Morphophylogenetic evidence reveals four new fungal species within Tetraplosphaeriaceae (Pleosporales, Ascomycota) from tropical and subtropical forest in China Author Tang, Xia https://orcid.org/0000-0003-2705-604X Engineering and Research Center for Southwest Biopharmaceutical Resource of National Education Ministry of China, Guizhou University, Guiyang, 550025, Guizhou Province, China & Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand Author Jeewon, Rajesh https://orcid.org/0000-0002-8563-957X School of Science, Mae Fah Luang University, Chiang Rai, 57100, Thailand & Department of Health Sciences, Faculty of Medicine and Health Sciences, University of Mauritius, Reduit, Mauritius r.jeewon@uom.ac.mu Author Lu, Yong-Zhong https://orcid.org/0000-0002-1033-5782 Engineering and Research Center for Southwest Biopharmaceutical Resource of National Education Ministry of China, Guizhou University, Guiyang, 550025, Guizhou Province, China & Department of Zoology, College of Science, King Saud University, P. O. Box 2455, Riyadh 11451, Saudi Arabia Author Alrefaei, Abdulwahed Fahad https://orcid.org/0000-0002-3761-6656 Department of Health Sciences, Faculty of Medicine and Health Sciences, University of Mauritius, Reduit, Mauritius Author Jayawardena, Ruvishika S. https://orcid.org/0000-0001-7702-4885 Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand Author Xu, Rong-Ju https://orcid.org/0000-0002-3968-8442 Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand Author Ma, Jian https://orcid.org/0009-0008-1291-640X Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai, 57100, Thailand Author Chen, Xue-Mei https://orcid.org/0009-0004-8631-0735 School of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang, Guizhou Province 550003, China Author Kang, Ji-Chuan https://orcid.org/0000-0002-6294-5793 Engineering and Research Center for Southwest Biopharmaceutical Resource of National Education Ministry of China, Guizhou University, Guiyang, 550025, Guizhou Province, China jckang@gzu.edu.cn text MycoKeys 2023 2023-12-06 100 171 204 http://dx.doi.org/10.3897/mycokeys.100.113141 journal article http://dx.doi.org/10.3897/mycokeys.100.113141 1314-4049-100-171 C062A9B98A2256E8B6D8E18531E0C50D Tetraploa hainanensis X. Tang, Jayaward., R. Jeewon & J.C. Kang sp. nov. Figs 6 , 7 Etymology. The specific epithet ' hainanensis ' refers to the place where the fungus was collected, Hainan Province, China. Holotype. GZAAS 23-0603. Description. Saprobic on unidentified decaying wood in forest. Teleomorph morph Not observed. Anamorph Hyphomycetous. Colonies effuse, gregarious on host substrate, brown to dark brown. Mycelium semi-immersed or immersed, pale brown, branched, septate. Conidiophores absent. Conidiogenous cells integrated, monoblastic, determinate. Conidia 30-46 x 18-36 μm (x̅ = 38 x 27 μm , n = 20), cylindrical with obtuse ends, pale brown to brown, verrucose, composed of four columns of cells, sometimes five columns of cells, 4-5-septate in each column, smooth, mostly with four apical appendages, some with one or two or five appendages. Appendages 52-209 x 3-6 μm (x̅ = 140 x 4 μm , n = 20) cylindrical, solitary, unbranched, guttulate, septate, wide at the base, divergent, pale brown to brown, 5-16-septate, straight or slightly flexuous, smooth-walled. Figure 6. Tetraploa hainanensis (GZAAS 23-0603, holotype) a, b colonies on natural substrates c-n conidia bearing 1-5 appendages o germinating conidium p colony on PDA (from reverse) q colony on PDA (from front). Scale bars 20 μm ( c ); 50 μm ( d-g ); 100 μm ( h-o ). Culture characteristics. Conidia germinated from both ends on PDA and incubated at room temperature (25 ˚C). Colonies circular, cottony, flat, slightly grey with an entire margin, contain a circular white mycelium in the centre. The reverse side is a pale brown in the centre that gradually extends outwards while the colour changes to pale grey, with a brown margin and no pigment. Figure 7. Tetraploa hainanensis (GZAAS 23-0604, paratype) a colonies on decay wood b, c colonies on natural substrates d-p conidia bearing 1-4 appendages q germinating conidium r colony on PDA (from front) s colony on PDA (from reverse). Scale bars: 20 μm ( d-g ); 100 μm (h-l, o, q ); 50 μm ( m, n, p ). Material examined. China , Hainan Province , Wuzhishan City , Wuzhishan National Nature Reserve , on unidentified decaying wood, 25 September 2021 , Zili Li, WZS59 (GZAAS 23-0603, holotype ), ex-type culture GZCC 23-0601; WZS66.2 (GZAAS 23-0604, paratype ), culture GZCC 23-0602 . Notes. Tetraploa hainanensis is morphologically similar to T. pseudoaristata . However, Tetraploa hainanensis can be distinguished from T. pseudoaristata in having larger conidia (30.5-46 x 18-36 μm vs. 22-31 x 15-20 µm ) with four columns of cells, sometimes five columns of cells, and longer appendages (52-209 x 3-6 μm vs. 23-107 x 2-5 μm ), commonly four in number, sometimes five. Based on the phylogenetic analysis, two of our Tetraploa collections which share similar morphology clustered together with high support (ML = 100, and BPP = 1 (Fig. 1 )). The base pair differences between the two strains were: LSU = 0.1% (1/806), ITS = 0% (0/516), and tub2 = 0% (1/633), respectively. Therefore, we considered them as the same species according to the guidelines for species delineation proposed by Jeewon and Hyde (2016) . Tetraploa hainanensis forms a distinct lineage but close to T. yakushimensis and T. tetraploa . However, Tetraploa hainanensis differs from T. yakushimensis by having four or five columns and appendages, while T. yakushimensis has only four columns and appendages; Tetraploa hainanensis differs from T. tetraploa in having four or five columns and shorter appendages (52-209 x 3-6 μm vs. 263-350 x 2-3 μm ), while T. tetraploa has only four columns and slender appendages. The comparison of pairwise nucleotide showed that Tetraploa hainanensis is different from T. yakushimensis in 31/620 bp (3%) in LSU, 7/814 (0.98%) in ITS, and 87/450 (19%) in tub2 and Tetraploa hainanensis is different from T. tetraploa in 31/620 bp (3%) in LSU, 7/814 (0.98%) in ITS, and 87/450 (19%) in tub2 . Based on the combination of morphological characters and multigene phylogeny, we describe Tetraploa hainanensis herein as a distinct species according to the guidelines of Jeewon and Hyde (2016) and Chethana et al. (2021) .