Potato Dextrose Medium Research Articles

5'-Methoxyarmillane, a Bioactive Sesquiterpenoid Aryl Ester from the Fungus Armillaria ostoyae.

Higher fungi of the genus Armillaria belonging to the phylum Basidiomycota produce bioactive sesquiterpenoid aryl esters called melleolides. A bioactivity-guided discovery process led to the identification of the new melleolide 5'-methoxyarmillane (1) in organic extracts from the mycelium of Armillaria ostoyae. Remarkably, supplementation of rapeseed oil to the culture medium potato dextrose broth increased the production of 1 by a factor of six during the course of the 35 days fermentation. Compound 1 was isolated and its structure elucidated by UHPLC-QTOF-HR-MS/MS and NMR spectroscopy. It showed toxicity against Madin-Darby canine kidney II (MDCK II, IC50 19.2 μg/mL, 44.1 μM) and human lung cancer Calu-3 cells (IC50 15.2 μg/mL, 34.9 μM) as well as moderate bioactivity against Mycobacterium tuberculosis (MIC 8 mg/mL, 18.4 μM) and Mycobacterium smegmatis (MIC 16 mg/mL, 36.8 μM), but not against Staphylococcus aureus, Escherichia coli, Candida albicans, and Septoria tritici. No inhibitory effects of 1 against the influenza viruses H3N2, H1N1pdm, B/Malaysia, and B/Massachusetts were observed.

In vitro Evaluation on Efficacy of Bioagents and Plant Extracts against Early Blight of Tomato (Alternaria alternata)

AIMS: In vitro evaluation on efficacy of Bioagents and plant extracts against Early blight of tomato (Alternaria alternata) Study Design: CRD (Completely Randomized Design). Place and Duration of Study: A trial was conducted in plant pathology laboratory at HRS Venkataramannagudem, Dr. Y.S.R. Horticultural University, Venkataramannagudem, during 2022. Methodology: The efficacy of bioagents were tested against isolates for radial growth inhibition on suitable media (Potato dextrose media and Nutrient agar media) using dual culture technique under in vitro conditions. The poisoned food technique was followed to evaluate theefficacy of botanicals in inhibiting the mycelial growth of test pathogen. Results: In the present investigation total nine bioagents and four plants extracts were tested against Alternariaalternata under in vitro. Results revealed that among the nine antagonists., Trichoderma harzianum, Tichoderma viride, Tichoderma longibraciatum (TCT4), Trichoderma reesi (TCT10), A10 (Tichoderma asperellum), A28 (Trichoderma harzianum), Trichoderma virens and two bacterial bioagents Pesudomonas fluorescens and Baciilus subtillis tested against A. alternata, maximum reduction in colony growth of A. alternata was observed in A10 (T. asperellum) (69.50%) and significantly superior over all other bioagents tested which was followed by A28 (T. harzianum) (62.96%) and the next best was T. virens (60.28%). Least inhibition was noticed in Bacillus subtillis (46.69%). Total four plant extracts neem leaves (Azadirachta indica), onion bulb (Allium cepa), garlic bulb (Allium sativum), and turmeric rhizome (Curcuma longa) tested at three concentrations (5, 7.5, 10 %) with suitable control by poisoned food technique against A. alternata, onion bulb extract (44.07%) which was found superior to all other tested botanicals. Least mean per cent inhibition was recorded with turmeric (21.34%).

Screening, characterization and optimization of potential dichlorodiphenyl trichloroethane (DDT) degrading fungi

Dichlorodiphenyltrichloroethane is an organo-chlorine insecticide used for malaria and agricultural pest control, but it is the most persistent pollutant, endangering both human and environmental health. The primary aim of the research is to screen, characterize, and assess putative fungi that degrade DDT for mycoremediation. Samples of soil and wastewater were gathered from Addis Ababa, Koka, and Ziway. Fungi were isolated and purified using potato dextrose media. Matrix-Assisted Laser Desorption, Ionization, and Flight Duration The technique of mass spectrometry was employed to identify fungi. It was found that the finally selected isolate, AS1, was Aspergillus niger. Based on growth factor optimization at DDT concentrations (0, 3500, and 7000 ppm), temperatures (25, 30, and 35 °C), and pH levels (4, 7, and 10), the potential DDT-tolerant fungal isolates were investigated. A Box-Behnken experimental design was used to analyze and optimize fungal biomass and sporulation. The highest biomass (0.981 ± 0.22 g) and spore count (5.60 ± 0.32 log/mL) of A. niger were found through optimization assessment, and this fungus was chosen as a potential DDT-degrader. For DDT degradation investigations by A. niger in DDT-amended liquid media, gas chromatograph-electron capture detector technology was employed. DDT and its main metabolites, DDE and DDD, were eliminated from both media to the tune of 96–99 % at initial DDT concentrations of 1750, 3500, 5250, and 7000 ppm. In conclusion, it is a promising candidate for detoxifying and/or removing DDT and its breakdown products from contaminated environments.

Chemical profiling of fungal metabolites via the OSMAC approach: Novel identification of Brianthein W from an endophytic fungus, Hypomontagnella monticulosa Zg15SU

In the previous report, we reported that Hypomontagnella monticulosa originating from the rhizome of Zingiber griffithii was known to produce a marine-derived natural product. An OSMAC-based approach was designed by modifying the nutritional composition of the growth medium to investigate any possible new metabolites produced by the strain. The culture filtrate and biomass were conditioned through the use of three basal media, namely, Czapek-dox, potato dextrose, and Wickerham broth medium. GC–MS and multivariate analysis was performed to distinguish the chemicals and determine their composition in the tested extracts. Antimicrobial activity was tested against selected human pathogenic microbes using the disk-diffusion method. The MeOH extract of both culture filtrate and biomass from different fermentation media revealed that the majority of identified compounds (n = 40) were found in Wickerham medium (n = 23), which is later termed as MeOH-Wi. The chemical composition of MeOH-Wi was fatty acids (21.74 %), followed by terpenoids (17.38 %), cyclosiloxane (13.04 %), aldehydes (13.04 %), alkenes (8.7 %), hydrocarbons (8.7 %), esters (8.7 %), alkaloids (4.35 %), and an unclassified compound, the 9,9-Dimethyl-3,7-diazabicyclo[3.3.1]nonane. The numerous chemical compounds in MeOH-Wi corresponded with its antimicrobial activities against Micrococcus luteus NBRC 13,867 (±11 mm), Candida maltosa NBRC 1977 (±9 mm), and Escherichia coli JM 109 (±5 mm) which were higher than the other tested extracts. Further purification using HPLC RP-C18 using NP-SiO2/n-Hex–EtOAc (10:2) to yield compound (1). Compound (1) was determined as an analog of briarian W—a diterpene mostly found in marine sponge, Briareum spp.—as a pure compound in the MeOH extract of H. monticulosa based on the 1D and 2D NMR analysis. The anticancer activity (IC50) of compound (1) was 0.077, 0.080, and 0.102 µg/mL against the growth of HCT-116, NBT-T2, and Panc-1, respectively. As far as we are aware, this is the first report on finding a briarian diterpene that originates from an endophytic fungus, especially by H. monticulosa.

Optimization and characterization of antileukemic l-asparaginase produced by Fusarium solani endophyte

l-asparaginase is an antileukemic enzyme that hydrolyzes l-asparagine into l-aspartic acid and ammonia, causing cell starvation and apoptosis in susceptible leukemic cell populations. Currently, l-asparaginase obtained from bacterial sources is constrained by several issues, including lesser productivity, stability, selectivity, and higher toxicity. The goal of this study is to provide fungal l-asparaginase with in-vitro effectiveness towards different human carcinomas. l-asparaginase from endophytic Fusarium solani (Gene Bank accession number MW209717) isolated from the roots of the medicinal plant Hedera helix L. was characterized and optimized experimentally for maximum l-asparaginase production in addition to evaluating its subsequent cytotoxicity towards acute monocytic leukemia and human skin fibroblast cell lines. The enzyme production was maximized using potato dextrose media (15.44 IU/ml/hr) at the 5th and 6th days of fermentation with incubation temperature 30 °C, 3% asparagine, 150–180 rpm agitation rate and a 250 ml flask. Enzyme characterization studies revealed that the enzyme maintained its thermal stability with temperatures up to 60 °C. However, its optimal activity was achieved at 35 °C. On measuring the enzymatic activity at various temperatures and different pH, maximum enzyme activity was recorded at 40 °C and pH 8 using 0.1 M asparagine concentration. Results also revealed promising cytotoxic activity against acute monocytic leukemia with IC50 = 3.66 µg/ml and low cytotoxicity against tested normal human skin fibroblast cell line which suggested that it might have selective toxicity, and consequently it could be used as a less toxic alternative to the current formulations.

Light effects on Lasiodiplodia theobromae metabolome cultured in vitro.

The present work identified and compared intracellular metabolites and metabolic networks in mycelial cultures of Lasiodiplodia theobromae grown under 12 natural light and 24 hours' dark using a 1H NMR-based metabolomics approach. Fungal cultures were grown in potato dextrose media, and metabolites were extracted by sonication with sodium phosphate-buffered saline (pH = 6.0, 10% D2O, 0.1 mM TSP) from mycelium samples collected every week over four weeks. Multivariate analyses revealed that the light exposure group showed a positive correlation within beta-hydroxybutyrate, acetoacetate, acetone, betaine, choline, glycerol, and phosphocholine. On the other hand, phenyl acetate, leucine, isoleucine, valine, and tyrosine were positively correlated with dark conditions. Light favored the oxidative degradation of valine, leucine, and isoleucine, leading to the accumulation of choline, phosphocholine, betaine, and ketone bodies (ketogenesis). Ketogenesis, gluconeogenesis, and the biosynthesis of choline, phosphocholine, and betaine, were considered discriminatory routes for light conditions. The light-sensing pathways were interlinked with fungal development, as verified by the increased production of mycelia biomass without fruiting bodies and stress signaling, as demonstrated by the increased production of pigments.

Chemical Resistance of Diaporthe citri against Systemic Fungicides on Citrus.

Citrus melanose, caused by Diaporthe citri, has been one of the serious diseases, and chemical fungicides were used for protection in many citrus orchards of Jeju Island. Establishing a disinfectant resistance management system and reducing pesticide usage would be important for contributing to safe agricultural production. In this study, monitoring of chemical resistance was performed with 40 representative D. citri isolates from many citrus orchards in Jeju Island. Four different fungicides, kresoxim-methyl, benomyl, fluazinam, and prochloraz manganese, with seven different concentrations were tested in vitro by growing the mycelium of the fungal isolates on the artificial medium potato dextrose agar. Among the 40 fungal isolates, 12 isolates were investigated as resistant to kresoxim-methyl which could not inhibit the mycelium growth to more than 50%. Especially isolate NEL21-2 was also resistant against benomyl, whose hyphae grew well even on the highest chemical concentration. However, any chemical resistance of fungal isolates was found against neither fluazinam nor prochloraz manganese. On the other hand, in vivo bio-testing of some resistant isolates was performed against both kresoxim-methyl and benomyl on young citrus leaves. Typical melanose symptoms developed on the citrus leaves pre-treated with both agrochemicals after inoculation with the resistant isolates. However, no or less symptoms were observed when the susceptible isolates were inoculated. Based on these results, it was suggested that some resistant isolates of D. citri occurred against both systemic fungicides, which may be valuable to build a strategy for protecting citrus disease.

Using food residues (potato peels) as an alternative to potato dextrose agar in the growth of edible food fungi

Due to the great importance of edible fungi, their rapid life cycle, and the possibility of their production throughout the year, their cultivation has become a vital resource that many countries depend on in their pursuit of economic growth and food security. The present study aimed to devise a local nutrient media for primary mycelium growth of nine species of fungi belonging to Pleurotus eryngii , Pleurotus floridanus , Pleurotus ostreatus (white strain) , Agaricus heterocystis , Agaricus bispous (brown strain ), Agaricus bispous (white strain), Lentinula edodesstrain No. 1, Lentinula edodes strain No. 2, Flammulina velutipe (yellow strain) using culture mycelium inoculation for reproduction and production of edible fungi. Different natural materials used for this purpose involved potato peels in a concentration ranging from 5 to 20 g/ L and compared with the commercial medium Potato Dextrose Agar at a rate of 39 g/Land according to the manufacturer's instructions. The study also focused on the effect of physical factors such as temperature and pH on the growth rate of fungal mycelium. The results showed that the media prepared from potato peels at 5 g/L and 10 g/L concentrations achieved the best growth rate for the studied fungi compared to Peptone Dextrose Agar (PDA). The increase in the growth rate of fungi on media prepared from potato peels was attributed to the effect of nutrients for the prepared medium because it contained effective and necessary compounds for the growth in addition to their ease of absorption and consumption to be ready for food for agricultural fungi. The effect on the growth and production of edible fungi or any other type of fungi has not previously been studied and compared to the commercial medium PDA.

Molecular and cultural characterization of Morchella spp. from disturbed environments of central-southern Chile

In Northwestern Patagonia (Chile), three species of Morchella from undisturbed environments have been identified to date: Morchella tridentina, Morchella andinensis and Morchella aysenina, all belonging to the Elata clade and associated mainly with Nothofagus forests. In this study, the search for Morchella specimens was extended to disturbed environments in Central-Southern Chile, to further explore Morchella species diversity in the country, which is still very limited. The Morchella specimens were identified through multilocus sequences analysis, and the mycelial cultures were characterized, establishing comparisons with specimens from undisturbed environments. To the best of our knowledge, these results reveal for the first time in Chile the presence of the species Morchella eximia and Morchella importuna, and in the case of the last one also the first record in South America. These species were found associated almost exclusively with harvested or burned coniferous plantations. The in vitro mycelial characterization revealed certain inter- and intra-specific patterns of the morphology, such as pigmentation, mycelium type, and development and formation of sclerotia, which varied according to growth media and incubation temperature. The growth rates (mm/day) and mycelial biomass (mg) were significantly influenced by the temperature (p < 0.05), with maximum rates (>10 mm/day) and biomass (approx. 20 mg) between 20 and 24 °C, while a significant growth reduction (70–90%) was observed at 28 °C, mainly in the species from undisturbed environments. Potato-dextrose (PDA) medium stimulated the greatest mycelial density and sclerotia formation in most of the isolates, mainly in M. eximia (UDEC-LAF 236 isolate) which recorded the best mycelial growth performance. Among isolates, UDEC-LAF 236 also showed the best performance in sclerotia production (>350 sclerotia/dish) in 10 days of growth. This study contributes to the knowledge of the diversity of Morchella species in Chile by broadening the species range to those from disturbed environments. It also provides molecular and morphological characterization of the in vitro cultures of different Morchella species. The report on M. eximia and M. importuna, species known as cultivable, adapted to local climatic and edaphic conditions could represent the first step to developing artificial Morchella cultivation methods in Chile.

Manipulation of the Global Regulator mcrA Upregulates Secondary Metabolite Production in Aspergillus wentii Using CRISPR-Cas9 with In Vitro Assembled Ribonucleoproteins.

Genome sequencing of filamentous fungi has demonstrated that most secondary metabolite biosynthetic gene clusters (BGCs) are silent under standard laboratory conditions. In this work, we have established an in vitro CRISPR-Cas9 system in Aspergillus wentii. To activate otherwise silent BGCs, we deleted the negative transcriptional regulator mcrA. Deletion of mcrA (mcrAΔ) resulted in differential production of 17 SMs in total when the strain was cultivated on potato dextrose media (PDA). Nine out of fifteen of these SMs were fully characterized, including emodin (1), physcion (2), sulochrin (3), physcion bianthrone (4), 14-O-demethylsulochrin (5), (trans/cis)-emodin bianthrone (6 and 7), and (trans/cis)-emodin physcion bianthrone (8 and 9). These compounds were all found to be produced by the same polyketide synthase (PKS) BGC. We then performed a secondary knockout targeting this PKS cluster in the mcrAΔ background. The metabolite profile of the dual-knockout strain revealed new metabolites that were not previously detected in the mcrAΔ parent strain. Two additional SMs were purified from the dual-knockout strain and were characterized as aspergillus acid B (16) and a structurally related but previously unidentified compound (17). For the first time, this work presents a facile genetic system capable of targeted gene editing in A. wentii. This work also illustrates the utility of performing a dual knockout to eliminate major metabolic products, enabling additional SM discovery.

Secondary metabolites produced by Macrophomina phaseolina, a fungal root endophyte of Brugmansia aurea, using classical and epigenetic manipulation approach.

Endophytic fungi are rich sources of structurally complex chemical scaffolds with interesting biological activities. However, their metabolome is still unknown, making them appealing for novel compound discovery. To maximize the number of secondary metabolites produced from a single microbial source, we used the "OSMAC (one strain-many compounds) approach." In potato dextrose medium, M. phaseolina produced phomeolic acid (1), ergosterol peroxide (2), and a volatile compound 1,4-benzene-diol. Incorporating an epigenetic modifier, sodium valproate, affected the metabolite profile of the fungus. It produced 3-acetyl-3-methyl dihydro-furan-2(3H)-one (3) and methyl-2-(methyl-thio)-butyrate (4), plus volatile chemicals: butylated hydroxy toluene (BHT), di-methyl-formamide, 3-amino-1-propanol, and 1,4-benzenediol, 2-amino-1-(O-methoxyphenyl) propane. The structure of compounds 1-4 was established with the help of spectroscopic data. This study revealed first-time compounds 1-4 in the fungus M. phaseolinausing a classical and epigenetic manipulation approach.

Populasi Sel Jamur Dalam Media Dextrose Kentang Dengan Penambahan Sumber Energi S

The growth of fungi can be influenced by nutrients is in the environment, With the additional two kinds of nutrients from sugar plant waste of molasses, and waste of the pepaya taken the filtrate. Both of these wastes contain sugar which is needed by fungal microorganisms to grow and reproduce. The fungus Aspergillus niger, Hansenulla sp, Trichoderma viride, and Candida sp have a synergy interaction in a media. This study aims to learn in the growth of fungal cell populations as cell masses in potato dextrose media with the addition of two kinds of nutrients. The method used in this study was a two-population mean test with 2 treatments, namely the first treatment with the addition of sugarcane drops, and the next with the addition of papaya fruit filtrate, consisting of 11 replications, with incubation for 3 days. The analysis used was the T test and the analysis results obtained were significant, which was shown from P&gt; 0.05. The mass of fungi cells in the addition of nutrients from sugar factory waste was greater than that of papaya fruit filtrate waste

First Report of Colletotrichum fructicola Causing Anthracnose on Camellia yuhsienensis Hu in China.

Camellia yuhsienensis Hu is an endemic species from China, where is the predominant oilseed crop due to its anthracnose resistance (Kuang 2015; J. Li et al. 2020; Nie et al. 2020). In April 2019, anthracnose symptoms were observed on C. yuhsienensis in a plantation in Youxian, Zhuzhou, Hunan Province, China (113.32°E, 26.79°N). It was detected approximately 10% anthracnose incidence in 500 two-year-old plants in a 5000 m2 cultivated area. Diseased leaves showed irregular grayish brown spots with dark brown edges and dark brown undersides. Symptomatic tissues (4 to 5 mm2) were surface-disinfected for 90 s in 75% ethanol, then rinsed twice with sterile water, and finally incubated on PDA (potato dextrose agar) at 28℃ (Jiang et al. 2018). Pure cultures were obtained by the single-spore isolation method. A total of 100 fungal isolates were obtained from 85 symptomatic leaves, from which 81 had similar colony morphology. Colonies on PDA were white, fluffy and cottony, and becoming dark gray after 5 days. The character of the reverse of the colony were similar to that of the upper of the colony, but the color was darker at the same time. The isolates produced a large number of single-celled, hyaline, straight and cylindrical conidia, with 10.35 to 17.58 length × 3.46 to 5.69 μm width (x=13.61 × 4.63 μm, n = 30). The isolates were preliminarily identified as Colletotrichum spp. according to morphological features (Weir et al. 2012). Representative isolate YX2-5-2 was used for molecular identification: internal transcribed spacer (ITS), partial actin (ACT), chitin synthase (CHS-1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genomic DNA regions were amplified by PCR (Weir et al. 2012). Gene sequences were deposited in GenBank (GenBank accession no. MW398863 for ACT, MW886232 for CHS-1, MW398864 for GAPDH, MW398865 for ITS). BLAST analysis revealed that DNA sequences of YX2-5-2 at the ITS, GAPDH, ACT, and CHS-1 loci showed 100%, 99.25%, 100%, and 99.33% sequence identity, respectively to their corresponding loci in strains ZH6 (GenBank accession no. MT476840.1), ICKP18B4 (LC494274.1), YN17 (MN525804.1), and ICKG4 (LC469131.1) of C. fructicola. A Maximum Likelihood phylogenetic tree based on the combined ACT, CHS-1, ITS and GAPDH sequences revealed that the representative isolate YX2-5-2 clustered with C. fructicola. In addition, the morphological features of YX2-5-2 were similar to C. fructicola which has been reported (Weir et al. 2012). Pathogenicity was tested using isolate YX2-5-2 by inoculating leaves of 2-year-old C. yuhsienensis. Four leaves of each healthy C. yuhsienensis were sprayed with a conidial suspension (105 conidial/mL) of isolate YX2-5-2, and the above steps were repeated three times. Two additional mock-inoculated control plants were sprayed with sterilized liquid potato dextrose medium. The plants were incubated in a greenhouse at 28℃ and 90% humidity with a 12 h photoperiod. Anthracnose-like symptoms were observed 5 days post-inoculation. The control plant tissues remained healthy. C. fructicola was re-isolated on PDA from lesions, and the morphological features were consistent with YX2-5-2, confirming Koch's postulates. To our knowledge, this is the first report of anthracnose of C. yuhsienensis caused by C. fructicola in China. Anthracnose of Camellia. oleifera has been reported for a long time (H. Li et al. 2016). C. yuhsienensis, as a wild relative of C. oleifera (commonly known as tea-oil tree), has been concerned about its resistance to anthracnose. Therefore, the occurrence of C. yuhsienensis anthracnose hindered the control of anthracnose tea-oil tree. This finding will lay the foundation for studying the pathogenesis of anthracnose of tea-oil tree and developing effective prevention methods.

Plant Growth Promotion by Two Volatile Organic Compounds Emitted From the Fungus Cladosporium halotolerans NGPF1.

Microbial volatiles have beneficial roles in the agricultural ecological system, enhancing plant growth and inducing systemic resistance against plant pathogens without being hazardous to the environment. The interactions of plant and fungal volatiles have been extensively studied, but there is limited research specifically elucidating the effects of distinct volatile organic compounds (VOCs) on plant growth promotion. The current study was conducted to investigate the impact of VOCs from Cladosporium halotolerans NGPF1 on plant growth, and to elucidate the mechanisms for the plant growth-promoting (PGP) activity of these VOCs. The VOCs from C. halotolerans NGPF1 significantly promoted plant growth compared with the control, and this PGP activity of the VOCs was culture medium-dependent. Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography–mass spectrometry (GC–MS) identified two VOC structures with profiles that differed depending on the culture medium. The two compounds that were only produced in potato dextrose (PD) medium were identified as 2-methyl-butanal and 3-methyl-butanal, and both modulated plant growth promotion and root system development. The PGP effects of the identified synthetic compounds were analyzed individually and in blends using N. benthamiana plants. A blend of the two VOCs enhanced growth promotion and root system development compared with the individual compounds. Furthermore, real-time PCR revealed markedly increased expression of genes involved in auxin, expansin, and gibberellin biosynthesis and metabolism in plant leaves exposed to the two volatile blends, while cytokinin and ethylene expression levels were decreased or similar in comparison with the control. These findings demonstrate that naturally occurring fungal VOCs can induce plant growth promotion and provide new insights into the mechanism of PGP activity. The application of stimulatory volatiles for growth enhancement could be used in the agricultural industry to increase crop yield.

Isolation and molecular identification of microorganisms isolated from soils contaminated with heavy metals in Mosul city

This research is concerned with organisms isolated from soils contaminated with heavy metals in industrial and residential areas in Mosul, the center of Nineveh Governorate, and the diagnosis of these organisms using molecular biology techniques. Samples were collected from four locations in the city between the industrial area and residential neighborhoods. Soil samples were analyzed, and dilutions were prepared, then the dilutions were grown on potato extract and dextrose (PDA) medium for the development of fungi and Nutrient agar for bacterial development. The dilutions were planted by the casting method by three replications, then the process of purifying the fungal and bacterial colonies was carried out using the traditional methods. To diagnose these pure colonies using PCR technique, colonies of fungi were grown on the medium of PDA, and bacteria were grown on the medium of nutritious broth. As a result, nine fungal species were diagnosed; two of them are new undiagnosed genera that have been registered in the gene bank, four of them contain genetic mutations, and three of them are known and previously diagnosed fungi. As for bacteria, two new strains were isolated and registered in the gene bank among the four diagnosed types. And some of these genera exhibited severe resistance to antibiotics, while others showed moderate resistance, in contrast to the control, which was very sensitive to antibiotics.

In vitro screening of new strains of predacious nematophagous fungi for biocontrol suitability when produced in liquid culture

Aim. To isolate, identify, and evaluate in vitro the predacious ability of nematophagous fungi strains, to determine their eligibility for mass production under submerged cultivation; to select the most efficient isolates of predacious nematopha- gous fungi as promising producers of the biopreparation to control plant-parasitic nematodes. Methods. Microbiological, microscopical, cultural-morphological, statistical. Results. The screening to determine the presence of predacious nema- tophagous fungi in different soils from different regions has been conducted for the first time in Ukraine. Out of 75 soil samples, isolated in Kyiv, Chernihiv, Dnipropetrovsk, Volyn and Odesa regions, 88 isolates of predacious nematophagous fungi were obtained which belong to 11 genera by their cultural-morphological characteristics: Arthrobotrys oligospo- ra – 39 isolates, A. musiformis – 25, A. conoides – 11, Drechslerella dactyloides – 3, A. artrobotryoides –2, A. superba – 2, A. megalospora – 2, A. sphaeroides –1, A. flagrans – 1, A. amerospora – 1, A. thaumasia – 1. High nematophagous activity in vitro (90–100 % trapping) regarding free-living nematodes (Rhabditis spp). was demonstrated by 4 isolates of A. oligospora, 18 isolates of A. musiformis, 5 isolates of A. conoides, 3 isolates of D. dactyloides, and one isolate of A. megalospora. Among the 47 most active isolates, 4 isolates of predacious fungi formed chlamydospores under submerged cultivation on two types of liquid media – wort medium and corn-molasses medium: A. musiformis-711, A. musiformis-911, A. conoides-90, D. dactyloides-19. It was found that isolate A. musiformis-911 and A. conoides-90 could form conidia in the submerged culture. Further experiments in determining the character of development for two latter isolates in different types of media under submerged cultivation established that the largest number of chlamydo- spores and conidia were formed by the isolates of A. conoides 90 and A. musiformis 911 on potato dextrose medium, 7.6·105 and 6.5·105 spores/ml respectively. In the wort medium, the productivity of spore formation for the isolate of A. conoides 90 was 35 times lower, amounting to 2.2·104 spores/ml, and in peptone-glucose medium – 90 times lower (8.5·103 spores/ml). The isolate of A. musiformis 911 in the wort medium produced 325 times fewer spores (2.0× ×103 spores/ml) than in potato-dextrose medium, while no chlamydospores and conidia were formed in the peptone- glucose medium. More detailed study of the ratio between isolates-producers and different sources of nutrition as the basis for the optimization of liquid media is required. Conclusions. According to the results of determining nematoph- agous activity regarding free-living nematodes, isolated from different soils in Ukraine and evaluating the character of development under submerged cultivation, two isolates of predacious fungi were selected – Arthrobotrys conoides-90 and A. musiformis-911, which are promising producers of a biological preparation eventually to be used in the biocon- trol of plant-parasitic nematodes. Conidia formation of the two isolates in liquid culture under mass production condi- tions was observed for the first time and could also contribute to their suitability for mass production and biocontrol.

In Vitro Antimicrobial Activity of Cinnabarin on Xanthomonas campestris Isolated from Bean Crops of Puebla, Mexico

Phaseolus vulgaris is the most legume cultivated in the world; in Mexico, it is considered the second most important crop after corn. The aim of this research was to determine the characteristics of Xanthomonas campestris strain “Xcf1-APJR” isolated from the leaves of bean crops, and determine the antimicrobial activity of cinnabarin on this strain. Bacterial cultures were obtained from leaves with necrotic leaf spot symptoms of bean plant variety “Flor de Mayo M38” in Puebla, Mexico. The antimicrobial activity of cinnabarin was tested at 7, 14 and 21 days on X. campestris pv. campestris. The Xcf1-APJR strain showed 100% identity with X. campestris pv. campestris as a causal agent of necrotic leaf spot. Treatment with a potato dextrose medium with a dehydrated sugar cane (PDA+C) showed a higher orange pigmentation than the other treatments after 7, 14 and 21 days of incubation and a higher concentration of cinnabarin (54.33 InU/g) with in vitro antimicrobial activity against X. campestris pv. campestris.

Characterization and Physical Properties of Mycelium Films Obtained from Wild Fungi: Natural Materials for Potential Biotechnological Applications

Fresh basidiomes of species belonging to the genera Aurantiporus, Ganoderma, Lentinus, and Panus species were collected from a cloud forest patch located in a natural protected area. Tissue portions of the fresh basidiomes were excised and grown in axenic culture with potato dextrose medium at 27 °C obtaining six strains. Additionally, a commercial strain of Pleurotus ostreatus was included for comparison with previous works. Mycelium films were obtained and morpho-anatomic characterized considering the main features. After that, the films were dried at room temperature and pressed at 7.5 g/cm2. SEM images were taken with a scanning electron microscope, and mechanical and thermo gravimetric analyses were carried out. The tensile modulus values obtained vary between 3.5 and 128.8 MPa and the tensile strength values between 0.27 and 1.5 MPa. The density was not uniform and is not related to the mechanical performance. The Maximum decomposition temperature of the mycelium films was reached at around 300 °C. The results showed significant differences in physic-mechanical behavior of mycelium films obtained with different fungal species, some of them with promissory characteristics for biotechnological applications.

Chromolaena laevigata (Asteraceae) as a source of endophytic non-aflatoxigenic Aspergillus flavus: chemical profile in different culture conditions and biological applications.

Endophytes are microorganisms that form symbiotic relationships with their host. These microorganisms can produce a variety of secondary metabolites, some of which have inhibitory effects on pests and pathogens or even act to promote plant growth. Due to these characteristics, these microorganisms are used as sources of biologically active substances for a wide range of biotechnological applications. Based on that, the aim of this study was to evaluate the production of metabolites of the endophytic Aspergillus flavus CL7 isolated from Chromolaena laevigata, in four different cultivation conditions, and to determine the antimicrobial, cytotoxic, antiviral, and antioxidant potential of these extracts. The multiphasic approach used to identify this strain was based on morphology and ITS gene sequence analysis. The chemical investigation of A. flavus using potato dextrose and minimal medium, using both stationary and agitated methods, resulted in the isolation of kojic acid, α-cyclopiazonic acid, and 20,25-dihydroxyaflavinine. Another 18 compounds in these extracts were identified by UHPLC-HRMS/MS, of which dideacetyl parasiticolide A has been described for the first time from A. flavus. Aflatoxins, important chemomarkers of A. flavus, were not detected in any of the extracts, thus indicating that the CL7 strain is non-aflatoxigenic. The biological potential of all extracts was evaluated, and the best results were observed for the extract obtained using minimal medium against Trichophyton rubrum and Mycobacterium tuberculosis.

Preparation and characterization of a novel packing material for the gas-phase fluidized-bed bioreactor

Gas-phase fluidized-beds reactor (FBR) can overcome the shortages of bed clogging and performance decline in packed-bed reactors on VOCs treatment. Ideal FBR packing materials with minimum fluidized velocity and surface characteristics are required. In this study, an expanded polystyrene (EPS) with particle size 1.0–2.0 mm was selected as the skeleton among six types of EPS and two types of expanded polypropylene (EPP). They were then physically coated with a combination of potato dextrose medium, fine wheat bran particles, and inorganic glue. Different glue concentrations and amounts of medium were tested and the optimal values of 2 % and 0.22 g∙cm−2 were achieved. The wheat-coated EPS obtained hydrophilic functional groups on the surface, and its contact angle decreased from 93° to 37° after coating. By scanning electronic microscopy, more microorganisms were observed on the surface of the coated EPS than that of the uncoated EPS. A lab-scale fluidized bioreactor was built to treat gaseous ethanol. The minimum fluidized velocity of coated packing materials was 0.2 m∙s−1 under a relative humidity of 50 ± 10 %, reaching an average elimination capacity of 681 g m-3 h−1 during the 28-days operation, while the bioreactor packed with uncoated EPS reached only 18 g m−3 h−1.