Botryosphaeria aterrima (Fuckel) Sacc. Syll. fung. (Abellini) 1: 458 (1882). Figure 1

Ascostromata not observed. Conidiostromata forming dense botryose aggregate, 2–7 mm diam., pseudothecial, aggregated into botryose clusters, 220–420 μm diam., spherical to globose with a central ostiole, ½ to ¾ emergent, rarely embedded, black. Peridium comprising 7–15 layers of textura angularis, outer region of dark brown cells, inner region of 3–7 layers of pale brown cells lining the locule. Paraphyses when present hyaline, septate, up to 70 μm long, 2–4 μm broad at the base, tapering to acutely rounded apices, 1.5–2 μm broad at the tip. Conidiogenous cells holoblastic, hyaline, sub-cylindrical, 8–20 × 3–5 μm. Conidia hyaline, narrowly fusiform, or irregularly fusiform, base subtruncate to bluntly rounded, (40–)42–60(–62) × (7–)9–11 μm (−x = 51.3 × 9.9 μm, n = 20), L/W = 5.2.

Specimen examined – GERMANY, Hessen, Ulmus sp. (Ulmaceae), Fuckel, K.W.G. (G 00266252, holotype).

Notes – Although phylogenetic analysis based on combined loci of ITS, tef1-a and LSU confirmed Botryosphaeria aterrima within Botryosphaeria, it cannot be distinguished from B. auasmontanum, B. berengeriana var. weigeliae, B. dothidea, B. minutispermatia, and B. mirabile (Supplementary Figure 4). The two isolates of B. dothidea used in this study are ex-epitype (CBS 115476) and a verified isolate (CBS 110302), respectively (Slippers et al., 2004; Phillips et al., 2013). Botryosphaeria dothidea is known as a cosmopolitan species associated with woody plants in numerous families (Slippers et al., 2004; Marsberg et al., 2017). The type of B. aterrima reassessed here was collected from Ulmus sp. in Germany, and only the asexual morph was observed. Conidia of B. aterrima examined in this study were much larger [(40–)42–60(–62) × (7–)9–11 μm] than those of B. dothidea [(17–)18–20(–22) × 4–5 μm, as reported by Slippers et al. (2004), Table 2]. A remarkable feature of B. aterrima was its multiloculate conidiomata, which was comparable with members of Aplosporellaceae and Melanopsaceae. The hyaline, fusiform conidia lacking mucous sheath, however, differed from these two families. Thus, we treat B. aterrima as a separate species within Botryosphaeria sensu stricto.

Botryosphaeria dothidea (Moug. : Fr.) Ces. and De Not., Comment. Soc. Crittog. Ital. 1:212. 1863. Figure 2

Ascostroma erumpent to nearly superficial, 1.5-4.5 mm diam. Ascomata 110-240 μm diam., pseudothecial, forming botryose clusters of up to 50 locules, globose with a central ostiole, papillate or not, black. Peridium comprising 6–15 layers of textura angularis, outer region of dark brown cells, inner region of 2–3 layers of hyaline cells lining the locule (Slippers et al., 2004). Pseudoparaphyses filiform, cellular, septate, 3–4 μm wide. Asci 8-spored, bitunicate, cylindric-clavate to clavate with a short pedicel, 72-130 × 17–27 μm, forming among pseudoparaphyses. Ascospores hyaline, broadly fusoid to ellipsoidal, smooth or with granular contents, sometimes become 1–2 septa with age, biseriate in the ascus, (18–)20–26(–28) × 7–10 μm (−x = 23.3 × 8.8 μm, n = 20), L/W = 2.6. Spermatia not observed.

Specimen examined – RUSSIA, Batum (i), Caucasus, on cortex Weigela sp. (Caprifoliaceae), Newodowski (MICH 13862, isotype of B. berengeriana var. weigeliae). United States, Washington, on bark of Acer macrophyllum (Aceraceae), June 1906, S. A. Harper (F C0003484F, holotype of Botryosphaeria berengeriana var. acerina).

Notes – The phylogeny based on ITS and tub2 nuDNA sequence analysis indicated that Botryosphaeria berengeriana var. weigeliae and B. dothidea cluster together with ITS [only one base-pair differences (of 202 base-pairs) and tub2 totally identical (of 344 base-pairs), Supplementary Figure 4]. All the morphological characteristics of B. berengeriana var. weigeliae agree with B. dothidea but the 1–2-septate mature ascospores (Slippers et al., 2004; Figure 2), which is insufficient to separate a species. Thus, we treated B. berengeriana var. weigeliae a synonym of B. dothidea. In addition, B. berengeriana var. acerina was reduced to synonymy with B. dothidea because of their morphological similarities.

Botryosphaeria mirabile (Fuckel) Cooke, Grevillea 13 (no. 68): 108 (1885). Figure 3

Ascostromata erumpent, clustered, black, with yellow globules on the top, 1–2 mm diam., globose, gregarious. Asci 8-spored, bitunicate, broadly clavate. Pseudoparaphyses not observed. Ascospores fusiform, irregularly biseriate to triseriate in asci, up to 10 × 5 μm. Spermatia numerous, ellipsoid to subglobose, hyaline, 3–4 μm (data of sexual stage are obtained from the label of the type specimen).

Specimen examined – SWITZERLAND, Genève, on trunk of Quercus sp. (Fagaceae), J.E. Duby (G 00266251, holotype).

Notes – Melanops mirabilis was introduced by Fuckel (1870), and was assigned to Botryosphaeria as B. mirabilis by Cooke (1885). Some large spermatogonia (up to 5 mm diam.) were found on the type specimen examined in this study, which contained numerous subglobose, pale brown spermatial-like cells (ca. 3–5 μm diam.) (Figure 3), from which the nuDNA was extracted in this study. Some immersed ascomata were also sectioned, but they were in ordinately old to enable a morphological study. Both spermatia and the sexual morph were considered in the original description (Fuckel, 1870). The sexual morph was described as having immersed pseudothecia, numerous, dark brown pseudoparaphyses, narrowly clavate asci, and oblong, hyaline, 2-septate ascospores which disagree with the concept of Botryosphaeria. Two distinct sexual morphs were illustrated on the envelope of the type specimen (Figure 3). Besides the one mentioned above, the other was illustrated as having broadly clavate asci, hyaline, aseptate ascospores (10 × 5 μm), which fits the concept of Botryosphaeria (Figure 3). Thus, the second illustrated sexual morph corresponds with B. mirabile (Figure 3). nuDNA sequence comparisons based on ITS and LSU indicated that Melanops mirabilis resides in Botryosphaeria (Supplementary Figure 4), although it cannot be distinguishable from B. aterrima, B. auasmontanum, B. berengeriana var. weigelae, B. dothidea, and B. minutispermatia. In addition, B. mirabile differs from B. dothidea by its nuDNA loci, e.g., 6 bp differences in ITS (1%) and 5 bp in LSU (1.2%). Based on its subglobose pale brown spermatia, small-sized ascospores as well as its unique nuDNA loci, we retain B. mirabile as a separate species in Botryosphaeria sensu stricto.

Neodeightonia mucosa (S.J. Kaur) Y. Zhang ter and Y.P. Zhou, comb. nov.

MycoBank number: 840943; Facesoffungi number: FoF 03579; Figure 4

Ascomata 170-320 μm diam., erumpent, pseudothecial, scattered, solitary or aggregated, globose with a central ostiole, ¼ to ½ emergent, almost embedded, papillate or not, black. Peridium comprising 5–15 layers of textura angularis, outer region of dark brown cells, inner region of 2–4 layers of hyaline cells lining the locule. Asci bitunicate, broadly clavate, pedicellate, 100-135 × 20–35 μm, 8-spored, with a long, narrowed and twisted pedicel, forming between pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 3–5 μm wide. Ascospores hyaline, aseptate, thin-walled, obovoid to obpyriform, with a distinct oil drop, biseriate in the ascus, 17–20 × 8–10 μm (−x = 18.8 × 8.9 μm, n = 20), L/W = 2.1. Conidiomata stromatic, pycnidial, solitary or aggregated, morphologically indistinguishable from the ascomata, walls composed of dark brown, thick-walled textura angularis, becoming thin-walled and hyaline toward the inner layer. Ostioles single, central, papillate or not. Paraphyses, hyaline, septate, 2–3 μm wide. Conidiophores reduced to conidiogenous cells. Conidiogenous cells hyaline, 8–9(–11) × (1–)2–3 μm. Conidia hyaline, aseptate, moderately thick-walled, ovoid or broadly ellipsoidal, surrounded by mucilaginous sheath, 1 μm thick, (7–)9–14 × 6–10(–12) μm (−x = 11.5 × 8.6 μm, n = 20), L/W = 1.3. Spermatia unicellular, hyaline, allantoid to rod-shaped, 4-6 × 1-2 μm.

Specimen examined – INDIA, Rajasthan, on dead bamboo wood, 25 May 1976 (K 204341, holotype).

Notes – The bambusicolous lifestyle, aggregated ascostroma, broadly clavate asci with long pedicel, hyaline, aseptate, thin-walled, obovoid to obpyriform ascospores and its hyaline, aseptate, moderately thick-walled, ovoid or broadly ellipsoidal conidia surrounded by mucilaginous sheath suggest Neodeightonia being an appropriate genus for this species. Neodeightonia was introduced by Booth (Punithalingam, 1969), and species are typically associated with monocotyledonous plants and especially bamboo (Punithalingam, 1969; Phillips et al., 2008, 2013; Liu et al., 2012; Adamčík et al., 2015; Dai et al., 2017). Morphologically, Neodeightonia mucosa is most similar to Neodeightonia microspora, while the smaller ascospores (10–12 × 4.5–6 μm) of N. microspora, can be readily distinguished from those of N. mucosa (Dai et al., 2017).

So far, six species have been assigned in Neodeightonia, namely Neodeightonia licuriensis, N. microspora, N. mucosa, Neodeightonia palmicola, Neodeightonia phoenicum, and Neodeightonia subglobosa. Of these, N. palmicola and N. phoenicum have been reported on palms (Phillips et al., 2008, 2013; Liu et al., 2012), N. subglobosa, N. microspora, and N. mucosa are reported as bambusicolous (Dai et al., 2017; this study), and N. licuriensis has been reported on Syagrus coronata (Adamčík et al., 2015). Neodeightonia subglobosa has also been reported causing keratomycosis in a human eyes (Phillips et al., 2008).

Neofusicoccum cruenta (Petr.) Y.P. Zhou, Y. Zhang ter., comb. nov.

MycoBank number: 840944; Facesoffungi number: FoF 03578; Figure 5

Ascomata erumpent, 70–140 μm diam., pseudothecial, solitary or gregarious, globose with a central ostiole, ¼ to ½ emergent, rarely embedded, black. Peridium composed of 6–10 layers of textura angularis, outer region of dark brown cells, inner region of 1–3 layers of hyaline cells lining the locule. Asci bitunicate, clavate, 38–65 × 17–22 μm. Pseudoparaphyses not observed. Ascospores hyaline, fusoid to ellipsoid, sometimes with tapered ends, bi- to triseriate, 13–20 × 5–9 μm (−x = 16.9 × 7 μm, n = 20), L/W = 2.4.

Specimen examined – CZECHIA, Prerov, on leaves of Polygonatum officinale (Liliaceae), April 1926, F. Petrak (W 1978-0010992/24018, holotype).

Notes – Only the sexual morph was observed on the type material, the morphology of which is consistent with members of Botryosphaeriaceae in having gregarious ascomata, broadly clavate asci and hyaline, aseptate ascospores as well as lacking pseudoparaphyses. Only LSU sequence was obtained for the type material of Melanops cruenta in this study, and a few other species of Neofusicoccum have LSU sequences available from GenBank as well. The phylogenetic analysis based on these LSU sequences suggested that M. cruenta resides in Neofusicoccum, being sibling to other species in the genus (Supplementary Figure 6). Thus, we have assigned M. cruenta to Neofusicoccum as a new combination, N. cruenta.

Neofusicoccum hamamelidis (Rehm) Y.P. Zhou, Y. Zhang ter., comb. nov.

MycoBank number: 840945; Facesoffungi number: FoF 03577; Figure 6

Ascomata erumpent, 170–400 μm diam., pseudothecial, scattered, solitary or aggregated, globose with a central ostiole, ¼ to ½ emergent, almost embedded, black. Peridium comprising 5–15 layers of textura angularis, outer region of dark brown cells, inner region of 4–6 layers of hyaline cells lining the locule. Asci bitunicate, clavate, 110–150 × 30–45 μm, 8-spored, forming among pseudoparaphyses. Pseudoparaphyses filiform, septate, constricted at the septa, rarely branched, 2–5 μm broad. Ascospores hyaline, thin-walled, ellipsoidal to ovoid, usually broadest in the middle, smooth, contents granular, biseriate in the ascus, 33–39(–48) × 13–16 μm (−x = 36.5 × 14.9 μm, n = 20), L/W = 2.4 (some data referred to Kobayashi, 1962).

Specimen examined – CANADA, near London, Ontario, on dead twigs of Hamamelidis virginiana (Hamamelidaceae), 18 May 1912, J. Dearness (W 07238/29850, type?). CHINA, Heilongjiang Province, Tieling, Langxiang, on twigs of Larix gmelinii (Rupr.) Kuzen. (Pinaceae), 10 July 2015, W. He (HMAS 246968, HMAS 246969).

Notes – The two isolates of Neofusicoccum hamamelidis (CGMCC3.18002/CGMCC3.18003) included in the phylogram (Supplementary Figure 5) were obtained from twigs of larch (Larix gmelinii) with shoot blight in Heilongjiang Province in China, which had been named as Physalospora laricina (on Larix kaempferi, Japan, Sawada, 1950), and subsequently combined to Guignardia as G. laricina (Sawada) W. Yamam. & Kaz. Itô and Botryosphaeria and B. laricina (Sawada) Y.Z. Shang (Yamamoto, 1961; Shang, 1987). Morphologically, “Botryosphaeria hamamelidis” and P. laricina were almost indistinguishable, but their ascospore broadness (9–12 vs. 13–16 μm, data from HMAS 246968 and HMAS 246969), which was insufficient to split them into two species. The phylogeny based on ITS, tef1-a, tub2 and LSU also supported the conspecific status of B. hamamelidis and P. laricina (Supplementary Figures 5, 6). Botryosphaeria hamamelidis, as an earlier epithet, had priority over Physalospora laricina. Thus, Physalospora laricina was reduced to a synonym of B. hamamelidis, which was assigned to Neofusicoccum as N. hamamelidis herein.

Botryosphaeria abuensis S.J. Kaur, 1996, Indian J. Mycol. Plant Path. 25(3): 334 (1996) [1995]. Figure 7

Ascomata 120-300 μm diam., pseudothecial, erumpent, scattered, solitary, globose with a central ostiole, ¼ to ½ emergent, rarely embedded, papillate or not, black. Peridium comprising 5–15 layers of textura angularis, outer region of dark brown cells, inner region of 2–4 layers of hyaline cells lining the locule. Asci bitunicate, clavate, pedicellate, up to 20 μm, 8-spored, 80-135 × 18–30 μm, forming among pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 2–5 μm broad. Ascospores oblong to narrowly fusoid, broadest in the upper third, biseriate to triseriate in the ascus, hyaline, smooth, sometimes granular to guttulate, aseptate, (17–)20–30(–32) × 6–11 μm (−x = 23.4 × 8.4 μm, n = 30), L/W = 2.8.

Specimen examined – INDIA, Rajasthan, on dead twigs of Lantana camara (Verbenaceae), 4 March 1975 (K 192142, type).

Notes – The solitary or scattered ascomata, clavate asci, hyaline, aseptate, oblong to narrowly fusiform ascospores fit Botryosphaeriales. Taxonomic status of this species, however, could not be determined due to a lack of morphological features of the asexual morph and DNA sequence data. Thus B. abuensis is tentatively kept in Botryosphaeria sensu stricto herein.

Botryosphaeria aesculi (Peck) M.E. Barr, Contr. Univ. Mich. Herb. 2: 561 (1972). Figure 8

Ascomata erumpent, 80-200 μm diam., black, immersed to semi-immersed in the host, becoming erumpent, scattered, sometimes in small groups of 2 locules, globose with a central ostiole, papillate or not. Peridium comprising 4–8 layers of textura angularis, outer region of dark brown cells, inner region of 1–2 layers of hyaline cells lining the locule. Pseudoparaphyses not observed. Asci bitunicate, subclavate to clavate, 8-spored, 46-66 × 12–22 μm. Ascospores hyaline, ellipsoidal-fusiform or fusiform, broadest in the upper third, irregularly biseriate in the ascus, 16–20(–23) × 6–11 μm (−x = 19 × 8.9 μm, n = 20), L/W = 2.1.

Specimen examined – United States, Albany, on petioles of Aesculus hippocastanum (Sapindaceae), May 1885, G. W. Clinton (NYS f93, holotype).

Notes – Botryosphaeria aesculi was introduced as Laestadia aesculi, which was subsequently assigned to Botryosphaeria as B. aesculi by Barr (1972). Morphologically, the scattered and erumpent ascomata, small-sized and ellipsoidal asci differ from the species of Botryosphaeria sensu stricto, while the hyaline, aseptate and ellipsoidal to fusiform ascospores are consistent with taxa in the Botryosphaeriales. Herein we tentatively retain it within Botryosphaeria with its taxonomical status remaining to be undetermined until further phylogenetic analysis is carried out on verified specimens.

Botryosphaeria dasylirii (Peck) Theiss. and Syd., Annls mycol. 13(5/6): 663 (1915). Figure 9

Ascostroma erumpent through the leave surface, 0.3-1.8 mm diam. Ascomata 90-150 μm diam., pseudothecial, usually forming botryose clusters of 3-4 locules, globose with a central ostiole, covered with epidermal leaf tissue, black. Peridium comprising 9–12 layers of textura angularis, outer region of dark brown cells, inner region of 2–3 layers of hyaline cells lining the locule. Asci 8-spored, bitunicate, ellipsoidal to subclavate, 100-115 × 23–40 μm, forming between pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 3–4 μm wide. Ascospores hyaline, ovoid to narrowly ellipsoid with granular content, aseptate, biseriate to triseriate, (24–)26–37(–40) × 10–15 μm (−x = 33 × 14 μm, n = 20), L/W = 2.3, sometimes becoming 0–2 septa with aging. Spermatia unicellular, hyaline, allantoid to rod-shaped, 4.5-6 × 1.5-2 μm.

Specimen examined – United States, Arizona, on leaves of Dasylirion sp. (Asparagaceae), May 1881, C. G. Pringle (NYS f950, holotype).

Notes – The erumpent botryose ascostroma, cellular pseudoparaphyses, hyaline, aseptate, large-sized ascospores suggest an affiliation in the Botryosphaeriaceae, while the ovoid to narrowly ellipsoid ascospores fit both Botryosphaeria and Neofusicoccum. Since there are no obvious morphological differences between Neofusicoccum and Botryosphaeria, the generic status of B. dasylirii remains uncertain until DNA sequence data can be obtained for it. Thus B. dasylirii is tentatively kept in Botryosphaeria sensu stricto herein until further phylogenetic analysis is carried out on verified specimens.

Botryosphaeria wisteriae (Rehm) Sacc., Syll. fung. (Abellini) 1: 459 (1882) Figure 10

Ascostroma erumpent from bark of host, 0.5–1.5 mm diam. Ascomata 140-220 μm diam., pseudothecial, botryose clustered, globose with a central ostiole, black. Peridium comprising 7–13 layers of textura angularis, outer region of dark brown cells, inner region of 2–3 layers of hyaline cells lining the locule. Asci 8-spored, bitunicate, clavate with a short pedicel, 70-118 × 17–27 μm, forming between pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 3–5 μm broad. Ascospores hyaline, ellipsoid to fusiform, granular content not sure, partially overlapping to biseriate in ascus, sometimes become 1-septate with aging, (17–)20–26(–30) × 7–10.5 μm (−x = 23.6 × 9.1 μm, n = 20), L/W = 2.6. Spermatia not observed.

Specimen examined – United States, South Carolina, Aiken, on dead twig of Wisteria chinensis (Fabaceae), Thuemen (MICH 15081, isotype).

Notes – The botryose ascomata, cellular pseudoparaphyses, and aseptate, hyaline ascospores are consistent with members of the Botryosphaeria sensu stricto. Thus B. wisteriae is tentatively kept in Botryosphaeria sensu stricto herein until further phylogenetic analysis is carried out on verified specimens.

Nothophoma ferruginea (Fuckel) Y.P. Zhou and Y. Zhang ter., comb. nov.

MycoBank number: 840946; Facesoffungi number: FoF 03576; Figure 11

Ascostroma immersed to erumpent, 2–8 mm diam. Ascomata 220-550 μm diam., densely gregarious, with elongate, rounded, obtuse or acute papilla, black. Peridium comprising 12–15 layers of textura angularis, outer region of dark brown cells, inner region of 3–5 layers of hyaline cells lining the locule. Asci 8-spored, bitunicate, cylindrical with a short, narrowed, twisted, furcate pedicel, 115-145 × 10–13 μm, forming between pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 3–4 μm wide. Ascospores uniseriate to partially overlapping, ellipsoidal to ovoid, muriform, with 3–4 transversal septa and 1–2 longitudinal septa in first, second or third cell(s), hyaline, (14–)17–20 × 6–9 μm (−x = 15.6 × 7.5 μm, n = 20), L/W = 2.5.

Specimen examined – SWITZERLAND, Neuchâtel, on the trunks of corruption Alnus glutinosa (Betulaceae), February 1872, Morthier (G 00127285, holotype).

Notes –The bitunicate asci, cellular pseudoparaphyses, the hyaline, broadly ellipsoid ascospores with 3–4 transversal septa and 1–2 longitudinal septa in central cells of Melanops ferruginea differ from members of Botryosphaeriales. Phylogeny based on ITS and LSU nuDNA sequences indicated that M. ferruginea closely related to Nothophoma Qian Chen and L. Cai (Didymellaceae, Pleosporales). Therefore, we assign it to Nothophoma as N. ferruginea.

Phyllachorella micheliae Syd., Annls mycol. 12(5): 489 (1914). Figure 12

Ascomata 140-250 μm diam., pseudothecial, aggregated forming a large botryose, irregularly rounded, 2–8 mm diam., lower side of the leaves, globose with a central white ostiole, ostiole 20–40 μm broad, ½ to ¾ emergent, rarely embedded, black. Peridium thick-walled, comprising 5–10 layers of textura angularis, outer region of dark brown cells, inner region of 4–6 layers of hyaline cells lining the locule. Asci 8-spored, bitunicate, broadly clavate with a long, narrow pedicel, which is up to 80 μm, with obvious apical chamber, 90-140 × 20–30 μm, forming between pseudoparaphyses. Pseudoparaphyses filiform, hyaline, cellular, septate, obviously constricted at the septa, rarely branched, 4–5 μm wide. Ascospores hyaline, aseptate, thin-walled, unequally gourd-shaped with upper part broader than the lower part, biseriate to triseriate in the ascus, (16–)18–21(–22) × 8–9(–11) μm (−x = 19.4 × 8.8 μm, n = 20), L/W = 2.2.

Specimen examined – INDIA, Kodaikanal, Tamil Nadu, on leaves of Michelia nilgarica (Magnoliaceae), 10 January 1987, L. N. Nair (K 316002, holotype).

Notes – Botryosphaeria foliicola was introduced by Sivanesan and Nair (1988) from the leaves of Michelia nilgarica in India, which distinguishes it from other species of Botryosphaeria by its obovoid and characteristically constricted ascospores. Phyllachorella micheliae, the generic type of Phyllachorella, was reported from the leaves of the same host in India. The strong morphological similarity of Botryosphaeria foliicola and Phyllachorella micheliae warranted their conspecific status (Liu et al., 2012). Based on priority, the later name Botryosphaeria foliicola is reduced to synonymy with Phyllachorella micheliae, which is retained in Botryosphaeriales genera incertae sedis (Wijayawardene et al., 2020).

Botryosphaeria gaubae Petr., Sydowia 21: 235 (1968). Supplementary Figure 1

Ascomata erumpent, 200-360 μm diam. Ascomata scattered, solitary, externally black, globose with a central ostiole, ¼ to ½ emergent, embedded in hairy seta on lower side of the leaves. Peridium comprising 5–15 layers of textura angularis, outer region of dark brown cells, inner region of 3–5 layers of hyaline cells lining the locule. Asci bitunicate, cylindrical or broadly cylindrical, pedicellate or not, pedicles up to 20 μm, 8-spored, 140-180 × 25–35 μm, forming among pseudoparaphyses. Pseudoparaphyses filiform, narrowly cellular, rarely branched, 2–3 μm broad. Ascospores hyaline, aseptate, fusiform to ellipsoid, with tapered ends and appearing spindle-shaped, sometimes slightly narrower at the middle, irregularly biseriate in the ascus, (23–)27–36(–46) × (6–)10–15(–17) μm (−x = 32.2 × 12.6 μm, n = 20), L/W = 2.6.

Specimen examined – AUSTRALIA, on leaves of Grevillea victoriae (Proteaceae), Mt Franklin, ca. 4500 ft., 27 January 1953, leg. E. Gauba (W 1992-05937, holotype).

Notes – Botryosphaeria gaubae was introduced and assigned in Botryosphaeria sensu lato, which included Botryosphaeria, Gibberella, and Lisea or even Melanops (Petrak, 1967). While the foliicolous habitation, scattered ascomata, cylindrical asci, fusiform to ellipsoid, aseptate ascospores with tapered ends and the presence of filiform pseudoparaphyses differ from these genera. We consequently treat it as a species in the Dothideomycetes incertae sedis.

Laestadia apocyni Ellis and Everh., Proc. Acad. nat. Sci. Philad. 42: 230 (1890). Supplementary Figure 2

Ascomata 100-215 μm diam., pseudothecial, scattered or clustered, globose with a central ostiole, papillate or not, black. Peridium thin, comprising 2–3 layers of textura angularis. Asci 8-spored, bitunicate, slightly obclavate, lack of pedicel, 47 × 27 μm (only a single complete mature ascus observed). Pseudoparaphyses not observed. Ascospores hyaline, fusiform with rounded ends, 1-septate, biseriate in the ascus, 14–20 × 4–8 μm (−x = 17.2 × 6.1 μm, n = 10), L/W = 2.8.

Specimen examined – CANADA, Ontario, Middlesex: London, on dead stems of Apocynum sp. (Apocynaceae), J. Dearness (MICH 14281, isotype).

Notes – The small-sized ascomata, bitunicate, slightly obclavate asci, lack of a pedicel, hyaline, 1-septate ascospores disagree with Botryosphaeria sensu stricto. Laestadia apocyni had been assigned to Guignardia as G. apocyni, while its 1-septate ascospores disagree with the non-septate ascospores of Guignardia (Vasyagina et al., 1987; Barr, 1993). Thus, its taxonomic status cannot be determined yet, and tentatively assigned in Dothideomycetes incertae sedis.

Sphaeria smilacinina Peck, Ann. Rep. N.Y. St. Mus. nat. Hist. 29: 62 (1878) [1876] Supplementary Figure 3

Ascomata 150-280 μm diam., pseudothecial, solitary or scattered, globose with a central ostiole, immersed or ¼ to ½ emergent, black. Peridium comprising 6–12 layers of textura angularis, outer region of dark brown cells, inner region of 2–4 layers of hyaline cells lining the locule. Asci bitunicate, broadly clavate to broadly cylindrical, 100-150 × 23–36 μm, forming among pseudoparaphyses. Pseudoparaphyses filiform, cellular, septate, 3–5 μm broad. Ascospores fusiform to narrowly fusiform, 1-septate, obviously constricted in the middle, biseriate in the ascus, 26–32 × 8–11 μm (−x = 29.6 × 9.4 μm, n = 10), L/W = 3.2.

Specimen examined – United States, Albany, on dead stems of Smilacina stellate (Liliaceae), Charles H. Peck (NYS f2818, holotype).

Notes – Sphaeria smilacinina has been assigned to Discochora as D. smilacinina (Phyllostictaceae, Botryosphaeriales) (Bissett, 1986). The 1-septate ascospore of S. smilacinina, however, disagree with the non-septate ascospores of Discochora. The immersed and scattered, ostiolate pseudothecia, clavate to broadly cylindrical bitunicate asci, 1-septate, constricted, hyaline ascospores suggest that this taxon probably resides in the Didymellaceae (Pleosporales), while its taxonomic status cannot be determined until further phylogenetic analysis is carried out on verified specimens (Zhang et al., 2012; Chen et al., 2015). Thus, we tentatively keep this species in Dothideomycetes incertae sedis.