Paecilomyces Marquandii Research Articles

Cultural Characteristics of Phylloplane Fungi and Leaf Spot Pathogens Isolated from Cajanus cajan

Four different isolates of fungi were discovered from the phylloplane of Arhar. They were recognized and identified to be Fusarium solani, Paecilomyces marquandii, Aspergillus niger and Aspergillus flavus. Similarly, from the diseased leaves of Arhar two pathogens were isolated, characterized and identified to be Curvularia lunata and Sterile. These fungal isolates were cultivated on seven different culture media viz., Czapek’s Dox Agar, Potato Dextrose Agar (PDA), Oat Meal Agar, Rose Bengal Agar, Richard’s Synthetic Agar, Sabouraud’s Dextrose Agar and Malt Extract Agar. Right after seven days of incubation at 250C, the mycelial growth rate, colony character and sporulation patterns of these fungi were evaluated. The type of growth media utilized had a significant impact on the diameter, cultural traits and sporulation of a few selected test fungi. On PDA and Richard’s synthetic agar media, Fusarium solani, Aspergillus niger and Curvularia lunata recorded the superlative radial growth. In PDA and Sabouraud’s dextrose agar Aspergillus flavus and Sterile recorded the highest growth. Paecilomyces marquandii recorded the maximal radial growth in Sabouraud’s dextrose agar.

Secondary metabolites with chemotaxonomic value from Paecilomyces marquandii

A systematic investigation on the secondary metabolites of Delphinium-associated fungus Paecilomyces marquandii afforded eight compounds, including six sorbicillinoid polyketides (1–6) and two ergosterol sterols (7–8). Among them, six compounds (1–5, 8) were identified for the first time from the genus Paecilomyces, and seven compounds (1–5, 7–8) have not previously been found in the fungus P. marquandii. The chemotaxonomic significance of these compounds was discussed. Some of the isolates (2, 5, and 6) showed limited distribution and could serve as chemotaxonomic markers to facilitate strain identification of P. marquandii.

Insights into the metabolic pathways and biodegradation mechanisms of chloroacetamide herbicides

Chloroacetamide herbicides are widely used around the world due to their high efficiency, resulting in increasing levels of their residues in the environment. Residual chloroacetamides and their metabolites have been frequently detected in soil, water and organisms and shown to have toxic effects on non-target organisms, posing a serious threat to the ecosystem. As such, rapid and efficient techniques that eliminate chloroacetamide residues from the ecosystem are urgently needed. Degradation of these herbicides in the environment mainly occurs through microbial metabolism. Microbial strains such as Acinetobacter baumannii DT, Bacillus altitudinis A16, Pseudomonas aeruginosa JD115, Sphingobium baderi DE-13, Catellibacterium caeni DCA-1, Stenotrophomonas acidaminiphila JS-1, Klebsiella variicola B2, and Paecilomyces marquandii can effectively degrade chloroacetamide herbicides. The degradation pathway of chloroacetamide herbicides in aerobic bacteria is mainly initiated by an N/C-dealkylation reaction, followed by aromatic ring hydroxylation and cleavage processes, whereas dechlorination is the initial reaction in anaerobic bacteria. The molecular mechanisms associated with bacterial degradation of chloroacetamide herbicides have been explored, with amidase, hydrolase, reductase, ferredoxin and cytochrome P450 oxygenase currently known to play a pivotal role in the catabolic pathways of chloroacetamides. The fungal pathway for the degradation of these herbicides is more complex with more diversified products, and the degradation enzymes and genes involved remain to be discovered. However, there are few reviews specifically summarizing the microbial degrading species and biochemical mechanisms of chloroacetamide herbicides. Here, we briefly summarize the latest progress resulting from research on microbial strain resources and enzymes involved in degradation of these herbicides and their corresponding genes. Furthermore, we explore the biochemical pathways and molecular mechanisms for biodegradation of chloroacetamide herbicides in depth, thereby providing a reference for further research on the bioremediation of such herbicides.

Enzymatic and Antimicrobial Activities in Polar Strains of Microscopic Soil Fungi.

Potential to produce inducible enzymes (several hydrolases and oxidases) and antibiotics as secondary metabolites was studied in soil micromycete strains from the Arctic (Franz Josef Land and Novaya Zemlya) and Antarctica (the oases Thala Hills, Larsemann Hills, Schirmacher, and Marie Byrd Land). Maximal esterase activity was observed in strains of two typical Antarctic species, Hyphozyma variabilis 218 and Thelebolus ellipsoideus 210 (51 and 29 nmol FDA/((g myceliumh), respectively). Cellulolytic activity was maximal (89 µmol glucose/mg biomass) in Ascochyta pisi 192. Extracellular phenol oxidase (laccase) and peroxidase activities were not detected in the strains examined. Antibacterial activity toward Bacillus subtilis ATCC 6633 was observed in 75% of the Antarctic micromycete strains. Higher-activity strains were isolated from organic-rich moist habitats with a moss or lichen cover. Maximal activities were displayed by Paecilomyces marquandii 166, Penicillium janczewskii 165, Penicillium roseopurpureum 169, and Thelebolus ellipsoideus 210. Antagonistic activity toward Antarctic bacterial strains was shown by 77% of the microfungal strains examined. Maximal inhibition was observed with strains of the typical Antarctic species Antarctomyces psychrotrophicus MT303855 and the eurytopic species Sarocladium kiliense MT303856. Antimycotic activity was observed in 42% of the strains. Both activities were detected in 38% of the Antarctic strains.

Aqueous Two-Phase System Extraction of Polyketide-Based Fungal Pigments Using Ammonium- or Imidazolium-Based Ionic Liquids for Detection Purpose: A Case Study.

Demand for microbial colorants is now becoming a competitive research topic for food, cosmetics and pharmaceutics industries. In most applications, the pigments of interest such as polyketide-based red pigments from fungal submerged cultures are extracted by conventional liquid–liquid extraction methods requiring large volumes of various organic solvents and time. To address this question from a different angle, we proposed, here, to investigate the use of three different aqueous two-phase extraction systems using either ammonium- or imidazolium-based ionic liquids. We applied these to four fermentation broths of Talaromyces albobiverticillius (deep red pigment producer), Emericella purpurea (red pigment producer), Paecilomyces marquandii (yellow pigment producer) and Trichoderma harzianum (yellow-brown pigment producer) to investigate their selective extraction abilities towards the detection of polyketide-based pigments. Our findings led us to conclude that (i) these alternative extraction systems using ionic liquids as greener extractant means worked well for this extraction of colored molecules from the fermentation broths of the filamentous fungi investigated here; (ii) tetrabutylammonium bromide, [N4444]Br-, showed the best pigment extraction ability, with a higher putative affinity for azaphilone red pigments; (iii) the back extraction and recovery of the fungal pigments from ionic liquid phases remained the limiting point of the method under our selected conditions for potential industrial applications. Nevertheless, these alternative extraction procedures appeared to be promising ways for the detection of polyketide-based colorants in the submerged cultures of filamentous fungi.

Preliminary study of the degradation of biocellulose based film using soil fungi Aspergillus unguis TP3 and Paecilomyces marquandii TP4 producing cellulose

Bacterial cellulose or biocellulose (BC) is cellulose produced from the activity of bacteria in the suitable growth media containing glucose as mainly carbon source. Due to its unique properties, BC is used for edible packaging. Many studies reported on anti bacterial activity on BC based-edible packaging against Escherichia coli and Staphylococcus aureus as as well as Listeria monocytogenes. This study reports the biodegradation of BC (sample A) based-film by using Aspergillus unguis and Paecilomyces marquandii. For comparison, BC film was added with CMC (sample B), with glycerol (sample C), with CMC and glycerol (sample D), respectively. Biodegradation was carried out using broth fermentation and solid substrate fermentation (SSF). BC films (1 cm x 1 cm) and fungi were fermented in 100 mL of sterile aquadest for 60 days with agitation (120 rpm at room temperature). For treatment in SSF, fungi were inoculated into BC films for 60 days at room temperature in petridish. Results showed that all compounds of films were still available based on FTIR results. The physical performance of films BC was in solid chewy (A), like chewy thread (B and C), and like powder (D) form. Results showed that both fungi biodegraded films through broth fermentation whereas no biodegradation activity on SSF. From SEM analysis, it showed that the film surfaces performed fine and smooth morphology.

Sixteen New Records of Ascomycetes from Crop Field Soil in Korea

The present study reports 16 species of Ascomycota that were previously unknown in Korea, namely Acremonium cellulolyticus (KNU14-25), Acremonium zonatum (KNU14-15), Chaetomium madrasense (KNU14-9), Cladosporium silenes (KNU 14-18-1), Humicolopsis cephalosporioides (KNU15-3), Leptosphaerulina chartarum (KNU14-16), Paecilomyces marquandii (KNU14- 8), Paecilomyces tenuis (KNU14-18-2), Paraphaeosphaeria sporulosa (KNU15-2), Penicillium rubidurum (KNU14-12), Pochonia suchlasporia (KNU15-6), Sporothrix inflata (KNU15-8), Thielavia hyrcaniae (KNU15-1), Thielavia terricola (KNU14-23-1), Xylogone sphaerospora (KNU15-7), and Zopfiella longicaudata (KNU15-5). These fungal species were isolated from soil samples collected from various regions of Korea and identified based on their morphological characteristics and rDNA internal transcribed spacer sequence data. Full descriptions and illustrations for each species are provided.

Nematicidal activity of Paecilomyces marquandii proteases on infective larvae of Ancylostoma spp

The present study aimed to evaluate the action of Paecilomyces marquandii proteases on Ancylostoma spp L3. White halos in the zymogram confirmed the proteolytic action. Difference (p <0.01) between the number of L3 in the differents groups was found, with 41.4% of reduction of Ancylostoma spp. L3 before 24 hours.

Pathogenicity of Paecilomyces marquandii on eggs of Meloidogyne incognita

Among fungi, species of the genus Paecilomyces are considered as promising biological control agents with high potential to reduce root-knot nematode (RKN) and other nematode populations. In this research we investigated a soil hyphomycetes, Paecilomyces marquandii and its pathogenicity on eggs of RKN in vitro. In greenhouse test, root weight, plant length, fresh weight and dry weight of plants, gall index and proportion of infected eggs were determined. P. marquandii, effectively promoted growth of plants inoculated with M. incognita by suppressing its pathogenesis as root galling and reducing egg mass production. At 25 °C, a great number of nematode eggs were parasitized by P. marquandii, inhibiting juvenile development.

Antibacterial, Antifungal and Anticancer Activity of Five Strains of Soil Microorganisms Isolated From Tangkuban Perahu Mountain by Fermentation

Microorganisms were isolated from soil taken from Tangkuban Perahu mountain. Five strains were investigated in this study, designated TP1, TP2, TP3, TP4, and TP5, respectively. Morphological, biochemical and molecular identifications were conducted for all five strains. These isolates were shown to be closely related to Nocardia sp. YIM 65630 (90%), Streptomyces galbus (99%), Aspergillus unguis (86%), Paecilomyces marquandii (100%) and Nocardia niigatensis (95%), respectively. Production of antibacterial, antifungal and anticancer metabolites was done by fermentation. Screening for bioactivity of five isolates was done by testing the fermentation broth against resistant and pathogenic bacteria, fungi and T47D breast cancer cell line. TP2 strain showed the best bioactivity; the metabolite was purified by extraction with ethyl acetate. Antibacterial, antifungal and anticancer activities from the ethyl acetate extract of TP2 strain were tested by agar diffusion, microdilution and MTT. The extract was shown to be active against methicillin resistant Staphylococcus, methicillin sensitive Staphylococcus aureus, methicillin resistant coagulase negative Staphylococcus, vancomycin resistant Enterococcus, Escherichia coli, Microsforum gypseum with the minimum inhibitory concentration (μg/mL) and diameter of inhibition (mm): 150, 35; 150, 30; 300, 35; 300, 35; 300, 29; 4.7, 36, respectively. The IC50 value of the T47D cell line was 457 μg/mL.

Efficient alachlor degradation by the filamentous fungus Paecilomyces marquandii with simultaneous oxidative stress reduction

The acceleration of alachlor degradation by Paecilomyces marquandii under controlled and optimized conditions of fungal cultivation in liquid batches was observed (by ca. 20% in comparison to the flask cultures). Acidic environment and oxygen limitation resulted in deterioration of herbicide elimination. Efficient xenobiotic degradation did not correlate with free radicals formation, but some conditions of bioreactor cultivation such as neutral pH and oxygen enriched atmosphere (pO2⩾30%) caused a decrease in the reactive oxygen species (ROS) accumulation in mycelia. The changes in the glutathione (GSH) and ascorbic acid (AA) levels, also in the dismutase (SOD) and catalase (CAT) activities showed active response of the tested fungus against alachlor induced oxidative stress. These results will contribute to the improvement of chloroacetanilides elimination by fungi and extend the knowledge concerning oxidative stress induction and fungal cellular defense.

Diversity of entomopathogenic Hypocreales in soil and phylloplanes of five Mediterranean cropping systems

The diversity of entomopathogenic Hypocreales from the soil and phylloplanes in five Mediterranean cropping systems with different degrees of management [organic olive orchard conventional olive orchard, holm oak reforestation, holm oak dehesa (a multifunctional agro-sylvo-pastoral system), and sunflower plantation] was studied during four seasons. A total of 697 entomopathogenic fungal isolates were obtained from 272 soil samples, 1608 crop phylloplane samples and 1368 weed phylloplane samples. The following nine species were identified: Beauveria amorpha, B. bassiana, B. pseudobassiana, B. varroae, Metarhizium brunneum, M. guizhoense, M. robertsii, Paecilomyces marquandii and lilacinum using EF-1α gene sequences. All the fungal entomopathogenic species were found in both the soil and phylloplane samples, with the exception of M. robertsii, which was only isolated from the soil. The species richness, diversity (Shannon–Wiener index) and evenness (Pielou index) were calculated for each cropping system, yielding the following species ranking, which was correlated with the crop management intensity: holm oak reforestation>organic olive orchard>conventional olive orchard>holm oak dehesa>sunflower plantation. The number of fungal species isolated was similar in both phylloplane habitats and dissimilar between the soil and the crop phylloplane habitats. The ISSR analysis revealed high genotypic diversity among the B. bassiana isolates on the neighbourhood scale, and the isolates were clustered according to the habitat. These results suggest that the entomopathogenic Hypocreales in the phylloplane could result from the dispersal of fungal propagules from the soil, which might be their habitat of origin; a few isolates, including EABb 09/28-Fil of Beauveria bassiana, inhabit only the phylloplane.

Mechanism study of alachlor biodegradation by Paecilomyces marquandii with proteomic and metabolomic methods

Alachlor is an herbicide that is widely used worldwide to protect plant crops against broadleaf weeds and annual grasses. However, due to its endocrine-disrupting activity, its application had been banned in the European Union. As described in our earlier work, Paecilomyces marquandii is a microscopic fungus capable of alachlor removal by N-acetyl oxidation. Our current work uses proteomics and metabolomics to gain a better understanding of alachlor biodegradation by the microscopic fungus P. marquandii. The data revealed that the addition of alachlor reduced the culture growth and glucose consumption rates. Moreover, the rates of glycolysis and the tricarboxylic acids (TCA) cycle increased during the initial stage of growth, and there was a shift toward the formation of supplementary materials (UDP-glucose/galactose) and reactive oxygen species (ROS) scavengers (ascorbate). Proteomic analysis revealed that the presence of xenobiotics resulted in a strong upregulation of enzymes related to energy, sugar metabolism and ROS production. However, the unique overexpression of cyanide hydratase in alachlor-containing cultures may implicate this enzyme as the key protein involved in the alachlor biodegradation pathway. The characterization of P. marquandii-mediated alachlor removal in terms of cell structure and function provides a deeper insight into the strategies of microorganisms toward xenobiotic biodegradation.

Alachlor oxidation by the filamentous fungus Paecilomyces marquandii

Alachlor, a popular chloroacetanilide herbicide, can be a potential health risk factor. Soil microorganisms are primarily responsible for conversion and migration of alachlor in natural environment, but knowledge concerning alachlor biodegradation is not complete. Therefore, we studied the ability of Paecilomyces marquandii, soil fungus tolerant to heavy metals, to eliminate alachlor and proposed a new pathway of its transformation. After 7 days of incubation only 3.3% of alachlor was detected from an initial concentration 50mgL−1 and 20.1% from a concentration 100mgL−1. The qualitative IDA LC–MS analysis showed the presence of ten metabolites. All of them were dechlorinated mainly through oxidation, but also reductive dechlorination was observed. The main route of alachlor conversion progressed via N-acetyl oxidation resulting in the formation of mono-, di- and trihydroxylated byproducts. N-acetyl oxidation as a dominant route of alachlor metabolism by fungi has not been described so far. The toxicity of alachlor tested with Artemia franciscana did not increase after treatment with P. marquandii cultures. Paecilomyces marquandii strain seems to be an interesting model for the research on alachlor conversion by soil microscopic fungi, due to its dechlorination and hydroxylation ability as well as high tolerance to heavy metals.

Solubilization of iron and calcium phosphates by soil fungi isolated from coffee plantations

Almost 900 fungal isolates were obtained from eight coffee plantations in Colombia and Mexico. Of these, 76 isolates showed activity to solubilize Ca3(PO4)2 (PCa) and FePO4·H2O (PFe), which had been added to agar in a plate test. Generally, PCa was better solubilized than PFe. Colombian isolates were generally somewhat less effective than Mexican isolates. The two most effective isolates from each country with apparent highest PFe, solubilization potential were selected and cultivated in liquid medium containing PFe, which is more prevalent in tropical soils. The pH value, solubilized P in the medium and P uptake in fungal biomass were determined. After 24 days, Cylindrocarpon didymum and C. obtusisporum (both from Colombia) had solubilized 9.9 and 6.4 mg PO4 3--P L−1 and took up 8.6 and 11.6 mg P in biomass. Penicillium janthinellum and Paecilomyces marquandii (both from Mexico) solubilized 7.0 and 1.9 mg PO4 3--P L−1 and took up 11.3 and 17.3 mg P in biomass. The potential practical use of the four fungal isolates for different strategies in making more P available for coffee growth is discussed.

Adaptive alterations in the fatty acids composition under induced oxidative stress in heavy metal-tolerant filamentous fungus Paecilomyces marquandii cultured in ascorbic acid presence

The ability of the heavy metal-tolerant fungus Paecilomyces marquandii to modulate whole cells fatty acid composition and saturation in response to IC50 of Cd, Pb, Zn, Ni, and Cu was studied. Cadmium and nickel caused the most significant growth reduction. In the mycelia cultured with all tested metals, with the exception of nickel, a rise in the fatty acid unsaturation was noted. The fungus exposure to Pb, Cu, and Ni led to significantly higher lipid peroxidation. P. marquandii incubated in the presence of the tested metals responded with an increase in the level of linoleic acid and escalation of electrolyte leakage. The highest efflux of electrolytes was caused by lead. In these conditions, the fungus was able to bind up to 100 mg g(-1) of lead, whereas the content of the other metals in the mycelium was significantly lower and reached from 3.18 mg g(-1) (Cu) to 15.21 mg g(-1) (Zn). Additionally, it was shown that ascorbic acid at the concentration of 1 mM protected fungal growth and prevented the changes in the fatty acid composition and saturation but did not alleviate lipid peroxidation or affect the increased permeability of membranes after lead exposure. Pro-oxidant properties of ascorbic acid in the copper-stressed cells manifested strong growth inhibition and enhanced metal accumulation as a result of membrane damage. Toxic metals action caused cellular modulations, which might contributed to P. marquandii tolerance to the studied metals. Moreover, these changes can enhance metal removal from contaminated environment.

N vitro larvicidal action of Paecilomyces marquandii crude extract

The aim of this present work is to produce Paecilomyces marquandii crude extract and its in vitro larvicidal action. Saccharose, glucose, coffee residue, soluble starch, insoluble starch, soybean meal and wheat bran were the carbon sources tested. NaNO3, NH4Cl, soybean meal, (NH4)2SO4, yeast extract, casein peptone, tryptone and NH4NO3 were the nitrogen sources tested. Central composite design was applied to determine the optimal concentration of the tested significant variables. The best carbon sources were glucose and soybean meal, while the best nitrogen source observed was tryptone. The greatest value for proteolytic activity was 246.58 U/ml due to the concentration of tryptone and pH value. Results showed that the optimum pH was 8.0, while the optimum temperature was 60°C. The optimized extract obtained from P. marquandii exhibited in vitro larvicidal activity with 60.1% reduction. Tryptone and pH optimized the production of proteases by fungus P. marquandii. Key words: Nematophagous fungi, Paecilomyces marquandii, crude extract, larvicidal action, surface response.

Efficient Zn 2+ and Pb 2+ uptake by filamentous fungus Paecilomyces marquandii with engagement of metal hydrocarbonates precipitation

Zinc and lead biosorption by living non-growing filamentous fungus Paecilomyces marquandii was examined for its potential application in heavy metals elimination from contaminated areas. Metal uptake by the studied fungus was pH dependent and reached the level of 308 mg of Zn 2+ g −1 and 505 mg of Pb 2+ g −1 at pH of 7.5 caused by microprecipitation in slightly alkaline environment. All other metal studies were cultivated with unregulated pH yielding the maximum of 186.2 mg of Zn 2+ g −1 and 305.8 mg of Pb 2+ g −1. Interestingly, zinc binding by mycelium increased intensively after 15 h of incubation, whereas the lead concentration in biomass extended gradually and proportionally to the initial concentration and the time of contact. The study showed that thermal pretreatment of mycelium led to a decline in metal uptake, especially in the case of zinc. The mycelium slightly digested by the cell wall lytic enzyme complex, could adsorb lead twice as well after 2 h of exposure whereas zinc loading did not differ from the metal uptake by mycelia without any digestion procedure. The release of potassium ions from the mycelium, concomitant with lead uptake was observed suggesting ion exchange participation in lead binding. Energy-dispersive X-ray analysis, X-ray diffraction and FTIR spectroscopy revealed the presence of both metals hydrocarbonates on the mycelium surface. Additionally, the contribution of carboxyl and amide groups, originating from the mycelium, in metal binding was confirmed by FTIR analysis. The obtained results suggest that the effective metals uptake by P. marquandii was due to a combined mechanism with a dominant role of metabolism dependent microprecipitation.

In vitro antagonistic activity of fungi isolated from sclerotia on potato tubers against Rhizoctonia solani

Forty-five fungal isolates were obtained from sclerotia of Rhizoctonia solani on potato tubers in Erzurum, Turkey. The interaction between fungal isolates and R. solani was studied in dual culture technique. Some fungal isolates affected R. solani by antibiosis and/or parasitism. Results of the antagonism tests showed that Acremonium sp., Gliocladium viride, Paecilomyces marquandii, Paecilomyces sulphurellus, Penicillium camemberti, Penicillium expansum, Penicillium frequentans (ME-50), Penicillium nigricans, Penicillium olsonii, Penicillium phialosporum, Sporothrix sp. (MCY-4), Sporothrix schenckii, and Verticillium dahliae isolates produced an inhibition zone in front of the R. solani colony to a varying degree. Trichoderma harzianum isolates were able to overgrow the mycelium of R. solani. Physical colony contact was observed between the remaining 21 fungal isolates and R. solani. Furthermore, coiling of hyphae of Acremonium sp., Acremonium strictum, Gliocladium catenulatum, G. viride, and T. harzianum around those of R. solani was commonly observed.

Optimization of medium composition for protease production by Paecilomyces marquandii in solid-state- fermentation using response surface methodology

The present work was aimed to evaluate the optimization of medium composition for protease production by Paecilomyces marquandii in solid-state-fermentation using response surface methodology. Wheat bran was used as substrate for protease production. The variables tested were as follows: pH, tryptone and days of incubation. The proteolytic activity was measured by caseinolytic method. The Central Composite Design (CCD) was applied to determine the optimal concentration of the tested significant variables. A full factorial central composite experimental was used with six replicates at the central point. A total of 20 experiments were used to investigate the variables. The best fermentation conditions found after optimization were pH 5.0, tryptone 0.02 g/l and 5 days of incubation which yielded about 118 U/ml. The enzyme production of submerged fermentation (46 U/ml) was 2.57 times lower than the highest enzyme production of solid state fermentation (118 U/ml). The results indicate that pH, tryptone and days of incubation optimized the production of proteases by fungus P. marquandii in solid state fermentation.This is the first work about optimizing the production of proteases in solid state fermentation by the fungus P. marquandii. Key words: Solid state fermentation, Paecilomyces marquandii, protease, optimization, response surface.