Inoculation effect of arbuscular mycorrhizal fungi on the growth of Vatica chinensis L.: a critically endangered species of Western Ghats

Pot experiments were conducted to assess the efficacy of three arbuscular mycorrhizal (AM) fungi viz. Glomus fasciculatum, Funneliformis mosseae and Acaulospora laevis on the growth of Vatica chinensis. Plants grown in the presence of AM fungi generally showed an increase in growth and biomass over those grown in the absence of AM fungal inoculation. It was observed that a consortium of AM fungi without organic manure was found to be the best treatment to enhance plant biomass. The plant height, number of leaves and plant dry and fresh matter, plant P and N contents varied significantly with treatments. The AM colonization was higher in the seedling roots inoculated with two or three AM fungi. This study indicated that the inoculation with AM fungi either alone or in combination improved the growth, biomass yield and phosphorus nutrition of V. chinensis compared to that of the control. This information may be helpful to improve the growth of these species in sustainable forestry program.     

Regulation of hyphal morphogenesis by Ras and Rho small GTPases

The fungal kingdom is extremely diverse – comprised of over 1.5 million species including yeasts, molds and mushrooms. Essentially, all fungi have cell walls that contain chitin and the cells of most fungi grow as tube-like filaments called hyphae. These filamentous fungi, such as the mold Neurospora crassa, develop branched radial networks of hyphae referred to as mycelium. In contrast, non-filamentous fungi do not form radial mycelia, but grow as single cells, which reproduce by either budding or fission such as Saccharomyces cerevisiae or Schizosaccharomyces pombe, respectively. Finally, there are fungi that are capable of switching between single cell, yeast form growth and filamentous growth such as Candida albicans. The switch from yeast to filamentous growth in these so-called dimorphic fungi is a virulence trait in many human and plant pathogens. Highly conserved master regulators of all three fungal growth modes – filamentous, non-filamentous and dimorphic – are the Ras and Rho small GTPases, which spatially and temporally control cell polarity establishment and maintenance. This review summarizes the key roles of the Ras and Rho GTPases during hyphal morphogenesis in a range of fungi.     

The arbuscular mycorrhizal fungus Rhizophagus intraradices and other microbial groups affect plant species in a copper-contaminated post-mining soil

Background and aimArbuscular mycorrhizal fungi (AMF) have an important role in plant-microbe interactions. But, there are few studies in which the combined effect of AMF with a stress factor, such as the presence of a metal, on plant species were assessed. This study investigated the effect of arbuscular mycorrhizal (AM) fungus Rhizophagus intraradices and other soil microbial groups in the presence of copper on three plant species in a microcosm experiment.MethodsTwo grass species Poa compressa and Festuca rubra and one herb species Centaurea jacea were selected as model plants in a pot-design test in which soils were artificially contaminated with copper. Treatments were bacteria (control), saprophytic fungi, protists, and a combined treatment of saprophytic fungi and protists, all in the presence or absence of the AM fungal species. After sixty days, plants were harvested and the biomass of grass and herb species and microbial respiration were measured.ResultsThe results showed almost equal above- and belowground plant biomass and microbial respiration in the treatments in the presence or absence of R. intraradices. The herb species C. jecea responded significantly to the soil inoculation with AM fungus, while grass species showed inconsistent patterns. Significant effect of AMF and copper and their interactions was observed on plant biomass when comparing contaminated vs. non-contaminated soils.ConclusionStrong effect of AMF on the biomass of herb species and slight changes in plant growth with the presence of this fungal species in copper-spiked test soils indicates the importance of mycorrhizal fungi compared to other soil microorganisms in our experimental microcosms.     

Production of sedoheptulose by Bacillus subtilis

Wild type strains of Bacillus subtilis produced sedoheptulose from d-ribose but not from d-glucose, B. subtilis mutants deficient in transketolase produced sedoheptulose when d-glucose was used as a carbon source. The addition of d-ribose to the culture medium increased the amount of sedoheptulose accumulated, reaching about 20 mg/ml of culture broth. The mutant strains reverted to wild type strains at a high frequency during cell growth, and therefore the accumulation of sedoheptulose was caused by the genetic instability of the mutant: d-ribose formed from d-glucose by the mutant strain was converted into sedoheptulose by revertant cells that appeared during cultivation.     

The FRP1 F-box gene has different functions in sexuality, pathogenicity and metabolism in three fungal pathogens

Plant‐pathogenic fungi employ a variety of infection strategies; as a result, fungi probably rely on different sets of proteins for successful infection. The F‐box protein Frp1, only present in filamentous fungi belonging to the Sordariomycetes, Leotiomycetes and Dothideomycetes, is required for nonsugar carbon catabolism and pathogenicity in the root‐infecting fungus Fusarium oxysporum. To assess the role of Frp1 in other plant‐pathogenic fungi, FRP1 deletion mutants were generated in Fusarium graminearum and Botrytis cinerea, and their phenotypes were analysed. Deletion of FgFRP1 in F. graminearum led to impaired infection of barley roots, but not of aerial plant parts. Deletion of BcFRP1 in B. cinerea did not show any effect on pathogenicity. Sexual reproduction, however, was impaired in both F. graminearum and B. cinerea FRP1 deletion mutants. The mutants of all three fungi displayed different phenotypes when grown on an array of carbon sources. The F. oxysporum and B. cinerea deletion mutants showed opposite growth phenotypes on sugar and nonsugar carbon sources. Replacement of FoFRP1 in F. oxysporum with the B. cinerea BcFRP1 resulted in the restoration of pathogenicity, but also in a switch from impaired growth on nonsugar carbon sources to impaired growth on sugar carbon sources. This effect could be ascribed in part to the B. cinerea BcFRP1 promoter sequence. In conclusion, the function of the F‐box protein Frp1, despite its high sequence conservation, is not conserved between different fungi, leading to differential requirements for pathogenicity and carbon source utilization.     

Life histories of two desert species of the bulbous genus Bellevalia

Summary Annual biomass increment and biomass partitioning to leaves, roots and reproduction, and biomass storage in the below-ground bulb was measured in plants of two species of the geophytic genus Bellevalia grown outdoors at three levels of soil moisture. The differences between the species were in accordance with the hypothesis that plants of more arid environments should rely more on internal reserves than plants of more productive environments. In Bellevalia desertorum, a shallow rooted species of the most arid habitats in the Central Negev, leaf and root development during outgrowth at the beginning of winter was rather variable, and followed soil moisture availability to a certain degree. A small portion of its biomass budget was committed to seed production, which varied little among the irrigation regimes. The rest of the biomass was stored in the bulb. The amount of biomass devoted annually to reproduction was mainly determined by the amount of reserves already present in the bulb. In contrast, in B. eigii, which grows in the more productive wadis with its bulb at a depth of 15 to 30 cm, leaf and root growth was not only determined by water availability, but also by initial bulb mass. This resulted in a greater potential relative growth rate than in B. desertorum, but also in a greater risk of accumulating less biomass than it spent in root and leaf construction under poor soil moisture conditions. In this species, reproductive biomass and seed yield were proportional to current biomass gain and, in contrast to B. desertorum, independent of initial bulb mass, provided that the bulb was large enough to initiate flowering.     

Interactions of Taxol-producing endophytic fungus with its host (Taxus spp.) during Taxol accumulation

Endophytic fungi (Fusarium mairei) culture broth (EFCB) was added to cell suspension cultures of Taxus cuspidata. After 5 days, cultures of T. cuspidata given 4 ml of EFCB produced a maximal yield of 6.11 mg/l paclitaxel, with a release ratio of 75%, 2- and 6.8-fold, respectively, greater than the controls. The active element in EFCB is an exopolysaccharide of ∼79 kD. Endophytic fungi produced 0.19 mg/l of paclitaxel in its producing medium. However, when the supernatant of Taxus cell suspension cultures from day 20 was added to the paclitaxel-producing medium, the biomass of fungi decreased by 24% and the yield of paclitaxel by 45%. In a co-culture system of plant and fungus, the yield of paclitaxel (12.8 mg/l) was >2-fold higher than that in the EFCB-treatment system.     

Factors affecting growth and urease production by Trichophyton spp.

Among Trichophyton spp. examined for urease production, T. rubrum was negative, whereas T. mentagrophytes appeared to be the most active species. Urease was not detected in cell-free culture fluids of the tested fungi. The endocellular urease of the test fungi was essentially constitutive. Moreover, addition of urea to the growth medium of these organisms markedly inhibited their mycelial biomass and ureolytic yield. Environmental factors showed variable effects on the test fungi and there was no correlation between mycelial growth and urease activity of these fungi.     

干旱条件下AM真菌对植物生长和土壤水稳定性团聚体的影响

采用分室培养系统,模拟正常水分和干旱胁迫两种环境条件,探讨不同丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)对紫花苜蓿(Medicago sativa L.)生长和土壤水稳性团聚体的影响.试验条件下,Glomus intraradices对苜蓿根系的侵染率均显著高于Acaulospora scrobiculata和Diversispora spurcum接种处理.正常水分条件下,供试AM真菌均能显著提高植株生物量及磷浓度.干旱胁迫显著抑制了植株生长和菌根共生体发育,总体上菌根共生体对植株生长没有明显影响,接种D.spurcum甚至趋于降低植株生物量;同时,仅有G.intraradices显著提高了植株磷浓度.AM真菌主要影响到＞2mm的水稳性团聚体数量,以G.intraradices作用效果最为显著.在菌丝室中,G.intraradices显著提高了总球囊霉素含量.研究表明AM真菌对土壤大团聚体形成具有积极作用,而菌根效应因土壤水分条件和不同菌种而异,干旱胁迫下仅有G.intraradices对土壤结构和植物生长表现出显著积极作用.在应用菌根技术治理退化土壤时,需要选用抗逆性强共生效率高的菌株,对于不同AM真菌抗逆性差异的生物学与遗传学基础尚需进一步研究.     

Expression and characterization of the Neurospora crassa endoglucanase GH5-1

Filamentous fungi secrete a wide range of enzymes, including cellulases and hemicellulases, with potential applications in the production of lignocellulosic biofuels. Of the cellulolytic fungi, Hypocrea jecorina (anamorph Trichoderma reesei) is the best characterized in terms of cellulose degradation, but other cellulolytic fungi, such as the model filamentous fungus Neurospora crassa, can serve a crucial role in building our knowledge about the fungal response to biomass due to the many molecular and genetic tools available for this organism. Here we cloned and expressed GH5-1 (NCU00762), a secreted endoglucanase in N. crassa. The protein was produced using a ccg-1 promoter under conditions in which no other cellulases are present. Native GH5-1 (nGH5-1) and this recombinant GH5-1 (rGH5-1) were purified to gauge differences in glycosylation and activity; both rGH5-1 and nGH5-1 were similarly glycosylated, with an estimated molecular weight of 52 kDa. On azo-carboxymethylcellulose, rGH5-1 activity was equal to that of nGH5-1, and on cellulose (Avicel) rGH5-1 was 20% more active. The activity of a GH5-1-GFP fusion protein (rGH5-1-GFP-6xHis) was similar to rGH5-1 and nGH5-1. To determine the binding pattern of catalytically active rGH5-1-GFP-6xHis to plant cell walls, Arabidopsis seedlings were incubated with rGH5-1-GFP-6xHis or Pontamine Fast Scarlet 4B (S4B), a cellulose-specific dye. Confocal microscopy showed that rGH5-1-GFP-6xHis bound in linear, longitudinal patterns on seedling roots, similar to S4B. The functional expression and characterization of rGH5-1 and its GFP fusion derivative set important precedents for further investigation of biomass degradation by filamentous fungi, especially N. crassa, with applications for characterization and manipulation of novel enzymes.     

Arbuscular mycorrhizal fungi in two mangroves in South China

The symbiosis between arbuscular mycorrhizal fungi (AMF) and mangrove plant species was investigated in two mangrove swamps in south China. AMF were mostly found in the form of hyphae and were commonly associated with all the mangrove species we investigated. Six AMF species belonging to the genera Glomus or Acaulospora were identified. Multiple step-wise linear regression analyses showed that hydrological conditions and phosphorus levels in the rhizosphere were the main abiotic factors affecting the colonization of mangrove species by AMF. A greenhouse experiment was conducted to evaluate the effects of AMF inoculation on the growth and nutrient uptake of a true mangrove plant species, Sonneratia apetala B. Ham. The inoculated AMF significantly improved growth, resulting in greater plant height, diameter at ground level and plant biomass, as well as increased absorption of N, P and K. These findings suggest that AMF play important roles in mangrove ecosystems.     

Production of quinoprotein d-glucose dehydrogenase in the culture medium of Acinetobacter calcoaceticus

On addition of low concentrations (0.005%) of Triton X-100 to a mineral medium supplemented with 0.5% heptadecane, a marked stimulation of growth rate was observed for Acinetobacter calcoaceticus strains able to grow on alkanes while appreciable amounts of soluble quinoprotein d-glucose dehydrogenase [d-glucose: (pyrroloquinoline-quinone) 1-oxidoreductase, EC 1.1.99.17] were found in the culture medium. At higher Triton X-100 concentrations (0.04%), still larger amounts of d-glucose dehydrogenase and also cytoplasmic enzyme activities appeared in the culture medium. Although combinations of other carbon sources plus non-ionic detergents also produced these enzymes in the medium, the combination of heptadecane and Triton X-100 gave higher levels and had a stabilizing effect on d-glucose dehydrogenase. Therefore, by using this combination and culturing within certain pH limits, a stable enzyme solution, having already a high specific activity, is produced while the cell harvesting and disruption steps can be circumvented. The results indicate that d-glucose dehydrogenase in this organism is a periplasmic enzyme, coupled to a cytochrome b.     

Disruption of the Eng18B ENGase gene in the fungal biocontrol agent Trichoderma atroviride affects growth, conidiation and antagonistic ability

The recently identified phylogenetic subgroup B5 of fungal glycoside hydrolase family 18 genes encodes enzymes with mannosyl glycoprotein endo-N-acetyl-β-D-glucosaminidase (ENGase)-type activity. Intracellular ENGase activity is associated with the endoplasmic reticulum associated protein degradation pathway (ERAD) of misfolded glycoproteins, although the biological relevance in filamentous fungi is not known. Trichoderma atroviride is a mycoparasitic fungus that is used for biological control of plant pathogenic fungi. The present work is a functional study of the T. atroviride B5-group gene Eng18B, with emphasis on its role in fungal growth and antagonism. A homology model of T. atroviride Eng18B structure predicts a typical glycoside hydrolase family 18 (αβ)(8) barrel architecture. Gene expression analysis shows that Eng18B is induced in dual cultures with the fungal plant pathogens Botrytis cinerea and Rhizoctonia solani, although a basal expression is observed in all growth conditions tested. Eng18B disruption strains had significantly reduced growth rates but higher conidiation rates compared to the wild-type strain. However, growth rates on abiotic stress media were significantly higher in Eng18B disruption strains compared to the wild-type strain. No difference in spore germination, germ-tube morphology or in hyphal branching was detected. Disruption strains produced less biomass in liquid cultures than the wild-type strain when grown with chitin as the sole carbon source. In addition, we determined that Eng18B is required for the antagonistic ability of T. atroviride against the grey mould fungus B. cinerea in dual cultures and that this reduction in antagonistic ability is partly connected to a secreted factor. The phenotypes were recovered by re-introduction of an intact Eng18B gene fragment in mutant strains. A putative role of Eng18B ENGase activity in the endoplasmic reticulum associated protein degradation pathway of endogenous glycoproteins in T. atroviride is discussed in relation to the observed phenotypes.     

Endophytic bacteria isolated from orchid and their potential to promote plant growth

Twelve endophytic bacteria were isolated from the meristem of in vitro Cymbidium eburneum orchid, and screened according to indole yield quantified by colorimetric assay, in vitro phosphate solubilization, and potential for plant growth promotion under greenhouse conditions. Eight strains with positive results were classified into the genus Paenibacillus by FAME profile, and evaluated for their ability to increase survival and promote the growth of in vitro germinated Cattleya loddigesii seedlings during the acclimatization process. The obtained results showed that all strains produced detectable indole levels and did not exhibit potential for solubilizing inorganic phosphate. Particularly, an increase of the total biomass and number of leaves was observed. Two strains of Paenibacillus macerans promoted plant growth under greenhouse conditions. None of the treatments had a deleterious effect on growth of inoculated plants. These results suggest that these bacterial effects could be potentially useful to promote plant growth during seedling acclimatization in orchid species other than the species of origin.     

Production of lytic enzymes by<Emphasis Type="Italic">Trichoderma</Emphasis> spp. and their effect on the growth of phytopathogenic fungi

The production of β-1,3-glucanases and chitinases by three strains ofTrichoderma in submerged cultures was determined. The synthesis of enzymes was induced by cell wall biopolymers of phytopathogenic fungi (Botrytis cinerea, Fusarium culmorum andF. oxysporum). T. hamatum produced the highest β-1,3-glucanase activity; the most effective inducer of enzyme synthesis was the biomass ofF. oxysporum. All examined strains ofTrichoderma inhibited phytopathogen growth in biotic tests. The diffusion tests showed that the lytic enzymes take part in growth inhibition of phytopathogenic fungi.     

Maize development and grain quality are differentially affected by mycorrhizal fungi and a growth-promoting pseudomonad in the field

Arbuscular mycorrhizal (AM) fungi and plant growth-promoting bacteria (PGPB) can increase the growth and yield of major crops, and improve the quality of fruits and leaves. However, little is known about their impact on seed composition. Plants were inoculated with AM fungi and/or the bacterial strain Pseudomonas fluorescens Pf4 and harvested after 7 months of growth in open-field conditions. Plant growth parameters were measured (biomass, length and circumference of spikes, number of grains per cob, grain yield, and grain size) and protein, lipid, and starch content in grains were determined. Plant growth and yield were increased by inoculation with the microorganisms. Moreover, spikes and grains of inoculated plants were bigger than those produced by uninoculated plants. Regarding grain composition, the bacterial strain increased grain starch content, especially the digestible components, whereas AM fungi-enhanced protein, especially zein, content. Plant inoculation with the fluorescent pseudomonad and mycorrhizal fungi resulted in additive effects on grain composition. Overall, results showed that the bacterial strain and the AM fungi promoted maize growth cultivated in field conditions and differentially affected the grain nutritional content. Consequently, targeted plant inoculation with beneficial microorganisms can lead to commodities fulfilling consumer and industrial requirements.     

Growth and sporulation of Beauveria bassiana and Metarhizium anisopliae in a basal medium containing various carbohydrate sources

Naturally occurring isolates of two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae, pathogenic to the pecan weevil, Curculio caryae, were cultured in flasks of a liquid synthetic culture medium containing one of 22 individual carbohydrate sources to determine growth and sporulation responses. B. bassiana grew best on d-melizitose but sporulated best on d-sucrose, d-trehalose, and d-glucose. M. anisopliae grew best on d-mannose but sporulated best on i-inositol and glycerol. B. bassiana grew least on l-rhamnose but M. anisopliae grew least on d-arabinose. Both fungi sporulated least on d-sorbose. Both fungi utilized d-trehalose, a major component of insect hemolymph.     

The effect of copper on the growth of wood-rotting fungi and a blue-stain fungus

The effect of copper (II) ions on the growth of three brown-rot fungi, six white-rot fungi and one blue-stain fungus in solid medium was evaluated. The fungi were grown in malt extract agar with different concentrations of copper added, and the radial growth rate was determined. At the end of the incubation period, the mycelial biomass and the media pH were determined. The white-rot and blue-stain fungus grew up to 3 mM and 6 mM copper, respectively and the brown-rot fungi were the only ones that grew up to 10 mM, with higher growth rates than those shown by the other fungi. In general, the brown-rot fungi produced greater acidification in the culture media than the white-rot fungi and blue-stain fungus, and the acidification increased when the amount of copper was increased. The biomass production for the different species, in the absence or presence of copper, was not related to the radial growth rate, and the fungal species that produced the greatest biomass amounts did not correspond to those that presented the highest growth rates. The brown-rot fungi Wolfiporia cocos and Laetiporus sulfureus and blue-stain fungus Ophiostoma sp. demonstrated greater tolerance to high copper concentrations in solid medium than the white-rot fungi, determined as radial growth rate. On the other hand, the highest biomass producers in solid medium with copper added were the white-rot fungi Ganoderma australe and Trametes versicolor and the brown-rot fungus Gloeophyllum trabeum.