Penicillium Oxalicum Research Articles

Studies on the synthesis, spectroscopy, thermal properties, docking, and biology of new Schiff base and its mono- and binuclear complexes.

A novel Schiff base has been synthesized from the condensation of the 3-formyl-2-hydroxybenzoic acid and 4-nitrobenzene-1,2-diamine. The new ligand was found to have two coordination sites. So, it has the capability to form mono- and binuclear complexes with different metal ions. The free ligand and its mono- and binuclear cobalt(II) complexes have been characterized by UV-Visible spectra, IR, elemental analyzes, H1 NMR, conductimetric, thermal, and magnetic measurements. Results indicated that the cobalt(II) ion is attached to the inside coordination site and the second metal ion attached to the outside coordination site. The complexes are all non-electrolytes, as demonstrated by the molar conductance tests. The thermodynamic parameters of the metal complexes are calculated using Horowitz Metzger and Coats-Redfern methods. The complexes' bonding properties have also been estimated. Molecular docking was employed to forecast the interaction of the prepared with the Candida-albicans receptor (1zap). The biological activities of these metal complexes were tested against some bacteria and fungi. It is evident from the biological screening data that the prepared Co(II) binuclear complexes exhibit predominant activity against Candida albicans, Penicillium oxalicum and Escherichia coli, while they have no activity against Micrococcus roseus and Micrococcus luteus.

Novel Transcription Factor CXRD Regulates Cellulase and Xylanase Biosynthesis in Penicillium oxalicum under Solid-State Fermentation.

Penicillium oxalicum produces an integrated, extracellular cellulase and xylanase system, strictly regulated by several transcription factors. However, the understanding of the regulatory mechanism of cellulase and xylanase biosynthesis in P. oxalicum is limited, particularly under solid-state fermentation (SSF) conditions. In our study, deletion of a novel gene, cxrD (cellulolytic and xylanolytic regulator D), resulted in 49.3 to 2,230% enhanced production of cellulase and xylanase, except for 75.0% less xylanase at 2 days, compared with the P. oxalicum parental strain, when cultured on solid medium containing wheat bran plus rice straw for 2 to 4 days after transfer from glucose. In addition, the deletion of cxrD delayed conidiospore formation, leading to 45.1 to 81.8% reduced asexual spore production and altered mycelial accumulation to various extents. Comparative transcriptomics and real-time quantitative reverse transcription-PCR found that CXRD dynamically regulated the expression of major cellulase and xylanase genes and conidiation-regulatory gene brlA under SSF. In vitro electrophoretic mobility shift assays demonstrated that CXRD bound to the promoter regions of these genes. The core DNA sequence 5'-CYGTSW-3' was identified to be specifically bound by CXRD. These findings will contribute to understanding the molecular mechanism of negative regulation of fungal cellulase and xylanase biosynthesis under SSF. IMPORTANCE Application of plant cell wall-degrading enzymes (CWDEs) as catalysts in biorefining of lignocellulosic biomass into bioproducts and biofuels reduces both chemical waste production and carbon footprint. The filamentous fungus Penicillium oxalicum can secrete integrated CWDEs, with potential for industrial application. Solid-state fermentation (SSF), simulating the natural habitat of soil fungi, such as P. oxalicum, is used for CWDE production, but a limited understanding of CWDE biosynthesis hampers the improvement of CWDE yields through synthetic biology. Here, we identified a novel transcription factor CXRD, which negatively regulates the biosynthesis of cellulase and xylanase in P. oxalicum under SSF, providing a potential target for genetic engineering to improve CWDE production.

Metabolite Chemical Composition of the Bletilla striata (Thunb.) Reichb. f. Endophyte Penicillium oxalicum

Penicillium oxalicum strain can be isolated from the Bletilla striata (Thunb.) Reichb. f. tubers. Its solid-state fermentation products are concentrated by percolation extraction. Separation and purification have been conducted to the ethyl acetate extracts by preparative HPLC. Based on the use of spectrometry, we have determined 17 known compounds, 12,13-dihydroxy-fumitremorgin C (1), pseurotin A (2), tyrosol (3), cyclo-(L-Pro-L-Val) (4), cis-4-hydroxy-8-O-methylmellein (5), uracil (6), cyclo-(L-Pro-L-Ala) (7), 1,2,3,4-tetrahydro-4-hydroxy-4-quinolin carboxylic acid (8), cyclo-(Gly-L-Pro) (9), 2'-deoxyuridine (10), 1-(β-D-ribofuranosyl)thymine (11), cyclo-(L-Val-Gly) (12), 2'-deoxythymidine (13), cyclo-(Gly-D-Phe) (14), cyclo-L-(4-hydroxyprolinyl)-D-leucine (15), cyclo-(L)-4-hydroxy-Pro-(L)-Phe (16), uridine (17). Here, we report compounds 1–3, 5, 7–8, 11–12, 14–17 are first found and isolated from this endophyte.

Kinase POGSK-3β modulates fungal plant polysaccharide-degrading enzyme production and development

The filamentous fungus Penicillium oxalicum secretes integrative plant polysaccharide-degrading enzymes (PPDEs) applicable to biotechnology. Glycogen synthase kinase-3β (GSK-3β) mediates various cellular processes in eukaryotic cells, but the regulatory mechanisms of PPDE biosynthesis in filamentous fungi remain poorly understood. In this study, POGSK-3β (POX_c04478), a homolog of GSK-3β in P. oxalicum, was characterised using biochemical, microbiological and omics approaches. Knockdown of POGSK-3β in P. oxalicum using a copper-responsive promoter replacement system led to 53.5 - 63.6%, 79.0 - 92.8% and 76.8 - 94.7% decreases in the production of filter paper cellulase, soluble starch-degrading enzyme and raw starch-degrading enzyme, respectively, compared with the parental strain ΔKu70. POGSK-3β promoted mycelial growth and conidiation. Transcriptomic profiling and real-time quantitative reverse transcription PCR analyses revealed that POGSK-3β dynamically regulated the expression of genes encoding major PPDEs, as well as fungal development-associated genes. The results broadened our understanding of the regulatory functions of GKS-3β and provided a promising target for genetic engineering to improve PPDE production in filamentous fungi. KEY POINTS: • The roles of glycogen synthase kinase-3β were investigated in P. oxalicum. • POGSK-3β regulated PPDE production, mycelial growth and conidiation. • POGSK-3β controlled the expression of major PPDE genes and regulatory genes.

Synergy among extracellular adsorption, bio-precipitation and transmembrane transport of Penicillium oxalicum SL2 enhanced Pb stabilization

As a potential bioremediation strain for Pb contamination, Penicillium oxalicum SL2 sometimes has secondary activation of Pb, so it is crucial to clarify its effect on Pb morphology and its intracellular response to Pb stress. We investigated the effect of P. oxalicum SL2 in medium on Pb2+ and Pb availability in eight minerals, and revealed the prioritization of Pb products. (i)Pb was stabilized within 30 days as Pb3(PO4)2 or Pb5(PO4)3Cl with sufficient phosphorus (P); (ii) under P deficiency but sulfur (S) sufficient, Pb was stabilized mainly in the form of PbSO4; (iii) under conditions of P and S deficiency, Pb was stabilized mainly in the form of PbC2O2. With the help of proteomic and metabolomics analysis, a total of 578 different proteins and 194 different metabolites were found to be matched in 52 pathways. Among them, the activation of chitin synthesis, oxalate production, sulfur metabolism and transporters improved the Pb tolerance of P. oxalicum SL2, and promoted the synergistic effect of extracellular adsorption, bio-precipitation and transmembrane transport on Pb stabilization. Our results fill the gap in the intracellular response of P. oxalicum SL2 to Pb and provide new insights into the development of bioremediation agent and technology for Pb contamination.

Analysis and phytochemical profile of Amaranthus tricolor L. extract with antioxidative and antimicrobial properties

Amaranthus tricolor L. is a cultivated green vegetable, commonly known as amaranth, which is similar to spinach, broccoli, and cabbage. In the present study, we investigated the antioxidant and antimicrobial activities of the methanol extract and various fractions from A. tricolor. Results show that A. tricolor ethyl acetate (EtOAc) extract displayed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity (IC50=16.43±1.53 μg/ml), which was higher than that of the positive control, butyl hydroxy anisd (IC50=19.42±0.91 μg/ml). This extract had a reducing power of 2.743 at 0.5 mg/ml and significantly attenuated production of reactive oxygen species in a dose-dependent manner. Further bioassay-monitored fractionation of the EtOAc extract yielded two flavonoids, kaempferol (1) and quercetin (2), and one phenolic acid, gallic acid (3). We found that the antimicrobial activity of compound 3 (at a dose of 63 μg/mL) was superior to that of the tetracycline control (at a dose of 250 μg/mL) against Escherichia coli. Additionally, compounds 1 and 2 (at a dose of 63 μg/mL each) displayed higher activities against Penicillium oxalicum and Staphylococcus aureus than the control. These results suggest that A. tricolor extract may represent a promising nutraceutical source due to the antioxidant and antimicrobial properties of its phenolic compounds.

A Pipeline to Investigate Fungal-Fungal Interactions: Trichoderma Isolates against Plant-Associated Fungi.

Soil fungi play essential roles in ecosystems, forming complex interaction networks with bacteria, yeasts, other fungi, or plants. In the framework of biocontrol strategies, Trichoderma-based fungicides are at the forefront of research as an alternative to synthetic ones. However, the impact of introducing new microbial strain(s) on the soil microbiome of a habitat is not well-explored. Aiming to identify a quantitative method to explore the complex fungal interactions, we isolated twelve fungi from three Italian vineyards and identified three strains of the Trichoderma genus in addition to nine more plant-associated fungi of different genera. Investigating in dual nucleation assay fungal-fungal interactions, we recognised two types of interaction: neutral or antagonistic. All three Trichoderma strains displayed a slight inhibitory behaviour against themselves. Trichoderma strains showed a mutually intermingling growth with Aspergillus aculeatus and Rhizopus arrhizus but antagonistic behaviour against the plant pathogens Alternaria sp., Fusarium ramigenum, and Botrytis caroliniana. Yet, in some cases, antagonistic behaviour by Trichoderma fungi was also observed against plant-promoting fungi (e.g., Aspergillus piperis and Penicillium oxalicum). Our study highlights the importance of studying the interactions between fungi, aiming to clarify better the impact of fungal-based biological fungicides in the soil communities, and offers a pipeline for further applications.

Safety evaluation and whole genome sequencing for revealing the ability of Penicillium oxalicum WX-209 to safely and effectively degrade citrus segments

The microbial potential of Penicillium has received critical attention. The present research aimed to elucidate the efficacy of crude enzyme secreted from Penicillium oxalicum WX-209 in degrading citrus segments and evaluate the safety of the process. Results showed that citrus segment membranes gradually dissolved after treatment with the crude enzyme solution, indicating good degradation capability. No significant differences in body weight, food ingestion rate, hematology, blood biochemistry, and weight changes of different organs were found between the enzyme intake and control groups. Serial experiments showed that the crude enzyme had high biological safety. Moreover, the whole genome of P. oxalicum WX-209 was sequenced by PacBio and Illumina platforms. Twenty-five scaffolds were assembled to generate 36 Mbp size of genome sequence comprising 11 369 predicted genes modeled with a GC content of 48.33 %. A total of 592 genes were annotated to encode enzymes related to carbohydrates, and some degradation enzyme genes were identified in strain P. oxalicum WX-209. Data availabilityThe whole genome sequencing data in this study can be found in GenBank database of NCBI website with the accession number PRJNA 715321. The link: http://www.ncbi.nlm.nih.gov/bioproject/715321.

First Report of Penicillium oxalicum Causing Postharvest Fruit Rot on Citrus in China.

Ougan (Citrus reticulata cv. Suavissima) is a distinct citrus cultivar local to Zhejiang province, China. (Guo et al. 2021). In November 2021, an unknown postharvest fruit rot was observed in the Sanyang wetland Ougan planting area, Wenzhou City of Zhejiang Province (27.96 °N, 120.69 °E). About 3% of diseased fruits with similar fruit rot symptoms were observed in 900 mandarin fruits from four commercial storages. Initially, the symptoms appeared as light brown lesions that turned deep brown as the lesions expanded. To identify the pathogen, segments (5 mm2) from margins of rotted tissue were excised from 5 symptomatic fruits, surface disinfested twice with 75% ethanol, rinsed three times with sterilized water, placed aseptically onto potato dextrose agar (PDA) medium and incubated for 7 days at 25℃ in darkness for identification. Five fungal isolates with the same morphology were obtained using the single spore method (Leslie and Summerell 2006). Initially, the fungus produced fluffy and white aerial mycelium that eventually turned green on PDA medium after 3 days. Conidiophores were broom-shaped (17.5 ± 2.5 μm) (n=50). Conidia were unicellular and ellipsoid (3.5 to 5.0 ×2.5 to 4.0 μm) (n=50). These morphological characteristics were consistent with Penicillium species (Wu et al. 2022). WZU-OG1 was chosen as a representative isolate for further study. For molecular identification, PCR amplification and DNA sequencing were performed using primers ITS1/ITS4 (White et al. 1990), bt2a/bt2b (Glass and Donaldson 1995), and RPB2-5F/RPB2-7R (Liu et al. 1999) to amplify the complete internal transcribed spacer (ITS) region, β-tubulin gene (TUB), and a portion of RNA polymerase second largest subunit (RPB2). The ITS, TUB, and RPB2 gene sequences of isolate WZU-OG1 were deposited in the GenBank database with acc. nos. ON332735, ON428245, and ON524171, respectively. BLASTn analysis respectively showed 561/561 (MH855125), 414/427 (KF296462), and 936/954 (JN121456) matching with Penicillium oxalicum CBS 219.30. A neighbor-joining phylogenetic analysis based on the concatenated nucleotide sequences (ITS, TUB, and RPB2) grouped this isolate in the Penicillium oxalicum species complex clade at 100% bootstrap support. To verify pathogenicity, 20 healthy mandarin fruit of cultivar Sanyang were superficially disinfested with 75% ethanol and then washed with distilled water. A conidial suspension of 1 × 105 conidia/ml from a 5-day-old culture of WZU-OG1 was injected into 10 fruits (10 μL per fruit). An equal number of fruits inoculated with sterile water were used as the negative control. The inoculated fruits were stored in a constant temperature incubator under the conditions of 28 ℃, 90% humidity, and incubated in a 12-h light/12-h dark cycle for 12 days. Symptoms similar to those on the naturally infected fruit began 5 days after inoculation, whereas no symptoms occurred on the controls. The experiment was repeated three times, and similar symptoms were observed in all diseased fruits. Then, the fungus was reisolated from these infected fruits and identified as P. oxalicum by the morphological and molecular methods described above. This is the first report of P. oxalicum causing postharvest mandarin decay and this study will enable us to rapidly diagnose this disease, identify the occurrence of this disease and develop adequate management strategies to control it in China.

Protein Kinase PoxMKK1 Regulates Plant-Polysaccharide-Degrading Enzyme Biosynthesis, Mycelial Growth and Conidiation in Penicillium oxalicum.

The ability to adapt to changing environmental conditions is crucial for living organisms, as it enables them to successfully compete in natural niches, a process which generally depends upon protein phosphorylation-mediated signaling transduction. In the present study, protein kinase PoxMKK1, an ortholog of mitogen-activated protein kinase kinase Ste7 in Saccharomyces cerevisiae, was identified and characterized in the filamentous fungus Penicillium oxalicum. Deletion of PoxMKK1 in P. oxalicum ΔPoxKu70 led the fungus to lose 64.4-88.6% and 38.0-86.1% of its plant-polysaccharide-degrading enzyme (PPDE) production on day 4 after a shift under submerged- and solid-state fermentation, respectively, compared with the control strain ΔPoxKu70. In addition, PoxMKK1 affected hypha growth and sporulation, though this was dependent on culture formats and carbon sources. Comparative transcriptomics and real-time quantitative reverse transcription PCR assay revealed that PoxMKK1 activated the expression of genes encoding major PPDEs, known regulatory genes (i.e., PoxClrB and PoxCxrB) and cellodextrin transporter genes (i.e., PoxCdtD and PoxCdtC), while it inhibited the essential conidiation-regulating genes, including PoxBrlA, PoxAbaA and PoxFlbD. Notably, regulons modulated by PoxMKK1 and its downstream mitogen-activated protein kinase PoxMK1 co-shared 611 differential expression genes, including 29 PPDE genes, 23 regulatory genes, and 16 sugar-transporter genes. Collectively, these data broaden our insights into the diverse functions of Ste7-like protein kinase, especially regulation of PPDE biosynthesis, in filamentous fungi.

Structural characterization of corn fiber hemicelluloses extracted by organic solvent and screening of degradation enzymes

An integrated treatment coupling peracetic acid delignification, dimethyl sulfoxide extraction, and ethanol precipitation were performed to isolate hemicellulose from de-starched corn fiber. Based on chemical composition, molecular weight distribution, methylation, and nuclear magnetic resonance spectroscopy, it is proposed that hemicelluloses in corn fiber were composed of two polysaccharides, glucuronoarabinoxylan (about 80 %) and xyloglucan (about 20 %). Xylose (about 46 %) and arabinose (about 32 %) were the main components in glucuronoarabinoxylan. More than half of the xylose units in the glucuronoarabinoxylan backbone chain were substituted at O-2 and/or O-3 by various monomers or oligomeric side chains. Based on structure analysis, five hemicellulases were selected and added to Penicillium oxalicum MCAX enzymes for enzymatic hydrolysis of corn fiber. The results showed that the addition of hemicellulases increased the sugar yield of corn fiber. These results demonstrate the effectiveness of enzyme consortium constructed by elucidating the chemical structure of hemicellulose in corn fiber for the degradation of corn fiber and also provide a general solution for the rational construction of targeted and efficient enzyme systems for the degradation of lignocellulosic biomass.

Interconvertible Pyridone Alkaloids from the Marine-Derived Fungus Penicillium oxalicum QDU1.

Eleven new pyridone alkaloids, penicipyridones A-K (1-11), and three new tetramic acids, tolypocladenols D-F (12-14), were isolated from rice media cultures of the marine-derived fungus Penicillium oxalicum QDU1. Their structures, including absolute configurations, were determined by comprehensive analyses of spectroscopic data, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction data. Interestingly, several of the penicipyridones undergo interconversions between hydroxy and methoxy groups at C-4 in acidic MeOH solution. Furthermore, in an acidic aqueous solution, OH-4 could be replaced by diverse substituent groups. Compounds 1, 4, 5, 8, 10, 11, and 14 exhibited moderate inhibitory effects on NO production in the LPS-induced RAW264.7 macrophages, with IC50 values ranging from 9.2 to 19 μM.

Inhibitory effect of three strains of biocontrol microbes on pathogens causing rhizome rot of Polygonatum cyrtonema

Rhizome rot is one of the main disease in the cultivation of Polygonatum cyrtonema, and it is also a global disease which seriously occurs on the perennial medicinal plants such as Panax notoginseng and P. ginseng. There is no effective control method at present. To identify the effects of three biocontrol microbes(Penicillium oxalicum QZ8, Trichoderma asperellum QZ2, and Brevibacillus amyloliquefaciens WK1) on the pathogens causing rhizome rot of P. cyrtonema, this study verified six suspected pathogens for their pathogenicity on P. cyrtonema. The result showed that Fusarium sp. HJ4, Colletotrichum sp. HJ4-1, and Phomopsis sp. HJ15 were the pathogens of rhizome rot of P. cyrtonema, and it was found for the first time that Phomopsis sp. could cause rhizome rot P. cyrtonema. Furthermore, the inhibitory effects of biocontrol microbes and their secondary metabolites on three pathogens were determined by confrontation culture. The results showed that the three tested biocontrol microbes significantly inhibited the growth of three pathogens. Moreover, the secondary metabolites of T. asperellum QZ2 and B. amyloliquefaciens WK1 showed significant inhibition against the three pathogens(P<0.05), and the effect of B. amyloliquefaciens WK1 sterile filtrate was significantly higher than that of high tempe-rature sterilized filtrate(P<0.05). B. amyloliquefaciens WK1 produced antibacterial metabolites to inhibit the growth of pathogens, and the growth inhibition rate of its sterile filtrate against three pathogens ranged from 87.84% to 93.14%. T. asperellum QZ2 inhibited the growth of pathogens through competition and antagonism, and P. oxalicum QZ8 exerted the inhibitory effect through competition. The research provides new ideas for the prevention and treatment of rhizome rot of P. cyrtonema and provides a basis for the di-sease control in other crops.

The interaction between the histone acetyltransferase complex Hat1-Hat2 and transcription factor AmyR provides a molecular brake to regulate amylase gene expression.

The chromatin structure is generally regulated by chromatin remodelers and histone modifiers, which affect DNA replication, repair, and levels of transcription. The first identified histone acetyltransferase was Hat1/KAT1, which belongs to lysine (K) acetyltransferases. The catalytic subunit Hat1 and the regulatory subunit Hat2 make up the core HAT1 complex. In this study, the results of tandem affinity purification and mass spectrometry and bimolecular fluorescence complementation proved that the Penicillium oxalicum PoHat1-Hat2 is the transcriptional cofactor of the sequence-specific transcription factor PoAmyR, a transcription activator essential for the transcription of amylase gene. ChIP-qPCR results demonstrated that the complex PoHat1-Hat2 is recruited by PoAmyR to the promoters of prominent amylase genes Poamy13A and Poamy15A and performs histone H4 lysine12 acetylation. The result of the yeast two-hybrid test indicated that PoHat2 is the subunit that directly interacts with PoAmyR. PoHat1-Hat2 acts as the molecular brake of the PoAmyR-regulating transcription of amylase genes. A putative model for amylase gene regulation by PoAmyR-Hat2-Hat1 was constructed. Our paper is the first report that the Hat1-Hat2 complex acts as a cofactor for sequence-specific TF to regulate gene expression and explains the mechanism of TF AmyR regulating amylase genes expression.

Screening of Dominant Cellulose-Degrading Microbe in Humus and Optimisation of its Enzyme Producing Conditions and Application Optimization of Enzyme Production Conditions and Evaluation of Efficient Straw Degradation by Penicillium oxalicum

Screening of Dominant Cellulose-Degrading Microbe in Humus and Optimisation of its Enzyme Producing Conditions and Application Optimization of Enzyme Production Conditions and Evaluation of Efficient Straw Degradation by <i>Penicillium oxalicum</i>

Endophytic Penicillium oxalicum CX-1 prevented Phytophthora cactorum blight on Salvia miltiorrhiza and promoted plant growth.

The main purpose of this study was to study the preventive effect of Penicillium sp. CX-1 on Phytophthora cactorum causing Salvia miltiorrhiza blight and its positive effect on plant growth. The endophytic strain CX-1 was isolated from the medicinal plant Corydalis saxicola Bunting and identified as Penicillium oxalicum. The growth inhibitory capacity of CX-1 against Ph. cactorum was 74.4% in the strain co-culture test and 86.2% in filtrate-modified plates. In the pot experiment, the in vivo control of CX-1 against Ph. cactorum in S. miltiorrhiza was 36.0%, which was higher than that of an anti-Phytophthora fungicide (23.4%). In addition, CX-1 had a potent ability to solubilize phosphate and also showed the ability to produce the plant hormone indole-3-acetic acid (IAA) and siderophores, which increase the bioavailability of iron to plants. It was demonstrated through pot experiments that CX-1 could significantly promote plant growth. As determined by real-time quantitative PCR, the expression of some S. miltiorrhiza tanshinone-related biosynthesis genes was significantly upregulated following colonization by CX-1. Strain CX-1 could effectively inhibit Ph. cactorum, the causative agent of S. miltiorrhiza blight, and significantly promoted the growth of plants through several different routes.

Different Maize Ear Rot Fungi Deter the Oviposition of Yellow Peach Moth (Conogethes punctiferalis (Guenée)) by Maize Volatile Organic Compounds

Yellow peach moth (Conogethes punctiferalis (Guenée), (Lepidoptera: Crambidae), YPM) and maize ear rot are important pests and diseases of maize (Zea mays L., (Poales: Poaceae)). In recent years, YPM has become the most destructive maize pest in the Huang-Huai-Hai summer maize region of China via the tunneling of larvae into maize ears. Interestingly, YPM infestation aggravates the occurrence of maize ear rot and causes heavier yield loss of maize in the field. However, few studies report whether maize ear rot would also affect the behavior of YPM. Here, we identified the effects of maize ear rot caused by four different fungi on maize ears’ volatile organic compounds (VOCs) and the cascading effects on the behavior of YPM. The current results found that mated YPM females showed a preference for mock-inoculated maize ears (MIM) or mechanically damaged maize ears (MDM) but showed repellence to Penicillium oxalicum (Eurotiales: Aspergillaceae)-infected maize ears (POM), Trichoderma asperellum (Hypocreales: Hypocreaceae)-infected maize ears (TAM), Aspergillus phoenicis (Eurotiales: Aspergillaceae)-infected maize ears (APM), Aspergillus flavus (Eurotiales: Aspergillaceae)-infected maize ears (AFM) in the oviposition selection and four-arm olfactometer experiments, indicating that VOCs emitting from fungi-infected maize ears were all repellent to mated YPM females. Further analyses showed that 57 VOCs were identified from all treatments. The partial least squares discriminant analysis (PLS-DA) displayed a separation between TAM, APM, AFM and POM, MDM, and MIM, with 24.3% and 19.1% explanation rates of the first two PLS components. Moreover, the relative quantities of eight common VOCs from different treatments were lower, and the other three common VOCs were higher in fungi-infected maize ears than those in MIM or MDM. There were also 17 unique VOCs in fungi-infected maize ears. In conclusion, these results suggested that maize ear rot negatively affected the behavior of YPM by changing both components and proportions of maize ears’ VOCs. These behavior-modifying VOCs may form the basis for the development of attractant or repellent formulations for YPM’s management in the future.

Late-stage cascade of oxidation reactions during the biosynthesis of oxalicine B in Penicillium oxalicum

Oxalicine B (1) is an α-pyrone meroterpenoid with a unique bispirocyclic ring system derived from Penicillium oxalicum. The biosynthetic pathway of 15-deoxyoxalicine B (4) was preliminarily reported in Penicillium canescens, however, the genetic base and biochemical characterization of tailoring reactions for oxalicine B (1) has remained enigmatic. In this study, we characterized three oxygenases from the metabolic pathway of oxalicine B (1), including a cytochrome P450 hydroxylase OxaL, a hydroxylating Fe(II)/α-KG-dependent dioxygenase OxaK, and a multifunctional cytochrome P450 OxaB. Intriguingly, OxaK can catalyze various multicyclic intermediates or shunt products of oxalicines with impressive substrate promiscuity. OxaB was further proven via biochemical assays to have the ability to convert 15-hydroxdecaturin A (3) to 1 with a spiro-lactone core skeleton through oxidative rearrangement. We also solved the mystery of OxaL that controls C-15 hydroxylation. Chemical investigation of the wild-type strain and deletants enabled us to identify 10 metabolites including three new compounds, and the isolated compounds displayed potent anti-influenza A virus bioactivities exhibiting IC50 values in the range of 4.0-19.9μmol/L. Our studies have allowed us to propose a late-stage biosynthetic pathway for oxalicine B (1) and create downstream derivatizations of oxalicines by employing enzymatic strategies.

Eco-friendly approach for control of fungal deterioration of archaeological skeleton dated back to the Greco-Roman period

The deterioration aspects of Papio Hamadryas skeleton dating back to the Greco-Roman period, and its associated fungal strains were investigated. Also, the efficacy of two green synthesized nanoparticles to suppress fungal growth was assessed. The results stated that the accumulated dust, erosion, holes, and colour change were the most significant deterioration aspects. The digital light microscope and scanning electron microscope (SEM) showed some micro-cracks and the presence of fauna identified as woodlice. Attenuated total reflection – Fourier transform infrared spectroscopy (ATR/FTIR) proved that the archaeological bone suffered from deterioration since the wavenumber of most bands and their intensities were lower compared to the control. The most common fungal strains associated with deteriorated bones were Aspergillus flavus, Aspergillus delicatus, Aspergillus parasiticus, Aspergillus niger, Aspergillus oryzae, Penicillium expansum, Penicillium oxalicum, Penicillium digitatum, Cladosporium sp., and Paecilomyces sp. These strains exhibited varied enzymatic activity, including protease, pectinase, cellulase, and gelatinase. Green synthesized silver nanoparticles (Ag-NPs) and magnesium nanoparticles MgO-NPs showed sufficient tools to suppress radial fungal growth. The growth inhibition percentages of Ag-NPs (69.1 ± 1.3–81.5 ± 2.01%) were more efficient than those recorded by MgO-NPs (59.5 ± 0.9 – 74.3 ± 2.8%) at a concentration of 300 ppm. The inhibition percentages were decreased at low NPs (200 and 100 ppm) concentrations, but it is still a promising tool to suppress fungal growth. This study recommended that Ag-NPs at 300 ppm may be used in the disinfection of the archaeological skeleton studied.

RESPONSE OF PH ON INDEGENOUS BIOCONTROL ACTIVITY OF FUNGAL AND BACTERIAL BIOLOGICAL AGENTS AGAINST RHIZOCTONIA SOLANI

Root rot incited by Rhizoctonia solani is one the major diseases of pea. As there are no long term management strategies for this soil borne pathogens therefore, the study was carried out to determine the effect of pH on the antagonistic activity of fungal and bacterial isolates (Th3 and Th5 (Trichoderma harzianum), An1 (Aspergillus niger), Po3 (Penicillium oxalicum) and PS1, PS2 and PS5 (Flourescent Pseudomonas) against Rhizoctonia solani at four different pH levels (5.0, 6.0, 7.0 and 8.0). All the seven indigenous fungal and bacterial isolates evaluated for their antagonistic potential against root-rot pathogen, significantly retarded the radial growth of Rhizoctonia solani in comparison to control at four different pH levels (5.0, 6.0, 7.0 and 8.0).The observations of the study revealed that all bio agents significantly inhibited the growth of the Rhizoctonia solani . Similarly, Trichoderma species had significant effects on the number and size of sclerotia. The pH range of 6.0-8.0 were found optimum for Trichoderma spp. while Flourescent Pseudomonas shows best antagonistic activity at pH 7.0. As far as the effect of pH levels is concerned, all the promising fungal and bacterial isolates caused maximum growth inhibition of test pathogens at pH 6.0 to 7.0.