Paraglomus Occultum Research Articles

Characterization of arbuscular mycorrhizal fungal species associating with Zea mays.

Taxonomic identification of arbuscular mycorrhizal (AM) fungal spores extracted directly from the field is sometimes difficult because spores are often degraded or parasitized by other organisms. Single-spore inoculation of a suitable host plant allows for establishing monosporic cultures of AM fungi. This study aimed to propagate AM fungal spores isolated from maize soil using single spores for morphological characterization. First, trap cultures were established to trigger the sporulation of AM fungal species. Second, trap cultures were established with individual morphotypes by picking up only one spore under a dissecting microscope and transferring it to a small triangle of sterilized filter paper, which was then carefully inoculated below a root from germinated sorghum seeds in each pot and covered with a sterile substrate. All pots were placed in sunbags and maintained in a plant growth room for 120 days. Spores obtained from single spore trap cultures from each treatment, maize after oats (MO), maize after maize (MM), maize after peas (MP), and maize after soybean (MS), were extracted using the sieving method. Healthy spores were selected for morphological analysis. Direct PCR was conducted by crushing spores in RNAlater and applying three sets of primer pairs: ITS1 × ITS4, NS31 × AML2, and SSUmcf and LSUmBr. Nucleotide sequences obtained from Sanger sequencing were aligned on MEGA X. The phylogenetic tree showed that the closest neighbors of the propagated AM fungal species belonged to the genera Claroideoglomus, Funneliformis, Gigaspora, Paraglomus, and Rhizophagus. The morphological characteristics were compared to the descriptive features of described species posted on the INVAM website, and they included Acaulospora cavernata, Diversispora spurca, Funneliformis geosporus, Funneliformis mosseae, Gigaspora clarus, Gigaspora margarita, Glomus macrosporum, Paraglomus occultum, and Rhizophagus intraradices. These findings can provide a great contribution to crop productivity and sustainable management of the agricultural ecosystem. Also, the isolate analyzed could be grouped into efficient promoters of growth and mycorrhization of maize independent of their geographical location.

Improved Waterlogging Tolerance in Roots of Cucumber Plants after Inoculation with Arbuscular Mycorrhizal Fungi

Mycorrhizal symbiosis enhances host plant resistance to various unfavorable environmental stresses, but whether and how it also enhances waterlogging tolerance in cucumber plants is not known. The objective of this study was to analyze the effect of Paraglomus occultum inoculation on biomass production, osmolyte levels, and the expression of 12 heat shock protein 70 (Hsp70) genes and 14 plasma membrane intrinsic protein (PIP) genes in the roots of cucumber plants under a short-term waterlogging stress (WS) (5 days) condition. Although the short-term WS treatment significantly inhibited the arbuscular mycorrhizal fungal colonization of roots, the inoculation with arbuscular mycorrhizal fungi (AMFs) significantly increased leaf, stem, and root biomass under WS. AMF inoculation also significantly increased root glucose, sucrose, betaine, and proline contents, along with decreased fructose levels, compared with the uninoculated control. More CsHsp70 and CsPIP genes were up-regulated in AMF-inoculated plants than in AMF-uninoculated plants in response to WS. AMF inoculation showed no significant effect on the expression of any of the examined CsHsp70 genes under no-waterlogging stress, but it did raise the expression of 11 of 12 CsHsp70 genes under WS. AMF colonization also down-regulated or had no effect on CsPIP expression under no-waterlogging stress, whereas it up-regulated the expression of 12 of the 14 CsPIP genes under WS. It is concluded that AMF inoculation enhances waterlogging tolerance in cucumber plants by increasing osmolyte levels and stress-responsive gene (CsPIP and CsHsp70) expression.

Mycorrhizas Affect Physiological Performance, Antioxidant System, Photosynthesis, Endogenous Hormones, and Water Content in Cotton under Salt Stress.

Saline-alkali stress seriously endangers the normal growth of cotton (Gossypium hirsutum). Arbuscular mycorrhizal fungi (AMF) could enhance salt tolerance by establishing symbiotic relationships with plants. Based on it, a pot experiment was conducted to simulate a salt environment in which cotton was inoculated with Paraglomus occultum to explore its effects on the saline-alkali tolerance of cotton. Our results showed that salt stress noticeably decreased cotton seedling growth parameters (such as plant height, number of leaves, dry weight, root system architecture, etc.), while AMF exhibited a remarkable effect on promoting growth. It was noteworthy that AMF significantly mitigated the inhibitory effect of salt on cotton seedlings. However, AMF colonization in root and soil hyphal length were collectively descended via salt stress. With regard to osmotic regulating substances, Pro and MDA values in roots were significantly increased when seedlings were exposed to salt stress, while AMF only partially mitigated these reactions. Salt stress increased ROS levels in the roots of cotton seedlings and enhanced antioxidant enzyme activity (SOD, POD, and CAT), while AMF mitigated the increases in ROS levels but further strengthened antioxidant enzyme activity. AMF inoculation increased the photosynthesis parameters of cotton seedling leaves to varying degrees, while salt stress decreased them dramatically. When inoculated with AMF under a salt stress environment, only partial mitigation of these photosynthesis values was observed. Under saline-alkali stress, AMF improved the leaf fluorescence parameters (φPSII, Fv'/Fm', and qP) of cotton seedlings, leaf chlorophyll levels, and root endogenous hormones (IAA and BR); promoted the absorption of water; and maintained nitrogen balance, thus alleviating the damage from salt stress on the growth of cotton plants to some extent. In summary, mycorrhizal cotton seedlings may exhibit mechanisms involving root system architecture, the antioxidant system, photosynthesis, leaf fluorescence, endogenous hormones, water content, and nitrogen balance that increase their resistance to saline-alkali environments. This study provide a theoretical basis for further exploring the application of AMF to enhance the salt tolerance of cotton.

Arbuscular mycorrhizal fungi and rhizobia accelerate plant growth and N accumulation and contribution to soil total N in white clover by difficultly extractable glomalin-related soil protein

Arbuscular mycorrhizal fungi and rhizobia are soil symbiotic microorganisms involved in plant nitrogen (N) acquisition, whereas it is unclear how single or combined inoculation of both contributes to soil total N in legume crops. This study analyzed the effects of single or combined inoculation with arbuscular mycorrhizal fungus (Paraglomus occultum) and rhizobium (Rhizobium leguminosarum bv. trifolii) on growth performance, root soluble protein, leghemoglobin, NH4+-N, NO3−-N, and N concentrations, nitrogenase activities, soil total N contents, N contents in glomalin-related soil protein (GRSP), as well as the contribution of N in GRSP to soil total N in white clover. Twelve weeks after inoculation, P. occultum promoted nodule formation, and R. leguminosarum bv. trifolii accelerated root mycorrhizal colonization. Single or combined inoculation with P. occultum and R. leguminosarum bv. trifolii improved biomass, root total length, surface area and volume, root soluble protein concentrations, nodule leghemoglobin concentrations, and nitrogenase activities to varying degrees, with the trend of improvement being more pronounced with combined inoculation. R. leguminosarum bv. trifolii and/or P. occultum significantly increased NO3−-N, NH4+-N, and total N concentrations in nodules and roots, as well as soil total N concentrations. The N in easily extractable GRSP (EE-GRSP) and difficultly extractable GRSP (DE-GRSP) was 4.37–5.64 mg/g and 6.69–7.83 mg/g, respectively, contributing 2.68–3.26 % and 2.28–3.10 % of soil total N. P. occultum and combined inoculation accelerated the production of EE-GRSP and DE-GRSP and N in EE-GRSP. R. leguminosarum bv. trifolii not only significantly increased DE-GRSP production but also reduced N in DE-GRSP. R. leguminosarum bv. trifolii or P. occultum significantly increased the contribution of N in DE-GRSP, not EE-GRSP, to soil total N. It was concluded that arbuscular mycorrhizal fungus and rhizobium accelerated N accumulation and contribution to soil total N in white clover by DE-GRSP.

The reduction of abiotic stress in food crops through climate-smart mycorrhiza-enriched biofertilizer.

Climate change enhances stress in food crops. Recently, abiotic stress such as metalloid toxicity, salinity, and drought have increased in food crops. Mycorrhizal fungi can accumulate several nutrients within their hyphae through a symbiotic relationship and release them to cells in the root of the food crops under stress conditions. We have studied arbuscular mycorrhizal fungi (AMF)-enriched biofertilizers as a climate-smart technology option to increase safe and healthy food production under abiotic stress. AMF such as Glomus sp., Rhizophagus sp., Acaulospora morrowiae, Paraglomus occultum, Funneliformis mosseae, and Claroideoglomus etunicatum enhance growth and yield in food crops grown in soils under abiotic stress. AMF also works as a bioremediation material in food crops grown in soil. More precisely, the arsenic concentrations in grains decrease by 57% with AMF application. In addition, AMF increases mineral contents, and antioxidant activities under drought and salinity stress in food crops. Catalase (CAT) and ascorbate peroxidase (APX) increased by 45% and 70% in AMF-treated plants under drought stress. AMF-enriched biofertilizers are used in crop fields like precision agriculture to reduce the demand for chemical fertilizers. Subsequently, AMF-enriched climate-smart biofertilizers increase nutritional quality by reducing abiotic stress in food crops grown in soils. Consequently, a climate resilience environment might be developed using AMF-enriched biofertilizers for sustainable livelihood.

Arbuscular Mycorrhizal Fungi Enhance Biomass Growth, Mineral Content, and Antioxidant Activity in Tomato Plants under Drought Stress

Arbuscular mycorrhizal fungi (AMF) are symbiotically associated with crops. They increase biomass production, nutritional elements, and antioxidant activities in food and vegetable crops grown in soil under stress conditions. The present study focused on the effects of AMF (Acaulospora morrowiae, Paraglomus occultum, Funneliformis mosseae, Rhizophagus clarus, and Rhizophagus intraradices) on biomass growth and yield, contents of chlorophyll and carotenoids, activities of catalase (CAT) and ascorbate peroxidase (APX), and contents of hydrogen peroxide (H2O2), malondialdehyde (MDA), and minerals (Na, K, Ca, Mg, and Fe) in Unnayan, LT896, and Minto super tomato (Solanum lycopersicum L.) varieties grown in soil under drought stress (<10% moisture). The results showed that root length and shoot mass in plants treated with R. clarus and P. occultum were significantly higher than those of the control (non-AMF) in Minto super tomato. Compared to the control, the shoot’s dry weight and yield were enhanced by 28% and 20% with AMF-treated tomatoes. The CAT activity in P. occultum-treated plants was statistically higher than that of the control in Unnayan tomatoes. H2O2 content was detected higher in the control than R. clarus-treated LT896 tomatoes. In plants treated with A. morrowiae and R. clarus, APX activity was significantly higher than that of the control in the Unnayan tomatoes. CAT and APX activity increased by 42% and 66% in AMF-treated leaves of tomatoes compared to non-AMF. Treatment with AMF reduced the content of MDA and H2O2 (ROS) in the leaves of tomato plants by 50% and 2% compared to the control, respectively. Potassium (K), calcium (Ca), magnesium (Mg), and iron (Fe) of tomato fruits increased by 2%, 13%, 24%, and 37% with AMF treatment compared to the control. These results suggested that biomass growth, yield, photosynthetic pigments, antioxidant enzyme activity, and mineral contents could be enhanced by AMF in food crops grown under drought stress. It is concluded that AMF might be used for the development of AMF-enriched biofertilizers that will improve the nutritional quality of food crops grown under stress conditions.

Arbuscular mycorrhizal fungi inoculation impacts expression of aquaporins and salt overly sensitive genes and enhances tolerance of salt stress in tomato

Arbuscular mycorrhizal fungi (AMF) that establish reciprocal symbiosis with plant roots can enhance resistance to various stresses, including salt stress, but relevant mechanisms, especially at the molecular level, are scarce. The objective of this study was to analyze the effect of an arbuscular mycorrhizal fungus Paraglomus occultum on plant growth, leaf gas exchange, and expression of plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs) and salt overly sensitive (SOS) genes in tomato under salt (150 mmol/L NaCl) and non-salt stress. Salt stress for 4 weeks inhibited root colonization rate of P. occultum and soil hyphal length by 0.21- and 0.57-fold, respectively. Salt stress also inhibited plant growth performance and leaf gas exchange, while inoculation with P. occultum significantly enhanced them under salt and non-salt stress conditions. AMF showed diverse regulation of root SlPIPs and SlTIPs expression, among which under salt stress, SlPIP1;2, SlPIP1;5, SlPIP2;1, SlPIP2;6, SlPIP2;9, SlPIP2;10, SlTIP2;2, SlTIP3;2, and SlTIP5;1 were up-regulated by AMF colonization, and SlPIP1;7, SlPIP2;5, SlPIP2;8, SlPIP2;11, SlPIP2;12, SlTIP2;3, and SlTIP3;1 were down-regulated, accompanied by no change in SlPIP1;1, SlPIP1;3, SlPIP2;4, SlTIP1;1, SlTIP1;2, SlTIP1;3, SlTIP2;1, and SlTIP2;5. Interestingly, salt stress inhibited the expression of SlSOS1 and SlSOS2 in non-mycorrhizal plants, while it increased the expression of SlSOS1 and SlSOS2 in mycorrhizal plants. AMF colonization down-regulated expression of SlSOS1 and SlSOS2 under non-salt stress while up-regulated expression of SlSOS1 and SlSOS2 under salt stress. It was concluded that AMF inoculation impacted the expression of stress-responsive genes, especially SOS1 and SOS2, and enhanced salt resistance of tomato.Graphical

Mycorrhizal Effects on Growth and Expressions of Stress-Responsive Genes (aquaporins and SOSs) of Tomato under Salt Stress.

Environmentally friendly arbuscular mycorrhizal fungi (AMF) in the soil can alleviate host damage from abiotic stresses, but the underlying mechanisms are unclear. The objective of this study was to analyze the effects of an arbuscular mycorrhizal fungus, Paraglomus occultum, on plant growth, nitrogen balance index, and expressions of salt overly sensitive genes (SOSs), plasma membrane intrinsic protein genes (PIPs), and tonoplast intrinsic protein genes (TIPs) in leaves of tomato (Solanum lycopersicum L. var. Huapiqiu) seedlings grown in 0 and 150 mM NaCl stress. NaCl stress severely inhibited plant growth, but P. occultum inoculation significantly improved plant growth. NaCl stress also suppressed the chlorophyll index, accompanied by an increase in the flavonoid index, whereas inoculation with AMF significantly promoted the chlorophyll index as well as reduced the flavonoid index under NaCl conditions, thus leading to an increase in the nitrogen balance index in inoculated plants. NaCl stress regulated the expression of SlPIP1 and SlPIP2 genes in leaves, and five SlPIPs genes were up-regulated after P. occultum colonization under NaCl stress, along with the down-regulation of only SlPIP1;2. Both NaCl stress and P. occultum inoculation induced diverse expression patterns in SlTIPs, coupled with a greater number of up-regulated TIPs in inoculated versus uninoculated plants under NaCl stress. NaCl stress up-regulated SlSOS2 expressions of mycorrhizal and non-mycorrhizal plants, while P. occultum significantly up-regulated SlSOS1 expressions by 1.13- and 0.45-fold under non-NaCl and NaCl conditions, respectively. It was concluded that P. occultum inoculation enhanced the salt tolerance of the tomato, associated with the nutrient status and stress-responsive gene (aquaporins and SOS1) expressions.

Early branching arbuscular mycorrhizal fungus Paraglomus occultum carries a small and repeat-poor genome compared to relatives in the Glomeromycotina.

The arbuscular mycorrhizal fungi (AMFs) are obligate root symbionts in the subphylum Glomeromycotina that can benefit land plants by increasing their soil nutrient uptake in exchange for photosynthetically fixed carbon sources. To date, annotated genome data from representatives of the AMF orders Glomerales, Diversisporales and Archaeosporales have shown that these organisms have large and highly repeated genomes, and no genes to produce sugars and fatty acids. This led to the hypothesis that the most recent common ancestor (MRCA) of Glomeromycotina was fully dependent on plants for nutrition. Here, we aimed to further test this hypothesis by obtaining annotated genome data from a member of the early diverging order Paraglomerales (Paraglomus occultum). Genome analyses showed this species carries a 39.6 Mb genome and considerably fewer genes and repeats compared to most AMF relatives with annotated genomes. Consistent with phylogenies based on ribosomal genes, our phylogenetic analyses suggest P. occultum as the earliest diverged branch within Glomeromycotina. Overall, our analyses support the view that the MRCA of Glomeromycotina carried hallmarks of obligate plant biotrophy. The small genome size and content of P. occultum could either reflect adaptive reductive processes affecting some early AMF lineages, or indicate that the high gene and repeat family diversity thought to drive AMF adaptability to host and environmental change was not an ancestral feature of these prominent plant symbionts.

Diversity of vam in soils used to cultivate sugarcane (Saccharum spp.) in Tabasco, Mexico

E Objective: A study was carried out in the Ingenio Santa Rosalía sugar mill supply area with the aim of identifying the soil subunits cultivated with sugar cane and to quantify the degree of mycorrhization of Saccharum officinarum. Design/Methodology/Approach: Rhizosphere samples were collected at a depth of 0-30 cm and root segments with diameters of ≤ 1 mm and 1-2.0 mm were studied. The intersect method and the fungal structures method were comparatively analyzed to determine the mycotrophic state of the roots. The study also quantified the number of spores in the soils and classified the morpho-species of vesicular-arbuscular mycorrhiza (VAM). Results: The results show that seven soil subunits exist in the area, but the intersect method did not detect differences in colonization by root diameter or by root colonization ( = 68.5%). For its part, the fungal structures method showed differences in colonization between soil subunits ( = 69.5%), being higher in thin roots, and statistical differences wer found for vesicles and spores. At the sites, an average of 696 spores was quantified per 100 g of soil, which indicates a high presence of vesicular-arbuscular mycorrhizal fungi in the study area. Finally, six species of mycorrhizae were identified, of which four are present in various soil subunits: Glomus aff. deserticola, Glomus etunicatum, Glomus viscosum, and Paraglomus occultum. Study Limitations/Implications: This was an exploratory study that indicates the potential of VAM. Findings/Conclusions: The presence of hyphae, vesicles, arbuscules, and spores indicates that there is an active mycotrophic process between VAM and sugarcane cultivation in the soils of the study area.

Propagation and characterization of viable arbuscular mycorrhizal fungal spores within maize plant (Zea mays L.).

The harmful effect of chemical fertilizer application on human health and the environment as a modern method of meeting the food demand of the increasing world population demands an urgent alternative that is environmentally friendly, which will pose no harm to human health and the environment. Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms that provide various ecological functions in increasing soil fertility and enhancing plant growth. This present study aimed to propagate, characterize and examine the effect of viable arbuscular mycorrhizal fungal spores on maize (Zea mays L) hosts using molecular methods. The propagation of AMF in the host plant using sterile soil and vermiculite was conducted in the greenhouse. The effect of AMF inoculation revealed a significant difference (P > 0.05) in maize growth, root colonization and AMF spore count when compared with the control. In all the parameters measured in this study, all the AMF spores propagated had a positive effect on the maize plant over the control, with the highest value mostly recorded in Rhizophagus irregularis AOB1. The molecular characterization of the spore using a specific universal primer for Glomeromycota established the success of the propagation process, which enhanced the classification of the AMF species into Rhizophagus irregularis OAB1, Glomus mosseae OAB2 and Paraglomus occultum OAB3. This finding will be a starting point in producing arbuscular mycorrhizal inoculum as a biofertilizer to enhance plant growth promotion. © 2021 Society of Chemical Industry.

Physiological responses of mycorrhizal symbiosis to drought stress in white clover

The aim of the present study was to analyze the effects of two arbuscular mycorrhizal fungi (AMF), Funneliformis mosseae and Paraglomus occultum, on leaf water status, root morphology, root sugar accumulation, root abscisic acid (ABA) levels, root malondialdehyde (MDA) content, and root antioxidant enzyme activities in white clover (Trifolium repens L.) exposed to well-watered (WW) and drought stress (DS) conditions. The results showed that root colonization by F. mosseae and P. occultum was significantly decreased by 7-week soil drought treatment. Under drought stress conditions, mycorrhizal fungal treatment considerably stimulated root total length, surface area and volume, as compared with non-mycorrhizal controls. In addition, inoculation with arbuscular mycorrhizal fungi also increased leaf relative water content and accelerated the accumulation of root glucose and fructose under drought stress. Mycorrhizal plants under drought stress registered higher activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) and ABA levels in roots, while lower MDA contents, relative to non-mycorrhizal plants. As a result, mycorrhiza-inoculated plants represented better physiological activities (e.g. antioxidant defense systems, root morphology, and sugar accumulation) than non-inoculated plants in response to soil drought, whilst P. occultum had superior effects than F. mosseae.

Single or dual inoculation of arbuscular mycorrhizal fungi and rhizobia regulates plant growth and nitrogen acquisition in white clover

The present work aimed to analyse whether and how single or dual inoculation with arbuscular mycorrhizal fungi (Funneliformis mosseae, Paraglomus occultum, and Rhizophagus intraradices) and rhizobia (Rhizobium trifolii) improved plant growth and stimulated nitrogen (N) acquisition of white clover. AMF inoculation significantly (P < 0.05) increased root nodule number by 117‒173%, and additional Rh considerably stimulated mycorrhizal growth. Single AMF or Rh treatment dramatically increased shoot by 36‒281% and root biomass by 16‒36% than non-inoculated control, and dual inoculation of Rh and P. occultum or R. intraradices further magnified the positive effect. Leaf and root N content, root total soluble protein content, root nitrogenase activity, and amino acid (e.g., alanine, arginine, asparagine, aspartate, phenylalanine, proline, and tryptophan) concentrations were significantly increased by single or dual inoculation, while dual inoculation of AMF and Rh had significantly superior roles than single corresponding AMF or Rh inoculation. These results suggested that AMF and Rh represented synergetic effects on accelerating N acquisition of white clover to some extent, while the combination of P. occultum and Rh was the best.

Small-scale soil heterogeneity affects the distribution of arbuscular mycorrhizal fungal species in a hot-spot field in a Mediterranean site

Arbuscular mycorrhizal fungi (AMF) establish mutualistic symbioses with the roots of most plant species, playing a key role in crop productivity and ecosystem functioning. The characterization of local AMF genotypes assemblages is a prerequisite for the reproduction, both ex situ and on farm, of efficient native AMF communities, in order to improve soil quality and fertility through agricultural management and to use selected AMF as inoculants in sustainable agriculture. Contrasting evidences have been found by studies addressing the influence of different environmental drivers, among which geographical distance, on the diversity and composition of AMF communities in agricultural soils.In the present study, a Mediterranean “hot-spot” of AMF species richness was studied to assess whether AMF display a spatial structure at a local scale and whether their spore distribution is affected by soil variables. AMF distribution patterns were estimated by spore analysis, in 12 field plots, which were characterized for their chemical and physical variables. Plots were distributed along a transect across the field, and sampling points distances ranged from 4 to 102 m. While most taxa displayed an aggregated distribution, no significant spatial correlation was detected among AMF communities. On the contrary, AMF communities were significantly correlated with the entire set of soil variables, and with distinct soil characteristics, such as pH and total N, but not P. Among the most abundant taxa, Funneliformis geosporus, Septoglomus constrictum, Funneliformis mosseae, Ambispora granatensis and Glomus badium, were positively affected by total N, and Paraglomus occultum negatively affected by pH. Multiple regression analysis revealed that distribution of most AMF species was significantly related to nitrogen, clay, cation exchange capacity and pH. Multivariate (RDA) analysis confirmed results of multiple regression, with the best model selecting three soil variables (N, clay and pH). Variance partitioning analysis also showed that AMF distribution was mostly explained by environmental variables (36.4%) and by environmental variables spatial structure (16.4%). Overall, the results of this work revealed the importance of soil variables in shaping AMF community composition and spatial heterogeneity, within a site characterized by high AMF richness.

Mycorrhiza-released glomalin-related soil protein fractions contribute to soil total nitrogen in trifoliate orange

Glomalin released from arbuscular mycorrhizal fungi (AMF) has important roles in soil nutrient cycles, whereas contributing to glomalin-related soil protein (GRSP) fractions to soil nitrogen (N) is unknown. In this study, a two-chambered root-box that was divided into root chamber (root and mycorrhizal fungi hypha) and hypha chamber (free of the root) was used, and three AMF species including Diversispora epigaea, Paraglomus occultum, and Rhizoglomus intraradices were separately inoculated into the root chamber. Plant growth, soil total N, N content of purified GRSP fractions, and its contribution to soil total N, and leaf and root N contents were analysed. After four months, total biomass and root total length, surface area, and volume were improved by all AMF inoculations. AMF inoculations dramatically increased soil total N content in two chambers. The N content of purified easily extractable GRSP (EE-GRSP) and difficultly extractable GRSP (DE-GRSP) was 0.10 ± 0.01 mg/g and 0.16 ± 0.02 mg/g, respectively, accounted for 15.6 ± 1.6% and 18.1 ± 1.8% of soil total N, respectively. AMF inoculations stimulated the N accumulation in EE-GRSP and DE-GRSP, especially in the hypha chamber. It concluded that GRSP, especially DE-GRSP, acts as a soil N pool accounting for 33.8 ± 1.9% of soil total N in orchards.

Contribution of glomalin-related soil proteins to soil organic carbon in trifoliate orange

Arbuscular mycorrhizal fungi (AMF) produce glomalin-related soil protein (GRSP) that influences organic carbon (C) storage in soil; however, how much purified GRSP fractions contribute to soil organic carbon (SOC) is yet not known. The present study evaluated the contribution of GRSP towards changes in SOC in trifoliate orange grown in a rootbox divided into a roots + hyphae chamber (roots colonized by AMF hyphae and AMF extraradical hyphae) and a hyphae chamber (only the presence of AMF extraradical hyphae, without roots). Three AMF species (Diversispora epigaea, Paraglomus occultum and Rhizoglomus intraradices) were inoculated into the roots + hyphae chamber. Following four months of plant growth, P. occultum showed higher AMF hyphal growth in roots, soils and nylon mesh than D. epigaea or R. intraradices. Mycorrhizal inoculation improved the plant growth performance and increased easily extractable GRSP (EE-GRSP) and difficultly extractable GRSP (DE-GRSP) concentrations in both chambers, regardless of AMF species. The C content observed in total GRSP of the soil after purification was 2.71 ± 0.49 mg g−1, while purified EE-GRSP and DE-GRSP showed the C content of 1.01 ± 0.19 mg g−1 and 1.70 ± 010.34 mg g−1, respectively. The C contribution by purified EE-GRSP and DE-GRSP accounted for 8.67 ± 0.95% and 14.59 ± 2.21%, respectively, of total SOC, with a total C contribution of purified GRSPs accounting for 23.26 ± 2.67% of total SOC. A significantly higher C content of GRSP and the C contribution of GRSP to SOC were observed in DE-GRSP than in EE-GRSP, as well as the soil of the roots + hyphae chamber than the soil of the hyphae chamber. The proportionate distribution of water-stable aggregate in 2–4 and 1–2 mm sizes and their stability were higher under AMF hyphae than under non-AMF hyphae. This study thus provided a database evidence of increased contribution of GRSP towards build-up of SOC in response to mycorrhizal symbiosis.

GROWTH OF Kielmeyera rubriflora INOCULATED WITH ARBUSCULAR MYCHORRIZAL FUNGI ON IRON MINING SUBSTRATES

Mining results in major landscape changes. Restoring areas after exploitation of natural resources can be accomplished through adopting techniques used to optimize the restoration process. Among such techniques, planting seedlings of species adapted to adverse edaphoclimatic conditions and which have interspecific associations with microorganisms stand out. One of these indigenous species which occur in the Southern Espinhaço Mountain Range with potential for use in restoring degraded areas is Kielmeyera rubriflora (Spreng) Mart. Thus, the objective of this study was to evaluate the influence of inoculating arbuscular mycorrhizal fungi on the survival and growth of Kielmeyera rubriflora Cambess. on two substrates from iron mining. The experiment was carried out in a nursery of the UFVJM Plant Species Propagation Center (CIPEF) in a randomized block design and treatments arranged in a 2 x 2 factorial design, using two iron mining substrates and inoculated and uninoculated plants with five repetitions. The seedlings received 25 g of the soil-inoculum mixture of Paraglomus occultum (C. Walker) JB Morton & D. Redecker and Claroideoglomus etunicatum (WN Becker & Gerd) C. Walker & A. Schüßler at the transplantation time to 17 dm3 pots. Stem diameter and leaf number were influenced only by the main effects of substrate and inoculation, while plant height was influenced by the interaction between substrate and inoculation. Inoculation with mycorrhizal fungi in Kielmeyera rubriflora ensures survival, species growth and root development on both iron mining substrates and is a suggested strategy for planting in areas degraded by iron mining.

Mycorrhiza Regulates Signal Substance Levels and Pathogen Defense Gene Expression to Resist Citrus Canker

Citrus canker is a quarantined disease, severely harming citrus plants. Soil beneficial arbuscular mycorrhizal fungi (AMF) can provide a biological control pathway to resist pathogens. This work was to test changes of signal substances including hydrogen peroxide (H2O2), nitric oxide (NO), calmodulin (CaM), salicylic acid (SA) and jasmonic acid (JA) and the pathogen defense gene expression in roots of AMF (Paraglomus occultum) and non-AMF trifoliate orange (Poncirus trifoliata) seedlings after infected by a expressions citrus canker pathogen (Xanthomonas axonopodis pv. Citri, Xac). AMF inoculation significantly improved plant height, stem diameter and leaf number. Xac infection dramatically decreased root H2O2, NO, and SA levels, but increased root CaM and JA concentrations in non-AMF seedlings. There were higher H2O2 and CaM levels and lower JA levels in Xac-infected seedlings than in non-Xac-infected seedlings under mycorrhization. Under non-Xac infection, mycorrhizal treatment reduced root H2O2, NO, and SA but increased CaM and JA levels. However, under Xac infection, mycorrhizal inoculation distinctly accelerated root H2O2, NO, CaM, and SA accumulation, accompanied with up-regulated expression levels of root PtEPS1 (enhance pseudomonas susceptibility 1) and PtPR4 (pathogenesis related gene 4), indicating that Xac stimulated mycorrhizal roles in enhancing resistance of citrus canker. Such results imply that citrus plants with pre-inoculated AMF had stronger resistance to Xac infection through increasing signal substrate accumulation and pathogen defense gene expressions.
 
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 In press - Online First. Article has been peer reviewed, accepted for publication and published online without pagination. It will receive pagination when the issue will be ready for publishing as a complete number (Volume 47, Issue 4, 2019). The article is searchable and citable by Digital Object Identifier (DOI). DOI link will become active after the article will be included in the complete issue.
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Effects of mycorrhizas on physiological performance and root TIPs expression in trifoliate orange under salt stress

ABSTRACTArbuscular mycorrhizal fungi (AMF) enhance salt tolerance of host plants, while it is unclear whether aquaporins are involved in the AMF event. This study analyzed the effects of inoculation with an arbuscular mycorrhizal fungus Paraglomus occultum on plant physiological performance and relative expression of root tonoplast intrinsic proteins (TIPs, a kind of aquaporins) genes in potted trifoliate orange (Poncirus trifoliata) seedlings subjected to 0 and 100 mM NaCl concentrations. AMF inoculation significantly increased plant growth performance, leaf water status (relative water content and water potential) and gas exchange (photosynthetic rate, transpiration rate and stomatal conductance), irrespective of non-salt and salt stress. Mycorrhizal seedlings exhibited significantly higher root salicylic acid concentrations and lower leaf and root jasmonic acid concentrations than non-mycorrhizal seedlings under non-salt and salt stress. Transcript levels of PtTIP1;1, PtTIP1;2 and PtTIP1;3 were down-regulated by AMF inoculation under non-salt and salt stress. PtTIP2;1 and PtTIP2;2 expression was unchanged under non-salt stress whereas was down-regulated by AMF colonization under salt stress. AMF inoculation considerably enhanced PtTIP4;1 transcript levels under non-salt and salt stress and PtTIP5;1 expression under non-salt stress. It is concluded that mycorrhizas mitigated salt stress of trifoliate orange through improving physiological performance and regulating root PtTIPs expression.

Diversity and distribution of Arbuscular Mycorrhizal Fungi in cassava (Manihot esculenta Crantz) croplands in Cameroon as revealed by Illumina MiSeq

Cassava, one of the most important crops in the tropics is positively affected by the association with Arbuscular Mycorrhizal Fungi (AMF). Such association is greatly influenced by the nature of the existing local AMF community. Here, we characterized AMF communities in soils from cassava fields in Andom and Bityili villages, located in Eastern and Southern Cameroon, respectively, by Illumina MiSeq of the ITS2 region. Soil characteristics involved marked changes in soil available P and slight differences in other nutrients availability and soil pH. Clustering based on a 97% sequence similarity generated 60 AMF Operational taxonomic units (OTU) grouped into the 12 genera Paraglomus, Claroideoglomus, Rhizophagus, Kamienskia, Funneliformis, Dominikia, Glomus, Ambispora, Archaeospora, Dentiscutata, Acaulospora, and Redeckera. Fifteen AMF species were identified from the 27 OTU, and they were dominated by Glomus sinuosum and Paraglomus occultum in both sites. The Andom soil which had less available P showed a higher AMF diversity and contained 10 site-specific species, compared to the Bityili soil where only 2 site-specific species were recovered. Multivariate analysis indicated that soil available P and C/N ratio were the main edaphic factors influencing the structure AMF in the study sites. This study provided useful information on the composition of AMF community in cassava field soils in two agro-ecologies of Cameroon.