Fungal Clusters and Their Uniqueness in Geographically Segregated Wetlands: A Step Forward to Marsh Conservation for a Wealth of Future Fungal Resources

Figures & data

Table 1. Geographic information of three representative wetlands in the Korea peninsula.

Sampling area		Administrative district including sampling area	GPS location (northern latitude, eastern longitude)	Average altitude (m)	National indications for conservation
Site 1	Mt. Gariwang’s primeval forest	Seoheung-ri, Seohwa-myeon, Inje-gun, Gangwon-do, Korea	37°25′58"N, 128°33′48"E	620	Ecological landscape protected Area of Korean nation
Site 2	Is. Jangdo	Bi-ri, Heuksan-myeon, Sinan-gun, Jeollanam-do, Korea	34°40′41.1"N, 25°22′15.4"E	169	Wetland conservation area of Korean nation
Site 3	Hanbando geology	Ongjeong-ri, Hanbando-myeon, Yeongwol-gun, Gangwon-do, Korea	37°13′5"N, 128°20′56"E	280	Wetland conservation area of Korean nation

Figure 1. Location of each wetland in the Korean Peninsula. Site 1: Mt. Gariwang’s primeval forest wetlands (37°25′58"N, 128°33′48"E, 620 m); Site 2, Is. Jangdo wetlands (34°40'41.1"N, 25°22'15.4"E, height above sea level 169 m); and Site 3, Hanbando geology wetland (37°13'5"N, 128°20'56"E, 280 m).

Table 2. Results of soil analysis.

Figure 2. Rarefaction curves for operational taxonomic units (OTUs) from each site. OTUs were clustered at 3% dissimilarity using CD-HIT. Curves indicate microbial community diversity in the Mt. Gariwang’s primeval forest wetlands (green, OTUs, 169; sequencing reads, 236,624), the Is. Jangdo wetlands (purple, OTUs, 283; sequencing reads, 157,649), and the Hanbando geology wetlands (red, OTUs, 145; sequencing reads, 179,870).

Table 3. Pyrosequencing results and statistical analyses of fungal clusters in each wetlands.

Sampling area	Mt. Gariwang’s primeval forest	Is. Jangdo	Hanbando geology
Number of total reads	277,927	209,609	229,098
Total bases	107,912,837	78,596,700	87,067,254
Number of valid sequences	236,624	157,649	179,870
GC (%)	43	47	50
OTUs	169	283	145
Phylum (identified)	3	5	3
Genus (identified)	32	42	25

1. Total Reads : The total number of sequence reads.

2. Total Bases : The total number of bases in identified reads.

3. GC (%) : The GC percentage in sequence reads.

4. OTUs : Operational Taxonomic Unit is an operational definition of a species or group of species often used when only DNA sequence data are available.

Table 4. Fungal phylum composition of each wetlands.

Fungal phyla	Mt. Gariwang’s primeval forest	Is. Jangdo	Hanbando geology
Ascomycota	2.8%	32.9%	37.4%
Basidiomycota	94.1%	6.7%	42.1%
Chytridiomycota	0.0%	0.4%	0.0%
Glomeromycota	0.0%	4.0%	0.0%
Zygomycota	1.4%	14.2%	4.6%

Figure 3. Fungal phyla distribution from each wetland (unit: %). Variance of phyla ratio was characteristic, and the uniqueness of fungal taxa could be expected.

Figure 4. Fungal diversity defined by Shannon, Simpson’s, and Chao richness indices. Variance of genera diversity was characteristic, and the Is. Jangdo wetlands showed higher evenness, diversity, and dominance of fungal genera. These results corroborate the variation of identified or unique genera.

Table 5. Fungal genera distributions in each wetland.

Genus	Wetland of the Gariwangsan primeval forest	Wetland of the Jangdo islands	Wetland of the Hanbando geology
Mortierella	0.60	4.19	3.98
Paecilomyces	0.02	1.12	0.00
Penicillium	0.30	0.04	8.55
Trichoderma	0.03	2.18	4.05
Phialocephala	0.03	1.45	–
Hypocrea	0.02	0.06	–
Chaunopycnis	0.01	1.18	–
Entoloma	0.00	2.65	–
Cryptosporiopsis	–	0.07	0.26
Lecanicillium	–	0.07	0.00
Ilyonectria	–	0.01	0.21
Fusarium	–	0.10	1.12
Neonectria	–	0.67	1.07
Amanita	0.14	–	0.29
Exophiala	0.01	–	0.81
Umbelopsis	0.74	–	0.43
Geminibasidium	0.17	–	32.65
Archaeorhizomyces	–	0.29	–
Ramularia	0.01	–	–
Devriesia	0.04	–	–
Cenococcum	0.96	–	–
Minutisphaera	–	0.70	–
Gymnostellatospora	0.06	–	–
Oidiodendron	0.35	–	–
Pseudeurotium	–	0.55	–
Didymosphaeria	–	0.28	–
Paraphoma	–	0.13	–
Pyrenochaeta	–	–	0.13
Subplenodomus	–	–	0.04
Lophiostoma	0.03	–	–
Epicoccum	–	0.15	–
Cyphellophora	–	–	0.23
Capronia	0.01	–	–
Cladophialophora	0.04	–	–
Cylindrocarpon	–	0.19	–
Aspergillus	0.00	–	–
Chalara	–	–	0.88
Staphylotrichum	–	–	0.36
Thelonectria	–	0.08	–
Carestiella	–	–	0.16
Acremonium	–	–	0.58
Neobulgaria	0.02	–	–
Lachnum	–	–	0.11
Scytalidium	0.01	–	–
Leohumicola	–	–	1.93
Meliniomyces	0.05	–	–
Leotia	0.01	–	–
Lophodermium	–	–	0.19
Tuber	0.04	–	–
Saccharomyces	0.00	–	–
Chaetosphaeria	–	0.37	–
Thozetella	–	0.10	–
Pochonia	–	0.07	–
Beauveria	–	0.65	–
Gibberella	–	0.31	–
Mariannaea	–	0.38	–
Nectria	–	0.15	–
Haptocillium	–	0.11	–
Tolypocladium	–	1.26	–
Arthrinium	–	0.21	–
Colletotrichum	–	0.46	–
Gaeumannomyces	–	0.05	–
Verticillium	0.06	–	–
Sphaerodes	–	0.02	–
Chaetomium	0.00	–	–
Podospora	–	0.54	–
Diatrypella	–	–	0.28
Clavaria	–	0.52	–
Cortinarius	1.55	–	–
Clitopilus	–	0.35	–
Laccaria	0.09	–	–
Tephrocybe	0.05	–	–
Hydropus	–	0.07	–
Mycena	–	0.39	–
Coprinellus	0.01	–	–
Tricholoma	88.94	–	–
Byssocorticium	0.04	–	–
Suillus	–	–	0.23
Ganoderma	–	0.05	–
Sebacina	–	–	0.20
Tremella	0.03	–	–
Archaeospora	–	0.74	–
Absidia	–	–	0.20
Mucor	–	0.03	–

Taxa shared with other wetlands and unique fungal genera are indicated. The term ‘0.00’ means a confirmation ratio not exceeding two decimal points; ‘-’means non-confirmed genera. Red font indicates ratio of genera above 1.0%. Common genera are indicated in blue highlight.

Figure 6. Uniqueness of fungal genera distributions in total microbiome samples. Wetlands sharing similar fungal taxa are below 6%, but uniquely distributed taxa were highly confirmed, especially in the Is. Jangdo wetlands. Percentages represent the proportion of unique or shared taxa.

Data availability statement

All the raw sequences obtained from this study have been deposited at the NCBI Sequence Read Archive (SRA metadata) under the project PRJNA616032 with sample accession numbers of SAMN14479947 (Mt. Gariwang’s primeval forest wetland), SAMN14479948 (Is. Jangdo wetland), and SAMN1447994 (Hanbando geology wetland).