Morphological and phylogenetic analyses reveal three new species of Apiospora in China Author Li, Shuji https://orcid.org/0009-0006-4734-8399 The Key Laboratory for Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China Author Peng, Cheng The Key Laboratory for Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China Author Yuan, Rong The Key Laboratory for Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China Author Tian, Chengming https://orcid.org/0000-0002-3352-7664 The Key Laboratory for Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China chengmt@bjfu.edu.cn text MycoKeys 2023 2023-10-20 99 297 317 http://dx.doi.org/10.3897/mycokeys.99.108384 journal article http://dx.doi.org/10.3897/mycokeys.99.108384 1314-4049-99-297 C99D3604D9ED56D79F74FB7A3277ACA0 Apiospora lophatheri S.J. Li & C.M. Tian sp. nov. Fig. 4 Type . China , Yunnan Province , Xishuangbanna Primeval Forest Park , on diseased leaves of Lophatherum gracile , 4 June 2022 , S.J. Li , holotype BJFC-S1917; ex-type living cultures CFCC 58975, CFCC 58976 . Etymology. Named after the host from which it was isolated. Description. Asexual morph : Sporulated on PDA, mycelium consisting of hyaline, smooth, branched, septate hyphae 1.0-5.2 µm in diam. (n = 20). Conidiophores reduced to conidiogenous cells. Conidiogenous cells aggregated in clusters on hyphae, hyaline to pale brown, smooth, doliiform, clavate to ampulliform, 2.2-11.9 x 2.2-4.9 µm , mean ( +/- SD): 6.4 ( +/- 2.5) x 3.4 ( +/- 0.6) µm (n = 50). Conidia globose, subglobose to lenticular, with a longitudinal germ slit, olive to dark brown, smooth to finely roughened and two or more conidia are produced on each conidiogenous cell, 5.1-8.9 x 4.6-7.7 µm , mean ( +/- SD): 6.5 ( +/- 0.8) x 5.9 ( +/- 0.7) µm , L/W = 1.0-1.4 (n = 50). Sexual morph : Undetermined. Figure 4. Apiospora lophatheri ( CFCC 58975, ex-holotype culture ) A leaf of host plant B colony on PDA C conidiomata formed in culture D conidiogenous cells giving rise to conidia E, F conidia. Scale bars: 1000 µm ( C ); 10 µm ( D-F ). Culture characteristics. On PDA, colonies flat, spreading, margin circular, thick, concentrically spreading with aerial mycelium, surface light greyish-brown, reverse tawny pigment diffused in media, mycelia white to grey and pale brown, sporulation on hyphae, reaching 9 cm in 7 days at 25 °C. Notes. Phylogenetic analysis indicated that Apiospora lophatheri is closely related to a clade comprising A. chromolaenae , A. euphorbiae , A. italicum , A. malaysiana , A. phyllostachydis , A. thailandica and A. vietnamense (Fig. 1 ). We compared the new species with phylogenetically similar taxa, based on morphological differences (Table 3 ) and base pair differences (Table 4 ). A. lophatheri can be differentiated from A. chromolaenae by its wider conidiogenous cells (2.2-11.9 x 2.2-4.9 µm vs. 6.5-12 x 1-2 µm ) (from Euphorbia sp.; collected in Zambia; Ellis (1965) ) and by 18 gene base pair differences (17/529 in ITS, 1/838 in LSU). A. lophatheri differs from A. euphorbiae by its larger olive to dark brown conidia (5.1-8.9 x 4.6-7.7 µm vs. 4-5.5 x 3-4 µm ) (from Euphorbia sp.; collected in Zambia; Ellis (1965) ), with nucleotide differences in ITS as 3/529, in LSU as 2/318, in tub2 as 22/801. A. italicum has smaller conidia (4-6 x 3-4 µm ) (from Arundo donax ; collected in Italy; Pintos et al. (2019) ) and has 125 nucleotides differences (41/552 in ITS, 2/828 in LSU, 27/432 in tef1 , 55/838 in tub2 ). Additionally, A. lophatheri is distinguished from A. malaysiana by having larger globose or subglobose conidia (5.1-8.9 x 4.6-7.7 µm vs. 5-6 x 3-4 µm ) (from Macaranga hullettii ; collected in Malaysia; Crous and Groenewald (2013) ), with 43 nucleotide differences (3/529 in ITS, 1/838 in LSU, 18/424 in tef1 , 21/801 in tub2 ). A. lophatheri differs from A. phyllostachydis by its relatively shorter conidiogenous cells (2.2-11.9 x 2.2-4.9 µm vs. 20-55 x 1.5-2.5 µm ) (from Phyllostachys heteroclada ; collected in China; Yang et al. (2019) ) and by 48 nucleotides differences (7/529 in ITS, 3/838 in LSU, 12/424 in tef1 , 26/795 in tub2 ). A. lophatheri can be differentiated from A. thailandica by having shorter conidiogenous cells (2.2-11.9 x 2.2-4.9 µm vs. 11.5-39 x 2-3.5 µm ) (from bamboo; collected in Thailand; Dai et al. (2017) ) and by 12 nucleotides differences (9/529 in ITS, 3/828 in LSU). The conidia of A. lophatheri are significantly wider and paler-coloured than those of A. vietnamense (5.1-8.9 x 4.6-7.7 µm vs. 5-6 x 3-4 µm ) (from Citrus sinensis ; collected in Vietnam; Wang et al. (2018) ) and there are 7 nucleotides differences between the two species (2/526 in ITS, 2/803 in LSU, 3/315 in tub2 ). Therefore, A. lophatheri is described as a new species, based on phylogeny and morphological comparison. Table 3. Summary of morphology of new Apiospora species and phylogenetic related species.

Species	Isolation source	Country	Conidiogenous cells (µm)	Conidia in surface view		Conidia in side view		References
				Shape	Diam (μm)	Shape	Diam (μm)
A. gaoyouense	Phragmites australis	China	1-2 x 2-3	globose to elongate ellipsoid	5-8	lenticular	4-8	Jiang et al. (2018)
A. hispanicum	Maritime sand	Spain	-	globose to ellipsoid	7.5-8.5 x 6-7.5	lenticular	6.5	Larrondo (1992)
A. locuta-pollinis	Brassica campestris	China	3-7.5 x 3-6	globose to elongate ellipsoid	8-15 x 5-9.5	-	-	Zhao et al. (2018)
A. longistroma	Bamboo	Thailand	-	asexual morph: Undetermined	-	-	-	Dai et al. (2017)
A. marii	Beach sand/ Poaceae	Spain	5-10 x 3-4.5	globose to elongate ellipsoid	8-10(-13)	lenticular	(5-)6(-8)	Crous and Groenewald (2013)
A. mediterranei	Airborn spore/ grass	Spain	-	lentiform	9-9.5 x 7.5-9	-	-	Larrondo (1992)
A. oenotherae	Oenothera biennis	China	2.0-14.2 x 1.1-4.9	globose, subglobose to lenticular	6.6-13.9 x 5.5-10.1	-	-	This study
A. piptatheri	Piptatherum miliaceum	Spain	6-27 x 2-5	globose to elongate ellips oid	6-8 x 3-5	lenticular	4.5-6	Pintos et al. (2019)
A. pseudomarii	Aristolochia debilis	China	8-13 x 2.5-5	subglobose to ellipsoid	6-9 x 4.5-6	-	-	Chen et al. (2021)
A. chromolaenae	Chromolaena odorata	Thailand	6.5-12 x 1-2	elongated, broadly fliform to ampulliform	4-6 x 4.5-6.5	-	-	Mapook et al. (2020)
A. euphorbiae	Bambusa	Bangladesh	-	circular or nearly circular	(4-)4.7(-5.5)	lenticular	(3-)3.2(-4)	Sharma et al. (2014)
A. italicum	Arundo donax	Italy	(3-)4-7(-9) x (1.5-)2-3(-5)	globose	4-6 x 3-4	lenticular	-	Pintos et al. (2019)
A. lophatheri	Lophatherum gracile	China	2.2-11.9 x 2.2-4.9	globose, subglobose to lenticular	5.1-8.9 x 4.6-7.7	-	-	This study
A. malaysiana	Macaranga hullettii	Malaysia	4-7 x 3-5	globose	5-6	lenticular	3-4	Crous and Groenewald (2013)
A. phyllostachydis	Phyllostachys heteroclada	China	20-55 x 1.5-2.5	globose to subglobose, oval or irregular	5-6 x 4-6	-	-	Yang et al. (2019)
A. thailandicum	Bamboo	Thailand	11.5-39 x 2-3.5	globose to subglobose, elongated to ellipsoidal	5-9 x 5-8	-	-	Dai et al. (2017)
A. vietnamense	Citrus sinensis	Vietnam	4-7 x 3-5	globose	5-6	lenticular	3-4	Wang et al. (2017)

Table 4. DNA base differences comparing Apiospora lophatheri sequences and sequences from related species.

Taxa	Loci	Nucleotides difference without gaps	Rates of base pair differences
A. chromolaenae	ITS	17/529 (40, 102, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122)	3.21%
	LSU	1/838 (426)	0.12%
A. euphorbiae	ITS	3/515 (26, 88, 89)	0.58%
	LSU	2/318 (146, 306)	0.63%
	tub2	22/801 (95, 96, 123, 151, 154, 163, 166, 182, 185, 193, 216, 237, 312, 347, 372, 429, 453, 454, 474, 559, 569, 574)	2.75%
A. italicum	ITS	41/552 (40, 82, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 132, 165, 177, 180, 205, 207, 213, 487, 529)	7.43%
	LSU	2/828 (406, 416)	0.24%
	tef1	27/432 (16, 18, 19, 20, 21, 22, 23, 24, 25, 27, 35, 46, 53, 60, 75, 80, 90, 102, 119, 123, 125, 172, 210, 211, 240, 248, 272)	6.25%
	tub2	55/838 (5, 29, 44, 45, 46, 92, 99, 119, 121, 122, 126, 155, 157, 171, 185, 188, 193, 194, 196, 198, 202, 297, 219, 229, 240, 265, 315, 338, 358, 363, 367, 368, 382, 384, 386, 390, 403, 407, 412, 430, 434, 454, 463, 465, 467, 480, 491, 499, 502, 556, 564, 580, 642, 756, 757)	6.56%
A. malaysiana	ITS	3/529 (40, 102, 103)	0.57%
	LSU	1/838 (426)	0.12%
	tef1	18/424 (15, 16, 19, 27, 29, 38, 52, 56, 82, 83, 91, 93, 95, 111, 115, 202, 203, 264)	4.25%
	tub2	21/801 (95, 96, 123, 151, 154, 163, 166, 182, 185, 193, 216, 237, 312, 347, 372, 429, 453, 474, 559, 569, 574)	2.62%
A. phyllostachydis	ITS	7/529 (40, 44, 85, 102, 106, 433, 500)	1.32%
	LSU	3/838 (7,8,9)	0.36%
	tef1	12/424 (16, 19, 26, 27, 51, 52, 53, 111, 197, 202, 203, 264)	2.83%
	tub2	26/795 (35, 52, 55, 84, 89, 112, 116, 147, 151, 175, 178, 186, 209, 211, 231, 329, 352, 354, 360, 462, 469, 489, 570, 572, 575, 608)	3.27%
A. thailandicum	ITS	9/529 (40, 82, 102, 107, 122, 175, 177, 183, 501)	1.70%
	LSU	3/828 (5, 416, 434)	0.36%
A. vietnamense	ITS	2/526 (37, 99)	0.38%
	LSU	2/803 (237, 391)	0.25%
	tub2	3/315 (72, 82, 87)	0.95%