of the arbuscular mycorrhizal fungi. The research was carried out on artichoke plants not inoculated and inoculated with two different AMF and irrigated with treated wastewater (TW), groundwater (GW) or GW spiked with different and selected PhACs (SGW). The inocula were a crude inoculum of Septoglomus viscosum (MSE) and a commercial inoculum of Glomus intraradices and Glomus mosseae (MSY). The results of the present study showed that carbamazepine and fluconazole were found in the artichoke only with SGW irrigation. The mycorrhizal plants showed a reduction of the pharmaceutical's uptake, and within the AMF, MSE was more effective in preventing their absorption and translocation.

Arbuscular Mycorrhizal Fungi Induce Tolerance to Salinity Stress in Taro Plantlets (Colocasia esculenta L. Schott) during Acclimatization.

Soil salinity is a problem that affects soil fertility and threatens agri-food crop production worldwide. Biotechnology, through plant micropropagation and the use of biofertilizers such as arbuscular mycorrhizal fungi (AMF), is an alternative to increase productivity and induce tolerance to salinity stress in different crops. This study aimed to evaluate the effect of different doses of the fungus Glomus intraradices on the ex vitro development of taro (Colocasia esculenta L. Schott cv. Criolla) plantlets under salinity stress during the acclimatization stage. In vitro-obtained C. esculenta plantlets were inoculated at different doses (0, 100, and 200 spores per plantlet) of G. intraradices during acclimatization. At 60 d of acclimatization in the greenhouse, plantlets were exposed to 100 mM NaCl salinity stress for 10 d. After the stress period, plantlet development, colonization percentage, and biomass were evaluated. In addition, the content of chlorophyll, carotenoids, proteins, proline, glycine-betaine, soluble phenols, and antioxidant capacity were quantified. The results showed differences in the developmental, physiological, and biochemical variables evaluated; however, no changes in total protein content were observed. Spore colonization showed that the symbiotic association has positive effects on the development of plantlets with or without salinity stress. This symbiotic interaction contributes to salinity stress tolerance in C. esculenta plantlets. The early application of AMF in in vitro-obtained taro plantlets is an alternative to increase or maintain the productivity of this crop in saline soils.

Assessment of mycorrhizal association of a threatened medicinal plant Clerodendrum indicum (L.) O. Kuntze (Verbenaceae) in different ecological variations.

Mycorrhizae association is reported to enhance the survivability of the host plant under adverse environmental conditions. The present study aims to explore the mycorrhizal association in the roots of different ecotypes of a threatened medicinal plant, Clerodendrum indicum (L.) O. Kuntze (Verbenaceae), collected from W.B., India, which correlates the degree of root colonization to the nutritional status of the native soil. Ten ecotypes of C. indicum having diverse morphological variations were collected. The mycorrhizae were characterized by both morphological and molecular methods. The nutritional status of the native soils was estimated. The study revealed that all the ecotypes have an association with mycorrhizal forms like hyphae, arbuscules, and vesicles. The molecular analysis showed Glomus intraradices and Rhizophagus irregularis as the associated arbuscular mycorrhizal fungi (AMF). A significant variation in arbuscule and vesicle formation was found growing in the varied nutritional statuses concerning soil parameters. The arbuscule was found negatively correlated with pH, conductivity, and potassium and positively correlated with organic carbon, nitrogen, and phosphorus. The vesicle was found positively correlated with pH, organic carbon, and potassium and negatively correlated with conductivity, nitrogen, and phosphorus. The interaction between conductivity: nitrogen, conductivity: phosphorus, organic-carbon: nitrogen, and pH: conductivity was significant in influencing vesicle formation. However, none of the interactions between parameters was found significant in influencing arbuscule formation. Thus, the study concludes that G. intraradices and R. irregularis are the principle mycorrhizae forming the symbiotic association with the threatened medicinal plant, C. indicum. They form vesicles and arbuscules based on their soil nutritive factors. Therefore, a large-scale propagation through a selective AMF association would help in the conservation of this threatened species from extinction.

Assessment of Silicon- and Mycorrhizae- Mediated Constitutive and Induced Systemic Resistance in Rice, Oryza sativa L., against the Fall Armyworm, Spodoptera frugiperda Smith.

Induced resistance provides protection in plants against insect herbivores. Silicon and mycorrhizae often prime plant defenses and thereby enhance plant resistance against herbivores. In rice, Oryza sativa L., insect injury has been shown to induce resistance against future defoliators. However, it is unknown if silicon and mycorrhizae treatments in combination with insect injury result in greater induced resistance. Using the fall armyworm (FAW), Spodoptera frugiperda Smith, two experiments were conducted to investigate whether (1) silicon or mycorrhizae treatment alters resistance in rice and (2) induced systemic resistance in response to insect injury is augmented in silicon- or mycorrhizae- treated plants. In the first experiment, silicon treatment reduced FAW growth by 20% while mycorrhizae increased FAW growth by 8%. In the second experiment, insect injury induced systemic resistance, resulting in a 23% reduction in FAW larval weight gains on injured compared to uninjured plants, irrespective of treatment. Neither silicon nor mycorrhizae enhanced this systemic resistance in insect-injured plants. Furthermore, mycorrhizae resulted in the systemic increase of peroxidase (POD) and polyphenol oxidase (PPO) activities, and injury caused a slight decrease in these enzyme activities in mycorrhizae plants. Silicon treatment did not result in a stronger induction of POD and PPO activity in injured plants. Taken together, these results indicate a lack of silicon and mycorrhizae priming of plant defenses in rice. Regardless of injury, silicon reduced FAW weight gains by 36%. Based on these results, it appears silicon-mediated biomechanical rather than biochemical defenses may play a greater role in increased resistance against FAW in rice.

Effect of Rhizophagus irregularis on Growth and Quality of Cannabis sativa Seedlings.

Rhizophagus irregularis is an arbuscular mycorrhiza fungus that can enhance plant nutrition and reduce transplant shock on seedlings. The present study aims to evaluate the effects of this fungus on the quality of cannabis (Cannabis sativa L.) seedlings.A greenhouse float system experiment was conducted in a completely randomized design with three treatments. The treatments included the application of 40, 80 and 120 fungus spores per L of nutrient solution (AMF1, AMF2 and AMF3, respectively). The evaluation was performed based on the agronomic characteristics of the seedlings (root and stem length and weight, stem diameter), N and P content, survival rate, and the Dickson's quality index (DQI). Results indicated that root length and stem dry weight were significantly increased (by 34.14% and 21.4%, respectively) in the AMF3 treatment. The biomass of the seedlings' roots, the fresh weight and the N content were not affected by the AMF. On the contrary, survival rate, P content and DQI were significantly increased in AMF3 (by 5%, 24.3% and 12.4% respectively). Overall, our findings suggest that the application of high doses of Rhizophagus irregularis (AMF3) on float system-produced cannabis seedlings results in a considerable increment of their quality.

Arbuscular mycorrhizal fungi (Glomus intraradices) and diazotrophic bacterium (Rhizobium BMBS) primed defense in blackgram against herbivorous insect (Spodoptera litura) infestation.

In the changing scenario of agriculture, plants are exposed to various biotic and abiotic stresses. Induction of both constitutive and inducible defense systems was noticed in plants exposed to stress. As a major defense response, production of phenolics and superoxide radicals quenching enzymes is accelerated in plants under stress. These metabolites production intensified further when arbuscular mycorrhizal fungi (AMF) infected plants are subjected to stress. With this background, we conducted experiments to explore the impacts of Glomus intraradices and Rhizobium on the stimulation of defense in blackgram against Spodoptera litura. Uninoculated plants accumulated considerable quantity of defense metabolites like phenolics, lignin and superoxide radicals quenching enzymes such as superoxide dismutase, peroxidase, catalase, phenylalanine ammonium lyase, and polyphenol oxidase constitutively. While production of these defense metabolites primed strongly due to G. intraradices inoculation. These defense responses augmented further when G. intraradices colonized plants were exposed to S. litura. Though the combined inoculation with G. intraradices and Rhizobium improved the defense response, the effect was more pronounced due to single inoculation with G. intraradices. Results of in vitro leaf feeding bioassay showed that the feeding capacity of S. litura reduced (36.32%) significantly due to feeding G. intraradices infected plants. These outcomes revealed that tolerance against S. litura in blackgram could be primed by mycorrhizal inoculation. This is the first report to state that G. intraradices besides improving nutrient use efficiency, also accelerates defense response in blackgram against S. litura. Hence, AMF could be recommended as a bioprotectant against S. litura in blackgram.

Spatial Effects and GWA Mapping of Root Colonization Assessed in the Interaction Between the Rice Diversity Panel 1 and an Arbuscular Mycorrhizal Fungus.

If water saving methods of rice management are to be adopted, the interaction between rice plants and arbuscular mycorrhizal (AM) fungi will grow in agronomic significance. As yet there are very few studies on the interaction between rice and AM fungi and none on host genetics. A subset 334 cultivars from the Rice Diversity Panel 1 were grown in 250 L boxes filled with phosphorus (P) deficient aerobic soil without addition, with added rock phosphate and with rock phosphate and the AM fungus Rhizophagus irregularis. Statistical analysis of position of plants revealed a positive effect of their neighbors on their dry weight which was stronger in the presence of rock phosphate and even stronger with rock phosphate and AM fungi. A weak but significant difference in the response of cultivars to AM fungus treatment in terms of shoot dry weight (SDW) was revealed. Neighbor hyphal colonization was positively related to a plant's hyphal colonization, providing insights into the way a network of AM fungi interact with multiple hosts. Hyphal colonization ranged from 21 to 89%, and 42% of the variation was explained by rice genotype. Colonization was slightly lower in aus cultivars than other rice subgroups and high in cultivars from the Philippines. Genome wide association (GWA) mapping for hyphal colonization revealed 23 putative quantitative trait loci (QTLs) indicating there is an opportunity to investigate the impact of allelic variation in rice on AM fungal colonization. Using published transcriptomics data for AM response in rice, some promising candidate genes are revealed under these QTLs being a calcium/calmodulin serine/threonine protein kinase at 4.9 Mbp on chromosome 1, two ammonium transporters genes at 24.6 Mbp on chromosome 2 and a cluster of subtilisin genes at 1.2 Mbp on chromosome 4. Future studies should concentrate on the biological significance of genetic variation in rice for AM colonization.

Lead uptake by the symbiotic Daucus carota L.-Glomus intraradices system and its effect on the morphology of extra- and intraradical fungal microstructures.

This work examines the strategies adopted by an arbuscular mycorrhizal symbiotic system to ameliorate environmental Pb stress by examining the concentrations of P, Fe, and Pb in the fungal microstructures and the host's root. In vitro cultures of Ri-T DNA-transformed carrot (Daucus carota L.) roots were inoculated with Glomus intraradices and treated with Pb(NO3)2 solution and the extraradical spores and mycelia (S/M) and the root with the vesicles, mycelia, and root cells were subsequently analyzed by polarized energy dispersive x-ray fluorescence (PEDXRF) spectrometry. Upon Pb treatment, within the root, the percentages of mycorrhizal colonization, the vesicles, and mycelia increased as well as the areas of the vesicles and the (extraradical) spores, although the number of spores and arbuscules decreased. The S/M and the mycorrhizal root showed enhanced concentrations of Pb, Fe, and P. These were particularly marked for Fe in the Pb-treated cultures. This indicates a synergistic relationship between the arbuscular mycorrhizal fungus and the host that confers a higher Pb tolerance to the latter by the induction of higher Fe absorption in the host. The intraradical vesicle, mycelia, and arbuscule numbers are interpreted as a "tactic to divert" the intraradical Pb traffic away from the root cells to the higher affinity cell walls of the arbuscular mycorrhizal fungi (AMF) microstructures in the apoplast. The results of this work show that the symbiosis between the AMF G. intraradices and the host plant D. carota distinctly improves the latter's Pb tolerance, and imply that the appropriate metal tolerant host-AMF combinations could be employed in process designs for the phytoremediation of Pb.

Integrated multi-omics analysis supports role of lysophosphatidylcholine and related glycerophospholipids in the Lotus japonicus-Glomus intraradices mycorrhizal symbiosis.

Interaction of plant roots with arbuscular mycorrhizal fungi (AMF) is a complex trait resulting in cooperative interactions among the two symbionts including bidirectional exchange of resources. To study arbuscular mycorrhizal symbiosis (AMS) trait variation in the model plant Lotus japonicus, we performed an integrated multi-omics analysis with a focus on plant and fungal phospholipid (PL) metabolism and biological significance of lysophosphatidylcholine (LPC). Our results support the role of LPC as a bioactive compound eliciting cellular and molecular response mechanisms in Lotus. Evidence is provided for large interspecific chemical diversity of LPC species among mycorrhizae with related AMF species. Lipid, gene expression and elemental profiling emphasize the Lotus-Glomus intraradices interaction as distinct from other arbuscular mycorrhizal (AM) interactions. In G. intraradices, genes involved in fatty acid (FA) elongation and biosynthesis of unsaturated FAs were enhanced, while in Lotus, FA synthesis genes were up-regulated during AMS. Furthermore, FAS protein localization to mitochondria suggests FA biosynthesis and elongation may also occur in AMF. Our results suggest the existence of interspecific partitioning of PL resources for generation of LPC and novel candidate bioactive PLs in the Lotus-G. intraradices symbiosis. Moreover, the data advocate research with phylogenetically diverse Glomeromycota species for a broader understanding of the molecular underpinnings of AMS.

An integrated functional approach to dissect systemic responses in maize to arbuscular mycorrhizal symbiosis.

Most terrestrial plants benefit from the symbiosis with arbuscular mycorrhizal fungi (AMF) mainly under nutrient-limited conditions. Here the crop plant Zea mays was grown with and without AMF in a bi-compartmented system separating plant and phosphate (Pi) source by a hyphae-permeable membrane. Thus, Pi was preferentially taken up via the mycorrhizal Pi uptake pathway while other nutrients were ubiquitously available. To study systemic effects of mycorrhizal Pi uptake on leaf status, leaves of these plants that display an increased biomass in the presence of AMF were subjected to simultaneous ionomic, transcriptomic and metabolomic analyses. We observed robust changes of the leaf elemental composition, that is, increase of P, S and Zn and decrease of Mn, Co and Li concentration in mycorrhizal plants. Although changes in anthocyanin and lipid metabolism point to an improved P status, a global increase in C versus N metabolism highlights the redistribution of metabolic pools including carbohydrates and amino acids. Strikingly, an induction of systemic defence gene expression and concomitant accumulation of secondary metabolites such as the terpenoids alpha- and beta-amyrin suggest priming of mycorrhizal maize leaves as a mycorrhiza-specific response. This work emphasizes the importance of AM symbiosis for the physiological status of plant leaves and could lead to strategies for optimized breeding of crop species with high growth potential.

Fatty acid synthesis and lipid metabolism in the obligate biotrophic fungus Rhizophagus irregularis during mycorrhization of Lotus japonicus.

Arbuscular mycorrhiza formation with fungi of the Glomeromycota represents a widespread symbiotic interaction of vascular plants. Different signaling events and metabolic adaptations are required for the close interaction between the two partners. Membrane lipid synthesis is a prerequisite for symbiosis, and membrane properties depend on lipid composition. Lipid profiling was performed by liquid chromatography mass spectrometry to study the role of triacylglycerol, diacylglycerol, phospholipids, galactolipids, sterols and sphingolipids during the colonization of Lotus japonicus roots with Rhizophagus irregularis (syn. Glomus intraradices). Mycorrhization leads to an increased phosphate supply and suppresses the increase in galactolipids commonly observed in phosphate-deprived plants. In addition to free sterols and sterol esters, R. irregularis contains sterol glucosides and acylated sterol glucosides. Glycosylated sphingolipids (glucosylceramide, dihexosylceramide) and inositolphosphorylceramide were detected in the fungus. Lyso-phosphatidylcholine, a lipid previously implicated in mycorrhiza signaling, is present in low amounts in mock-infected and mycorrhized roots. The composition of fungal phospholipids changes after mycorrhization because molecular species with palmitvaccenic (di-16:1) or tetracosenoic (24:1) acyl groups decrease in intraradical mycelium. This adaptation of lipid metabolism during intraradical growth is likely a prerequisite for symbiosis, achieving functional compatibility between the fungal and the periarbuscular membrane. Data mining in genomic and transcript databases revealed the presence of genes encoding enzymes of lipid biosynthesis in R. irregularis. However, no gene encoding multidomain fatty acid de novo synthase was detected in the genome sequence of this obligate biotrophic fungus.

Multitrophic interactions among Western Corn Rootworm, Glomus intraradices and microbial communities in the rhizosphere and endorhiza of maize.

The complex interactions among the maize pest Western Corn Rootworm (WCR), Glomus intraradices (GI-recently renamed Rhizophagus intraradices) and the microbial communities in both rhizosphere and endorhiza of maize have been investigated in view of new pest control strategies. In a greenhouse experiment, different maize treatments were established: C (control plants), W (plants inoculated with WCR), G (plants inoculated with GI), GW (plants inoculated with GI and WCR). After 20 days of WCR root feeding, larval fitness was measured. Dominant arbuscular mycorrhizal fungi (AMF) in soil and maize endorhiza were analyzed by cloning of 18S rRNA gene fragments of AMF, restriction fragment length polymorphism and sequencing. Bacterial and fungal communities in the rhizosphere and endorhiza were investigated by denaturing gradient gel electrophoresis of 16S rRNA gene and ITS fragments, PCR amplified from total community DNA, respectively. GI reduced significantly WCR larval development and affected the naturally occurring endorhiza AMF and bacteria. WCR root feeding influenced the endorhiza bacteria as well. GI can be used in integrated pest management programs, rendering WCR larvae more susceptible to predation by natural enemies. The mechanisms behind the interaction between GI and WCR remain unknown. However, our data suggested that GI might act indirectly via plant-mediated mechanisms influencing the endorhiza microbial communities.

Screening of efficient arbuscular mycorrhizal fungi for Azadirachta indica under nursery condition: a step towards afforestation of semi-arid region of western India.

To optimize nursery practices for efficient plant production procedures and to keep up to the ever growing demand of seedlings, identification of the most suitable species of arbuscular mycorrhizal fungi (AMF), specific for a given tree species, is clearly a necessary task. Sixty days old seedlings of Neem (Azadirachta indica A. Juss) raised in root trainers were inoculated with six species of AMF and a mixed inoculum (consortia) and kept in green house. Performances of the treatments on this tree species were evaluated in terms of growth parameters like plant height shoot collar diameter, biomass and phosphorous uptake capabilities. Significant and varied increase in the growth parameters and phosphorous uptake was observed for most of the AMF species against control. Consortia culture was found to be the best suited AMF treatment for A. indica, while Glomus intraradices and Glomus mosseae were the best performing single species cultures. It is the first time in the state of Gujarat that a wide variety of AMF species, isolated from the typical semi-arid region of western India, were tested for the best growth performance with one of the most important tree species for the concerned region.

Aquaporin genes GintAQPF1 and GintAQPF2 from Glomus intraradices contribute to plant drought tolerance.

Arbuscular mycorrhizal (AM) symbiosis, established between AM fungi (AMF) and roots of higher plants, occurs in most terrestrial ecosystems. It has been well demonstrated that AM symbiosis can improve plant performance under various environmental stresses, including drought stress. However, the molecular basis for the direct involvement of AMF in plant drought tolerance has not yet been established. Most recently, we cloned two functional aquaporin genes, GintAQPF1 and GintAQPF2, from AM fungus Glomus intraradices. By heterologous gene expression in yeast, aquaporin localization, activities and water permeability were examined. Gene expressions during symbiosis in expose to drought stress were also analyzed. Our data strongly supported potential water transport via AMF to host plants. As a complement, here we adopted the monoxenic culture system for AMF, in which carrot roots transformed by Ri-T DNA were cultured with Glomus intraradices in two-compartment Petri dishes, to verify the aquaporin gene functions in assisting AMF survival under polyethylene glycol (PEG) treatment. Our results showed that 25% PEG significantly upregulated the expression of two aquaporin genes, which was in line with the gene functions examined in yeast. We therefore concluded that the aquaporins function similarly in AMF as in yeast subjected to osmotic stress. The study provided further evidence to the direct involvement of AMF in improving plant water relations under drought stresses.

Screening of efficient arbuscular mycorrhizal fungi for Azadirachta indica under nursery condition: a step towards afforestation of semi-arid region of western India

To optimize nursery practices for efficient plant production procedures and to keep up to the ever growing demand of seedlings, identification of the most suitable species of arbuscular mycorrhizal fungi (AMF), specific for a given tree species, is clearly a necessary task. Sixty days old seedlings of Neem (Azadirachta indica A. Juss) raised in root trainers were inoculated with six species of AMF and a mixed inoculum (consortia) and kept in green house. Performances of the treatments on this tree species were evaluated in terms of growth parameters like plant height shoot collar diameter, biomass and phosphorous uptake capabilities. Significant and varied increase in the growth parameters and phosphorous uptake was observed for most of the AMF species against control. Consortia culture was found to be the best suited AMF treatment for A.indica, while Glomus intraradices and Glomus mosseae were the best performing single species cultures. It is the first time in the state of Gujarat that a wide variety of AMF species, isolated from the typical semi-arid region of western India, were tested for the best growth performance with one of the most important tree species for the concerned region.