Showing papers on "Mycelium published in 2022"

Control of Fungal Diseases and Fruit Yield Improvement of Strawberry Using Bacillus velezensis CE 100

Abstract: Due to the increasing health and environmental risks associated with the use of fungicides in agriculture, alternatives—such as using plant growth-promoting bacteria (PGPB) to suppress phytopathogens—that simultaneously improve plant yield, are important. This study evaluated the biocontrol efficiency of Bacillus velezensis CE100 against Macrophomina phaseolina and Fusarium oxysporum f. sp. fragariae, the respective causal agents for charcoal rot and fusarium wilt diseases in strawberry, and its potential to enhance strawberry growth and fruit production. B. velezensis CE 100 produced fungal cell wall-degrading enzymes, chitinases, and β-1,3-glucanases; and inhibited the mycelial growth of M. phaseolina and F. oxysporum f. sp. fragariae by 64.7% and 55.2%, respectively. The mycelia of both phytopathogenic fungi showed severe swelling and rupturing of the hyphae compared to the smooth, normal growth in the control group. Moreover, B. velezensis CE100 produced up to 2.8 units/mL of indole-3-acetic acid (IAA) during incubation and enhanced root biomass in strawberries. Consequently, B. velezensis CE 100 not only increased the fruit yield of strawberries by controlling the fungal diseases but also through enhancing plant growth. The findings of this study indicate that B. velezensis CE100 could be a safe, ecofriendly biocontrol alternative to chemical fungicides in strawberry production.

Design, Synthesis, and Antifungal Activity of Novel Chalcone Derivatives Containing a Piperazine Fragment.

Abstract: In an attempt to find the biorational pesticides, 20 novel chalcone derivatives containing a piperazine fragment were designed and synthesized. Their fungicidal activities and preliminarily action mechanism against Rhizoctonia solani were evaluated. Strikingly, the biological activity of compound D2 was obtained by optimizing the structure of the system. Subsequently, the practical value of compound D2 was ascertained by the relative surveys on in vivo anti-R. solani and anti-Colletotrichum gloeosporioides. The results revealed by scanning electron microscopy demonstrated that compound D2 could induce irregular and shrivelled growth of mycelium and rupture of the mycelium surface. This study indicates that chalcone derivatives containing a piperazine skeleton had better inhibitory effect on plant fungi, providing further complementary research on new pesticides.

Antifungal application of biosynthesized selenium nanoparticles with pomegranate peels and nanochitosan as edible coatings for citrus green mold protection

Abstract: Citrus production and trading are seriously affected by fungal decays worldwide; the green mold infection by Penicillium digitatum could be the most disastrous. The substitutions of chemical and synthetic fungicides with effectual natural alternatives are global demands; plant extract from pomegranates peels (PPE), biosynthesized selenium nanoparticles with PPE (PPE/SeNPs) and chitosan nanoparticles (NCT) were suggested as efficacious fungicidal agents/nanocomposites to control P. digitatum strains.PPE from Punica granatum was extracted and employed directly for synthesizing SeNPs, whereas NCT was produced using ionic gelation method of chitosan extracted from white prawn (Fenneropenaeus indicus) shells. The physiochemical, biochemical and structural characterization of generated molecules were conducted using infra-red spectroscopy, particles' size (Ps) and charge assessment and electron microscopes imaging. Antifungal potentialities were investigated in vitro and in infected fruits with P. digitatum by applying NCT nanocomposites-based edible coating.The synthesis of PPE-synthesized SeNPs and NCT was successfully achieved, the molecular bonding in synthesized agents/composites were proved with infrared spectroscopy to have both biochemical and physical interactions. The nanoparticles had 82.72, 9.41 and 85.17 nm mean diameters for NCT, PPE/SeNPs and NCT/PPE/SeNPs nanocomposites, respectively. The nanoparticles had homogenous spherical shapes and good distribution attributes. The entire agents/nanocomposites exhibited potent fungicidal potentialities toward P. digitatum isolates; NCT/PPE/SeNPs nanocomposite was the most forceful and significantly exceeded the fungicidal action of standard fungicide. The direct treatment of fungal mycelia with NCT/PPE/SeNPs nanocomposite led to remarkable lysis and deformations of P. digitatum hyphae within 12 h of treatment. The coating of infected orange with NCT-based edible coatings reduced the green mold infection signs by 91.7, 95.4 and 100%, for NCT, NCT/PPE and NCT/PPE/SeNPs based coating solutions, respectively.NCT, PPE-synthesized SeNPs, and their innovative nanocomposites NCT/PPE/SeNPs are convincingly recommended for formulating effectual antifungal and edible coatings to eliminate postharvest fungal pathogen, both with protection from their invasion or with destructing their existing infections.

Fungi in Mycelium-Based Composites: Usage and Recommendations

Abstract: Mycelium-Based Composites (MBCs) are innovative engineering materials made from lignocellulosic by-products bonded with fungal mycelium. While some performance characteristics of MBCs are inferior to those of currently used engineering materials, these composites nevertheless prove to be superior in ecological aspects. Improving the properties of MBCs may be achieved using an adequate substrate type, fungus species, and manufacturing technology. This article presents scientifically verified guiding principles for choosing a fungus species to obtain the desired effect. This aim was realized based on analyses of scientific articles concerning MBCs, mycological literature, and patent documents. Based on these analyses, over 70 fungi species used to manufacture MBC have been identified and the most commonly used combinations of fungi species-substrate-manufacturing technology are presented. The main result of this review was to demonstrate the characteristics of the fungi considered optimal in terms of the resulting engineering material properties. Thus, a list of the 11 main fungus characteristics that increase the effectiveness in the engineering material formation include: rapid hyphae growth, high virulence, dimitic or trimitic hyphal system, white rot decay type, high versatility in nutrition, high tolerance to a substrate, environmental parameters, susceptibility to readily controlled factors, easy to deactivate, saprophytic, non-mycotoxic, and capability to biosynthesize natural active substances. An additional analysis result is a list of the names of fungus species, the types of substrates used, the applications of the material produced, and the main findings reported in the scientific literature.

AoPEX1 and AoPEX6 Are Required for Mycelial Growth, Conidiation, Stress Response, Fatty Acid Utilization, and Trap Formation in Arthrobotrys oligospora

Abstract: Nematode-trapping (NT) fungi are important resources for the biological control of plant-parasitic nematodes. They are widely distributed in various ecological environments and capture nematodes by producing unique predatory organs (traps). ABSTRACT Arthrobotrys oligospora (A. oligospora) is a typical nematode-trapping (NT) fungus that can capture nematodes by producing adhesive networks. Peroxisomes are single membrane-bound organelles that perform multiple physiological functions in filamentous fungi. Peroxisome biogenesis proteins are encoded by PEX genes, and the functions of PEX genes in A. oligospora and other NT fungi remain largely unknown. Here, our results demonstrated that two PEX genes (AoPEX1 and AoPEX6) are essential for mycelial growth, conidiation, fatty acid utilization, stress tolerance, and pathogenicity in A. oligospora. AoPEX1 and AoPEX6 knockout resulted in a failure to produce traps, conidia, peroxisomes, and Woronin bodies and damaged cell walls, reduced autophagosome levels, and increased lipid droplet size. Transcriptome data analysis showed that AoPEX1 and AoPEX6 deletion resulted in the upregulation of the proteasome, membranes, ribosomes, DNA replication, and cell cycle functions, and the downregulation of MAPK signaling and nitrogen metabolism. In summary, our results provide novel insights into the functions of PEX genes in the growth, development, and pathogenicity of A. oligospora and contribute to the elucidation of the regulatory mechanism of peroxisomes in trap formation and lifestyle switching in NT fungi. IMPORTANCE Nematode-trapping (NT) fungi are important resources for the biological control of plant-parasitic nematodes. They are widely distributed in various ecological environments and capture nematodes by producing unique predatory organs (traps). However, the molecular mechanisms of trap formation and lifestyle switching in NT fungi are still unclear. Here, we provided experimental evidence that the AoPEX1 and AoPEX6 genes could regulate mycelial growth and development, trap formation, and nematode predation of A. oligospora. We further analyzed the global transcription level changes of wild-type and mutant strains using RNA-seq. This study highlights the important role of peroxisome biogenesis genes in vegetative growth, conidiation, trap formation, and pathogenicity, which contribute to probing the mechanism of organelle development and trap formation of NT fungi and lays a foundation for developing high-efficiency nematode biocontrol agents.

Antifungal Activity of Volatile Organic Compounds from Bacillus velezensis CE 100 against Colletotrichum gloeosporioides

Abstract: Colletotrichum gloeosporioides is the most prevalent phytopathogen, causing anthracnose disease that severely affects the production of various fruit trees, including walnut and jujube. In this study, the volatile organic compounds (VOCs) from Bacillus velezensis CE 100 disrupted the cell membrane integrity of C. gloeosporioides and reduced the spore germination by 36.4% and mycelial growth by 20.0% at a bacterial broth concentration of 10%, while the control group showed no antifungal effect. Based on the headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS) analysis, seven VOCs were identified from the headspace of B. velezensis CE 100. Out of the seven VOCs, 5-nonylamine and 3-methylbutanoic acid were only detected in the headspace of B. velezensis CE 100 but not in the control group. Both 5-nonylamine and 3-methylbutanoic acid showed significant antifungal activity against the spore germination and mycelial growth of C. gloeosporioides. Treatment with 100 µL/mL of 5-nonylamine and 3-methylbutanoic acid suppressed the spore germination of C. gloeosporioides by 10.9% and 30.4% and reduced mycelial growth by 14.0% and 22.6%, respectively. Therefore, 5-nonylamine and 3-methylbutanoic acid are the potential antifungal VOCs emitted by B. velezensis CE 100, and this is the first report about the antifungal activity of 5-nonylamine against C. gloeosporioides.

Language of fungi derived from their electrical spiking activity

Abstract: Fungi exhibit oscillations of extracellular electrical potential recorded via differential electrodes inserted into a substrate colonized by mycelium or directly into sporocarps. We analysed electrical activity of ghost fungi (Omphalotus nidiformis), Enoki fungi (Flammulina velutipes), split gill fungi (Schizophyllum commune) and caterpillar fungi (Cordyceps militaris). The spiking characteristics are species specific: a spike duration varies from 1 to 21 h and an amplitude from 0.03 to 2.1 mV. We found that spikes are often clustered into trains. Assuming that spikes of electrical activity are used by fungi to communicate and process information in mycelium networks, we group spikes into words and provide a linguistic and information complexity analysis of the fungal spiking activity. We demonstrate that distributions of fungal word lengths match that of human languages. We also construct algorithmic and Liz-Zempel complexity hierarchies of fungal sentences and show that species S. commune generate the most complex sentences.

Dense and continuous networks of aerial hyphae improve flexibility and shape retention of mycelium composite in the wet state

Abstract: The mycelium composites composed of fungal mycelium and plant substrate are a practical alternative to petroleum plastic-based foam materials. However, the effects of the physiological traits of fungus on the composites’ microscopic structure and mechanical properties remain poorly understood. Here, we compared two basidiomycetes with distinct mycelium morphology and white-decay modes. Cross-sectional observation revealed that the mycelium composites possess a core/shell structure with the shell formed of dense aerial mycelium and the core of plant particles and spongy aerial mycelium. Dense and continuous mycelium networks provided by Trametes hirsuta strengthen the mechanical properties of the composite compared to the coarse mycelium networks of Pleurotus ostreatus. In particular, the firm mycelial shell skeleton confers high flexibility and shape-retention to the composite in the wet state. This unique characteristic of the mycelium composite indicates its potentials in new industrial applications.

Dense and continuous networks of aerial hyphae improve flexibility and shape retention of mycelium composite in the wet state

Abstract: The mycelium composites composed of fungal mycelium and plant substrate are a practical alternative to petroleum plastic-based foam materials. However, the effects of the physiological traits of fungus on the composites’ microscopic structure and mechanical properties remain poorly understood. Here, we compared two basidiomycetes with distinct mycelium morphology and white-decay modes. Cross-sectional observation revealed that the mycelium composites possess a core/shell structure with the shell formed of dense aerial mycelium and the core of plant particles and spongy aerial mycelium. Dense and continuous mycelium networks provided by Trametes hirsuta strengthen the mechanical properties of the composite compared to the coarse mycelium networks of Pleurotus ostreatus. In particular, the firm mycelial shell skeleton confers high flexibility and shape-retention to the composite in the wet state. This unique characteristic of the mycelium composite indicates its potentials in new industrial applications.

First Report of <i>Lasiodiplodia theobromae</i> Causing Leaf Spot on <i>Aloe vera</i> var. <i>chinensis</i> in Malaysia

Abstract: Aloe vera L. var. chinensis (Haw.) Berg. (family Asphodelaceae), locally known as 'Lidah Buaya', is an economically important plant as the gel from the leaves possesses anti-inflammatory, anti-arthritic, antibacterial, and hypoglycemic properties and is used for cosmetic, pharmaceutical and healing purpose in Malaysia. In July 2021, irregular black sunken spots (3- to 10-mm in diameter) were observed on the leaves of 'Lidah Buaya' plants under leaf development stage in the field located in the district Penampang of Sabah province (N5°56'37.1" E116°04'21.5"). The disease severity was about 30% with 10% incidence. The tissues surrounding the black spots became brown and dry when the plants grew older. No gel contained in the sunken zones. Symptomatic leaf tissues (5 x 5 mm) were cut from the infected margin, surface sterilised with 75% ethanol for 1 minute, washed with 2% sodium hypochlorite solution for 1 minute, rinsed, and air dried before plating on five potato dextrose agar (PDA) plates (pH 7). Plates were incubated at 25°C for 3 days in the dark. Greyish-white fluffy mycelia were observed, and then became dark grey with age. Dark pigmentation in each plate was produced after a week of incubation at 25°C. A representative isolate Penampang was further characterized morphologically and molecularly. Immature conidia were single-celled, aseptate, ellipsoid and hyaline, measuring 19.4 × 24.5 µm (n = 30). Mature conidia were brown, thick-walled and one-septate with longitudinal striations, 22.5 × 28.3 µm (n = 30). Genomic DNA was extracted from fresh mycelia of isolate Penampang based on the extraction method described by Khoo et al. (2021) with additional of mechanical disruption using micro pestle before heating. KOD One PCR master mix (Toyobo, Japan) containing hot-start modified KOD DNA polymerase was used for PCR amplification. The PCR condition were 94°C for 10 s, 55°C for 5 s and 72°C for 2 s, for 30 cycles, and initial denaturation of 94°C for 3 min and a final extension step of 72°C for 5 min. The internal transcribed spacer (ITS) region of rDNA and tubulin (TUB) genes were amplified using ITS1/ITS4 and T10/Bt2b primer sets, respectively (O'Donnell et al. 1997; White et al. 1990). The products were then sent to Apical Scientific Sdn. Bhd. for sequencing. The generated ITS (OK209451) and TUB (OL660667) were 100% identical to L. theobromae isolate MRR-161 and CPC:27690 (GenBank MW282884 and MT592639, respectively) in BLASTn analysis. Phylogenetic analysis using maximum likelihood based on the combined ITS and TUB sequences indicated that the isolates formed a supported clade (91% bootstrap value) to the related L. theobromae. The morphological and molecular characterization of the fungus matched L. theobromae described by Pečenka et al. (2021). Mycelial agar plugs (5-mm-diameter) from 7-day-old PDA culture of Penampang isolate were placed onto pinpricked leaves of three 2-month-old 'Lidah Buaya' plants. Pinpricked leaves of three 2-month-old 'Lidah Buaya' plants received sterile 5-mm-diameter PDA agar plugs to serve as controls. The inoculated 'Lidah Buaya' plants were covered with plastics for 48 h, and were incubated at 25°C. All inoculated leaves developed symptoms as described above 6 to 7 days post-inoculation, whereas no symptoms occurred on controls, thus fulfilling Koch's postulates. The experiments were repeated twice. The reisolated fungus was identical to representative isolate Penampang morphologically and molecularly. L. theobromae was reported previously on A. vera in Cuba (Urtiaga 1986) and India (Mathur 1979). To our knowledge, this is the first report of L. theobromae causing leaf spot on A. vera in Malaysia. The occurrence of this disease emphasizes the importance of disease surveillance in the region. Plant disease management strategies need to be established to reduce the losses.

AoSsk1, a Response Regulator Required for Mycelial Growth and Development, Stress Responses, Trap Formation, and the Secondary Metabolism in Arthrobotrys oligospora

Abstract: Ssk1, a response regulator of the two-component signaling system, plays an important role in the cellular response to hyperosmotic stress in fungi. Herein, an ortholog of ssk1 (Aossk1) was characterized in the nematode-trapping fungus Arthrobotrys oligospora using gene disruption and multi-phenotypic comparison. The deletion of Aossk1 resulted in defective growth, deformed and swollen hyphal cells, an increased hyphal septum, and a shrunken nucleus. Compared to the wild-type (WT) strain, the number of autophagosomes and lipid droplets in the hyphal cells of the ΔAossk1 mutant decreased, whereas their volumes considerably increased. Aossk1 disruption caused a 95% reduction in conidial yield and remarkable defects in tolerance to osmotic and oxidative stress. Meanwhile, the transcript levels of several sporulation-related genes were significantly decreased in the ΔAossk1 mutant compared to the WT strain, including abaA, brlA, flbC, fluG, and rodA. Moreover, the loss of Aossk1 resulted in a remarkable increase in trap formation and predation efficiency. In addition, many metabolites were markedly downregulated in the ΔAossk1 mutant compared to the WT strain. Our results highlight that AoSsk1 is a crucial regulator of asexual development, stress responses, the secondary metabolism, and pathogenicity, and can be useful in probing the regulatory mechanism underlying the trap formation and lifestyle switching of nematode-trapping fungi.

Biological Control and Plant Growth Promotion by Volatile Organic Compounds of Trichoderma koningiopsis T-51

Abstract: Trichoderma spp. are widely used in plant disease control and growth promotion due to their high efficacy and multiple biocontrol mechanisms. Trichoderma koningiopsis T-51 is an effective biocontrol agent against gray mold disease by direct contact. However, the indirect physical contact biocontrol potential of Trichoderma spp. is not clear. In this study, the volatile organic compounds (VOCs) produced by T-51 showed high inhibitory activity against plant pathogenic fungi Botrytis cinerea and Fusarium oxysporum. The percentage of B. cinerea and F. oxysporum mycelial growth inhibition by T-51 VOCs was 73.78% and 43.68%, respectively. In both B. cinerea and F. oxysporum, conidial germination was delayed, and germ tube elongation was suppressed when exposed to T-51 VOCs, and the final conidial germination rate of B. cinerea decreased significantly after T-51 treatment. The VOCs from T-51 reduced the Botrytis fruit rot of tomato compared with that noted when using the control. Moreover, the T-51 VOCs significantly increased the size and weight of Arabidopsis thaliana seedlings. Twenty-four possible compounds, which were identified as alkenes, alkanes, and esters, were detected in VOCs of T-51. These results indicate that T. koningiopsis T-51 can exert biological control by integrating actions to suppress plant disease and promote plant growth.

Comparative Extraction of Phenolic Compounds from Olive Leaves Using a Sonotrode and an Ultrasonic Bath and the Evaluation of Both Antioxidant and Antimicrobial Activity

Abstract: A sonotrode ultrasound-assisted extraction of phenolic compounds from olive leaves has been developed using a Box–Behnken design to optimize the effects of solvent composition and ultrasound parameters. The determination of single phenolic compounds was performed by HPLC–MS and the highest recovery in total compounds, oleuropein and hydroxytyrosol was achieved using EtOH/H2O (55:45, v/v), 8 min and 100% of amplitude. The optimal conditions were applied on leaves from seven olive cultivars grown under the same conditions and the results were compared with those found by using a conventional ultrasonic bath, obtaining no statistical differences. Moreover, antioxidant activity by FRAP, DPPH and ABTS in these olive leaf extracts was evaluated and they exhibited a significant correlation with oleuropein and total phenolic content. All cultivars of olive leaf extracts were found to be active against S. aureus and methicillin-resistant S. aureus with minimum bactericidal concentration (MBC) values) that ranged from 5.5 to 22.5 mg mL−1. No extracts showed antimicrobial activity against C. albicans. The percentages of mycelium reduction in B. cinerea ranged from 2.2 and 18.1%. Therefore, sonotrode could be considered as an efficient and fast extraction technique that could be easily scaled-up at industrial level, thus allowing for olive leaves to be revalorized.