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The silk tree (Albizia lebbeck (L.) Benth) is one of the ornamental plants in Ahvaz (southwest of Iran). Two endophytic fungal strains, isolated from the leaves of this plant, were characterized. Molecular phylogenetic analyses based on a combined matrix of the internal transcribed spacer regions 1 and 2 including the intervening 5.8S nuclear ribosomal DNA (ITS), and partial regions of the β-tubulin (tub2) and RNA polymerase II second largest subunit (rpb2) indicated that both strains represent a new Chaetomium species, C. albiziae sp. nov. in the family Chaetomiaceae. Chaetomium albiziae formed a sister lineage with sterile species C. cucumericola. This new species can be characterized by ostiolate perithecia with septate lateral and terminal setae, clavate asci and aseptate ellipsoidal to ovoid ascospores with an apical germ pore. In addition, this is the first report of the endophytic association of the genus Chaetomium with Albizia lebbeck worldwide.

The multiprotein Fab1p/PIKfyve-complex regulating the abundance of the phospholipid phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2) is highly conserved among eukaryotes. In yeast/mammals, it is composed of the phosphatidylinositol 3-phosphate 5-kinase Fab1p/PIKfyve, the PtdIns(3,5)P2 phosphatase Fig4p/Sac3 and the scaffolding subunit Vac14p/ArPIKfyve. The complex is located to vacuolar membranes in yeast and to endosomal membranes in mammals, where it controls the synthesis and turnover of PtdIns(3,5)P2. In this study, we analyzed the role and function of the Fab1p/PIKfyve-complex scaffold protein SmVAC14 in the filamentous ascomycete Sordaria macrospora (Sm). We generated the Smvac14 deletion strain ∆vac14 and performed phenotypic analysis of the mutant. Furthermore, we conducted fluorescence microscopic localization studies of fluorescently labeled SmVAC14 with vacuolar and late endosomal marker proteins. Our results revealed that SmVAC14 is important for maintaining vacuolar size and appearance as well as proper sexual development in S. macrospora. In addition, SmVAC14 plays an important role in starvation stress response. Accordingly, our results propose that the turnover of PtdIns(3,5)P2 is of great significance for developmental processes in filamentous fungi.

During a study of the diversity of soilborne fungi from Spain, a strain belonging to the family Chaetomiaceae (Sordariales) was isolated. The multigene phylogenetic inference using five DNA loci showed that this strain represents an undescribed species of the genus Amesia, herein introduced as A. hispanica sp. nov. Investigation of its secondary metabolome led to the isolation of two new derivatives (2 and 3) of the known antifungal antibiotic dactylfungin A (1), together with the known compound cochliodinol (4). The planar structures of 1–4 were determined by ultrahigh performance liquid chromatography coupled with diode array detection and ion mobility tandem mass spectrometry (UHPLC-DAD-IM-MS/MS) and extensive 1D and 2D nuclear magnetic resonance (NMR) spectroscopy after isolation by HPLC. All isolated secondary metabolites were tested for their antimicrobial and cytotoxic activities. Dactylfungin A (1) showed selective and strong antifungal activity against some of the tested human pathogens (Aspergillus fumigatus and Cryptococcus neoformans). The additional hydroxyl group in 2 resulted in the loss of activity against C. neoformans but still retained the inhibition of As. fumigatus in a lower concentration than that of the respective control, without showing any cytotoxic effects. In contrast, 2500-dehydroxy-dactylfungin A (3) exhibited improved activity against yeasts (Schizosaccharomyces pombe and Rhodotorula glutinis) than 1 and 2, but resulted in the appearance of slight cytotoxicity. The present study exemplifies how even in a well-studied taxonomic group such as the Chaetomiaceae, the investigation of novel taxa still brings chemistry novelty, as demonstrated in this first report of this antibiotic class for chaetomiaceous and sordarialean taxa.

Chaetomium albiziae M. Mehrabi-Koushki & A. Safi, sp. nov. (Fig. 1) MycoBank: MB 841114 Holotype: IRAN, Khuzestan Province, Ahvaz, isolated from a living leaf of Albizia lebbeck (Fabaceae), Oct. 2018, A. Safi (holotype, IRAN 18081F; ex-type cultures, IRAN 4137C = SCUA-Saf-B10). Etymology. Species epithet refers to the host genus Albizia from which the holotype was isolated. Sexual morph: Hyphae hyaline to pale brown, septate, branched. Ascomata perithecioid, superficial, brown to dark brown in reflected light, globose to subglobose, (79–)118–170(–184) × (71–)100–145(–158) μm, 95 % confidence limits = 134–148.5 × 112.2–124.8 μm, (± SD = 141.3 ± 25.5 × 118.5 ± 22.1 μm, n = 50). Ascomatal wall brown, textura angularis in surface view. Terminal hairs seta-like, septate, unbranched, smooth, brown, tapering towards the tips, 2.5–3.8 μm diam near the base. Lateral hairs similar but more flexuous, narrower and shorter. Asci fasciculate, clavate, spore bearing part (20–)22.5–35(–38.75) × 7.5–16.5 μm, stalks 11.25–22.5 μm long, with 8 irregularly-arranged ascospores, evanescent. Ascospores brown to olivaceous brown, ellipsoidal to ovoid, aseptate, biapiculate, bilaterally flattened, with an apical germ pore, (7.5–)10–11.25(–12.5) × 7.5–10 μm, 95 % confidence limits = 10.4–10.9 × 8.1–8.6 μm, (± SD = 10.7 ± 0.8 × 8.3 ± 0.7 μm, n = 50). Asexual morph: not observed. Culture characteristics—Colonies on OA reaching 60–66 mm after 4 d of incubation at 28 ± 0.5 °C, circular with regular margin, initially white, with age becoming creamy white or slightly pinkish, highly floccose; reverse pinkish white. Colonies on PDA reaching 40–45 mm d after 4 d of incubation at 28 ± 0.5 °C, circular with regular margin, initially pale buff with creamy margin, with age becoming buff with paler margin and sectors, floccose; reverse buff with paler sectors. Additional materials examined. IRAN, Khuzestan Province; Karoon, isolated from a living leaf of A. lebbeck, Sep 2020, A. Safi (SCUA-Saf-B10-2). Notes: In the phylogenetic tree, C. albiziae is closely related to C. cucumericola, but can be distinguished by single nucleotide polymorphism (SNP) analysis of three ITS, tub2 and rpb2 loci (Fig. 2). This new species and the extype strain of C. cucumericola (CBS 378.71) showed 1 bp difference (0.2 %) across 496 nucleotides of the ITS region, 6 bp difference (1.1 %) across 547 nucleotides of the tub2 region, and 5 bp difference (1 %) across 481 nucleotides of the rpb2 region. Chaetomium cucumericola was established by Wang et al. (2016) to accommodate two sterile strains of the genus Chaetomium (CBS 378.71 and CBS 126777), which formed a well-supported clade within phylogenetic group III, including the related species C. olivaceum Cooke & Ellis, C. subglobosum Sergeeva, and C. undulatulum Asgari & Zare. C. albiziae can be distinguished from these three species by smaller ascomata (C. olivaceum: 260–440 × 200–360 μm, C. subglobosum: 300–450 × 265–355 μm, C. undulatulum: 230–280 × 185–250 μm) (Asgari & Zare 2011, Wang et al. 2016b)., Published as part of Mehrabi-Koushki, Mehdi & Safi, Atena, 2023, Chaetomium albiziae, a new endophytic species from Albizia lebbeck in Iran, pp. 137-146 in Phytotaxa 591 (2) on pages 140-142, DOI: 10.11646/phytotaxa.591.2.5, http://zenodo.org/record/7797484, {"references":["Asgari, B. & Zare, R. (2011) The genus Chaetomium in Iran, a phylogenetic study including six new species. Mycologia 103: 863 - 882. https: // doi. org / 10.3852 / 10 - 349","Wang, X. W., Lombard, L., Groenewald, J. Z., Li, J., Videira, S. I. R., Samson, R. A., Liu, X. Z. & Crous, P. W. (2016 b) Phylogenetic reassessment of the Chaetomium globosum species complex. Persoonia 36: 83 - 133. https: // doi. org / 10.3767 / 003158516 X 689657"]}

Sordaria Ces. & De Not., Commentario della Società Crittogamologica Italiana 1 (4): 225 (1863) Index Fungorum number: IF 5061 Type species: Sordaria fimicola (Roberge ex Desm.) Ces. & De Not., Commentario della Società Crittogamologica Italiana 1 (4): 226 (1863) Notes: Sordaria was introduced with the type species S. fimicola, and 264 records are listed in Index Fungorum (2023), but only 23 species have available sequence data (Huang et al. 2021). The sexual morph of Sordaria was characterized by having sub-immersed to superficial, perithecioid, coriaceous ascomata, contain with cylindrical paraphyses, oblong, upright to slightly curved asci, with a lobate pedicel and distinct apical ring, with eight-spored, uniseriate, ellipsoidal to ovoid, verruculose ascospore, immature ascospore enclosed in a hyaline gelatinous sheath, however, sometimes thick and conspicuous to even difficult to detect (Ivanová et al. 2015; Phukhamsakda et al. 2020). Species of Sordaria are commonly found in dung, soil, and seed pods (Furtado 1969; Watanabe 1989; Mungai et al. 2012). Notably, the filamentous ascomycete Sordaria macrospora was considered a model organism to study the molecular mechanisms that regulate fruiting bodies’ development (Teichert et al. 2020).

Boothiella Lodhi & Mirza, Mycologia 54: 217 (1962) Index Fungorum number: IF 627 Type species: Boothiella tetraspora Lodhi & Mirza, Mycologia 54: 217 (1962) Notes: Boothiella was a monotypic genus and established by the type species B. tetraspora, which was isolated from soil in Lahore City, Pakistan. Boothiella is similar to Thielavia but differs by having a colourless ascomatal wall (Lodhi & Mirza 1962). Eriksson et al. (2004) and Kirk et al. (2008) proposed Boothiella should be accommodated in the family Sordariaceae, and this phylogenetic placement was confirmed by other researchers later (Vu et al. 2019, Wang et al. 2019). The sexual morph was described by having a superficial to immersed, globose to subglobose, cleistothecial ascomata, solitary to aggregated, non-ostiolate, hyaline ascomatal walled, clavate to cylindrical asci with four-spored, short pedicellate, uniseriate, evanescent. Ascospore ellipsoidal to broad ovoid, hyaline to yellow to olivaceous brown with the maturity, one-celled, with apical germ pores, however, no asexual morphs have been reported (Huang et al. 2021). Thielaviella humicola and Thielavia tetraspora were recently re-identified as Boothiella tetraspora based on evidences of morphology and phylogeny (Wang et al. 2019). This genus has a single species (B. tetraspora), and most collections were isolated from soil (Wang et al. 2019)., Published as part of Yang, Erfu, Lu, Wenhua, Tibpromma, Saowaluck, Dai, Dongqin, Gao, Ying, Promputtha, Itthayakorn & Karunarathna, Samantha C., 2023, Three interesting fungi from American bullfrog larvae (Rana catesbeiana) in Yunnan, China, pp. 251-268 in Phytotaxa 587 (3) on page 257, DOI: 10.11646/phytotaxa.587.3.4, http://zenodo.org/record/7744408, {"references":["Lodhi, S. A. & Mirza, R. F. (1962) A new genus of the Eurotiales. Mycologia 54: 217 - 219. https: // doi. org / 10.1080 / 00275514.1962.12024993","Eriksson, O. E., Baral, H. O., Currah, R. S., Kurtzman, C. P., Rambold, G. & Laessoe, T. (2004) Outline of Ascomycota - 2004. Myconet 10: 1 - 99.","Kirk, P. M., Cannon, P. F., Minter, D. W. & Stalpers, J. A. (2008) Ainsworth & Bisby's Dictionary of the Fungi, 10 th ed. CABI, UK. pp. 1 - 771. https: // doi. org / 10.1079 / 9780851998268.0000","Vu, D., Groenewald, M., de Vries, M. D., Gehrmann, T., Stielow, B., Eberhardt, U., Al-Hatmi, A., Groenewald, J. Z., Cardinali, J. & Houbraken, J. (2019) Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation. Studies in Mycology 92: 135 - 154. https: // doi. org / 10.1016 / j. simyco. 2018.05.001","Wang, X. W., Bai, F. Y., Bensch, K., Meijer, M., Sun, B. D., Han, Y. F., Crous, P. W., Samson, R. A., Yang, F. Y. & Houbraken, J. (2019) Phylogenetic re-evaluation of Thielavia with the introduction of a new family Podosporaceae. Studies in Mycology 93: 155 - 252. https: // doi. org / 10.1016 / j. simyco. 2019.08.002","Huang, S. K., Hyde, K. D., Mapook, A., Maharachchikumbura, S. S. N., Bhat, J. D., McKenzie, E. H. C., Jeewon, R. & Wen, T. C. (2021) Taxonomic studies of some often over-looked Diaporthomycetidae and Sordariomycetidae. Fungal Diversity 111: 443 - 572. https: // doi. org / 10.1007 / s 13225 - 021 - 00488 - 4"]}

Boothiella tetraspora Lodhi & J.H. Mirza, Mycologia 54(2): 217 (1962) (Figure 4) Index Fungorum number: IF 327083 Isolated from intestinal contents of dead American bullfrog larvae. Sexual morph on PDA: Mycelium 3–5 μm wide, hyaline to yellow pale, thick-walled, branched, septate, with granules. Ascomata 260–370 × 240–330 μm (x̅ =245 × 285 μm, n=10), globose to subglobose, solitary or gregarious, superficial to immersed on PDA, brown to dark brown or black, membranaceous, glabrous, without ostiolate. Ascomatal wall composed of membranaceous, subhyaline to pale brown cells of textura angularis. Hamathecium not observed. Asci 80–100 × 15–20 μm (x̅ =87.5 × 15.5 μm, n=20), unitunicate, four-spored, clavate to broadly cylindrical, apically round, short pedicellate, furcate pedicel, evanescent. Ascospores 20–30 × 10–20 μm (x̅ = 24 × 14.5 μm, n=20), ovoid to ellipsoidal, hyaline to yellowish brown to dark brown, aseptate, rough to verruculose, sometimes visible a germ pore at one or each end, without sheath. Asexual morph: Undetermined. Culture characteristics: Colonies growing on PDA reach 30–40 mm in diameter after one week at 27 °C, forming the brown to dark brown fruiting bodies within one month in PDA. Obverse: flat, velvety, hairy, pale brown, entire edge. Reverse: yellowish brown. Without pigments produced in PDA. Known substratum: Soil (Lodhi & Mirza 1962, Wang et al. 2019); Sand (Wang et al. 2019); Intestinal contents of dead American bullfrog larvae (this study). Known Distribution: Pakistan (Lodhi & Mirza 1962); Spain, India (Wang et al. 2019), China (This study). Material examined: China, Yunnan Province, Qujing Normal University, intestinal contents of dead American bullfrog larvae, GPS: 103°44’35”E, 25°30’46”N, 1856.6 m, Wen-hua Lu, ER 4, (Herb. HKAS 125767), living culture KUNCC22-12510. Notes: Based on detailed descriptions of Boothiella tetraspora, our isolate KUNCC 22-12510 fits with previous strains in morphology, forming brown to dark brown ascomata, with globose to subglobose, superficial to immersed, non-ostiolate, additionally, both strains almost have the same size of asci and ascospores, with four-spored asci, ovoid to ellipsoidal conidia (Lodhi & Mirza 1962, Wang et al. 2019). Phylogenetically, our isolate KUNCC 22-12510 clustered with Boothiella tetraspora (CBS 334.67 and CBS 887.97) with reliable statistical supports (99% ML /1 BYPP, Figure 2). The BLASTn results of ITS, LSU, tub2, and rpb2 region demonstrate 99–100% similarity with Boothiella tetraspora strains (CBS 334.67 and CBS 887.97). Therefore, our isolate KUNCC 22-12510 is identified as B. tetraspora, a new host and country record based on morphological features combined with phylogeny evidence., Published as part of Yang, Erfu, Lu, Wenhua, Tibpromma, Saowaluck, Dai, Dongqin, Gao, Ying, Promputtha, Itthayakorn & Karunarathna, Samantha C., 2023, Three interesting fungi from American bullfrog larvae (Rana catesbeiana) in Yunnan, China, pp. 251-268 in Phytotaxa 587 (3) on pages 258-259, DOI: 10.11646/phytotaxa.587.3.4, http://zenodo.org/record/7744408, {"references":["Lodhi, S. A. & Mirza, R. F. (1962) A new genus of the Eurotiales. Mycologia 54: 217 - 219. https: // doi. org / 10.1080 / 00275514.1962.12024993","Wang, X. W., Bai, F. Y., Bensch, K., Meijer, M., Sun, B. D., Han, Y. F., Crous, P. W., Samson, R. A., Yang, F. Y. & Houbraken, J. (2019) Phylogenetic re-evaluation of Thielavia with the introduction of a new family Podosporaceae. Studies in Mycology 93: 155 - 252. https: // doi. org / 10.1016 / j. simyco. 2019.08.002"]}

Sordaria macrospora Auersw., Fungi Europaei exsiccati, Klotzschii herbarii vivi mycologici continuatio. Editio nov. Series secunda. Cent. 10: no. 954 (1866) (Figure 5) Index Fungorum number: IF 237763 Isolated from intestinal contents of dead American bullfrog larvae. Sexual morph on PDA: Ascomata 340–420 × 270–380 μm (x̅ = 330 × 380 μm, n=20), semi-immersed to superficial, perithecial, solitary or gregarious, ampulliform or pyriform, globose, coriaceous, brown to black, ostiolate. Neck 140–180 × 85–120 μm (x̅ = 160 × 99 μm, n=10) central, oblong, straight to bent, extended, blunt ends. Peridium consist of dark-brown, thick-walled cells of textura angularis. Hamathecium 6–15 wide (x̅ = 9.5 μm, n=20), numerous, wide, branched, septate, markedly constricted at the septa, hyaline, paraphyses. Asci 130–180 × 17–22 μm (̄x̅ = 155×19 μm, n=20), unitunicate, eight-spored, slenderly cylindrical, with a lobate pedicel and a prominent J- ring. Ascospores 20–30 × 15–20 μm (x̅ =25.5 × 15.5 μm, n=20), one-celled, ellipsoidal to ovoid, uniseriate, hyaline to golden to brown to black when mature, guttulate, with some prominent oil droplets, with a thin gelatinous sheath, acute to round ends, with a basal germ pore. Asexual morph: Undetermined. Culture characteristics: Colonies growing on PDA reach around 60 mm in diameter after one week at 27 °C, forming dark brown fruiting bodies within one month in PDA. Obverse: flat, white, entire edge, peripheral fertile. Reverse: pale brown. Without pigments produced in PDA. Known substratum: Pinus sylvestris, Fagus sylvatica (Petrini & Fisher 1988); Salix fragilis, Quercus robur (Petrini & Fisher 1990); Olea europaea (Fisher et al. 1992); Eucalyptus nitens (Fisher et al. 1993); Dactylis glomerata (Sanchez Marquez et al. 2007); Herbivore dung (Lytvynenko & Hayova 2018); Intestinal contents of dead American bullfrog larvae (this study). Known Distribution: England (Petrini & Fisher 1998,1990); Balearic Islands (Fisher et al. 1992); Australia (Fisher et al. 1993); Spain (Sanchez Marquez et al. 2007); Ukraine (Lytvynenko & Hayova 2018); China (This study). Material examined: China, Yunnan Province, Qujing Normal University, on intestinal contents of dead American bullfrog larvae, GPS: 103°44’35”E, 25°30’46”N, 1856.6 m, Wen-hua Lu, ER3, (Herb. HKAS 125766), living culture 22-12509. Notes: Sordaria macrospora is a coprophilic homothallic pyrenomycete, a fungal model organism in biology that was first described in 1866 by Auerswald. Our isolate fits the concept of S. macrospora by having elongated neck, oblong asci with a “J-” apical ring, enwrap eight hyaline to golden to dark brown ascospores, ascospores uniseriate, visible distinct oil droplets when its young, present verruculose surface when it is mature, without gelatinous sheath (Lord & Read 2011). The BLASTn results of ITS, and LSU showed our isolate (KUNCC 22-12509) are highly similar to Sordaria species (Sordaria macrospora CBS:346.62, S. humana CBS:416.82, S. fimicola CBS:566.67, S. lappae CBS:154.97, S. tamaensis and S. nodulifera NBRC:32551), with 99–100% similarity. However, the tub2 gene is mostly similar to Sordaria macrospora (FGSC 4818) and S. superba (CBS 784.96) with 99% similarity, the tub2 gene of other species is not available (Table 1). From morphology, Sordaria superba is different, as the ascospores are wrapped by a distinct gelatinous sheath with a single basal germ pore (Yul et al. 2010), our isolate is more similar to S. macrospora (ascomata: 370–400 (500) × 250–300 µm VS. 340–420 × 270–380 μm; and asci: 160–175 ×20 VS. 20–30 × 15–20 μm) (Ivanová et al. 2015). The phylogenetic tree indicated that our Sordaria macrospora KUNCC 22-12509 is complex with the other species that have high similarity in ITS, and LSU genes (Figure 2), and this problem may be caused by lacking tub2 genes, therefore, our isolate KUNCC 22-12509 is identified based on morphological features (Table 2) coupled with BLASTn results and phylogenetic analyses as Sordaria macrospora which is a new host and country record., Published as part of Yang, Erfu, Lu, Wenhua, Tibpromma, Saowaluck, Dai, Dongqin, Gao, Ying, Promputtha, Itthayakorn & Karunarathna, Samantha C., 2023, Three interesting fungi from American bullfrog larvae (Rana catesbeiana) in Yunnan, China, pp. 251-268 in Phytotaxa 587 (3) on pages 259-260, DOI: 10.11646/phytotaxa.587.3.4, http://zenodo.org/record/7744408, {"references":["Petrini, O. & Fisher, P. J. (1988) A comparative study of fungal endophytes in xylem and whole stem of Pinus sylvestris and Fagus sylvatica. Transactions of the British Mycological Society 91: 233 - 238. https: // doi. org / 10.1016 / S 0007 - 1536 (88) 80210 - 9","Petrini, O. & Fisher, P. J. (1990) Occurrence of fungal endophytes in twigs of Salix fragilis and Quercus robur. Mycology Research 94: 1077 - 1080. https: // doi. org / 10.1016 / S 0953 - 7562 (09) 81336 - 1","Fisher, P. J., Petrini, O., Petrini, L. E. & Descals, E. (1992) A preliminary study of fungi inhabiting xylem and whole stems of Olea europaea. Sydowia 44: 117 - 121.","Fisher, P. J., Petrini, O. & Sutton, B. C. (1993) A comparative study of fungal endophytes in leaves, xylem and bark of Eucalyptus in Australia and England. Sydowia 45: 338 - 345.","Sanchez Marquez, S., Bills, G. F. & Zabalgogeazcoa, I. (2007) The endophytic mycobiota of the grass Dactylis glomerata. Fungal Diversity 27: 171 - 195.","Lytvynenko, Y. I. & Hayova, V. P. (2018) New and noteworthy records of coprophilous species of Coniochaeta and Sordaria (Sordariomycetes, Ascomycota) from Ukraine. Ukrainian Botanical Journal 75: 538 - 551. https: // doi. org / 10.15407 / ukrbotj 75.06.538","Lord, K. M. & Read, N. D. (2011) Perithecium morphogenesis in Sordaria macrospora. Fungal Genetics and Biology 48: 388 - 399. https: // doi. org / 10.1016 / j. fgb. 2010.11.009","Yul, G., Mikos, I. G. & OYu, A. (2010) New records of coprophilous ascomycetes in the Crimea. Chornomorski Botanical Journal 6: 67 - 83. https: // doi. org / 10.14255 / 2308 - 9628 / 10.61 / 6","Ivanova, H., Pristas, P. & Ondruskova, E. M. I. L. I. A. (2015) Comparison of two Coniochaeta species (C. ligniaria and C. malacotricha) with a new pathogen of black pine needles - S ordaria macrospora. Plant Protection Science 52: 18 - 25. https: // doi. org / 10.17221 / 45 / 2014 - PPS"]}

Sordaria Ces. & De Not., Commentario della Società Crittogamologica Italiana 1 (4): 225 (1863) Index Fungorum number: IF 5061 Type species: Sordaria fimicola (Roberge ex Desm.) Ces. & De Not., Commentario della Società Crittogamologica Italiana 1 (4): 226 (1863) Notes: Sordaria was introduced with the type species S. fimicola, and 264 records are listed in Index Fungorum (2023), but only 23 species have available sequence data (Huang et al. 2021). The sexual morph of Sordaria was characterized by having sub-immersed to superficial, perithecioid, coriaceous ascomata, contain with cylindrical paraphyses, oblong, upright to slightly curved asci, with a lobate pedicel and distinct apical ring, with eight-spored, uniseriate, ellipsoidal to ovoid, verruculose ascospore, immature ascospore enclosed in a hyaline gelatinous sheath, however, sometimes thick and conspicuous to even difficult to detect (Ivanová et al. 2015; Phukhamsakda et al. 2020). Species of Sordaria are commonly found in dung, soil, and seed pods (Furtado 1969; Watanabe 1989; Mungai et al. 2012). Notably, the filamentous ascomycete Sordaria macrospora was considered a model organism to study the molecular mechanisms that regulate fruiting bodies’ development (Teichert et al. 2020)., Published as part of Yang, Erfu, Lu, Wenhua, Tibpromma, Saowaluck, Dai, Dongqin, Gao, Ying, Promputtha, Itthayakorn & Karunarathna, Samantha C., 2023, Three interesting fungi from American bullfrog larvae (Rana catesbeiana) in Yunnan, China, pp. 251-268 in Phytotaxa 587 (3) on page 259, DOI: 10.11646/phytotaxa.587.3.4, http://zenodo.org/record/7744408, {"references":["Huang, S. K., Hyde, K. D., Mapook, A., Maharachchikumbura, S. S. N., Bhat, J. D., McKenzie, E. H. C., Jeewon, R. & Wen, T. C. (2021) Taxonomic studies of some often over-looked Diaporthomycetidae and Sordariomycetidae. Fungal Diversity 111: 443 - 572. https: // doi. org / 10.1007 / s 13225 - 021 - 00488 - 4","Ivanova, H., Pristas, P. & Ondruskova, E. M. I. L. I. A. (2015) Comparison of two Coniochaeta species (C. ligniaria and C. malacotricha) with a new pathogen of black pine needles - S ordaria macrospora. Plant Protection Science 52: 18 - 25. https: // doi. org / 10.17221 / 45 / 2014 - PPS","Phukhamsakda, C., McKenzie, E. H. C., Phillips, A. J. L., Gareth Jones, E. B., Jayarama Bhat, D., Marc, S., Bhunjun, C. S., Wanasinghe, D. N., Thongbai, B., Camporesi, E., Ertz, D., Jayawardena, R. S., Perera, R. H., Ekanayake, A. H., Tibpromma, S., Doilom, M., Xu, J. C. & Hyde, K. D. (2020) Microfungi associated with Clematis (Ranunculaceae) with an integrated approach to delimiting species boundaries. Fungal Diversity 102: 1 - 203. https: // doi. org / 10.1007 / s 13225 - 020 - 00448 - 4","Furtado, J. S. (1969) Ascal cytology of Sordaria sclerogenia. Protoplasma 67: 473 - 478. https: // doi. org / 10.1007 / BF 01254908","Watanabe, T. (1989) Three species of Sordaria, and Eudarluca biconica from cherry seeds. Transactions of the British Mycological Society 30: 395 - 400.","Mungai, P. G., Chukeatirote, E., Njogu, J. G. & Hyde, K. D. (2012) Studies of coprophilous ascomycetes in Kenya: Sordariales from wildlife dung. Mycosphere 3: 437 - 448. https: // doi. org / 10.5943 / mycosphere / 3 / 4 / 7","Teichert, I., Poggeler, S., Nowrousian, M. (2020) Sordaria macrospora: 25 years as a model organism for studying the molecular mechanisms of fruiting body development. Applied Microbiology and Biotechnology 104: 3691 - 3704. https: // doi. org / 10.1007 / s 00253 - 020 - 10504 - 3"]}

American bullfrog (Rana catesbeiana) is an alien invasive species in southwest China native to the central and eastern United States and southeastern Canada. After the 19th century, they extensively appear in aquaculture and natural environments worldwide as a delicious food but also creating a serious threat to the survival and development of native species. In the early rainy season, dead American bullfrog larvae floating on the water of unnamed ponds in Qujing Normal University, Yunnan Province, China were collected and brought to the mycology laboratory, and three interesting fungal strains were isolated from their intestinal contents. Phylogenetic analyses were carried out on the resultant isolates based on multiple gene sequences (ITS, LSU, rpb2, tub2, tef1-α), and results confirmed that the three strains belong to three species, namely; Boothiella tetraspora, Sordaria macrospora and Trichoderma virens. The morphological characteristics were also used to describe the fungal taxa. Photographic plates, descriptions, and phylogenetic trees that show the placements of the fungal species are reported herein.

Cytochalasans (CYTs), as well as their polycyclic (pcCYTs) and polymerized (meCYTs) derivatives, constitute one of the largest families of fungal polyketide-nonribosomal peptide (PK-NRP) hybrid natural products. However, the mechanism of chemical conversion from mono-CYTs (moCYTs) to both pcCYTs and meCYTs remains unknown. Here, we show the first successful example of the reconstitution of the CYT core backbone as well as the whole pathway in a heterologous host. Importantly, we also describe the berberine bridge enzyme (BBE)-like oxidase AspoA, which uses Glu538 as a general acid biocatalyst to catalyse an unusual protonation-driven double bond isomerization reaction and acts as a switch to alter the native (for moCYTs) and nonenzymatic (for pcCYTs and meCYTs) pathways to synthesize aspochalasin family compounds. Our results present an unprecedented function of BBE-like enzymes and highly suggest that the isolated pcCYTs and meCYTs are most likely artificially derived products., Cytochalasans are a large family of fungal polyketide-nonribosomal peptide hybrid natural products that exhibit important pharmaceutical activities, but the mechanism of conversion of the monocytochalasans to the polycyclic, fused analogues is unclear. Here the authors reconstitute the core backbone of the cytochalasin family and describe an oxidase that catalyzes an unusual double-bond isomerization reaction.

Objectives This study aimed to evaluate endophytic fungi isolated from Tocoyena bullata and Humiria balsamifera plant species for their antimycobacterial and anti-inflammatory activities, focusing on severe pulmonary tuberculosis cases which are often associated with exacerbated inflammation. Methods Mycobacterium suspensions were incubated with the samples for 5 days. RAW 264.7 macrophages stimulated with LPS were also incubated with them for 24 h to assess the inhibition of inflammatory mediator production and cytotoxicity. C57BL/6 mice were infected with Mtb M299 and treated for 15 days with lasiodiplodin (Lasio). Key findings Endophytic fungus Sordaria tamaensis, obtained from T. bullata, was the most promising. Its ethanolic extract impaired mycobacterial growth with MIC50 (µg/ml): 1.5 ± 0.6 (BCG), 66.8 ± 0.1 (H37Rv) and 80.0 ± 0.1 (M299). (R)-(+)-Lasio showed MIC50 92.2 ± 1.8 µg/ml (M299). In addition, Lasio was able to inhibit NO, IL-1β and TNF-α production and was not cytotoxic for macrophages. M. tuberculosis-infected C57BL/6 animals treated by Lasio reduced the number of acid-fast bacilli, lung pathology, leucocyte influx and proinflammatory cytokine production in the lungs. The class IIa fructose 1,6-bisphosphate aldolase was the predicted hypothetical target of Lasio. Conclusions (R)-(+)-Lasio stood out as a promising anti-TB compound, exhibiting anti-inflammatory and antimycobacterial effects, as well as low cytotoxicity.

The thermophilic fungus

Sporothrix (Sordariales, Ascomycota) is a well-supported monophyletic lineage within the Ophiostomatales, species of which occur in a diverse range of habitats including on forest trees, in the soil, associated with bark beetles and mites as well as on the fruiting bodies of some Basidiomycota. Several species have also been reported as important human and animal pathogens. During surveys of insect- and wound-associated Ophiostomatales from hardwood trees in Poland, many isolates with affinity to Sporothrix were recovered. In the present study, six undescribed Sporothrix spp. collected during these surveys are characterized based on their morphological characteristics and multi-locus phylogenenetic inference. They are described as Sporothrix cavum, Sporothrix cracoviensis, S. cryptarchum, S. fraxini, S. resoviensis, and S. undulata. Two of the Sporothrix spp. reside in the S. gossypina-complex, while one forms part of the S. stenoceras-complex. One Sporothrix sp. is a member of lineage F, and two other species grouped outside any of the currently defined species complexes. All the newly described species were recovered from hardwood habitats in association with sub-cortical insects, wounds or woodpecker cavities. These species were morphologically similar, with predominantly asexual states having hyaline or lightly pigmented conidia, which produce holoblastically on denticulate conidiogenous cells. Five of the new taxa produce ascomata with necks terminating in long ostiolar hyphae and allantoid ascospores without sheaths. The results suggest that Sporothrix species are common members of the Ophiostomatales in hardwood ecosystems of Poland.

β-Galactosidases are key enzymes in the food industry. Apart from the hydrolysis of the saccharide bond of lactose, they also catalyze transgalactosylation reactions, producing galactooligosaccharides (GOS) with prebiotic activity. Here we report the heterologous production in Pichia pastoris of a novel β-galactosidase from the fungus Thermothielavioides terrestris. The enzyme (TtbGal1) was purified and characterized, showing optimal activity at 60 °C and pH 4. TtbGal1 is thermostable, retaining almost full activity for 24 h at 50 °C. It was applied to the production of GOS from defined lactose solutions and acid whey, a liquid waste from the Greek yoghurt industry, reaching yields of 19.4 % and 14.8 %, respectively. HILIC-ESI-QTOF-MS analysis revealed the production of GOS with up to 4 saccharide monomers. The results demonstrate efficient GOS production catalyzed by TtbGal1, valorizing acid whey, a waste with a heavy polluting load from the dairy industry.

Fungi are eukaryotic organisms that have the ability to synthesize an enormous number of metabolites with a vast diversity of chemical structures and bioactivities. Chaetomiaceae species are a wealthy source of enzymes with diverse biotechnological and industrial applications such as PMO (polysaccharide monooxygenase), L-methioninase, β-1,3-glucanase, laccase, dextranase, lipolytic, pectinolytic, amylolytic, chitinolytic, and proteolytic enzymes. Different classes of secondary metabolites have been reported from this family are derived from various biosynthetic pathways such as alkaloids, polyketides, peptides, terpenes, and polyketide-amino acid hybrid secondary metabolites. These metabolites have attracted research interest due to their fascinating structural frameworks and bioactivities. Therefore, these compounds can be taken into account as candidates for the development of effective and novel lead compounds for medicine, as well as for plant protection. The current review represents the relevant information for Chaetomiaceae species, in particular, its new and first reported secondary metabolites and their pharmacological activities, as well as the biotechnological applications published from 2016 until the beginning of 2021.

Background: Molecular phylogenetic algorithms frequently disagree with the approaches considering reproductive compatibility and morphological criteria for species delimitation. The question stems if the resulting species boundaries from molecular, reproductive and/or morphological data are definitively not reconcilable; or if the existing phylogenetic methods are not sensitive enough to agree morphological and genetic variation in species delimitation. Objectives : We propose DISTATIS as an integrative framework to combine alignment-based (AB) and alignment-free (AF) distance matrices from ITS2 sequences/structures to shed light whether Gelasinospora and Neurospora are sister but independent genera. Methods: We aimed at addressing this standing issue by harmonizing genus-specific classification based on their ascospore morphology and ITS2 molecular data. To validate our proposal, three phylogenetic approaches: i) traditional alignment-based, ii) alignment-free and iii) novel distance integrative (DI)-based were comparatively evaluated on a set of Gelasinospora and Neurospora species. All considered species have been extensively characterized at both the morphological and reproductive levels and there are known incongruences between their ascospore morphology and molecular data that hampers genus-specific delimitation. Results: Traditional AB phylogenetic analyses fail at resolving the Gelasinospora and Neurospora genera into independent monophyletic clades following ascospore morphology criteria. In contrast, AF and DI approaches produced phylogenetic trees that could properly delimit the expected monophyletic clades. Conclusions: The DI approach outperformed the AF one in the sense that it could also divide the Neurospora species according to their reproduction mode.

Xyloglucan is closely associated with cellulose and still retained with some modification in pretreated lignocellulose; however, its influence on lignocellulose biodegradation is less understood.

The present investigation primarily focusses on enhancement of resveratrol production by endophytic production from the endophytic fungus, Arcopilus aureus via one variable at a time approach (OVAT) followed by statistical approach using response surface methodology (RSM). The paper also highlights the characterization of fungal resveratrol using spectroscopic techniques. Further the tyrosinase inhibitory property was also explored in this communication for its possible use as a cosmeceutical ingredient. Optimization of physicochemical and nutritional parameters using OVAT approach exhibited 1.23-fold enhancement in production of resveratrol when compared to initial yield, 89.1 ± 0.08 µg/mL. Further RSM resulted in 1.49-fold enhancement in resveratrol production i.e. 133.53 µg/ml. Further, 25 mg of fungal resveratrol in pure form was obtained from the spent broth of Arcopilus aureus by column chromatography using a mobile phase comprising of MeOH: DCM in a ratio of 1.75:98.25. Further its purity on TLC was checked using 5% MeOH: DCM as mobile phase. Symmetrical peak with Rt of 3.36 min using a C18 reverse phase column confirmed the homogeneity of the purified fungal resveratrol with standard resveratrol and further corroborated by 1H-NMR, 13C-NMR and HR-MS analysis. Fungal resveratrol exhibited a good tyrosinase inhibition with an IC50 of 2.654 ± 0.432 µg/mL as compared to Kojic acid (1.329 ± 0.333). The present study has provided sufficient lead that process optimization techniques can complement each other for optimized production of bioactive compounds by microorganisms apart from strain improvement techniques which are generally adopted in the industry. The enhancement of resveratrol production by Arcopilus aureus by process optimization further opens up avenues for strain improvement for commercial resveratrol production through fermentation for nutraceutical and cosmeceutical applications.

The filamentous fungus Podospora anserina has been used as a model organism for more than 100 years and has proved to be an invaluable resource in numerous areas of research. Throughout this period, P. anserina has been embroiled in a number of taxonomic controversies regarding the proper name under which it should be called. The most recent taxonomic treatment proposed to change the name of this important species to Triangularia anserina. The results of past name changes of this species indicate that the broader research community is unlikely to accept this change, which will lead to nomenclatural instability and confusion in literature. Here, we review the phylogeny of the species closely related to P. anserina and provide evidence that currently available marker information is insufficient to resolve the relationships amongst many of the lineages. We argue that it is not only premature to propose a new name for P. anserina based on current data, but also that every effort should be made to retain P. anserina as the current name to ensure stability and to minimise confusion in scientific literature. Therefore, we synonymise Triangularia with Podospora and suggest that either the type species of Podospora be moved to P. anserina from P. fimiseda or that all species within the Podosporaceae be placed in the genus Podospora.

Abstract Arabinofuranose substitutions on xylan are known to interfere with enzymatic hydrolysis of this primary hemicellulose. In this work, two novel α-l-arabinofuranosidases (ABFs), TtABF51A from Thielavia terrestris and EpABF62C from Eupenicillium parvum, were characterized and functionally analyzed. From sequences analyses, TtABF51A and EpABF62C belong to glycoside hydrolase (GH) families 51 and 62, respectively. Recombinant TtABF51A showed high activity on 4-nitrophenyl-α-l-arabinofuranoside (83.39 U/mg), low-viscosity wheat arabinoxylan (WAX, 39.66 U/mg), high-viscosity rye arabinoxylan (RAX, 32.24 U/mg), and sugarbeet arabinan (25.69 U/mg), while EpABF62C preferred to degrade arabinoxylan. For EpABF62C, the rate of hydrolysis of RAX (94.10 U/mg) was 2.1 times that of WAX (45.46 U/mg). The optimal pH and reaction temperature for the two enzymes was between 4.0 and 4.5 and 65 °C, respectively. Calcium played an important role in the thermal stability of EpABF62C. TtABF51A and EpABF62C showed the highest thermal stabilities at pH 4.5 or 5.0, respectively. At their optimal pHs, TtABF51A and EpABF62C retained greater than 80% of their initial activities after incubation at 55 °C for 96 h or 144 h, respectively. 1H NMR analysis indicated that the two enzymes selectively removed arabinose linked to C-3 of mono-substituted xylose residues in WAX. Compared with the singular application of the GH10 xylanase EpXYN1 from E. parvum, co-digestions of WAX including TtABF51A and/or EpABF62C released 2.49, 3.38, and 4.81 times xylose or 3.38, 1.65, and 2.57 times of xylobiose, respectively. Meanwhile, the amount of arabinose released from WAX by TtABF51A with EpXYN1 was 2.11 times the amount with TtABF51A alone. Key points • Two novel α-l-arabinofuranosidases (ABFs) displayed high thermal stability. • The thermal stability of GH62 family EpABF62C was dependent on calcium. • Buffer pH affects the thermal stability of the two ABFs. • Both ABFs enhance the hydrolysis of WAX by a GH10 xylanase.

به منظور شناسایی قارچ‌های متحمل به گرما و گرمادوست، طی سال‌های ۹8-1396، نمونه‌برداری از خاک، کمپوست ضایعات شهری و کمپوست قارچ خوراکی در شهرستان‌های استان کرمانشاه (ماهیدشت، هرسین، کرند، اسلام‌آباد، قصر شیرین، سرپل‌ذهاب، جوانرود و گیلانغرب) انجام شد. جداسازی این قارچ‌ها از خاک و کمپوست به روش سری رقت روی محیط‌کشت عصاره سیب‌زمینی-دکستروز-آگار در دمای 45 و 50 درجه سلسیوس انجام شد. در مجموع، ۲۴ جدایه قارچ متحمل به گرما و گرمادوست به دست آمد. شناسایی اولیه جدایه‌ها براساس ویژگی‌های رشدی و ریخت‌شناسی و با استفاده از کلیدهای تاکسونومیکی معتبر صورت گرفت. برای شناسایی مولکولی، دی.ان.ای.، جدایه‌ها با استفاده از کیت خالص‌سازی دی.ان.ای. ژنومی استخراج و ناحیه ITS rDNA (ITS1-5.8S-ITS2) با استفاده از آغازگرهای عمومی ITS1و ITS4تکثیر شد. قطعات ۷۰۰–۵۰۰ جفت بازی به دست آمده از واکنش زنجیره‌ای پلیمراز، خالص‌سازی، توالی‌یابی و ویرایش شده و در بانک ژن ثبت شدند. با استفاده از ابزار جستجوی BLAST، توالی‌های ITS rDNA به دست آمده در این بررسی با سایر آرایه‌های معتبر مستخرج از بانک ژن مقایسه شد. در نهایت نه گونه شامل: Aspergilus fumigatus، A. nidulans، A. niger، A. terrus،Melanocarpus albomyces*، Malbranchea cinnamomea*، Thermomyces dupontii*، Th. lanuginosus* و Thielavia arenaria*شناسایی شد که گونه‌های ستاره‌دار برای فلور قارچی ایران جدید می‌باشند. فراوانی قارچ‌های گرمادوست در کمپوست ضایعات شهری بیش‌تر از خاک بود و گونه‌های آسپرژیلوس فراوان‌ترین قارچ‌های شناسایی شده در این تحقیق بودند.

The production of fuels and chemicals from renewable plant biomass has been proposed as a feasible strategy for global sustainable development. However, the economic efficiency of biorefineries is low. Here, through metabolic engineering, Myceliophthora thermophila, a cellulolytic thermophilic fungus, was constructed into a platform that can efficiently convert lignocellulose into important bulk chemicals—four carbon 1, 4-diacids (malic and succinic acid), building blocks for biopolymers—without the need for extra hydrolytic enzymes. Titers of >200 g/L from crystalline cellulose and 110 g/L from plant biomass (corncob) were achieved during fed-batch fermentation. Our study represents a milestone in consolidated bioprocessing technology and offers a new and promising system for the cost-effective production of chemicals and fuels from biomass.

Made available in DSpace on 2020-12-12T01:35:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Abstract: Arabinanases from glycoside hydrolase family GH93 are enzymes with exo-activity that hydrolyze the α-1,5 bonds between arabinose residues present on arabinan. Currently, several initiatives aiming to use byproducts rich in arabinan such as pectin and sugar beet pulp as raw material to produce various compounds of interest are being developed. However, it is necessary to use robust enzymes that have an optimal performance under pH and temperature conditions used in the industrial processes. In this work, the first GH93 from the thermophilic fungus Thermothielavioides terrestris (Abn93T) was heterologously expressed in Aspergillus nidulans, purified and biochemically characterized. The enzyme is a thermophilic glycoprotein (optimum activity at 70 °C) with prolonged stability in acid pHs (4.0 to 6.5). The presence of glycosylation affected slightly the hydrolytic capacity of the enzyme, which was further increased by 34% in the presence of 1 mM CoCl2. Small-angle X-ray scattering results show that Abn93T is a globular-like-shaped protein with a slight bulge at one end. The hydrolytic mechanism of the enzyme was elucidated using capillary zone electrophoresis and molecular docking calculations. Abn93T has an ability to produce (in synergism with arabinofuranosidases) arabinose and arabinobiose from sugar beet arabinan, which can be explored as fermentable sugars and prebiotics. Key points: • Thermophilic exo-arabinanase from family GH93 • Molecular basis of arabinan depolymerization. Department of Biotechnology Lorena School of Engineering University of São Paulo Departamento de Física e Ciências Aplicadas Instituto de Física de São Carlos Universidade de São Paulo Departamento de Bioquímica Instituto de Biologia Universidade Estadual de Campinas Programa de Processos Tecnológicos e Ambientais Universidade de Sorocaba Instituto de Pesquisas Tecnológicas do Estado de São Paulo Diretoria de Operações e Negócios Núcleo de Bionanomanufatura Department of Physics – Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Department of Physics – Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) FAPESP: 2014/06923-6 FAPESP: 2014/18714-2 FAPESP: 2017/00525-0 FAPESP: 2019/01165-0 CNPq: 2019/06663-8 FAPESP: 2019/06663-8 FAPESP: 2019/22284-7 CNPq: 302627/2018-9 CNPq: 443916/2014-4

Organismal interactions within microbial consortia and their responses to harmful intruders remain largely understudied. An important step toward the goal of understanding functional ecological interactions and their evolutionary selection is the study of increasingly complex microbial interaction systems. Here, we discovered a tripartite biosystem consisting of the fungus Aspergillus nidulans, the unicellular green alga Chlamydomonas reinhardtii, and the algicidal bacterium Streptomyces iranensis. Genetic analyses and MALDI-IMS demonstrate that the bacterium secretes the algicidal compound azalomycin F upon contact with C. reinhardtii. In co-culture, A. nidulans attracts the motile alga C. reinhardtii, which becomes embedded and surrounded by fungal mycelium and is shielded from the algicide. The filamentous fungus Sordaria macrospora was susceptible to azalomycin F and failed to protect C. reinhardtii despite chemotactically attracting the alga. Because S. macrospora was susceptible to azalomycin F, this data imply that for protection the fungus needs to be resistant. Formation of the lichen-like association between C. reinhardtii and A. nidulans increased algal growth. The protection depends on the increased amounts of membrane lipids provided by resistant fungi, thereby generating a protective shelter against the bacterial toxin. Our findings reveal a strategy whereby algae survive lethal environmental algicides through cooperation with fungi.

Fruiting bodies are among the most complex multicellular structures formed by fungi, and the molecular mechanisms that regulate their development are far from understood. However, studies with a number of fungal model organisms have started to shed light on this developmental process. One of these model organisms isSordaria macrospora, a filamentous ascomycete from the orderSordariales. This fungus has been a genetic model organism since the 1950s, but its career as a model organism for molecular genetics really took off in the 1990s, when the establishment of a transformation protocol, a mutant collection, and an indexed cosmid library provided the methods and resources to start revealing the molecular mechanisms of fruiting body development. In the 2000s, “omics” methods were added to theS. macrosporatool box, and by 2020, 58 developmental genes have been identified in this fungus. This review gives a brief overview of major method developments forS. macrospora, and then focuses on recent results characterizing different processes involved in regulating development including several regulatory protein complexes, autophagy, transcriptional and chromatin regulation, and RNA editing.Key points•Sordaria macrospora is a model system for analyzing fungal fruiting body development.•More than 100 developmental mutants are available for S. macrospora.•More than 50 developmental genes have been characterized in S. macrospora.

Trehalase catalyzes the hydrolysis of the non-reducing disaccharide trehalose. The highly active trehalase MthT from Myceliophthora thermophila was screened from the trehalase genes of six species of filamentous fungi. An ingenious multi-copy knock-in expression strategy mediated by the CRISPR/Cas9 tool and medium optimization were used to improve MthT production in Aspergillus niger, up to 1698.83 U/mL. The protein background was dramatically abated due to insertion. The recombinant MthT showed optimal activity at pH 5.5 and 60 °C, and exhibited prominent thermal stability between 50 and 60 °C under acid conditions (pH 4.5–6.5). The ethanol conversion rate (ethanol yield/total glucose) was significantly improved by addition of MthT (51.88%) compared with MthT absence (34.38%), using 30% starch saccharification liquid. The results of this study provided an effective strategy, established a convenient platform for heterologous expression in A. niger and showed a potential strategy to decrease production costs in industrial ethanol production.

The economical production of pectin oligosaccharides with a specific degree of polymerization and structure from agro-food waste is an industrially important process. This study identified a novel pectate lyase gene (plhy1) from the thermophilic cellulolytic fungus H. insolens Y1 and tested its ability to produce pectin oligosaccharides. The recombinant PLHY1 produced in Pichia pastoris was superior to other similar enzymes due to its high thermal and pH stability. PLHY1 demonstrated optimal enzymatic activity at 55°C and pH 10.0 in the presence of 0.4 mM Ca2+, and preferred methyl esterified substrates for digestion. High performance anion exchange chromatography-pulsed amperometric detector and ultra high performance liquid chromatography in combination with electrospray ionization tandem mass spectrometry analysis showed that galacturonic acid-oligosaccharides with a small degree of polymerization (4–6) were the major hydrolysates produced by the degradation of apple peel pectin by PLHY1. The properties of PLHY1 make it valuable for application in the agro-food industry for the production of pectin oligosaccharides.

Myceliophthora thermophila, a thermophilic fungus that can degrade and utilize all major polysaccharides in plant biomass, has great potential in biotechnological industries. Here, the first manually curated genome-scale metabolic model iDL1450 for M. thermophila was reconstructed using an autogenerating pipeline with thorough manual curation. The model contains 1450 genes, 2592 reactions, and 1784 unique metabolites. High accuracy was shown in predictions related to carbon and nitrogen source utilization based on data obtained from Biolog experiments. Besides, metabolism profiles were analyzed using iDL1450 integrated with transcriptomics data of M. thermophila at various growth temperatures. The refined model provides new insights into thermophilic fungi metabolism and sheds light on model-driven strain design to improve biotechnological applications of this thermophilic lignocellulosic fungus.

Lytic polysaccharide monooxygenases (LPMOs) play a key role in enzymatic conversion of plant cell wall polysaccharides. Continuous discovery and functional characterization of LPMOs highly contribute to the tailor-made design and improvement of hydrolytic-activity based enzyme cocktails. In this context, a new MtLPMO9F was characterized for its substrate (xyloglucan) specificity, and MtLPMO9H was further delineated. Aided by sodium borodeuteride reduction and hydrophilic interaction chromatography coupled to mass spectrometric analysis, we found that both MtLPMOs released predominately C4-oxidized, and C4/C6-double oxidized xylogluco-oligosaccharides. Further characterization showed that MtLPMO9F, having a short active site segment 1 and a long active site segment 2 (−Seg1+Seg2), followed a “substitution-intolerant” xyloglucan cleavage profile, while for MtLPMO9H (+Seg1−Seg2) a “substitution-tolerant” profile was found. The here characterized xyloglucan specificity and substitution (in)tolerance of MtLPMO9F and MtLPMO9H were as predicted according to our previously published phylogenetic grouping of AA9 LPMOs based on structural active site segment configurations.

Four new depsidones, mollicellins V-Y (

The purpose of this study was to report the first case of keratitis caused by Cladorrhinum samala and review of the literature.This was a case report and literature review.A 35-year-old immunocompetent man presented with pain, redness, and watering in the right eye 7 days after trauma with some foreign body. He was diagnosed with infectious keratitis, and a thorough microbiological workup was performed. Corneal scrapings were subjected to a potassium hydroxide (KOH) examination, Gram staining, bacterial (blood agar and Robertson cooked meat broth), and fungal culture (Sabouraud dextrose agar and brain-heart infusion agar). The KOH mount revealed septate fungal hyphae with irregular margins. Yellow-white nonsporulating mycelial growth was noted on the Sabouraud dextrose agar, which was identified as C. samala by sequencing. The patient responded to 5% natamycin and 1% voriconazole eye drops, and there was a formation of a corneal opacity in a period of 3 weeks.We report the first case of keratitis by C. samala, highlighting the emergence of a rare dematiaceous fungi causing keratitis and the role of molecular modalities in the diagnosis of nonsporulating fungi in suspected cases.

Cellulolytic fungi usually have multiple genes for C1-oxidizing auxiliary activity 9 (AA9) lytic polysaccharide monooxygenases (LPMOs) in their genomes, but their potential functional differences are less understood. In this study, two C1-oxidizing AA9 LPMOs, SbLPMO9A and SbLPMO9B, were identified from Sordaria brevicollis, and their differences, particularly in terms of thermostability, reducing agent specificity, and synergy with cellulase, were explored. The two enzymes exhibited weak binding to cellulose and intolerance to hydrogen peroxide. Their oxidative activity was influenced by cellulose crystallinity and surface morphology, and both enzymes tended to oxidize celluloses of lower crystallinity and high surface area. Comparably, SbLPMO9A had much better thermostability than SbLPMO9B, which may be attributed to the presence of a carbohydrate binding module 1 (CBM1)-like sequence at its C-terminus. In addition, the two enzymes exhibited different specificities and responsivities toward electron donors. SbLPMO9A and SbLPMO9B were able to boost the catalytic efficiency of endoglucanase I (EGI) on physically and chemically pretreated substrates but with different degrees of synergy. Substrate- and enzyme-specific synergism was observed by comparing the synergistic action of SbLPMO9A or SbLPMO9B with commercial Celluclast 1.5L on three kinds of cellulosic substrates. On regenerated amorphous cellulose and PFI (Papirindustriens Forskningsinstitut)-fibrillated bleached eucalyptus pulp, SbLPMO9B showed a higher synergistic effect than SbLPMO9A, while on delignified wheat straw, the synergistic effect of SbLPMO9A was higher than that of SbLPMO9B. On account of its excellent thermostability and boosting effect on the enzymatic hydrolysis of delignified wheat straw, SbLPMO9A may have high application potential in biorefineries for lignocellulosic biomass. KEY POINTS: • C1-oxidizing SbLPMO9A displayed higher thermostability than SbLPMO9B, probably due to the presence of a CBM1-like module. • The oxidative activity of the two SbLPMO9s on celluloses increased with decreasing cellulose crystallinity or increasing beating degree. • The two SbLPMO9s boosted the catalytic efficiency of cellulase, but the synergistic effect was substrate- and enzyme-specific.

In our continuous search for antibacterial agents against Pseudomonas syringae pv. actinidiae (Psa) from kiwi-associated fungi, two pairs of epimeric cytochalasins, zopfiellasins A–D (1–4), were characterized from the fungus Zopfiella sp. The structures were established on the basis of spectroscopic data analysis, while the absolute configurations were determined by single-crystal X-ray diffraction. Compounds 1 and 3 exhibited antibacterial activity against Psa with MIC values of 25 and 50 μg/mL, respectively. This is the first report of anti-Psa activity of cytochalasin derivatives.

BACKGROUND A growing number of non-dermatophytic moulds and yeasts with the ability to act as human pathogens are reported every year. Dematiaceous fungi cause phaeohyphomycosis which encompasses a broad spectrum of diseases ranging from superficial (cutaneous and subcutaneous) to disseminated infections. Such fungal infections are responsible for causing significant morbidity and mortality, frequently in immunocompromised patients and rarely in immunocompetent patients. OBJECTIVES To investigate the prevalence of cutaneous mycosis in Jammu district (India) and to isolate and identify the recovered causal agents from the affected skin of the patients. METHODS For direct microscopy, 10% KOH was used. Skin samples were collected carefully from the affected areas of suspected patients, followed by the isolation and identification of the causal agents by cultural examination, morphological examination and ITS sequencing. RESULTS Herein, we report and describe three new cases of cutaneous phaeohyphomycosis from District Jammu of Union Territory Jammu and Kashmir, India. The age of the patients under study ranged from 17 to 42 years and the duration of infection from 1 to 2 years. The etiological agents that were recovered from the patients under study were Alternaria alstromeriae, Epicoccum tritici and Phialemonium obovatum. These dematiaceous fungal species were isolated from the skin specimen of immunocompetent hosts. CONCLUSION Among the three isolated etiological agents, two (Alternaria alstromeriae, Epicoccum tritici) represent new global records and one (Phialemonium obovatum) new record to India as causal agents of cutaneous phaeohyphomycosis. Careful microscopic and mycological examination form the basis of correct diagnosis of such fungal infections in the absence of simple and reliable laboratory tests (serologic or antigen tests).

We report a case of a 56-year-old Brazilian woman, with relapsing lepromatous leprosy, and onychomycosis caused by a non-dermatophyte filamentous fungi. The pathogenic fungi was identified as Arthrinium arundinis and treated with chemical abrasion of the nail with 40% urea and application of terbinafine cream. Onychomycosis caused by Arthrinium species is rare, and this is the second reported case.

Interest in the use of natural non-toxic pigments by the food industry has grown. Some filamentous fungi are producers of natural pigments that are more stable at temperature and pH than other pigments also classified as natural, such as those produced by plants. Production potential of natural pigments by endophytic fungi from grapevines was evaluated. Arcopilus aureus was selected as a potential source for a yellow pigment, which was characterized and tested for stability to variations in temperature and pH. Components, cochlioquinol II and riboflavin, were detected, which has not previously been reported in A. aureus. The pigment was stable and showed increased absorption at lower / acidic pH. These results provide information on the potential of this fungus and a yellow pigment for the first time, which can be used for further development and industrial application.

The current research was designed to reach extracellular protease production potential in different strains of Sordaria fimicola which were previously obtained from Dr. Lamb (Imperial College, London) from North Facing Slope and South Facing Slope of Evolution Canyon. After initial and secondary screening, two hyper-producers strains S2 and N6 were selected for submerged fermentation and cultural conditions including temperature, pH, incubation period, inoculum size, substrate concentration, and different carbon and nitrogen sources were optimized for enzyme production. S2 strain showed maximum protease production of 3.291 U/mL after 14 days of incubation at 30 °C with 7 pH, 1% substrate concentration and 1 mL inoculum, While N6 strain showed maximum protease production of 1.929 U/mL under fermentation optimized conditions. Another aim of the present research was to underpin the biodiversity of genetics and post-translational modifications (PTMs) of protease DPAP (peptidyl-aminopeptidase) in Sordaria fimicola. Five polymorphic sites were observed in amino acid sequence of S. fimicola strains with reference to Neurospora crassa. PTMs prediction from bioinformatics tools predicted 38 phosphorylation sites on serine residues for protease peptidyl-aminopeptidase in S1 strain of S. fimicola while 45 phosphorylation sites on serine in N7 strain and 47 serine phosphorylation modifications were predicted in N. crassa. Current research gave an insight that change in genetic makeup effected PTMs which ultimately affected the production of protease enzyme in different strains of same organism (S. fimicola). The production and molecular data of the research revealed that environmental stress has strong effects on the specific genes through mutations which may cause genetic diversity. S. fimicola is non- pathogenic fungus and has a short life cycle. This fungus can be chosen to produce protease enzyme on a commercial scale. Resumo A pesquisa atual foi projetada para alcançar o potencial de produção de protease extracelular em diferentes cepas de Sordaria fimicola que foram previamente obtidas do Dr. Lamb (Imperial College, Londres) de North Facing Slope e South Facing Slope de Evolution Canyon. Após a triagem inicial e secundária, duas cepas hiperprodutoras S2 e N6 foram selecionadas para fermentação submersa e condições culturais, incluindo temperatura, pH, período de incubação, tamanho do inóculo, concentração de substrato, e diferentes fontes de carbono e nitrogênio foram otimizadas para produção de enzima. A cepa S2 apresentou produção máxima de protease de 3,291 U/mL após 14 dias de incubação a 30 °C com pH 7, concentração de substrato de 1% e inóculo de 1 mL, enquanto a cepa N6 apresentou produção máxima de protease de 1,929 U/mL em condições otimizadas de fermentação. Outro objetivo da presente pesquisa foi sustentar a biodiversidade da genética e modificações pós-tradicionais (PTMs) da protease DPAP (peptidil-aminopeptidase) em Sordaria fimicola. Cinco sítios polimórficos foram observados na sequência de aminoácidos de cepas de S. fimicola com referência a Neurospora crassa. A previsão de PTMs a partir de ferramentas de bioinformática previu 38 locais de fosforilação em resíduos de serina para protease peptidil-aminopeptidase na cepa S1 de S. fimicola, enquanto 45 locais de fosforilação em serina na cepa N7 e 47 modificações de fosforilação de serina foram previstas em N. crassa. A pesquisa atual deu uma ideia de que a mudança na composição genética afetou os PTMs que, em última análise, afetaram a produção da enzima protease em diferentes cepas do mesmo organismo (S. fimicola). A produção e os dados moleculares da pesquisa revelaram que o estresse ambiental tem fortes efeitos sobre genes específicos por meio de mutações que podem causar diversidade genética. S. fimicola é um fungo não patogênico e tem um ciclo de vida curto. Esse fungo pode ser escolhido para produzir enzima protease em escala comercial.

In this study, we compared the properties and structures of three fungal GH12 enzymes: the strict endoglucanase Bgh12A and the xyloglucanase Xgh12B from Aspergillus cervinus, and the endoglucanase Egh12 from Thielavia terrestris combining activity on linear β-glucan and branched xyloglucan. Egh12 from T. terrestris was produced in Pichia pastoris, purified, and characterized as a thermostable enzyme with maximal activity at 70 ºC and a half-life time of 138 min at 65 °C. We for the first time demonstrated that the GH12 endoglucanases Egh12 and Bgh12A, but not the strict xyloglucanase Xgh12B, hydrolyzed (1,3)-β-linkages in (1,3;1,4)-β-D-glucooligosaccharides and had transglycosylase activity on (1,3)-β-D-glucooligosaccharides. Phylogenetic analysis indicated that Egh12 from T. terrestris and Bgh12A from A. cervinus are more related than Bgh12A and Xgh12B isolated from one strain. The X-ray structure of Bgh12A was determined with 2.17 Å resolution and compared with 3D-homology models of Egh12 and Xgh12B. The enzymes have a β-jelly roll structure with a catalytic cleft running across the protein. Comparative analysis and a docking study demonstrated the importance of endoglucanase-specific loop 1 partly covering the catalytic cleft for correct placement of the linear substrates. Variability in substrate specificity between the GH12 endoglucanases is determined by non-conservative residues in structural loops framing the catalytic cleft. A residue responsible for the thermostability of Egh12 was predicted. The key structural elements and residues described in this study may serve as potential targets for modification aimed at the improvement of enzymatic properties. KEY POINTS: • Thermostable endoglucanase Egh12 from T. terrestris was produced in P. pastoris, purified, and characterized • The X-ray structure of GH12 endoglucanase Bgh12A from A. cervinus was resolved • GH12 endoglucanases, but not GH12 xyloglucanases, hydrolyze (1,3)-β-linkages in (1,3;1,4)-β-D-glucooligosaccharides.

Resveratrol is an important stilbene, initially identified from red wine possessing immense therapeutic, cosmeceutical and nutraceutical applications. In the present study, endophytic fungus

The formation of fruiting bodies is one of the most complex developmental processes in filamentous ascomycetes. It requires the development of sexual structures that give rise to meiosporangia (asci) and meiotic spores (ascospores) as well as surrounding structures for protection and dispersal of the spores. Previous studies have shown that these developmental processes are accompanied by significant changes of the transcriptome, and comparative transcriptomics of different fungi as well as the analysis of transcriptome changes in developmental mutants have aided in the identification of differentially regulated genes that are themselves involved in regulating fruiting body development. In previous analyses, we used transcriptomics to identify the genes asm2 and spt3, which result in developmental phenotypes when deleted in Sordaria macrospora. In this study, we identified another gene, asm3, required for fruiting body formation, and performed transcriptomics analyses of Δasm2, Δasm3, and Δspt3. Deletion of spt3, which encodes a subunit of the SAGA complex, results in a block at an early stage of development and drastic changes in the transcriptome. Deletion mutants of asm2 and asm3 are able to form fruiting bodies, but have defects in ascospore maturation. Transcriptomics analysis of fruiting bodies revealed a large overlap in differentially regulated genes in Δasm2 and Δasm3 compared to the wild type. Analysis of nuclear distribution during ascus development showed that both mutants undergo meiosis and postmeiotic divisions, suggesting that the transcriptomic and morphological changes might be related to defects in the morphogenesis of structural features of the developing asci and ascospores.

Nuankaew, Salilaporn, Chuaseeharonnachai, Charuwan, Somrithipol, Sayanh, Kwantong, Papichaya, Srihom, Charisa, Ros, Chandapheakdey, Boonyuen, Nattawut (2021): Contribution to Chaetosphaeriaceae-Kionochaeta setosimplicia sp. nov. from Thailand. Phytotaxa 508 (2): 175-186, DOI: 10.11646/phytotaxa.508.2.6, URL: http://dx.doi.org/10.11646/phytotaxa.508.2.6

Lytic Polysaccharide Monooxygenases (LPMOs) are powerful redox enzymes able to oxidatively cleave recalcitrant polysaccharides. Widely conserved across biological kingdoms, LPMOs of the AA9 family are deployed by phytopathogens to deconstruct cellulose polymers. In response, plants have evolved sophisticated mechanisms to sense cell wall damage and thus self-triggering Damage Triggered Immunity responses. Here, we show that Arabidopsis plants exposed to LPMO products triggered the innate immunity ultimately leading to increased resistance to the necrotrophic fungus Botrytis cinerea. We demonstrated that plants undergo a deep transcriptional reprogramming upon elicitation with AA9 derived cellulose- or cello-oligosaccharides (AA9_COS). To decipher the specific effects of native and oxidized LPMO-generated AA9_COS, a pairwise comparison with cellobiose, the smallest non-oxidized unit constituting cellulose, is presented. Moreover, we identified two leucine-rich repeat receptor-like kinases, namely STRESS INDUCED FACTOR 2 and 4, playing a crucial role in signaling the AA9_COS-dependent responses such as camalexin production. Furthermore, increased levels of ethylene, jasmonic and salicylic acid hormones, along with deposition of callose in the cell wall was observed. Collectively, our data reveal that LPMOs might play a crucial role in plant-pathogen interactions., Zarattini et al. confirm the capacity of Lytic Polysaccharide Monooxygenases (LPMO) active on cellulose to trigger immune responses in Arabidopsis. These results bring insight to the field of cell wall modifying enzymes and their roles in plant defense mechanisms.

Fungal peroxygenases (UPOs) have emerged as oxyfunctionalization catalysts of tremendous interest in recent years. However, their widespread use in the field of biocatalysis is still hampered by their challenging heterologous production, substantially limiting the panel of accessible enzymes for investigation and enzyme engineering. Building upon previous work on UPO production in yeast, we have developed a combined promoter and signal peptide shuffling system for episomal high throughput UPO production in the industrially relevant, methylotrophic yeast

Structure-selective endonucleases cleave branched DNA substrates. Slx1 is unique among structure-selective nucleases because it can cleave all branched DNA structures at multiple sites near the branch point. The mechanism behind this broad range of activity is unknown. The present study structurally and biochemically investigated fungal Slx1 to define a new protein interface that binds the non-cleaved arm of branched DNAs. The DNA arm bound at this new site was positioned at a sharp angle relative to the arm that was modeled to interact with the active site, implying that Slx1 uses DNA bending to localize the branch point as a flexible discontinuity in DNA. DNA binding at the new interface promoted a disorder-order transition in a region of the protein that was located in the vicinity of the active site, potentially participating in its formation. This appears to be a safety mechanism that ensures that DNA cleavage occurs only when the new interface is occupied by the non-cleaved DNA arm. Models of Slx1 that interacted with various branched DNA substrates were prepared. These models explain the way in which Slx1 cuts DNA toward the 3′ end away from the branch point and elucidate the unique ability of Slx1 to cleave various DNA structures.

Lytic polysaccharide monooxygenases (LPMOs) have attracted vast attention because of their unique mechanism of oxidative degradation of carbohydrate polymers and the potential application in biorefineries. This study characterized a novel LPMO from Myceliophthora thermophila, denoted MtLPMO9L. The structure model of the enzyme indicated that it belongs to the C1-oxidizing LPMO, which has neither an extra helix in the L3 loop nor extra loop region in the L2 loop. This was confirmed subsequently by the enzymatic assays since MtLPMO9L only acts on cellulose and generates C1-oxidized cello-oligosaccharides. Moreover, synergetic experiments showed that MtLPMO9L significantly improves the efficiency of cellobiohydrolase (CBH) II. In contrast, the inhibitory rather than synergetic effect was observed when combining used MtLPMO9L and CBHI. Changing the incubation time and concentration ratio of MtLPMO9L and CBHI could attenuate the inhibitory effects. This discovery suggests a different synergy detail between MtLPMO9L and two CBHs, which implies that the composition of cellulase cocktails may need reconsideration.

D-glucuronic acid (GlcUA) is an important intermediate with numerous applications in the food, cosmetics, and pharmaceutical industries. Its biological production routes which employ myo-inositol oxygenase (MIOX) as the key enzyme are attractive. In this study, five diverse MIOX-encoding genes, from Cryptococcus neoformans, Chaetomium thermophilum, Arabidopsis thaliana, Thermothelomyces thermophila, and Mus musculus were overexpressed in Escherichia coli, respectively. A novel MIOX from Thermothelomyces thermophila (TtMIOX) exhibited high specific activity, and efficiently converted myo-inositol to GlcUA. Meanwhile, the degradation of GlcUA was inhibited by inactivation of uxaC from the Escherichia coli genome. Finally, the BWΔuxaC whole-cell biocatalyst harboring TtMIOX resulted in the production of 106 g/L GlcUA within 12 h in a 1-L bioreactor, corresponding to a conversion of 91% and productivity of 8.83 g/L/h. This study provides a feasible method for the industrial production of GlcUA.

The eye arose during the Cambrian explosion from pre-existing proteins that would have been recruited for the formation of the specialized components of this organ, such as the transparent lens. Proteins suitable for the role of lens crystallins would need to possess unusual physical properties and the study of such earliest analogs of ocular crystallins would add to our understanding of the nature of recruitment of proteins as lens/corneal crystallins. We show that the Abundant Perithecial Protein (APP) of the fungi Neurospora and Sordaria fulfils the criteria for an early crystallin analog. The perithecia in these fungal species are phototropic, and APP accumulates at a high concentration in the neck of the pitcher-shaped perithecium. Spores are formed at the base of the perithecium, and light contributes to their maturation. The hydrodynamic properties of APP appear to exclude dimer formation or aggregation at high protein concentrations. APP is also deficient in Ca2+ binding, a property seen in its close homolog, the calcium-binding cell adhesion molecule (DdCAD-1) from Dictyostelium discoidum. Comparable to crystallins, APP occurs in high concentrations and seems to have dispensed with Ca2+ binding in exchange for greater stability. These crystallin-like attributes of APP lead us to demonstrate that it is a primitive form of ocular crystallins.

Corn bins in the midwestern United States can reach temperatures up to 52 C. High temperatures combined with sufficient moisture and humidity in bins provide the perfect environment to promote the growth of thermophilic and thermotolerant fungi. In this article, we characterize for the first time thermophilic and thermotolerant fungi in corn grain bins using culture-based methods and pyrosequencing techniques. Corn samples were collected from local farms in western Illinois. Samples were plated and incubated at 50 C using a variety of approaches. Of several hundred kernels examined, more than 90% showed colonization. Species identified using culture methods included Thermomyces lanuginosus, Thermomyces dupontii, Aspergillus fumigatus, Thermoascus crustaceus, and Rhizomucor pusillus. Pyrosequencing was also performed directly on corn grain using fungal-specific primers to determine whether thermophilic fungi could be detected using this technique. Sequences were dominated by pathogenic fungi, and thermophiles were represented by less than 2% of the sequences despite being isolated from 90% of the grain samples using culturing techniques. The high abundance of previously undocumented viable fungi in corn could have negative implications for grain quality and pose a potential risk for workers and consumers of corn-derived products in the food industry. Members of the Sordariales were absent among thermophile isolates and were not represented in nuc rDNA internal transcribed spacer (ITS) sequences. This is in striking contrast with results obtained with other substrates such as litter, dung, and soils, where mesophilic and thermophilic members of the Sordariaceae and Chaetomiaceae are common. This absence appears to reflect an important difference between the ecology of Sordariales and other orders within the Ascomycota in terms of their ability to compete in microhabitats rich in sugars and living tissues.