Fungi Dyfrolomycetales Pleurotremataceae XuRui-FangKarunarathnaSamantha C.PhukhamsakdaChayanardDaiDong-QinElgorbanAbdallah M.SuwannarachNakarinKumlaJaturongWangXiao-YanTibprommaSaowaluckFour new species of Dothideomycetes (Ascomycota) from Pará Rubber (Heveabrasiliensis) in Yunnan Province, ChinaMycoKeys22032024103719510.3897/mycokeys.103.117580 2A8B05E7-481A-522A-B739-D3BF341077D4 Melomastia puerensis Index Fungorum number: IF901419 Facesoffungi number: FoF15195 R.F. Xu & Tibprommasp. nov.Fig. 2Etymology.

The name refers to the location “Pu’er, Yunnan, China”, where the holotype was collected.

Holotype.

ZHKU 23–0106.

Description.

Saprobic on a dead branch of Heveabrasiliensis. Sexual morph: Ascomata 260–720 μm high, 225–850 μm diam. (x‾ = 540 × 520 μm, n = 10), visible as black dots on the host surface, solitary or gregarious, immersed to slightly erumpent, subglobose or pyriform, carbonaceous, dark brown to black, ostiolate, papillate. Ostioles 205–220 × 195–258 µm (x‾ = 233 × 207 μm, n = 5), central, carbonaceous, dark brown to black. Peridium 40–120 µm wide, two-layered, outer layer, thick, carbonaceous, inner layer composed of several layers, brown to pale brown cells of textura angularis. Hamathecium comprises 2–4.5 μm wide, filiform, unbranched, hyaline, aseptate, guttulate, pseudoparaphyses, longer than asci. Asci 175–205 × 6–10 μm (x‾ = 190 × 8, n = 15), 8-spored, hyaline, bitunicate, cylindrical, flexuous, apically obtuse, with an ocular chamber, smooth-walled, short pedicellate. Ascospores 20–30 × 5–8 μm (x‾ = 24 × 7, n = 30), uniseriate, hyaline, fusiform, obtuse or conical ends, narrow towards the apex, 3-septate, constricted at the central septum, with guttulate in each cell. Asexual morph: Undetermined.

10.3897/mycokeys.103.117580.figure2E87DE527-7D71-5A91-8C85-9FAE846C1270

Melomastiapuerensis (ZHKU 23–0106, holotype) a–c appearance of ascomata on host surface d vertical section of an ascoma e vertical section of ostiole f section of peridium g hamathecium h–l asci m–q ascospores r asci stained in Lugol’s iodine s germinated ascospore t, u colonies on PDA (t-front and u-reverse views). Scale bars: 100 µm (d–f); 50 µm (g–l); 10 µm (m–q); 30 µm (s).

https://binary.pensoft.net/fig/1007721Culture characteristics.

Colonies on PDA that grow at 28 °C, flat, rough surface, entire edges, culture from above, brownish-grey, forming zonate grey, reverse dark brown, brown at the edge, turning reddish-brown.

Material examined.

China, Yunnan Province, Pu'er on a dead branch of Heveabrasiliensis, 16 September 2021, Rui-Fang Xu, XPER–14 (ZHKU 23–0106, holotype); ex-type ZHKUCC 23–0802, ZHKUCC 23–0803.

GenBank numbers.

ZHKUCC 23–0802 = ITS: OR941077, LSU: OR922309, SSU: OR922340, tef1-α: OR966284; ZHKUCC 23–0803 = ITS: OR941078, LSU: OR922310, SSU: OR922341, tef1-α: OR966285.

Notes.

The phylogenetic analyses showed that Melomastiapuerensis clustered basal to M.distoseptata, M.fusispora, M.maomingensis, M.oleae, M.pyriformis, M.thamplaensis and M.winteri with 99% MP, 1.00 PP support (Fig. 1). We compared the morphology of our collection with closely-related species and the differences are mentioned in Table 2. Our collection has slight differences from other closely-related species by having larger ascomata and wider peridium, but the phylogenetic tree shows that they are different species (Fig. 1, Table 2). Therefore, we introduce M.puerensis as a new species, based on morphology and phylogenetic analyses.

Morphological comparison of M.puerensis and closely-related species viz. M.distoseptata, M.fusispora, M.maomingensis, M.oleae, M.pyriformis, M.thamplaensis and M.winteri.

SpeciesAscomataPeridiumPseudoparaphysesAsciAscosporesReferences
M.distoseptata 550–630 × 450–600 μm, perithecial, immersed, erumpent neck with pseudoparaphyses, clypeate40 μm, with two strata, outer thick, and inner brown and hyaline cells of textura angularis to epidermoidea cells1.8–2.1 μm, flamentous, septate, unbranched, dense, longer than asci126.7–146.2 × 4.7–6.3 μm, apical ends obtuse, short pedicellate19.7–24.9 × 4.3–5 μm, fusoid, obtuse ends, apical ends slightly bent Hongsanan et al. (2020)
M.fusispora 432–624 × 527–618 μm, cone-shaped structures on the host surface, immersed to erumpent through host tissue, pyriform25.5–61.5 µm, two-layered, outer layer of cells of textura intricata, inner layer of textura angularis2–2.6 µm, dense, filiform, unbranched, hyaline, aseptate200–231 × 7.6–9.2 µm, slightly flexuous, apically round, with well-developed ocular chamber, cylindrical pedicellate27.5–32 × 6.5–7.5 µm, fusiform, with rounded to acute ends, narrow towards apex, constricted at the central septum, surrounded by an irregular and thin gelatinous sheath Li et al. (2022)
M.maomingensis 300–550 µm high × 250–500 µm diam., solitary, semi-immersed to immersed, visible on the host surface as black, obvious, raised spots, black, uni-loculate, globose to subglobose35–100 µm wide, comprising dense, thick, brown to dark brown cells of textura angularis, fusion with host tissue1.5–3.5 µm wide, comprising numerous, filamentous, hyaline, septate, sometimes branched, longer than asci, attached at the base and between the asci175–195 × 7–9 µm, cylindrical pedicel, rounded in apex, J-(23–)24.5–29 × 6–8 µm, fusiform with acute ends, constricted at the septum, with a large guttule in each cell when mature Du et al. (2024)
M.oleae 410–440 × 493–520 µm, cone-shaped structures on host surface, semi-immersed, globose to compressed globose54–65 µm, two-layered, outer thick and inner composed of 5–6 layers of textura angularis to textura prismatica2–2.5 µm, dense, filiform, unbranched, aseptate209–237 × 7.5–9 µm, slightly flexuous, apically rounded with ocular chamber, cylindrical pedicellate28–34 × 6–7 µm, fusiform with obtuse ends, slightly constricted at the septa Li et al. (2022)
M.puerensis 260–720 × 225–850 μm, black dot on the host surface, immersed to erumpent to superficial, pyriform 40–120 µm, two-layered, outer thick, carbonaceous, inner composed of several layers, pale brown to brown cells of textura angularis 2–4.5 μm, filiform, unbranched, guttulate, pseudoparaphyses, longer than asci 175–205 × 6–10 μm, flexuous, apical ends obtuse, with ocular chamber, smooth-walled, short pedicellate 20–30 × 5–8 μm, fusiform, obtuse or conical ends, narrow towards apex, constricted at the central septum, with guttules in each cell This study
M.pyriformis 330–640 × 275–420 μm, erumpent to superficial when mature, pyriform, papillate, ostiolate20–50 μm, thin at the base and become thick towards sides, comprised of brown, thick-walled, cells of textura intricata in sides; and thin-walled, pale brown, cells of textura angularis in base1.8–2.5 μm wide, dense, filiform, unbranched, septate, anastomosing between and above the asci135–160 × 5.5–7.5 μm, fissitunicate, apically round, with an indistinct ocular chamber, short pedicellate20–25 × 4.5–7 μm, , fusiform with acute ends, not constricted at the septa, with guttules in each cell Kularathnage et al. (2023)
M.thamplaensis 550–630 × 450–600 μm, black spots on the host surface, immersed, clypeate, subglobose to obpyriform, some with a broad, flattened base14–49 μm, composed of three strata, an outer stratum, dense, amorphous, thick-walled cells fusing with host tissue, a middle layer of thick-walled, black cells of textura angularis and an inner layer of thin-walled black cells of textura angularis1.8–2.1 μm, attached at the base and between the asci, embedded in a gelatinous matrix126.7–146.2 × 4.7–6.3 µm, long cylindrical, short-pedicellate, apically rounded with an obvious apical ring19.7–24.9 × 4.3–5 μm, fusiform with acute angular ends, constricted at the septum, smooth-walled, containing several guttules when young Zhang et al. (2017)
M.winteri 340–365 × 364–410 µm, semi-immersed to immersed, globose55–62.5 µm, two-layered, outer thick, and inner composed of 3–4 layers of hyaline to lightly brown cells of textura angularis to textura prismatica1.5–3.5 µm, dense, filiform, unbranched, septate165–189 × 7–8.5 µm, slightly flexuous, apically round, with a distinct ocular chamber, cylindrical pedicellate25–30 × 5–6.5 µm, partially overlapping, fusiform with acute ends, deeply constricted at the median septum Li et al. (2022)
<tp:taxon-name><tp:taxon-name-part taxon-name-part-type="order">Pleosporales</tp:taxon-name-part></tp:taxon-name> Luttrell ex M.E. Barr10.3897/mycokeys.103.117580.figure1ABFD69FB-55C9-560C-A822-159F918AFDDE

Phylogram generated from Maximum Likelihood analysis, based on combined LSU, SSU and tef1-α sequence data of 41 taxa, which comprised 2836 base pairs (LSU = 902 bp, SSU = 1031 bp, tef1-α = 903 bp). The best scoring RAxML tree with a final likelihood value of -14798.632437 is presented. The matrix had 1013 distinct alignment patterns, with 24.90% of undetermined characters or gaps. Estimated base frequencies were as follows: A = 0.241740, C = 0.258134, G = 0.292403, T = 0.207722; substitution rates: AC = 0.834723, AG = 2.021967, AT = 1.126143, CG = 1.032150, CT = 7.231944, GT = 1.000000; gamma distribution shape parameter α = 0.320795. Bootstrap support values for ML equal to or greater than 60% and Bayesian Inference analysis values equal to or greater than 0.90 PP are labelled at each node. The tree is rooted with Anisomeridiumphaeospermum (MPN539) and A.ubianum (MPN94). Related sequences were collected following Li et al. (2022), Kularathnage et al. (2023) and Dong et al. (2023). The new isolates are indicated in red and the ex-type strains are in bold. Group A indicates the taxa used to compare the morphology with our new species (Melomastiapuerensis).

https://binary.pensoft.net/fig/1007720LiW-LMaharachchikumburaSSNCheewangkoonRLiuJ-K (2022) Reassessment of Dyfrolomyces and four new species of Melomastia from Olive (Oleaeuropaea) in Sichuan Province, China.Journal of Fungi (Basel, Switzerland)8(1): 76. https://doi.org/10.3390/jof8010076KularathnageNTennakoonDZhuXZhouJSuBXieYChenQCalabonMKirkPSenanayakeIDoilomMXuBDongWSongJ (2023) Reinstating Dyfrolomyces and introducing Melomastiapyriformis sp. nov. (Pleurotremataceae, Dyfrolomycetales) from Guangdong Province, China.Current Research in Environmental & Applied Mycology13(1): 13. https://doi.org/10.5943/cream/13/1/16DongWHydeKDJeewonRLiaoCFZhaoHJKularathnageNDLiHYangYHPemDShuYXGafforovYManawasingheISDoilomM (2023) Mycosphere notes 449–468: Saprobic and endophytic fungi in China, Thailand, and Uzbekistan.Mycosphere: Journal of Fungal Biology14(1): 2208–2262.HongsananSHydeKDPhookamsakRWanasingheDNMcKenzieEHCSarmaVVLückingRBoonmeeSBhatJDLiuN-GTennakoonDSPemDKarunarathnaAJiangS-HJonesGEBPhillipsAJLManawasingheISTibprommaSJayasiriSCSandamaliDJayawardenaRSWijayawardeneNNEkanayakaAHJeewonRLuY-ZPhukhamsakdaCDissanayakeAJZengX-YLuoZ-LTianQThambugalaKMDaiDSamarakoonMCChethanaKWTErtzDDoilomMLiuJ-KPérez-OrtegaSSuijaASenwannaCWijesingheSNNiranjanMZhangS-NAriyawansaHAJiangH-BZhangJ-FNorphanphounCde SilvaNIThiyagarajaVZhangHBezerraJDPMiranda-GonzálezRAptrootAKashiwadaniHHarishchandraDSérusiauxEAbeywickramaPDBaoD-FDevadathaBWuH-XMoonKHGueidanCSchummFBundhunDMapookAMonkaiJBhunjunCSChomnuntiPSuetrongSChaiwanNDayarathneMCYangJRathnayakaARXuJ-CZhengJLiuGFengYXieN (2020) Refined families of Dothideomycetes: Orders and families incertae sedis in Dothideomycetes.Fungal Diversity105(1): 17–318. https://doi.org/10.1007/s13225-020-00462-6DuTYTibprommaSHydeKDDaiD-QMapookAZhangG-QStephensonSLSuwannarachNElgorbanAMRajeshkumarKCMaharachchikumburaSSNLiQKarunarathnaSC (2024) The polyphasic approach reveals twelve novel ascomycota taxa from terrestrial agarwood-producing trees. [Not published]ZhangJLiuJ-KHydeKDChenY-YLiuY-XLiuZ-Y (2017) Two new species of Dyfrolomyces (Dyfrolomycetaceae, Dothideomycetes) from karst landforms. Phytotaxa 313: 267. https://doi.org/10.11646/phytotaxa.313.3.4