Four New Species of Jelly Fungi from Northeastern China

TL;DR

This paper introduces four new jelly fungi species found in northeastern China, identified using both physical and genetic analysis.

Contribution

The discovery and classification of four new jelly fungi species in four different genera from northeastern China.

Findings

Four new jelly fungi species were identified and classified into four different genera.

Phylogenetic trees were constructed using molecular data from ITS and nrLSU regions.

Morphological and ecological details of the new species were documented.

Abstract

Four new species of jelly fungi were described from northeastern China based on morphological and molecular evidence. These new species were classified into the four genera Sirobasidium (Sirobasidium jilinense), Calocera (Calocera velutina), Dacrymyces (Dacrymyces jauensis), and Dacryopinax (Dacryopinax manghanensis). Maximum likelihood and Bayesian analyses were performed using a combined nuc rDNA internal transcribed spacer region (ITS) and nuc 28S rDNA (nrLSU) dataset for the construction of phylogenetic trees. Morphological descriptions, line illustrations, and the ecological habits of these new species are provided.

Figures

Tables

Table 1. Sequence information from phylogenetic trees.

Species	Locality	Sample No.	GenBank No.		References
			ITS	nrLSU
Bullera alba	USA	CBS 501T	AF444368	AF075500	[40]
Bullera hannae	USA	CBS 8286T	AF444486	AF363661	[40]
Bullera penniseticola	USA	CBS 8623T	AF444471	AF363649	[40]
Bullera unica	USA	CBS 8290T	AF444441	AF075524	[40]
Calocera bambusicola	China	Wu9910 12	FJ195751	-	[23]
Calocera cornea	USA	AFTOL ID 438	AY789083	AY701526	[41]
Calocera cornea	Sweden	UPS F 940774	MN595626	MN595626	[41]
Calocera cornea	Germany	MW 55	-	AF291302	[42]
Calocera cornea	Canada	CBS 124 84	AB712437	AB472738	[43]
Calocera furcata	Russia	H Spirin 10949	MW191975	MW159088	[43]
Calocera fusca	New Zealand	PDD 107972	LC131406	LC131365	[20]
Calocera glossoides	Ukraine	CWU6247	MW191968	MW159084	[44]
Calocera lutea	New Zealand	PDD 107841	LC131413	LC131372	[20]
Calocera palmata	Japan	CBS 127 51	MH856777	MH868295	[32]
Calocera pedicellata	New Zealand	PDD 107830	LC131415	LC131374	[20]
Calocera sinensis	China	FJAU68949	PP749224	-	This study
Calocera sinensis	China	Dai22645	OL518939	OL518948	[32]
Calocera tibetica	China	Dai20171	MW549777	MW750403
Calocera velutina	China	FJAU68950T	PP776476	PP776564	This study
Calocera velutina	China	FJAU68951	PP776543	PP776565	This study
Calocera viscosa	Japan	TUFC12873	AB712439	-	[43]
Calocera viscosa	China	FJAU68917	-	PP749235	This study
Calocera viscosa	Japan	175	-	AB299048	[18]
Calocera viscosa	Germany	AFTOL ID 1679T	DQ520102	DQ520102	[41]
Calocera viscosa	Sweden	UPS F 940773	MN595628	MN595628	[41]
Cerinomyces cokeri	Canada	TU135089	MW191983	MW191983	[44]
Cerinomyces concretus	Colombia	O F 919450T	MW191933	-	[44]
Cerinomyces fugax	USA	HHB 8856T	MW191905	MW159051	[44]
Cerinomyces hesperidis	USA	NY01782362T	MW191921	MW159065	[44]
Cerinomyces inermis	New Zealand	PDD87816T	MW191887	-	[44]
Cerinomyces lipoferus	Netherlands	ENZ20001	MZ147626	MZ147626	[44]
Cerinomyces nepalensis	Nepal	O F 904088	MW191896	-	[44]
Cerinomyces neuhoffii	Sweden	UPS F 941020T	MN595625	MN595625	[41]
Cerinomyces pallidus	USA	ARIZ AN09245	MZ147624	-	[44]
Cerinomyces paulistanus	Brazil	O Ryvarden 24759T	MW191934	-	[44]
Cerinomyces pinguis	Nepal	O F 904085T	MW191907	MW159043	[44]
Cerinomyces tortus	Sweden	UPS F 946515	MW191999	MW191999	[44]
Cerinomyces volaticus	Russia	LE295748	MW191901	MW159047	[44]
Dacrymyces adpressus	Japan	554	-	AB472729	[14]
Dacrymyces ancyleus	Japan	MAFF 241177T	AB712448	AB472713	[43]
Dacrymyces capitatus	China	FJAU68952	PP749225	PP749236	This study
Dacrymyces cerebriformis	China	Dai 19832T	OM955202	OM955197	[25]
Dacrymyces chiangraiensis	Thailand	MFLU 16 0572	KY498587	-	[45]
Dacrymyces chrysocomus	Japan	CBS 280 84	AB712451	AB712427	[43]
Dacrymyces chrysospermus	China	FJAU68956	PP749226	PP749237	This study
Dacrymyces citrinus	New Zealand	PDD 107915T	LC131417	LC131376	[20]
Dacrymyces corticioides	USA	NY02686162	MW191944	MW159068	[44]
Dacrymyces cylindricus	New Zealand	PDD 105052T	LC131419	LC131378	[20]
Dacrymyces cyrtosporus	New Zealand	PDD 107980T	LC131422	LC131381	[20]
Dacrymyces dictyosporus	Japan	HHB 8618	AB712454	AB712429	[43]
Dacrymyces fennicus	Finland	H Miettinen 21174	MW191957	MW159071	[44]
Dacrymyces flabelliformis	New Zealand	HHB 18308T	AB712455	AB712430	[43]
Dacrymyces grandinioides	Kenya	K M 237139T	MW191980	MW191980	[44]
Dacrymyces intermedius	New Zealand	PDD 107851	-	LC131384	[20]
Dacrymyces invisibilis	Chile	14597MDT	MH230100	MH230102	[46]
Dacrymyces jauensis	China	FJAU68959	PP776532	PP776577	This study
Dacrymyces jauensis	China	FJAU68960	PP776533	PP776578	This study
Dacrymyces jauensis	China	FJAU68961T	PP776534	PP776579	This study
Dacrymyces lacrymalis	Japan	TUFC13327	AB712456	AB299069	[43]
Dacrymyces longistipitatus	New Zealand	PDD 107997T	LC131426	LC131387	[20]
Dacrymyces microsporus	China	Wu561	AB712458	OL546779	[32]
Dacrymyces minor	Japan	TUFC12836	AB712458	-	[43]
Dacrymyces minutus	Japan	648	AB712459	AB472733	[43]
Dacrymyces novae-zelandiae	New Zealand	PDD 107953	LC131428	LC131391	[20]
Dacrymyces ovisporus	Norway	H Spirin 11145	MW191960	MW159073	[44]
Dacrymyces pachysporus	New Zealand	PDD 105004T	LC131429	LC131392	[20]
Dacrymyces parastenosporus	New Zealand	PDD 104963T	LC131432	LC131395	[20]
Dacrymyces pezizoides	Japan	TNS F 54909	LC386890	LC386894	[47]
Dacrymyces pinacearum	Japan	UPS F 593533	MN595637	MN595637	[41]
Dacrymyces puniceus	China	FJAU68955	PP749227	PP749238	This study
Dacrymyces san-augustinii	China	Wu544	OL587813	OL546781	[32]
Dacrymyces sinostenosporus	China	Dai20003T	MW540888	MW540890	[25]
Dacrymyces stenosporus	New Zealand	PDD 105018T	LC131433	LC131396	[20]
Dacrymyces stillatus	Sweden	UPS F 939814 1	MN595677	MN595677	[41]
Dacrymyces subalpinus	Japan	228	-	AB299060	[18]
Dacrymyces subantarcticensis	New Zealand	PDD 107948	LC131435	LC131399	[20]
Dacrymyces subarcticus	Japan	HNo 544	AB712467	-	[43]
Dacrymyces variisporus	Japan	TUFC14203	AB712470	-	[43]
Dacrymyces venustus	Ethiopia	O Adane 150	MW191949	MW159075	[44]
Dacryomitra pusilla	Sweden	UPS F 176774	MN595639	MN595639	[41]
Dacryopinax elegans	USA	HHB 18731	AB712471	AB712433	[43]
Dacryopinax indacocheae	Venezuela	CRM 72	AB712472	AB712434	[43]
Dacryopinax lowyi	Mexico	27618	MN733712	MN733723	[22]
Dacryopinax manghanensis	China	FJAU68940	PP749230	PP749241	This study
Dacryopinax manghanensis	China	FJAU68938	PP749228	PP749239	This study
Dacryopinax manghanensis	China	FJAU68941	PP749231	PP749242	This study
Dacryopinax manghanensis	China	FJAU68943T	PP749233	PP749244	This study
Dacryopinax manghanensis	China	FJAU68942	PP749232	PP749243	This study
Dacryopinax manghanensis	China	FJAU68939	PP749229	PP749240	This study
Dacryopinax martinii	Mexico	682	MN733715	MN733726	[22]
Dacryopinax maxidorii	Brazil	RLMA306	OK257531	OK257536	[48]
Dacryopinax primogenitus	Costa Rica	MIN 862738	KT251038	KT251039	[49]
Dacryopinax sp.	Kenya	H7008759	MW191959	MW159091	[44]
Dacryopinax spathularia	China	FJAU68908	PP749234	PP749245	This study
Dacryopinax spathularia	USA	H Miettinen 16740	MW191973	MW159085	[44]
Dacryopinax spathularia	Indonesia	H Miettinen 20559	MW191976	MW159092	[44]
Dacryopinax spathularia	Chinna	Wu 331	OL614833	OL616184	[32]
Dacryopinax sphenocarpa	Japan	534	-	AB472726	[14]
Dendrodacrys brasiliense	Brazil	INPA 241458T	AB744230	AB723514	[50]
Dendrodacrys ciprense	Cyprus	UPS F 946590T	OM519385	OM519385	[51]
Dendrodacrys concrescens	Sweden	UPS F 946602T	OM519390	OM519390	[51]
Dendrodacrys dendrocalami	Japan	TUFC13914	AB712453	AB712428	[43]
Dendrodacrys ellipsosporum	Spain	UPS F 946604T	OM519392	OM519392	[51]
Dendrodacrys kennedyae	Panama	TAAM192134T	OP529836	OP529830	[51]
Dendrodacrys laetum	Kenya	H7008757T	OP529841	OP529841	[51]
Dendrodacrys oblongisporum	Norway	UPS F 946599	OM519399	OM519399	[51]
Dendrodacrys rigoratum	Japan	TUFC12845T	AB712447	-	[14]
Ditiola haasii	Czech Republic	PRM 944647	MW557322	MW557322	[52]
Ditiola peziziformis	Finland	H Haikonen 2426	MW191972	MW159070	[18]
Fellomyces horovitziae	USA	CBS 7515T	-	AF189856	[53]
Femsjonia monospora	Thailand	MFLU 16 0608	KY498588	-	[45]
Femsjonia uniseptata	Japan	TNS F 54019T	NR_164095	NG_060435	[20]
Fibulobasidium inconspicuum	USA	CBS 8237T	AF444318	AF363641	[40]
Fibulobasidium murrhardtense	China	CBS 9109T	GU327540	AF416648	[54]
Fibulobasidium sirobasidioides	Germany	RJB 12787	-	AF416644	[55]
Fonsecazyma mujuensis	China	CBS 10308T	KF036595	DQ333884	[12]
Genolevuria amylolytica	China	CBS 10048T	KF036585	AY562134	[12]
Genolevuria armeniaca	China	CBS 10050	KF036587	AY562140	[12]
Genolevuria bromeliarum	Brazil	BI20	EU386359	DQ784566	[56]
Genolevuria tibetensis	China	AS 2 2653T	EF363146	EF363143	[57]
Guepiniopsis alpina	USA	HAY F 002724	OR767855	-	[32]
Guepiniopsis buccina	USA	AFTOL ID 888	DQ206986	AY745711	[41]
Guepiniopsis estonica	Sweden	UPS F 940137	MN595632	MN595632	[41]
Heterotextus miltinus	New Zealand	PDD 107924	LC131438	LC131402	[20]
Kockovaella imperatae	USA	CBS 7554T	AB054091	AF189862	[53]
Pseudotremella allantoinivorans	USA	CBS 9604T	AY315664	AY315662	[58]
Pseudotremella moriformis	USA	CBS 7810	AF444331	AF075493	[40]
Sirobasidium apiculatum	Japan	MY62 1	LC203425	LC203426	[13]
Sirobasidium apiculatum	Japan	MY62 4	LC203428	LC203429	[13]
Sirobasidium brefeldianum	Spain	AM71	JN053472	JN043578	[9]
Sirobasidium brefeldianum	USA	CBS 7805	AF444330	AF075492	[40]
Sirobasidium japonicum	Japan	MY111 05	LC203420	LC016573	[13]
Sirobasidium japonicum	Japan	MY111 09	LC203422	LC203423	[13]
Sirobasidium jilinense	China	FJAU68667	PP749214	-	This study
Sirobasidium jilinense	China	FJAU68670T	PP749217	PP749222	This study
Sirobasidium jilinense	China	FJAU68668	PP749215	PP749220	This study
Sirobasidium jilinense	China	FJAU68669	PP749216	PP749221	This study
Sirobasidium jilinense	China	FJAU68672	PP749219	-	This study
Sirobasidium jilinense	China	FJAU68671	PP749218	PP749223	This study
Sirobasidium magnum	Spain	AM70	JN053497	JN043603	[9]
Sirobasidium magnum	Netherlands	CBS 6803	AF444314	KY109657	[40]
Sirobasidium magnum	Netherlands	CBS 6964	KY105421	KY109658	[59]
Sirobasidium magnum	China	FJAU68996	PP741731	PP741733	This study
Sirobasidium magnum	China	FJAU68997	PP741732	PP741734	This study
Tremella giraffa	Germany	CCJ 1553	AF042453	AF042271	[60]

Full text

1. Introduction

Sirobasidium Lagerh. & Pat. was established in 1892, with the type species being Sirobasidium sanguineum Lagerh. & Pat [1]. Due to the presence of hypobasidia that are divided vertically, the genus was first assigned to Tremellaceae Fr. In 1895, Möller proposed the establishment of Sirobasidiaceae Lindau due to the unique chain-like hypobasidia of this species [2]. Two years later, Sirobasidiaceae was officially published by Lindau [3]. Lowy then discovered that the French record of S. cerasi Bourdot & Galzin did not match the requirements of the genus concerning the size of its basidiomata, the quantity of chain-shaped basidia, and the size of its basidiospores. Lowy also provided a key for the recognized species S. brefeldianum Möller, S. magnum Boedijn, S. albidum, and S. sanguineum [4]. It was not until Bandoni examined the basidia and basidiospores of Sirobasidium that the spindle-shaped epiprobasidia shed by species in this genus were recognized as basidiospores [5]. In 2015, Roedel and Putzmann compiled a key for the Sirobasidium in their study of S. albidum, which included eight species [6]. Unfortunately, molecular information was not provided. At present, the genus contains ten species [3], three of which have been identified in China: S. japonicum, S. magnum, and S. sanguineum [7]. The chain-like hypobasidia and exfoliated epiprobasidia are the two primary characteristics of members of this genus. Phylogenetic analyses have shown the genus to be polyphyletic [8,9,10,11,12,13]. It is currently challenging to confirm the taxonomic status of Sirobasidium, however, as there are currently no molecular sequences available for the type species.

Dacrymycetaceae J. Schröt. (1888) belongs to Basidiomycota [3]. This family includes a variety of small-sized, morphologically diverse, and gelatinous species that are hard to find when dry. The majority of the basidiomata are yellow and contain carotene. The majority of basidiospores are often separated transversely, occasionally they can also be separated longitudinally or obliquely, or not septate when immature, septate by thin or thick walls; germination produces conidia or not; basidia and basidiospores contain oil droplets; hyphidia are present or absent; and with or without clamp connection [14].

In the 1960s and 1970s, McNabb conducted a detailed study on the genera of Dacrymycetaceae based on their morphology and compiled a key [15,16,17]. Since the 21st century, with the development of molecular systematics, researchers have conducted more in-depth research on Dacrymycetaceae species. In 2007, Shirouzu et al. found, in their phylogenetic analysis of Dacrymyces Nees based on 28S, that Dacrymyces was polyphyletic. While some species of Dacrymyces nest in Cerinomycetaceae Jülich or form a separate clade within Dacrymycetes, the majority of Dacrymyces species are found nested in Dacrymycetaceae [18]. In their 2009 study of Dacrymycetes in Japan, Shirouzu et al. described five new species and provided line illustrations [14]. Based on morphological observations, Malysheva (2013) identified the species of Calocera (Fr.) Fr. in Russia, described eight species of the genus, and provided line illustrations [19]. Later, Shirouzu et al. published eight new species of Dacrymycetes from New Zealand [20]. Dacryopinax G.W. Martin species from Brazil and Mexico were studied by Renato et al. and Castro-Santiuste et al., respectively [21,22]. The above studies show that most genera in Dacrymycetaceae are polyphyletic, and species are scattered in various branches of Dacrymycetaceae.

“Flora Fungorum Sinicorum Vol. 2 Tremellales et Dacrymycetales” by Liu was the primary focus of early Chinese research on Dacrymycetaceae species [7]. At the time, only morphological research was included due to the restricted conditions. Chinese researchers have recently studied species such as Calocera and Dacrymyces using multi-gene fragments [23,24,25]. However, most of the specimens used in these studies were collected from southern China, while there are relatively few specimens from northeastern China.

To enrich the species resources of jelly fungi in Northeast China. We have discovered four new species through morphological research and phylogenetic analysis. This study provided a detailed description and discussion of these species.

2. Materials and Methods

2.1. Specimen Collection

The specimens for this study were collected from Jilin Province and the Inner Mongolia Autonomous Region in China and deposited in the herbarium Fungarium of Jilin Agricultural University (FJAU).

2.2. Morphological Studies

Field records of fresh basidiomata, photographs of the habitat, and records after drying were used for macromorphological descriptions. The basidiomata’s color description was derived from Kornerup and Wanscher [26]. Using the technique of freehand slicing, utilizing water and 5% KOH solution as the floating carrier, and dyeing with 1% Phloxine B solution, the structures of basidia, basidiospores, hyphidia, and hyphae were examined under an optical microscope (Olympus CX33, Olympus, Tokyo, Japan). Twenty mature microstructures were chosen at random, examined under a microscope, and measured. “a–b × c–d” indicates the measurement of basidia, basidiospores, and hyphidia; “e–f” indicates the measurement of hyphae. Moreover, we determined the basidiospore’s Q value (Q = length/width). The extreme values of length, width, and diameter are denoted by the letters “a–b”, “c–d”, and “e–f”, respectively. For a given species, “n” denotes the ratio of the total number of measured basidiospores to the total number of measured specimens. All data were collected within a measurement range of 90%. The width was measured by choosing the area that was the widest. The apiculum was not taken into account when measuring the length of basidiospores. Along with the macroscopic morphology and microstructure features, a description and line charts are provided.

2.3. DNA Extraction, PCR and Sequencing

To extract DNA from dry specimens, the plant genome extraction kit from Kangwei Century (CWBIO, Beijing, China) was used. The ITS1F/ITS4 [27] and LR0R/LR7 [28,29] primers were used to amplify and sequence the ITS and nrLSU fragments, respectively. Sangon Bioengineering (Shanghai, China) Co., Ltd. was entrusted with the sequencing work, and the PCR reaction procedure was based on the method of Wang and Bau [30].

2.4. Molecular Phylogenetic Analyses

Sequencher 5.4.5 was used to observe the peak maps, further screen and sort the self-test sequences, and perform blast alignment [31]. ITS and nrLSU sequences were downloaded from GenBank based on the alignment outcomes and morphological similarity [32] (Table 1). The ITS and nrLSU sequences were aligned using the GINS-i algorithm with two iterative cycles only, using the online Mafft tool version 7 [33]. The alignment was manually adjusted and trimmed using MEGA 7.0.

The concatenated alignment of ITS (1–646) and nrLSU (647–1248) of Sirobasidium comprised 64 sequences. The concatenated alignment of ITS (1–682) and nrLSU (683–1517) of Dacrymycetaceae comprised 197 sequences. The best-fit model (edge-unlinked) was selected using the BIC criterion with ModelFinder [34]. The best-fit model of Sirobasidium and Dacrymycetaceae, according to BIC, was both SYM + I + G4. With the use of IQ-TREE [35] and the Shimodaira–Hasegawa-like approximate likelihood ratio test [36], maximum likelihood phylogenies were inferred under the edge–unlinked partition model for 10,000 ultrafast [37] bootstraps. The best-fit partition model (edge-unlinked) was again selected using the BIC criterion with ModelFinder. The best-fit model of Sirobasidium according to BIC was SYM + I + G4 (ITS) and K2P + I + G4 (nrLSU). The best-fit model of Dacrymycetaceae according to BIC was SYM + I + G4 (ITS) and GTR + F + I + G4 (nrLSU). Under the partition model, in which the first 25% of sampled data were discarded as burn-in, Bayesian inference phylogenies were inferred using MrBayes 3.2.6 [38]. Fig Tree v1.4.3 [39] and Photoshop 2021 (Adobe, Sam Jose, CA, USA) were then used to visualize the phylogenetic tree.

3. Results

3.1. Phylogenetic Analyses

This study generated a total of 46 new sequences, including 24 ITS sequences and 22 nrLSU sequences. These new sequences have been uploaded to GenBank. The multi-locus dataset (ITS + nrLSU) of Sirobasidium had an aligned length of 1248 total characters), including gaps. The multi-locus dataset (ITS + nrLSU) of Dacrymycetaceae had an aligned length of 1517 total characters, including gaps. Only the topological structures of Bayesian inference are displayed, as the topological structures of ML and BI are very similar. Bootstrap support (BS) values ≥ 75% and Bayesian posterior probability (PP) values ≥ 0.75 are indicated on branches (BS/PP) (Figure 1 and Figure 2).

From the phylogenetic tree of Sirobasidium and its related species constructed with Fellomyces horovitziae Spaaij, G. Weber & Oberw. and Kockovaella imperatae Nakase, I. Banno & Y. Yamada as outgroups (highlighted in blue), it can be seen that Sirobasidium is polyphyletic (Figure 1). High support rates (PP = 1.00, MLbs = 100%) were attained by the six specimens of the new species Sirobasidium jilinense T. Bau et X. Wang sp. nov., which were grouped in an independent branch. S. jilinense and S. magnum share a sister relationship and received high support (PP = 1.00, MLbs = 100%). The novel species could also be easily distinguished from other species, thanks to its distinct placement in the phylogenetic tree (Figure 1).

From the phylogenetic tree of Dacrymycetaceae constructed with Cerinomycetaceae as an outgroup (highlighted in blue), it can be seen that Dacrymyces, Dacryopinax, and Calocera are polyphyletic. The species belonging to each genus are dispersed across different branches of Dacrymycetaceae (Figure 2).

The two specimens of the recently discovered species Calocera velutina T. Bau et X. Wang sp. nov. were grouped into a separate branch and had a high support rate (PP = 1.00, MLbs = 100%). C. velutina forms a sister clade with C. cornea (Batsch) Fr. and C. furcata (Fr.) Fr., which is highly supported (PP = 1.00, MLbs = 100%). At the same time, the new species can be distinguished from other species in the genus in the phylogenetic tree (Figure 2).

A high support rate (PP = 1.00, MLbs = 100%) was obtained for the three specimens of the new species, Dacrymyces jauensis T. Bau et X. Wang sp. nov. They were grouped into an independent branch. D. jauensis forms a sister relationship with the clade consisting of D. chiangraiensis Ekanayaka, Karun., Q. Zhao & KD Hyde, and D. san-augustinian Kobayasi and gained a lower support rate (PP = 0.75, MLbs = 78%). This suggests that other species among them have not yet been identified. The new species and other species could also be well distinguished in the phylogenetic tree (Figure 2).

The six specimens of the new species Dacryopinax manghanensis T. Bau et X. Wang sp. nov. were clustered into an independent branch and obtained a high support rate (PP = 1.00, MLbs = 100%). It is a sister clade to Dacryopinax sp. (H7008759), with a high support rate (PP = 1.00, MLbs = 100%), and which is well distinguished from other species.

3.2. Taxonomy

Sirobasidium jilinense T. Bau et X. Wang, sp. nov. (Figure 3 and Figure 4)

MycoBank number: 853841

Diagnosis: Sirobasidium jilinense is characterized by cerebriform basidiomata, which are reddish brown to dark reddish brown when fresh and dark brown when dry. The hypobasidia consist of three to nine chains and are divided vertically or obliquely into four cells, whereas the basidiospores are subglobose to broadly ellipsoid, and the hyphae are thick-walled.

Etymology: “jilinense” refers to the discovery of a type specimen in Jilin Province, China.

Type: CHINA. Jilin Province, Jilin, Jiaohe, Shansong Ridge, on rotten wood of Quercus, elev. 577.2 m, 43°75′ N, 127°33′ E, 26 August 2023, T. Bau and Xia Wang, (FJAU68870, holotypus!). Same location, 29 July 2023, Xia Wang, (FJAU68668, paratypus!).

Basidiomata are soft gelatinous when fresh, easily rotting, cerebriform, reddish brown (8E5–E8) to dark reddish brown (8F5–F8), usually remaining coalescing, occasionally separate, 1–3 cm long and 1 cm thick. The hymenium surface is smooth, with full edges, folded, and relatively blunt. Shrinkage when dry, dark brown (8F1–F5), fragile.

Cross section without medulla, composed of the hymenium and intertwined jelly hyphae. Hymenium peripheral, pale yellow, composed of probasidia and hypobasidia. Probasidia 12–20 × 11–16 μm, globose to ellipsoid, light yellow, containing oil droplet-like substances, with clamp connection at the base; hypobasidia, 14–22 × 10–15 μm, ellipsoid to ovoid, the vertical or diagonal partitions are 4-cells, 3–9 chain-like structures, wrinkled at maturity, light yellow, containing oil droplet-like substances, with clamp connection at the base; epibasidia, 13–18 × 5–8 μm, spindle-shaped, detached, containing oil droplet-like substances. Basidiospores 6.7–9.4 × 6.7–7.2 μm, Q = 1.04–1.37 (n = 100/5), subglobose to broadly ellipsoid, apiculate, colorless, smooth, with oil droplets, germinating to produce regenerated spores or germination tubes. Hyphae with rich clamp connection, thick-walled, 2.3–3.6 µm in diameter.

Habitat: In summer and autumn, it grows on fallen trees or branches such as Acer and Quercus in broad-leaved forests.

Distribution: Currently, only known in the Jilin Province, China, Asia.

Additional specimens examined: China. Jilin Province: Jilin, Jiaohe, Shansong Ridge, on fallen branches of Acer, July 24, 2022, Liyang Zhu, FJAU68667 (Z22072428). Same location, Shien Wang, FJAU68669 (E2307209); Jilin, Huandian, Hongshi National Forest Park, on fallen branches of Acer, August 27, 2023, T. Bau & Xia Wang FJAU68671 (W23082706), T. Bau & Mu Liu, FJAU68672 (lm230890).

Calocera velutina T. Bau et X. Wang, sp. nov. (Figure 5 and Figure 6)

MycoBank number: 854036

Diagnosis: Calocera velutina is characterized by stipitate and pileate when young, needle-like when mature, light yellow, with a white disc-shaped villus at the base, simple or dichotomously branched. Hyphae thick-walled, without clamp connection. In summer, they grow on decaying trees in coniferous forests.

Etymology: “velutina” refers to the base of the basidiomata with villus.

Type: CHINA. Jilin Province, Yanbian Korean Autonomous Prefecture, Antu County, Erdaobaihe Town, Forest Industry Cultural Park, on rotten wood of Pinus L., elev. 708 m, 42°43′ N, 128°12′ E, 31 July 2022, T. Bau and Xia Wang, (FJAU68950, holotypus!). Jilin Province, Huadian City, Hongshi National Forest Park, 27 August 2023, T. Bau and Mu Liu, (FJAU68951, paratypus!).

Basidiomata are gelatinous when fresh, stipitate and pileate when young, needle-like when mature, light yellow (4A4–A5), slightly lighter at the tip, with white (4A1) disc-shaped villus at the base, simple or dichotomously branched, up to 15 mm high. The hymenium is located in the upper middle part, and the base is sterile. When dry, it is greyish yellow (4B3–B6, 4C5), and the surface under the body mirror is fine and granular.

The transverse section of the apex of the basidiomata is composed of the hymenium and interwoven hyphae. The transverse section of the basidiomata cylinder presents three annular zones. The central hyphae are densely arranged vertically, surrounded by loosely interlaced hyphae, and the periphery is the hymenium. Hymenium amphigenous is composed of probasidia, basidia, and hyphidia. Probasidia 18–25 × 2.9–4.4 μm, sub-clavate, forked when mature, containing oil droplet-like substances, with septate at the base; basidia 21–33 × 3.5–4.8 μm, clavate, containing oil droplet-like substances, with septate at the base; sterigmata 10–17 × 2.3–3.7 μm, cylindrical, the top gradually becomes pointed and gradually weakens when mature, containing oil droplet-like substances. Hyphidia 19–28 × 1.8–2.7 μm, narrowly cylindrical, simple or slightly curved, with septum at the base. Basidiospores 9.1–10.4 × 4.2–5.3 μm, Q = 1.90–2.27 (n = 40/2), curved cylindrical, narrow top, smooth, thin-walled, mature 1 septate, septate thin-walled, contains oil droplet-like substances. No germination was observed. Central hyphae 4.9–6.8 μm in diameter, septate, slightly rough, thick-walled. Interlaced hyphae, 3.4–5.3 μm in diameter, septate, thick-walled, basal hyphae 2.5–3.8 μm in diameter. All structures without clamp connection.

Habitat: In summer, it grows on decaying trees in coniferous forests.

Distribution: Currently, only known in the Jilin Province, China, Asia.

Dacrymyces jauensis T. Bau et X. Wang, sp. nov. (Figure 7 and Figure 8)

MycoBank number: 854037

Diagnosis: Dacrymyces jauensis is characterized by basidiomata flat to cushion-shaped when young, cinnamon, golden yellow to brownish yellow; basidiospores sausage-shaped, separated horizontally when mature, usually divided into 7 septate; germination produces rod-shaped conidia; hyphidia absent.

Etymology: “jauensis” refers to the discovery of a type specimen at Jilin Agricultural University.

Type: CHINA. Jilin Province, Changchun city, wild plantation garden of the campus of Jilin Agricultural University, grows on highly decayed wood of broad-leaved trees in broad-leaved forests, 43°81′ N, 125°40′ E, 235 m, 16 July 2022, Xia Wang & Tolgor Bau, (FJAU68961, 22071605W, holotypus!). Same location, 4 July 2022, Xia Wang & Tolgor Bau, (FJAU68959, T22070437W; FJAU68960, T22070438W, paratypus!).

Basidiomata are soft gelatinous when fresh, flat to cushion-shaped when young, cinnamon, golden yellow (4C6), up to 0.5 cm, usually remaining coalescing when mature, brownish yellow (5D6, 5E5), sessile, up to 2.5 cm. Hymenium surface with wrinkles and grooves, edge curved, folded, wavy, blunt. When dry, it shrinks and collapses in a brown (6E6), making it difficult to observe.

The transverse section is composed of hymenium and context hyphae. Hymenium amphigenous. Probasidia 32–47 × 3.7–6.3 μm, cylindrical, subclavate to broadly clavate, thin-walled, transparent, forked when mature, containing oil-dripping substance, basally septate; sterigmata 11–33 × 3.0–4.4 μm, cylindrical, apically acuminate when mature, gradually weakening. Hyphidia absent. Basidiospores 15.9–20.9 × 5.1–6.3 μm, Q = 2.80–3.49 (n = 90/3), sausage-shaped, smooth, thin-walled, transparent, separated horizontally when mature, usually divided into 7 septate, occasionally 0–3 septate, thin-walled, containing oil drop-like substance, and conidia germinate at the septate. Conidia 4.8–12.3 × 1.4–1.8 μm, long cylindrical to rod-shaped. Context hyphae 1.6–3.2 μm in diameter, thin-walled, septate, septate not swollen. All structures without clamp connection.

Habitat: In summer, it grows on highly decayed wood of broad-leaved trees in broad-leaved forests.

Distribution: Currently, only known in the Jilin Province, China, Asia.

Dacryopinax manghanensis T. Bau et X. Wang, sp. nov. (Figure 9 and Figure 10)

MycoBank number: 853843

Diagnosis: Dacryopinax manghanensis is characterized by small basidiomata, 1–5 mm wide, 1–8 mm high, spathulate and stipitate to substipitate, light yellow, cross section without medulla, the short and fine villi on the sterile surface. Basidiospores curved cylindrical to navicular, with 0–1 septate at maturity, thin-walled septate, and no germination observed; cortical hyphae curved, thick-walled, swollen, and branched.

Etymology: “manghanensis” is a Mongolian word for “sandy land”, which means that type specimens are collected from rotten wood in sandy land.

Type: CHINA. Inner Mongolia Autonomous Region, Tongliao, in the later stage of the left wing of Horqin, Wudantara Forest Farm, fallen branches of broad-leaved trees, 43°01′ N, 122°44′ E, 363 m, 16 July 2022, Weinan Hou & Tolgor Bau, (FJAU68943, H2207159, holotypus!). Same location, 15 July 2023, Hong Cheng & Tolgor Bau, (FJAU68940, C2371524, paratypus!).

When the basidiomata are fresh, soft gelatinous, spathulate, and stipitate to substipitate when young, the stipitate is light yellow (4A4–A5) and slightly hyaline, petaloid when mature, surface with longitudinal ridges, 1–5 mm wide, and 1-8 mm high. Hymenial surface smooth, with entire edges, blunt. The short and fine villi on the sterile surface are visible under a stereomicroscope. When dry, it shrinks, with a white (4A1) base, a dark yellow (4C8) surface on the hymenial, and obvious longitudinal edges on the stipitate.

Cross section without medulla, composed of the hymenium, context hyphae, and cortical hyphae. Hymenium unilateral, occasionally bilateral, composed of basidia and hyphidia. Probasidia 18–27 × 2.5–3.5 μm, cylindrical to clavate, thin-walled, hyaline, forked when mature, containing oil droplet-like substances, with septate at the base; sterigmata 7–13 × 1.8–2.9 μm, the top gradually becomes pointed and gradually weakens when mature. Hyphidia 15–27 × 1.4–2.2 μm, narrow cylindrical, thin-walled, hyaline, simple or slightly curved, with 1 septum at the base. Basidiospores 7.9–9.0 × 3.7–4.9 μm, Q = 1.73–2.07 (n = 80/4), curved cylindrical to navicular, apiculate, smooth, thin-walled, hyaline, mature 0–1 septate, septate thin-walled, contains oil droplet-like substances. No germination was observed. Cortical hyphae 4.5 μm in diameter, cylindrical branching at the end, curved, septum, constriction at intervals, thick-walled. All structures without clamp connection.

Habitat: In summer and autumn, it decays on fallen branches in broad-leaved forests or mixed coniferous and broad-leaved forests.

Distribution: Currently, it is only distributed in China, Inner Mongolia Autonomous Region, and Jilin Province.

Additional specimens examined: CHINA. Jilin Province, Tonghua, Yuhuangshan Park, broad-leaved forest rotting wood, 14 August 2022, Xia Wang, FJAU68941 (2281405W); Changchun, Jilin Agricultural University Campus Back Mountain, 16 September 2022, Xia Wang, FJAU68942 (2291601W); Tonghua, Yushan Park in Ji’an City, 6 July 2023, Zhengqing Chen, FJAU68938 (Q237603); Changchun, Jingyue Pool National Forest Park, 12 July 2023, Xia Wang, FJAU68939 (W23071206).

4. Discussion

Sirobasidium jilinense is easily confused with the type species S. sanguineum in the wild; however, the hypobasidia of the latter are solitary or in chains of 2–4, without a clamp connection at the base, and the clamp connection of hyphae is enlarged (5.5–8 μm). S. jilinense and S. rubrofuscum (Berk.) P. Roberts are similar; the basidiomata are all cerebriform and dark reddish brown, but they can be distinguished by the number of hypobasidia in the chains and the size of the basidiospores. Other species in Sirobasidium are white, off-white, or yellowish and are easily distinguished from S. jilinense (Table 2). Due to its brain-like, reddish-brown basidiomata, S. jilinense is easily confused with individual species of Tremella Pers. and Exidia Fr. in the wild, but it is easily distinguished from the latter under a microscope according to the basidium morphology.

Molecular phylogeny analysis showed that members of Fibulobasidium Bandoni and Sirobasidium jilinense, S. magnum, S. apiculatum, and S. japonicum form a separate branch (PP = 0.99, MLbs = 91%), while S. brefeldianum is located in another branch. This suggests that Sirobasidium is polyphyletic, which is consistent with the results of recent studies [9,10,11,13] (Figure 1). The new species S. jilinense and S. magnum have a sister relationship and obtained a high support rate (PP = 1, MLbs = 100%), which is consistent with the results of morphological studies.

Calocera velutina is characterized by basidiomata that are stipitate and pileate when young, needle-like when mature, yellow with a white disc-shaped villus at the base, and simple or dichotomously branched. The hyphae are thick-walled, without clamp connection. In summer, they grow on decaying trees in coniferous forests. In the wild, C. velutina is easy to be confused with C. cornea (Batsch) Fr., C. sinensis McNabb, and C. tibetica F. Wu, L.F. Fan & Y.C. Dai. They all have fresh yellow basidiomata with sharp, unbranched, or even-branched cylindrical upward-narrowing tops; however, the base of the latter three does not have a white disc-shaped villus [7,23,24]. The basidiomata of C. sinensis are small (1–5 mm), and the hyphae have clamp connections [23]. The basidiospores of C. tibetica are divided into three transverse septates when mature [24]. In the phylogenetic tree, they were also found to have a distant relationship.

Dacrymyces jauensis is characterized by basidiomata flat to cushion-shaped when young, cinnamon, golden yellow to brownish yellow; basidiospores sausage-shaped, separated horizontally when mature, usually divided into seven septate; germination produces rod-shaped conidia; hyphidia absent. Dacrymyces jauensis, Dacrymyces san-augustinii Kobayasi, and Dacrymyces chiangraiensis Ekanayaka, Karun., Q. Zhao & K.D. Hyde present multiple transverse septates when basidiospores mature. In the phylogenetic tree, a close relationship can be observed (Figure 2). However, the basidiomata of Dacrymyces san-augustinii are yellow, and the basidia (38–58 × 5.5–7 µm) and basidiospores (16–27.5 × 6–10 µm) are larger, with hyphidia [14]. The basidiomata of Dacrymyces chiangraiensis are yellow to orange, and the basidia (32–53 × 7–10 µm) and basidiospores (19–23 × 6.5–8 µm) are also large, with hyphidia [45].

There are two kinds of basidiomata of Dacryopinax manghanensis, most of which are clustered and spoon-shaped in the sand of the Inner Mongolia Autonomous Region but petal-shaped in Jilin Province (Figure 9). However, their microstructure is the same, as they are clustered in the same branch in the phylogenetic tree, and the shape of the basidiomata may be related to the environment. There are seven species of Dacryopinax distributed in China. The mature basidiospores of Dacryopinax aurantiaca and Dacryopinax spathularia are 0–1 septate, and they are easily confused with Dacryopinax manghanensis in the wild. However, the basidiospores of Dacryopinax aurantiaca are longer (10.4–13 × 3.5–5 μm) [7]. Basidiospores germinate as conidia or germ tubes and are mainly distributed in South China. Dacryopinax spathularia had obvious villi (visible to the naked eye when dry), with medulla on the transverse section and a bulbous hyphae septum [7]. In the phylogenetic tree, they are not closely related (Figure 2). In addition, the basidiospores of Dacryopinax indacocheae Lowy, Dacryopinax primogenitus, and Dacryopinax sphenocarpa Shirouzu & Tokum. are also divided into 0–1 septate at maturity. However, the basidiomata of Dacryopinax indacocheae are yellowish brown and larger (30 mm wide, 20 mm high). The cortical hyphae of the stipe have chain-like enlarged cells, the basidiospores are longer (8–11.5 × 3–3.5 μm), and the hymenium has abundant conidia [15]. Dacryopinax primogenitus has longer basidia (23–49 × 2.5–4 μm) and longer hyphidia (45.5–48.5 × 2.5–3 μm) and germinates as conidia or germ tubes [49]. The basidiomata of Dacryopinax sphenocarpa are yellow-white or light amber, the hyphae have clamp connections, the base of the probasidia has clamp connections, and the basidiospores are larger (10–16 × 4.5–8.5 μm) [14]. Dacryopinax is polyphyletic, with a total of eight species currently supported by molecular data, located in five distinct clades, consistent with recent findings [6,13,49]. In addition, in the phylogenetic tree constructed in this study, the branch composed of Dacryopinax manghanensis and Dacryopinax sp. (H7008759) was closely related to the branch composed of Calocera species (Figure 2); however, the species of Dacryopinax manghanensis and Calocera were different in morphology, so, it was placed in Dacryopinax.

Regarding jelly fungi, due to their gelatinous texture, researchers tend to focus more on damp environments when collecting specimens. In addition to adding three new species from the humid environment of Jilin Province—S. jilinense, C. velutina, and Dacrymyces jauensis—this study also added a new species from the Horqin Sandy Land—Dacryopinax manghanensis. This shows that jelly fungi have a wide geographical distribution and are well-adapted to different conditions and environments. As such, this type of fungus is worthy of further exploration.