Eutypa lata (Pers.) Tul. & C. Tul., Select. fung. carpol. (Paris) 2: 56 (1863)

Index Fungorum number: IF 140211; MycoBank number: MB 140211; Facesoffungi number: FoF 09963; Fig. 1

Saprobic on the dead aerial branch of Corylus avellana. Sexual morph Ascostromata 400–600 μm high, 350–450 μm diam. (x̅=380×490 μm, n=10) length with ostiole neck, solitary, scattered semi-immersed, erumpent on the bark, subglobose to irregular, black, ostiolate. Ostiole individual, central, ostiole neck 150–200 μm high (x̅=165 μm, n=5). Peridium 20–30 μm wide, composed of two layers, outer layer comprising 4–5 layers of thick-walled, brown to pale brown cells of textura angularis to textura prismatica, thin inner layer comprising 3–5 layers of thinwalled, hyaline cells of textura prismatica. Hamathecium 1.8–3 μm wide, filiform, longer than the asci, septate, guttulate, unbranched, hyaline paraphyses. Asci 40–65×4–5 μm (x̅=53×4.5 μm, n=10), 8-spored, unitunicate, thin-walled, clavate, long pedicillate, apically truncate. Ascospores 5–9×1–2 μm (x̅=7×1.5 μm, n=30), overlapping bi-seriate, aseptate, hyaline, ellipsoidal to cylindrical or allantoid, slightly curved, smooth-walled, mostly with small guttules at the edges. Asexual morph sees (Moyo et al. 2018).

Culture characteristics – Ascospores germinating on PDA within 24 h and germ tubes arising from both ends of the ascospore. Colonies on PDA, reaching 4–5 cm diam. after 7 days at 25 °C, colonies medium dense, flat or effuse, slightly raised, cottony, white, margin rough.

Material examined – ITALY, Province of Forlì-Cesena, Teodorano—Meldola, on the dead aerial branch of Corylus avellana (Betulaceae), 20 January 2019, E. Camporesi, IT 4212 (MFLU 19-0501, new record), living culture, MFLUCC 21-0068.

GenBank numbers – ITS = MZ453405, TUB2=MZ476772.

Notes – The multigene phylogeny (ITS and TUB2) of this study, our strain (MFLUCC 21-0068) and Eutypa lata (EP18, RGA01 and ATCC 28120) strains clustered together with 89% MLBS, 0.96 BYPP support (Clade B; Fig. 2). Comparing the ITS (ITS1-5.8S-ITS2) and TUB2 rDNA regions between EP18 and RGA01 strains, only 1 bp (0.19%) difference was found from 526 nucleotides in the ITS and no base pair differences in the TUB2. Comparison of the same gene regions between our strain and RGA01 strain, there are only 1 bp (0.18%) difference that could in the ITS and within the 350 bp of the TUB2 region, there are 7 bp (2.0%) difference. In comparison of the same gene regions between our strain and ATCC 28,120 strain there were no base pair differences in the ITS and 7 bp (1.88%) of difference from 372 bp in the TUB2. When morphological characters of our species were examined, they were similar with the genus description provided by Vasilyeva and Stephenson (2006) and Senanayake et al. (2015). The species description of Eutypa lata provided by Moyo et al. (2018) is morphologically similar to our isolate MFLUCC 21-0068 (Fig. 1). According to morpho-molecular analyses we conclude that our new collection is another record of Eutypa lata and also a new host record on Corylus avellana (Betulaceae) in Italy.

Figure 1 – Eutypa lata (MFLU 19-0501, new record). a, b Stromata on substrate. c Cross-section of ascoma. d Ostiole. e Peridium. f Paraphyses. g–i Asci. j–l Ascospores. m, n Culture on PDA from above and below. Scale bars: a=200 μm, b, c=100 μm, d=50 μm, f, h, i=20 μm, e, g=10 μm, j–l=5 μm

Figure 2 – Phylogram generated from maximum likelihood analysis based on combined ITS and TUB2 sequence data representing Diatrypaceae in Xylariales. Related sequences are taken from Konta et al. (2020) and additions according to the BLAST searches in NCBI. Hundred and twenty-nine strains are included in the combined analyses which comprised 1180 characters (665 characters for ITS and 515 characters for TUB2) after alignment. Kretzschmaria deusta (CBS 826.72) and Xylaria hypoxylon (CBS 122620) in Xylariaceae (Xylariales) were used as the outgroup taxa. The best scoring RAxML tree with a final likelihood value of − 17362.853779 is presented. The matrix had 906 distinct alignment patterns, with 37.13% of undetermined characters or gaps. Estimated base frequencies were as follows: A=0.224946, C=0.263658, G=0.237000, T=0.274396; substitution rates: AC=1.093989, AG=3.037146, AT=1.238822, CG=0.814851, CT=4.048082, GT=1.000000; gamma distribution shape parameter α=0.183977. Bootstrap support values for ML equal to or greater than 75% are given above the nodes (left side). Bayesian posterior probabilities (BYPP) equal to or greater than 0.95 are given above the nodes (right side). Ex-type strains are in bold and newly generated sequences are in blue

Figure 2 – (continued)