Neoeutypella baoshanensis M. Raza, Q.J. Shang, Phookamsak & L. Cai, in Phookamsak et al., Fungal Diversity: https://doi.org/10.1007/s13225-019-00421-w, [168] (2019)

Index Fungorum number: IF 555372; MycoBank number: MB 555372; Facesoffungi number: FoF 04928; Fig. 1

Saprobic on dead wood of Robinia pseudoacacia L. Sexual morph: Ascomstromata 200–300 μm diam., carbonaceous, black, solitary to gregarious, globose to long irregular in shape on host surface, erumpent through host epidermis. Ascomata perithecial 500–770 μm high, 450–530 μm diam., black, immersed to semi-immersed, solitary to aggregated, globose or subglobose, ostiolate. Ostiole distinct, papillate, slightly conspicuous, filled with periphyses. Peridium 145–250 μm wide, thickened, composed of two layers; outer layer unequally thicken, brown to dark brown cells of textura angularis; inner layer thin-walled, hyaline to brown cells of textura prismatica. Hamathecium comprising filamentous paraphyses and asci. Paraphyes 0.6–2 μm thickness, numerous, anastomosing, not branched, aseptate, hyaline, filamentous, generally exceeding the length of asci. Asci 60–80 × 10–24 µm (x = 70 × 17 μm, n = 20), 8-spored, unitunicate, spindle-shaped, tip blunted, long pedicellate, amyloid. Ascospores 10–17 × 6–9 µm (x=13.5 × 7.5 μm, n=40), 8-spored, ellipsoid, overlapping 1–3 seriate, initially hyaline, becoming pale brown at maturity, allantoid, slightly or moderately curved, aseptate, mostly with 1–2 small guttules. Asexual morph: Undetermined.

Culture characteristics – Colonies on PDA, 16–20 mm diam., after three weeks, colonies from above: medium dense, circular, flat, slightly raised, surface smooth with crenate edge, with smooth aspects, grey to dark brown at the margin, light brown in the center; reverse: dark brown to black at the margin, brown to grey in the center.

Material examined – Italy, Robinia pseudoacacia  (Fabaceae), Erio Camporesi, 24 June 2021, D27 (MFLU 21-4687).

Hosts and distribution – China (Pinus armandii) (Phookamsak et al. 2019), Italy (R. pseudoacacia).

GenBank numbers – ITS: OR053733; β-tubulin: OR079742.

Notes – Our collection (MFLU 21-4687) grouped with Neoeutypella baoshanensis (HMAS 255436) with ML/BI=100%/1.00 statistical support forming a sister clade to Diatrype. Our collection obtained from a dead stem of Robinia pseudoacacia from Italy shares similar morphological characteristics with N. baoshanensis in the size and shape of ascomata, asci and pale brown ascospores. They also share similar morphological characteristics such as solitary to gregarious, globose to long irregular and carbonaceous ascomata, papillate ostiole, periphyses, a two-layered peridium and aseptate, filamentous paraphyses. However, our collection differs from the holotype material of N. baoshanensis in the absence of the yellow pigment and asexual structures. In the base pair comparison between our collection and N. baoshanensis (HMAS 255436) there are 1% and 1.3% differences for ITS and β-tubulin respectively. Based on the recommendation from Jeewon and Hyde (2016) with morphological characteristics, we provide our collection as new geographical record from Italy as well as a new host record from Robinia pseudoacacia (Fig. 2).

Figure 1 – Neoeutypella baoshanensis (MFLU 21-4687). a–c Appearance of ascomata on substrate. d–f Vertical cross-sections through ascomata. g Vertical cross-sections through ostiole. h Peridium. i Paraphyses. j–m Asci. n1–n10 Ascospores. o A germinating ascospore. p Colony from above and below (on PDA). Scale bars: c–g=100 μm, h=20 μm, i=10 μm, j–m=30 µm, n1–n10, o=10 µm

Figure 2 – The best scoring RAxML tree with a final likelihood value of − 14,228.890517 for combined dataset of ITS and β-tubulin sequence data. The topology and clade stability of the combined gene analyses was compared to the single gene analyses. The tree is rooted with Pestalotiopsis macadamiae (BRIP 63738b). The matrix had 743 distinct alignment patterns with 31.15% undetermined characters and gaps. Estimated base frequencies were as follows; A=0.228059, C=0.263930, G=0.237101, T=0.270909; substitution rates AC=0.991638, AG=2.523032, AT=0.315447, CG=0.674576, CT=3.904225, GT=1.0; gamma distribution shape parameter α=0.582489. Ex-type strains are in bold and newly generated sequences are in red. Bootstrap support for ML equal to or greater than 65% and BI equal to or greater than 0.90 are given above the nodes

Figure 2 – (continued)