A New Lichen-Forming Fungus, Aspicilia humida, from a Forested Wetland in South Korea, with a Taxonomic Key for Aspicilioid Species of Korea

Figures & data

Figure 1. Specific collection site for the new species Aspicilia humida (black star mark).

Table 1. Species list and DNA sequence information employed for phylogenetic analysis.

No.	Species	ID (ITS)	ID (mtSSU)	Voucher
1	Aspicilia abbasiana	KM609324		WDLC-20111154
2	Aspicilia aschabadensis	GU289916		Borisova1934 (LE)
3	Aspicilia berntii	EU502747		Nordin 6392 (UPS)
4	Aspicilia bicensis	KU341407		Anderson 16123 (NY)
5	Aspicilia blastidiata	KX129963		AGP20111009-01
6	Aspicilia brucei	JF703111		UPS:Owe-Larsson 9147
7	Aspicilia calcitrapa	JF703113		MARSSJ:Roux 24309
8	Aspicilia cinerea	EU057899	HM060695	Hermansson 13275 (UPS)
9	Aspicilia cinerea	HQ406799	HM060696	Nordin 5542 (UPS)
10	Aspicilia cinerea	HQ650637	DQ986890	AFTOL-ID 647
11	Aspicilia coronata	EU057901		Lattman 13.V.2006 (UPS)
12	Aspicilia desertorum	HQ406802	HM060689	Owe-Larsson 9814 (UPS)
13	Aspicilia dudinensis	EU057906	HM060710	Nordin 6036 (UPS)
14	Aspicilia dudinensis	MN906265		McCune 36017 (OSC)
15	Aspicilia dudinensis		HM060719	UPS:Nordin 5971
16	Aspicilia epiglypta	MH248866	MH248867	Nordin 7037 (UPS)
17	Aspicilia fluviatilis	HQ259264		UPS:Nordin 6188
18	Aspicilia granulosa	HQ259265		UPS:Nordin 6174
19	Aspicilia humida	MW832805	MW832823	BDNA-L-0000703
20	Aspicilia humida	MW832806	MW832824	BDNA-L-0000711
21	Aspicilia indissimilis	EU057909	HM060708	Nordin 5943 (UPS)
22	Aspicilia laevata	EU057910	HM060692	Tibell 23659 (UPS)
23	Aspicilia prestensis	JF703122		UPS:Roux 24883a
24	Aspicilia pseudoabbasiana	KY249599		KoLRI 018579
25	Aspicilia pseudovulcanica	KY249601		KoLRI 012338
26	Aspicilia pseudovulcanica	KY249602		KoLRI 012420
27	Aspicilia pseudovulcanica	KY249603		KoLRI 023829
28	Aspicilia subdepressa	JF703123		MARSSJ:Roux 24653
29	Aspicilia subepiglypta	KY249607		KoLRI 012535_2
30	Aspicilia subfarinosa	MN989235		Sipman & Raus 63720
31	Aspicilia subgeographica	KY249611		KoLRI 018704
32	Aspicilia subgoettweigensis	KY249616		KoLRI 007090
33	Aspicilia submamillata	KY249620		KoLRI 011146
34	Aspicilia subradians	HQ259267		UPS:Nordin 5984
35	Aspicilia verrucigera	EU057939		Tibell 22669 (UPS)
36	Aspiciliella intermutans	MH248863	MH248869	Roux 25790 (CR)
37	Circinaria affinis	HQ389194		Abbas 20081364 (H)
38	Circinaria caesiocinerea	HQ650636	DQ986892	AFTOL-ID 653
39	Circinaria calcarea	EU057898		Nordin 5888 (UPS)
40	Circinaria calcarea		AY853310	Wedin 6500 (UPS)
41	Circinaria cerebroides	JQ797534		Ringel 5138 (H)
42	Circinaria contorta	HQ650638	DQ986876	AFTOL-ID 1358
43	Circinaria cupreogrisea	EU057903		Nordin 6046 (UPS)
44	Circinaria esculenta	HQ406803		Owe-Larsson 9824 (UPS)
45	Circinaria esculenta	JQ797511	JQ797485	Owe-Larsson 9796 (UPS)
46	Circinaria fruticulosa	HQ389199		Vondrak 5188 (GBFS)
47	Circinaria fruticulosa		KC020253	ASPFRU 2361
48	Circinaria gibbosa	EU057908	HM060702	Nordin 5878 (UPS)
49	Circinaria gyrosa	JQ797528	JQ797487	Sohrabi 10401 A
50	Circinaria hispida	HQ171233		Sohrabi 15099
51	Circinaria jussuffii	JQ797518		Esnault 2033 (GZU)
52	Circinaria lacunosa	JQ797517		Abbas 940003 (H)
53	Circinaria leprosescens	EU057911	HM060711	Nordin 5906 (UPS)
54	Circinaria sphaerothallina	JQ797545		Sohrabi 9369
55	Lobothallia alphoplaca	JQ797516	JQ797480	Sohrabi 3677
56	Lobothallia alphoplaca	KT456207	KT456211	SK A20
57	Lobothallia melanaspis	JF825524		Owe-Larsson 8943a (UPS)
58	Lobothallia melanaspis		HM060688	UPS:Nordin 6622
59	Lobothallia praeradiosa	MK347501	MK348229	UFU L-1264
60	Lobothallia recedens	HQ406807		Nordin 6035 (UPS)
61	Lobothallia recedens		HM060724	UPS:Nordin 6582
62	Oxneriaria dendroplaca	HQ259259	HM060706	UPS:Nordin 5952
63	Oxneriaria mashiginensis	EU057912	HM060694	Nordin 5790 (UPS)
64	Oxneriaria permutata	EU057918	HM060709	Nordin 6027 (UPS)
65	Oxneriaria rivulicola	EU057922	HM060715	Nordin 5957 (UPS)
66	Oxneriaria supertegens	EU057936	HM060704	Owe-Larsson 9002 (UPS)
67	Oxneriaria verruculosa	EU057940	HM060703	Owe-Larsson 9007 (UPS)
68	Oxneriaria virginea	HQ259270		UPS:Nordin 6017a
69	Sagedia mastrucata	EU057913	HM060698	Nordin 5708 (UPS)
70	Sagedia nunatakkorum	KT630250		Malicek-228
71	Sagedia simoensis	EU057926	HM060701	Owe-Larsson 9000 (UPS)
72	Sagedia zonata	EU057946	HM060700	Owe-Larsson 8942 (UPS)
73	Teuvoa tibetica	GU289915		Obermayer 04386
	Overall	68	34

DNA sequences for the new species Aspicilia humida (in bold) were generated in this study. All others were obtained from GenBank. The species names are followed by GenBank accession numbers and voucher information. ITS, internal transcribed spacer; mtSSU, mitochondrial small subunit; Voucher, voucher information.

Figure 2. Phylogenetic relationship among available species in the genus Aspicilia based on a maximum likelihood analysis of the dataset of ITS sequences. The tree was rooted with six Lobothallia and Teuvoa sequences. Maximum likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Aspicilia humida is presented in bold, and all species names are followed by the Genbank accession numbers. Reference Table 1 provides the species related to the specific GenBank accession numbers and voucher information.

Figure 3. Phylogenetic relationships among available species in the genus Aspicilia based on a maximum likelihood analysis of the dataset of the mitochondrial small subunit (mtSSU) sequences. The tree was rooted with five Lobothallia sequences. Maximum-likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Aspicilia humida is presented in bold, and all species names are followed by the GenBank accession numbers. Reference Table 1 provides the species related to the specific GenBank accession numbers and voucher information.

Figure 4. Aspicilia humida (BDNA-L-0000703, holotype for A–K; BDNA-L-0000711, paratype for L & M) in morphology. (A–D): Habitus and apothecia emerging single to several per an areole; (E): Adnate apothecia without constriction at the base in section; (F): Epihymenium in olive-brown pigment; (G–J): Clavate asci with eight spores; (K): Ellipsoid or globose ascospores with no septation; (L): Immersed pycnidia; (M): Thread-like pycnoconidia. Bars: A–D 1 mm; E 200 μm; F 50 μm; G–K 10 μm; L 100 μm; M 10 μm.

Table 2. Comparison of Aspicilia humida with closely-related species.

Species	Aspicilia humida	Aspicilia aquatica	Aspicilia pseudovulcanica	Aspicilia vulcanica
Thallus growth form	mainly areolate, partially rimose	areolate to rimose	areolate or slightly cracked to continuous	rimose-areolate
Thallus color	pale gray to white	white to gray, often with a yellow or olive tinge	whitish gray to light gray	whitish to white-gray
Thallus thickness (μm)	175–300	200–400	150–200	120–160
Prothallus	inconspicuous	black to gray	–	–
Apothecia per areole	one to several	1 to 2(–4)	–	1 to 2
Apothecia (mm in diam.)	0.2–1.7	0.2–0.5	0.5–0.9	0.3–0.4
Pruina	absent	absent	white pruina on disk	white pruina on disk
Disk color	black without green in water	black with translucent green in water	gray to gray-black	(gray to gray-black)
Epihymenium color	olive green to brown	green to olive	dirty greenish brown	colorless
Hymenium height (μm)	50–60	150–200	110–130	(110–140)
Hymenium in Iodine	yellowish blue-green	blue, or turning dark red-brown	–	–
Paraphyses	2–2.5	1.5–2	–	c. 4
Paraphysial tip width (μm)	4.5–6	2–5	5–6	5–7
Paraphysial tip pigment	inconspicuous	blackened	brownish	colorless
Asci (μm)	64–72 × 17–27	85–140 × 25–35	–	80–90 × 14–12
Ascospores (μm)	10.5–23 × 6–13.5	20–35 × 13–20	18–23 × 10–12	17–20 × 7–10
Substance	stictic acid	no substance	stictic acid	–
Substrate	siliceous rocks nearby streams	siliceous rocks nearby streams	siliceous rocks	volcanic rock
Reference	BDNA-L-0000703 (holotype), BDNA-L-0000711 (paratype)	[33,34]	[16]	[13,16]

The morphological and chemical characteristics for several species close to the new species are referenced from the previous literature. All information on the new species is produced from type specimens (BDNA-L-0000703 and BDNA-L-0000711) in this study. The brackets for Aspicilia vulcanica, i.e. disk color and hymenium height, are assumed from previous literature.

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