Short Communication New accounts on Hypoxylaceae and Xylariaceae from Brazil

This work aims to bring new reports of Hypoxylaceae and Xylariaceae from Brazil. The collections were performed in cocoa plantations in Ilhéus, Bahia, Brazil. Six new occurrences of Hypoxylaceae and two Xylariaceae species are reported to Brazil, Northeast and/or Bahia. A dichotomous key to the species treated

Southern Bahia is a region of huge importance and considered a hotspot of biodiversity due to some well preserved fragments of Atlantic Rain Forest (ARF) within this area, presenting high levels of diversity of plants and animals. Despite that, there is still a need for studies with other organisms such as fungi, which, except for pathogens, are poorly known in Bahia, even with their huge importance as decomposers of organic matter, endophytes and mycorrhizal symbionts with plants. Cocoa culture is one of the pillars of economy in Bahia, and the fact the cocoa have been cultivated inside or nearby ARF fragments, denotes these areas might be of great importance to biodiversity. Cassano et al. (2009) and Faria & Baumgarten (2007), e.g., highlighted the importance of Shaded Cocoa Plantations (SCP) as refuge for species from ARF such as mammals, including threatened species. Even though, studies with fungi in these areas are scarce or focused primarily on pathogenic fungi. Xylariaceae is one of the iconic families of Ascomycota, being highly diverse through the Tropics. In Brazil, the xylariaceous fungi are among the richest in number of species, and despite it is still ranked as Order in the Flora do Brasil database (BFG 2018), the family Xylariaceae is probably the most diverse, with the genus Xylaria, e.g., having more than 100 species known so far to Brazil (Maia et al. 2015). The diversity of morphology on the Xylariaceae, lead to attempts to divide the family in the Subfamilies Xylarioideae, Hypoxyloideae and Thamnomycetoideae, which has been proposed by Dennis (1961), though not supported, as it was invalidly erected (Stadler et al. 2013). Moreover the third subfamily, Thamnomycetoideae, was as well not validated since the genus Thamnomyces is closely related to Daldinia, supporting its affinity with the Hypoxylaceae (Stadler et al. 2010). Some studies (Hsieh et al. 2005(Hsieh et al. , 2010Wendt et al. 2018) reinforced the need of a rearrangement of the Xylariaceae based on phylogeny and chemotaxonomy, with the last being the focus of studies from Stadler (2011), Stadler et al. (2014 and Kuhnert et al. (2017), corroborating the importance Rodriguésia 71: e03012018. 2020 of more studies to a better comprehension and resolution of the subgroups within the Xylariaceae. The diversity of xylariaceous fungi is poorly known in Brazil probably due to its large territory, which becomes an issue to explore farther locations. A few studies such as Pereira et al. (2008aPereira et al. ( , b, 2009Pereira et al. ( , 2010, Cruz & Cortez (2015a, b, 2016 and Trierveiler-Pereira et al. (2009)  The specimens were collected randomly, with substrate whenever possible, and packed in paper bags, tagged with location and date of collection, then taken to the Phytopathology and Nematology Laboratory at UESC and dried at 50 °C for 24 hours. To identification of species, morphological characters of stromata, KOH-extractable pigments, perithecia, asci and ascospores were analyzed. The specimens were deposited at the Tropical Fungarium (TFB) in UESC. Due to the most recent adjustments in the Xylariaceae, we decided to follow the modifications proposed by Wendt et al. (2018)  Description according to Wendt et al. (2018): stromata varying from erect to effused-pulvinate; solitary or confluent; surface colored or black, pruinose or polished, planar or with perithecial mounds; waxy or carbonaceous tissue immediately beneath surface and between perithecia, with or without KOH-extractable pigments; the tissue below the perithecial layer inconspicuous, conspicuous, or massive, most often dark brown to black, persistent or loculate; Some genera presents peculiar features, such as alternating zones under perithecial layer, or hollow and filled with liquid, observed in the Daldinia and Entonaema, respectively. Perithecia embedded in the stroma, spherical, obovoid, tubular, or long tubular, monostichous, with or without carbonaceous stromatal material surrounding individual perithecia. Ostioles can be umbilicate, at the same level or higher than the level of stromatal surface, with or without discs. The asci are typically eight-spored, cylindrical, stipitate; with apical ring discoid, amyloid or infrequently inamyloid, distinct, highly reduced, or apparently lacking. Ascospores brown, ellipsoid or shorty fusoid, inequilateral, slightly inequilateral or nearly equilateral, with acute, narrowly rounded, or broadly rounded ends, in most species bearing a germ slit; perispore dehiscent or indehiscent in 10% KOH. The anamorph Nodulisporiumlike with branching patterns varying from "regular" nodulisporium, periconiella, virgariella or sporothrix-like.  Fig. 1a-d Stromata turbinate, brown vinaceous, blackened in age, 3.2 cm diam, KOH-extractable pigment yellowish, becoming vinaceous after a few minutes, ostioles inconspicuous. Perithecia tubular, 1-2 × 0.3-0.5 mm, tissue beneath perithecia fibrous, composed of alternating zones, the darker zones brown, 0.3-0.4 mm thick, pithy to woody, the lighter zones grayish, 0.3-0.6 mm thick, pithy to woody. Asci not seen. Ascospores unicellular, brown to dark brown, ellipsoid-inequilateral, with narrowly rounded ends, 12-13 × 5-6 μm, with straight germ slit spore-length, perispore dehiscent in 10% KOH. Daldinia starbaeckii was previously collected in Bahia by Camille Torrend in 1915, however was identified as D. eschscholtzii (see Child 1932 andStadler et al. 2014), and apparently there were no other reports for the species after that. This might be probably due to the fact that many specimens collected in the country are misidentified as D. eschscholtzii. Daldinia starbaeckii differs from D. eschscholtzii for yielding yellowish olivaceous pigment in 10% KOH, instead of purplish. Stadler et al. (2014) mentioned that D. starbaeckii has ascospores smaller than those of D. eschscholtzii, although we have not found significant difference in ascospore size. The previous authors mentioned yet that the specimens collected by Starbäck in 1901 in Brazil as D. concentrica var. eschscholtzii corresponded very much with this species, considering the teleomorphic features and stromatal metabolites. The fact that this species co-occur with D. eschscholtzii in the Americas may have caused them not to be properly identified back then, especially the specimens which pigments were not taken into account.

Key to
Known distribution: the Americas and Africa. Rogers, Mycologia Memoirs, 20: 99 (1996). Fig. 1e-h Stromata effused-pulvinate, surface plane, apricot, reddish granules immediately beneath the surface and between perithecia, ostioles lower than stromatal surface, KOH-extractable pigments orange to rust, grayish to black tissue beneath perithecia up to 2 mm thick. Perithecia tubular, 0.7-1 × 0.3-0.4 mm. Asci damaged. Ascospores brown to dark brown, unicellular, ellipsoid nearly equilateral, narrowly rounded ends, some slightly citriform, 11-12(-13) × 6-7 μm, with germ slit almost to spore-length, perispore indehiscent in 10% KOH. This specimen was collected on dead tree of Citrus sp., what seems to be the first report on this host. Apparently there is no evidence of Hypoxylon cinnabarinum exhibiting pathogenic behavior so far, but the typical saprophytic behavior observed in most species of Hypoxylaceae and Xylariaceae, or perhaps a weak pathogen, invading the host when it is already affected by a previous pathogen or is under abiotic stress conditions, which is as well not unusual in both Families. According to Ju & Rogers (1996) H. cinnabarinum is closely related to H. crocopeplum, with the former having ascospores with perispore usually indehiscent in 10% KOH, which was corroborated with our specimen.
potent Nodulisporic-acids with insecticidal effects from its asexual morph. , reported this species from Martinique as being the sexual morph of the asexual morph isolated by Bills et al. (2012). Known distribution: probably Pantropical.
1.6. Phylacia bomba (Mont.) Pat. in Duss, Crypt. des Antilles, Champ., 74 (1903). Fig. 2e-g Stromata gregarious, erumpent, sessile, black, carbonaceous, hemispherical, 2.5-8 mm diam, older stromata with large hole at the apex communicating a single locule containing the mass of ascospores with exterior, perithecia not seen, asci not seen, ascospores oblong, translucent yellowish brown, 9-11 × 5-6 μm, germ slit inconspicuous. Phylacia is quite a different genus, presenting some aberrant features for a typical xylariaceous fungus, such as the unusually subglobose to globose asci, which tend to deliquesce prematurely. Rodrigues & Samuels (1989), mentioned the stromata of Phylacia species seems to be cleistothecial, as it do not present the typical ostiolar canal observed in perithecial stromata, releasing the ascospores through a large hole observed on the top of mature stromata. The genus share some features with Camillea, such as its translucent ascospores apparently lacking germ slit and pigments not extractable in 10% KOH. A specimen was collected in Bahia in 2006, which is deposited in the CEPLAC Herbarium (CEPEC-Fungi 2182) but we have found no report of the species to Bahia or Northeast.

Xylariaceae
Known distribution: Tropical and Subtropical.
We have brought in this study the first report of Daldinia starbaeckii, Hypoxylon pulicicidum and Nemania immersidiscus to Brazil; Hypoxylon cinnabarinum, H. haematostroma and Phylacia bomba to Northeast, Hypomontagnella monticulosa and Xylaria schweinitzii to Bahia. Hypoxylon was the most dominant genus in all sampled areas, colonizing different substrates. Xylaria has a greater number of species and is found quite easily specially in high humidity rates. Despite that, we focused primarily on the Hypoxylaceae, due to the diverse morphological features of some Xylaria species, which makes identification more difficult, with X. schweinitzii being the only species reported in this work.  observed that some species of Phylacia, including P. bomba, yielded KOHextractable pigments and the ascospores presents an inconspicuous germ slit less than spore-length, characters we did not observed on the specimens we collected, with both being important diagnostic aspects on the majority of Hypoxylaceae species.
Although the collections were performed in cocoa plantations, the only species we collected in Theobroma cacao were Hypomontagnella monticulosa and Hypoxylon pulicicidum which does not necessarily means the Hypoxylaceae and Xylariaceae fungi do not colonize this host. Costa (2008) isolated some species of Xylaria as endophytic from T. cacao, although this kind of interaction between the xylariaceous fungi and plants is hitherto poorly comprehended. Several species of Hypoxylaceae and Xylariaceae are deposited in many herbaria in Brazil and Bahia, such as the CEPEC-Fungi, even though many specimens are identified until generic level, what becomes an issue to assess the diversity of these fungi in Brazil. This uphold the need of more studies towards the comprehension of diversity of Hypoxylaceae and Xylariaceae fungi in Brazil, particularly in North and Northeast regions, from where new species, even genera, of Ascomycota and Basidiomycota have been described in the last decade, evidencing this region can be promising for fungal diversity.