Nitrogen sources alter ligninase and cellulase activities of thermophilic fungi isolated from compost and vermicompost.

Fungi harboring lignocellulolytic activity accelerate the composting process of agricultural wastes; however, using thermophilic fungal isolates for this process has been paid little attention. Moreover, exogenous nitrogen sources may differently affect fungal lignocellulolytic activity. A total of 250 thermophilic fungi were isolated from local compost and vermicompost samples. First, the isolates were qualitative assayed for ligninase and cellulase activities using Congo red (CR) and carboxymethyl cellulose (CMC) as substrates, respectively. Then, twenty superior isolates harboring higher ligninase and cellulase activities were selected and quantitatively assayed for both enzymes in basic mineral (BM) liquid medium supplemented with the relevant substrates and nitrogen sources including (NH4)2SO4 (AS), NH4NO3 (AN), urea (U), AS + U (1:1), or AN + U (1:1) with final nitrogen concentration of 0.3 g/L. The highest ligninase activities of 99.94, 89.82, 95.42, 96.25, and 98.34% of CR decolorization were recorded in isolates VC85, VC94, VC85, C145, and VC85 in the presence of AS, U, AS + U, AN, and AN + U, respectively. Mean ligninase activity of 63.75% in superior isolates was achieved in the presence of AS and ranked the highest among other N compounds. The isolates C200 and C184 exhibited the highest cellulolytic activity in the presence of AS and AN + U by 8.8 and 6.5 U/ml, respectively. Mean cellulase activity of 3.90 U/mL was achieved in AN + U and ranked the highest among other N compounds. Molecular identification of twenty superior isolates confirmed that all of them are belonging to Aspergillus fumigatus group. Focusing on the highest ligninase activity of the isolate VC85 in the presence of AS, the combination can be recommended as a potential bio-accelerator for compost production.

Biofilm forming rhizobacteria affect the physiological and biochemical responses of wheat to drought.

Plant growth promoting rhizobacteria (PGPR) can attenuate the adverse effects of water deficit on plant growth. Since drought stress tolerance of bacteria has earlier been associated to biofilm formation, we aimed to investigate the role of bacterial biofilm formation in their PGPR activity upon drought stress. To this end, a biofilm-forming bacterial collection was isolated from the rhizospheres of native arid grassland plants, and characterized by their drought tolerance and evaluated on their plant growth promoting properties. Most bacterial strains formed biofilm in vitro. Most isolates were drought tolerant, produced auxins, showed 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity and solubilized mineral phosphate and potassium, but at considerably different levels. Greenhouse experiments with the most promising isolates, B1, B2 and B3, under three levels of water deficit and two wheat varieties led to an increased relative water content and increased harvest index at both moderate and severe water deficit. However, the bacteria did not affect these plant parameters upon regular watering. In addition, decreased hydrogen peroxide levels and increased glutathione S-transferase activity occurred under water deficit. Based on these results, we conclude that by improving root traits and antioxidant defensive system of wheat, arid grassland rhizospheric biofilm forming bacilli may promote plant growth under water scarcity.

Role of lignin and thermophilic lignocellulolytic bacteria in the evolution of humification indices and enzymatic activities during compost production.

This study aimed to evaluate the effect of lignin content and thermophilic lignocellulolytic bacteria bioaugmentation on composting process. Treatments including bioaugmentation with thermophilic lignocellulolytic bacteria isolates such as Paenibacillus validus, Paenibacillus koreensis, Bacillus nealsonii, a mixture of the three mentioned bacterial isolates and control were compared at two level of organic media (high lignin content and low lignin content) in the form of nested factorial design. Several indices such as humification and enzymatic activities were monitored to evaluate the composting rate. The results revealed that high lignin treatments displayed higher ligninase, xylanase, protease and urease enzymatic activities compared to low lignin treatments. On the other hand, low lignin treatments showed higher level of humification indices, cellulase, beta-glucosidase and alkaline phosphomonoesterase enzymatic activities in comparison with high lignin treatments. Also, all measured enzymatic activities are at their highest between the second and the tenth weeks; however, this trend decreased to reach a steady point from the 18th weeks to the 24th weeks, but for urease enzymatic activity, a totally different trend in high and low lignin treatments was observed. Moreover, the highest humification indices as well as the cellulase and ß-glucosidase enzymatic activities were associated to the Bacillus nealsonii isolate and the full consortium. They also displayed the highest ligninase, xylanase, protease, and urease and phosphatase activities. The efficient isolates shortened the time required for completing the composting process for about 2 to 4 weeks compared to the control treatments. For all measured indices, the control treatment had the lowest values.

The arbuscular mycorrhizal mycelium from barley differentially influences various defense parameters in the non-host sugar beet under co-cultivation.

The interactions between arbuscular mycorrhizal fungi (AMF) and non-host species are poorly studied. Particularly scarce is information on members of the Amaranthaceae/Chenopodiaceae family. Sugar beet (Beta vulgaris) plants were co-cultivated with a host species (Hordeum vulgare) in the presence (+AMF) or absence of Rhizophagus intraradices to explore the hypothesis that the presence of an active, pre-established AMF mycelium induces defense responses in the non-host species. Biomass of sugar beet did not respond to the +AMF treatment, while its root exudation of organic acids and phenolic acids was drastically decreased upon co-cultivation with +AMF barley. The most conspicuous effect was observed on a wide range of potential defense parameters being differentially influenced by the +AMF treatment in this non-host species. Antioxidant defense enzymes were activated and the level of endogenous jasmonic acid was elevated accompanied by nitric oxide accumulation and lignin deposition in the roots after long-term +AMF treatment. In contrast, significant reductions in the levels of endogenous salicylic acid and tissue concentration and exudation of phenolic acids indicated that AM fungus hyphae in the substrate did not induce a hypersensitive-type response in the sugar beet roots and downregulated certain chemical defenses. Our results imply that the fitness of this non-host species is not reduced when grown in the presence of an AMF mycelium because of balanced defense costs. Further studies should address the question of whether or not such modulation of defense pattern influences the pest resistance of sugar beet plants under field conditions.

Positive effects of co-inoculation with Rhizophagus irregularis and Serendipita indica on tomato growth under saline conditions, and their individual colonization estimated by signature lipids.

Tomato roots can be colonized by both mycorrhizal fungi and the endophytic fungus Serendipita indica. This study was aimed at assessment of the impact of single or dual inoculation with R. irregularis and S. indica on tomato growth under saline conditions. We used signature compounds to estimate the abundance of each of these two fungi (fatty acid 16:1ω5 for R. irregularis and ergosterol for S. indica) in roots. A randomized block design was applied with four types of inoculation (no fungus, R. irregularis, S. indica or S. indica + R. irregularis) at different levels of salinity (1.2, 5, and 10 dS/m) with four replications per treatment. The plant dry weight was slightly higher in R. irregularis- than S. indica-inoculated plants, and the highest plant biomass was achieved with dual inoculation. R. irregularis increased the content of the neutral lipid fatty acid 16:1ω5 from 97 to 5300 nmol/g and phospholipid fatty acid 16:1ω5 from 8 to 141 nmol/g in roots (at a salinity level of 1.2 dS m-1), but the increases were lower at higher levels of salinity. Moreover, both these arbuscular mycorrhizal fungal markers were slightly decreased in the presence of S. indica. Root ergosterol increased from 7 to 114 µg g-1 with S. indica inoculation. With increasing salinity, the concentration of ergosterol in roots decreased. Inoculation with R. irregularis caused a decrease in root ergosterol. Increasing salinity resulted in an increase of Cl and Na in tomato shoots, but the increase was significantly lower in single- or dual-inoculated plants in contrast to the control plants.

Silicon influences growth and mycorrhizal responsiveness in strawberry plants.

Effect of silicon (Si) on the response of strawberry (Fragaria × ananassa var. Parus) plants to arbuscular mycorrhizal fungus (AMF) was studied under growth chamber conditions. Plants were grown in perlite irrigated with nutrient solution without (- Si) or with (+ Si) 3 mmol L-1 Si (~ 84 mg L-1 Si as Na2SiO3) in the absence (- AMF) or presence (+ AMF) of fungus. Dry matter production, root colonization rate, photosynthesis rate and water relation parameters were all improved by both Si and AMF, and the highest amounts were achieved by + Si + AMF treatment. Mycorrhizal effectiveness increased by Si treatment associated with higher Si concentration in the + AMF plants. Leaf concentrations of total soluble and cell wall-bound phenolics were increased by Si accompanied by the enhanced activity of phenylalanine ammonia lyase, but not polyphenol oxidase. Profile of phenolics compound revealed that gallic acid, caffeic acid, epicatechin, chlorogenic acid, ellagic acid and kaempferol increased by both Si and AMF treatments, while p-coumaric acid decreased. In addition to vegetative growth, both treatments improved fruit yield and its quality parameters. Our results showed that Si and AMF acted in a synergistic manner and improved growth and biochemical parameters in strawberry plants. However, the mechanism for Si-mediated increase of mycorrhizal effectiveness is not known, thereby needing further elucidation.

Cd-induced production of glomalin by arbuscular mycorrhizal fungus (Rhizophagus irregularis) as estimated by monoclonal antibody assay.

Glomalin is a specific fungal glycoprotein produced by arbuscular mycorrhizal (AM) fungi belonging to the Glomerales which could efficiently sequestrate heavy metals. The glomalin has been introduced as a heat shock protein and there are evidences that increasing levels of heavy metals could enhance its production. We examined the influence of Cd concentrations on glomalin production by AM fungus, as well as its contribution to the sequestration of Cd in both pot and in vitro culture conditions. Pot experiment was carried out using pure sand with Trifolium repens L. as host plant, mycorrhized by Rhizophagus irregularis and treated with Cd levels of 0, 15, 30, and 45 µM. In vitro experiment was performed in two-compartment plates containing the transformed carrot roots mycorrhized with the same fungus and treated with Cd levels of 0, 0.001, 0.01, and 0.1 mM. The immunoreactive and Bradford reactive glomalin contents in both experiments increased as so raising Cd concentration. Total Cd sequestrated by hyphal glomalin in both cultures was significantly increased as the levels of Cd increased. The highest contents of Cd sequestration in pot (75.78 µg Cd/mg glomalin) and in vitro (11.44 µg Cd/mg glomalin) cultures were recorded at the uppermost levels of Cd, which significantly differed with other levels. Our results suggested that under Cd-induced stress, stimulated production of glomalin by AM fungus may be a protective mechanism against the toxic effect of Cd.

Carbon dynamics in mycorrhizal symbioses is linked to carbon costs and phosphorus benefits.

The nutrient and carbon (C) allocation dynamics in mycorrhizal hyphal networks cause variation in costs and benefits for individual plants and fungi and influence the productivity, diversity and C cycling in ecosystems. We manipulated light and phosphorus (P) availability in a pot experiment with Trifolium subterraneum colonised by the arbuscular mycorrhizal (AM) fungus Glomus intraradices. Stable (13)C-labelling was used to trace assimilated CO(2) to the mycorrhizal fungus in roots and soil using compound-specific isotope ratio mass spectrometry. We used the neutral lipid fatty acid 16:1omega5 as a signature for AM fungal storage lipids. Both P and shading reduced the AM fungal lipid accumulation in the intraradical mycelium, while only P reduced the amount of lipids in the extraradical mycelium. Recently assimilated plant C was only allocated to the mycorrhizal fungus to a small extent when plant mycorrhizal benefit was reduced by P fertilization, while increasing the plant C cost by shading did not reduce the C flow to the fungus. These results are of importance for our conception of mycorrhizal dynamics during periods of shade in nature.

Effects of mycorrhizal colonization on growth parameters of onion under different irrigation and soil conditions.

The effects of three Arbuscular Mycorrhizal Fungi (AMF), Glomus versiforme, G. intraradices and G. etonicatum) and three irrigation intervals (7, 9 and 11 days) on growth of onion (Allium cepa L.) cv. Red Azar Shahr were studied under two soil conditions (sterilized and non-sterilized). The results indicated that, AMF colonization improved plant height, Leaf Area Index (LAI), total biomass, bulb dry mass and diameter, Harvest Index (HI) and chlorophyll content (p < 0.001). Bulbing occurred 10-15 days earlier in mycorrhizal plants. Irrigation interval decreased biomass, LAI, Leaf Area Ratio (LAR), bulb diameter and dry mass and chlorophyll content (b and total) at 11 day irrigation interval. In term of interaction, G. versiforme at 9 day and non-mycorrhizal plants at 11 day produced the greatest and the lowest LAI (8.56 vs. 1.57), respectively. Mycorrhizal onions in contrary to non-mycorrhizal ones produced more LAI and biomass in sterilized soil and inoculation with G. etonicatum and the non-mycorrhizal onions in sterilized soil had the highest and the lowest biomass, respectively.