Studies on the Rapid and Simple DNA Extraction Method, Antibacterial Activity and Enzyme Activity Involved in Plant Biomass Conversion by Cookeina sulcipes and C. tricholoma (Cup Fungi)

Cookeina sulcipes and C. tricholoma are a cup fungi (Ascomycota) collected in Saraburi, Thailand. The fungi have been isolated, cultured and confirmed as respective species. For morphology, both Cookeina sp. are white mycelium and the growth rate on potato dextrose agar (PDA) result present C. sulcipes is faster than C. tricholoma. The molecular characterization from a rapid and simple DNA extraction method that is modified based on thermolysis method, The DNA extraction is finish in thirty minutes and efficiency to continuous with polymerase chain reaction (PCR) amplification to fungi species level identification. The DNA sequence from internal transcribed spacer (ITS) gene regions by universal primer pairs ITS5/ITS4 is effective to confirm Cookeina species level that C. sulcipes has 616 bp and C. tricholoma has 570 bp. Including, DNA sequence of large subunit (LSU) gene regions by universal primer pairs LROR/LR5 is generate that C. sulcipes has 912 bp and C. tricholoma has 906 bp. The cultures are screened for antibacterial activity by agar plug diffusion method and found that both isolates have been no activity against test strains (Bacillus subtilis, Escherichia coli, Kocuria rhizophila (Micrococcus luteus), Pseudomonas aeruginosa, Staphylococcus aureus and S. epidermidis). In a preliminary screening test of enzymes involved in plant biomass breakdown by agar plate method, both Cookeina sp. show cellulolytic and hemicellulolytic enzymatic activity, and manganese peroxidase (MnP) productivity. In contrast, only C. sulcipes had additional laccase activity. Neither isolate generate pectinolytic and lignin peroxidase (LiP) activities. Thus, Cookeina spp. proved the potentiality to break down lignocelluloses

Hidden diversity of Pestalotiopsis and Neopestalotiopsis (Amphisphaeriales, Sporocadaceae) species allied with the stromata of entomopathogenic fungi in Taiwan

Pestalotiopsis sensu lato, commonly referred to as pestalotiopsis-like fungi, exhibit a broad distribution and are frequently found as endophytes, saprobes and pathogens across various plant hosts. The taxa within pestalotiopsis-like fungi are classified into three genera viz. Pestalotiopsis, Pseudopestalotiopsis and Neopestalotiopsis, based on the conidial colour of their median cells and multi-locus molecular phylogenies. In the course of a biodiversity investigation focusing on pestalotiopsis-like fungi, a total of 12 fungal strains were identified. These strains were found to be associated with stromata of Beauveria, Ophiocordyceps and Tolypocladium in various regions of Taiwan from 2018 to 2021. These strains were evaluated morphologically and multi-locus phylogenetic analyses of the ITS (internal transcribed spacer), tef1-α (translation elongation factor 1-α) and tub2 (beta-tubulin) gene regions were conducted for genotyping. The results revealed seven well-classified taxa and one tentative clade in Pestalotiopsis and Neopestalotiopsis. One novel species, Pestalotiopsis manyueyuanani and four new records, N. camelliae-oleiferae, N. haikouensis, P. chamaeropis and P. hispanica, were reported for the first time in Taiwan. In addition, P. formosana and an unclassified strain of Neopestalotiopsis were identified, based on similarities of phylogeny and morphology. However, the data obtained in the present study suggest that the currently recommended loci for species delimitation of pestalotiopsis-like fungi do not deliver reliable or adequate resolution of tree topologies. The in-vitro mycelial growth rates of selected strains from these taxa had an optimum temperature of 25 °C, but growth ceased at 5 °C and 35 °C, while all the strains grew faster under alkaline than acidic or neutral pH conditions. This study provides the first assessment of pestalotiopsis-like fungi, associated with entomopathogenic taxa

FIGURE 3 in Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand

FIGURE 3. Greeneria saprophytica: conidiogenous cells and conidia.Published as part of <i>Tangthirasunun, Narumon, Silar, Philippe, Bhat, Darbhe Jayarama, Maharachchikumbura, Sajeewa S.N. & Hyde, Kevin D., 2014, Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand, pp. 275-282 in Phytotaxa 184 (5)</i> on page 280, DOI: 10.11646/phytotaxa.184.5.3, <a href="http://zenodo.org/record/10089683">http://zenodo.org/record/10089683</a&gt

FIGURE 2 in Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand

FIGURE 2. Greeneria saprophytica (MFLUCC 12-0298, holotype). A. Specimen on dead leaf of Syzygium cumini. B. Conidiomata on the host surface. C. L.S. of a conidioma. D–H. Phialidic conidiogenous cells with developing conidia; in G. note proliferating conidiogenous cell. I–L. Conidia. M. Germinating conidium. N–O. Colonies on PDA; N. From top, O. From reverse. Scale bars: C = 50 μm, D–M = 10 μm.Published as part of <i>Tangthirasunun, Narumon, Silar, Philippe, Bhat, Darbhe Jayarama, Maharachchikumbura, Sajeewa S.N. & Hyde, Kevin D., 2014, Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand, pp. 275-282 in Phytotaxa 184 (5)</i> on page 279, DOI: 10.11646/phytotaxa.184.5.3, <a href="http://zenodo.org/record/10089683">http://zenodo.org/record/10089683</a&gt

Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand

Tangthirasunun, Narumon, Silar, Philippe, Bhat, Darbhe Jayarama, Maharachchikumbura, Sajeewa S.N., Hyde, Kevin D. (2014): Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand. Phytotaxa 184 (5): 275-282, DOI: 10.11646/phytotaxa.184.5.

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina

International audienceThe filamentous fungus Podospora anserina is a good model to study the breakdown of lignocellulose, owing to its ease of culture and genetical analysis. Here, we show that the fungus is able to use a wide range of lignocellulosic materials as food sources. Using color assays, spectroscopy and pyrolysis–gas chromatography mass spectrometry, we confirm that this ascomycete is able to degrade lignin, primarily by hydrolyzing β–O-4 linkages, which facilitates its nutrient uptake. We show that the limited weight loss that is promoted when attacking Miscanthus giganteus is due to a developmental blockage rather than an inefficiency of its enzymes. Finally, we show that lignin, and, more generally, phenolics, including degradation products of lignin, greatly stimulate the growth and fertility of the fungus in liquid cultures. Analyses of the CATΔΔΔΔΔ mutant lacking all its catalases, pro-oxidants and antioxidants indicate that improved growth and fertility of the fungus is likely caused by augmented reactive oxygen species levels triggered by the presence of phenolics

FIGURE 1 in Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand

FIGURE 1. Maximum likelihood (ML) majority rule 28S nuclear large subunit (nuLSU) consensus tree for Greeneria saprophytica, G. uvicola and other representatives in order Diaporthales and genera incertae sedis. RAxML bootstrap support values are given at the nodes. The tree is rooted to Coniochaeta velutina (Coniochaetales).Published as part of <i>Tangthirasunun, Narumon, Silar, Philippe, Bhat, Darbhe Jayarama, Maharachchikumbura, Sajeewa S.N. & Hyde, Kevin D., 2014, Greeneria saprophytica sp. nov. on dead leaves of Syzygium cumini from Chiang Rai, Thailand, pp. 275-282 in Phytotaxa 184 (5)</i> on page 278, DOI: 10.11646/phytotaxa.184.5.3, <a href="http://zenodo.org/record/10089683">http://zenodo.org/record/10089683</a&gt

Plant biomass degrading ability of the coprophilic ascomycete fungus Podospora anserina

International audienceThe degradation of plant biomass is a major challenge towards the production of bio-based compounds and materials. As key lignocellulolytic enzyme producers, filamentous fungi represent a promising reservoir to tackle this challenge. Among them, the coprophilous ascomycete Podospora anserina has been used as a model organism to study various biological mechanisms because its genetics are well understood and controlled. In 2008, the sequencing of its genome revealed a great diversity of enzymes targeting plant carbohydrates and lignin. Since then, a large array of lignocellulose-acting enzymes has been characterized and genetic analyses have enabled the understanding of P. anserina metabolism and development on plant biomass. Overall, these research efforts shed light on P. anserina strategy to unlock recalcitrant lignocellulose deconstruction

Camarosporium sensu stricto in Pleosporinae, Pleosporales with two new species

Wijayawardene, Nalin N., Bhat, D. Jayarama, Hyde, Kevin D., Camporesi, E., Chethana, K.W.T., Tangthirasunun, Narumon, Wang, Y. (2014): Camarosporium sensu stricto in Pleosporinae, Pleosporales with two new species. Phytotaxa 183 (1): 16-26, DOI: 10.11646/phytotaxa.183.1.2, URL: http://dx.doi.org/10.11646/phytotaxa.183.1.

Inactivation of cellobiose dehydrogenases modifies the cellulose degradation mechanism of Podospora anserina

Conversion of biomass into high-value products, including biofuels, is of great interest to developing sustainable biorefineries. Fungi are an inexhaustible source of enzymes to degrade plant biomass. Cellobiose dehydrogenases (CDHs) play an important role in the breakdown through synergistic action with fungal lytic polysaccharide monooxygenases (LPMOs). The three CDH genes of the model fungus Podospora anserina were inactivated, resulting in single and multiple CDH mutants. We detected almost no difference in growth and fertility of the mutants on various lignocellulose sources, except on crystalline cellulose, on which a 2-fold decrease in fertility of the mutants lacking P. anserina CDH1 (PaCDH1) and PaCDH2 was observed. A striking difference between wild-type and mutant secretomes was observed. The secretome of the mutant lacking all CDHs contained five beta-glucosidases, whereas the wild type had only one. P. anserina seems to compensate for the lack of CDH with secretion of beta-glucosidases. The addition of P. anserina LPMO to either the wild-type or mutant secretome resulted in improvement of cellulose degradation in both cases, suggesting that other redox partners present in the mutant secretome provided electrons to LPMOs. Overall, the data showed that oxidative degradation of cellulosic biomass relies on different types of mechanisms in fungi.IMPORTANCE:Plant biomass degradation by fungi is a complex process involving dozens of enzymes. The roles of each enzyme or enzyme class are not fully understood, and utilization of a model amenable to genetic analysis should increase the comprehension of how fungi cope with highly recalcitrant biomass. Here, we report that the cellobiose dehydrogenases of the model fungus Podospora anserina enable it to consume crystalline cellulose yet seem to play a minor role on actual substrates, such as wood shavings or miscanthus. Analysis of secreted proteins suggests that Podospora anserina compensates for the lack of cellobiose dehydrogenase by increasing beta-glucosidase expression and using an alternate electron donor for LPMO