Molecular phylogenetic results
Eight sequences (3 of nrLSU, 2 of TEF1-α, 3 of RPB2) from 3 collections were newly generated.
The Maximum likelihood and Bayesian phylograms have no conflict in topology, so only the ML trees with both BS and PP values are shown (Fig.1). The nrLSU, TEF1-α and RPB2 datasets consisted of 48 sequences (including 45 from GenBank and 3 new generated), 40 sequences (including 38 from GenBank and 2 new generated), 24 sequences (including 21 from GenBank and 3 new generated), respectively. The combined nuclear dataset (nrLSU+ TEF1-α+RPB2) contains 112 sequences with 2182 nucleotide sites after reducing gaps by Gblocks.
In the three-gene phylogenetic analysis, R. huanggangensis was sister to R. fuscus with bootstrap support (BS = 73%) (Fig. 1) but was independented from the collection from southwest China. And it formed a large clades with R. ater, R. griseus, R. nigerrimus, R. nigrogriseus, R. pseudogriseus, R. sinogriseus and R. zhangfeii with higher statistical support (BS = 67%, PP = 1).
Table 1. Specimens used in molecular phylogenetic study and their GenBank accession numbers.
Species
|
Voucher
|
Locality
|
Accession
|
Reference
|
nrLSU
|
TEF1-α
|
RPB2
|
Leccinum duriusculum
|
HKAS101160
|
Austria
|
MZ675541
|
MZ707785
|
MZ707779
|
Meng et al. 2021
|
L. parascabrum
|
HKAS:99903T
|
China
|
MW413911
|
MW439271
|
MW439264
|
Meng et al. 2021
|
L. versipelle
|
HKAS97997
|
China
|
MZ675545
|
MZ707789
|
MZ707781
|
Meng et al. 2021
|
L. versipelle
|
HKAS99380
|
China
|
MZ675546
|
MZ707790
|
MZ707782
|
Meng et al. 2021
|
Retiboletus ater
|
Li1215T
|
SW China
|
MT010611
|
MT010621
|
—
|
Liu et al. 2020
|
R. ater
|
Li1224
|
SW China
|
MT010612
|
MT010622
|
—
|
Liu et al. 2020
|
R. brevibasidiatus
|
OR0570
|
Thailand
|
—
|
MT085476
|
MT085479
|
Chuankid Boontiya et al. 2021
|
R. brunneolus
|
LC_LJW23
|
SW China
|
MT010615
|
MT010625
|
—
|
Liu et al. 2020
|
R. brunneolus
|
HKAS 52680
|
SE China
|
KF112424
|
KF112179
|
KF112690
|
Wu et al. 2016
|
R. flavoniger
|
RH7247T
|
Costa Rica
|
AF456828
|
—
|
—
|
Binder and Bresinsky 2002
|
R. flavoniger
|
RH7189
|
Costa Rica
|
AF456829
|
—
|
—
|
Binder and Bresinsky 2002
|
R. fuscus
|
FHMU1403
|
Yunnan, SW China
|
MH367473
|
MH367486
|
—
|
Zeng et al. 2018
|
R. fuscus
|
HKAS63624
|
China
|
KT990635
|
KT990829
|
KT990466
|
Wu et al. 2016
|
R. fuscus
|
OR0738
|
Thailand
|
—
|
MT085472
|
MT085477
|
Chuankid Boontiya et al. 2021
|
R. fuscus
|
OR0231
|
China
|
—
|
MG212600
|
MG212642
|
Vadthanarat et al. 2018
|
R. fuscus
|
HKAS74756
|
SW China
|
KT990636
|
KT990830
|
KT990467
|
Wu et al. 2016
|
R. fuscus
|
HKAS:63590
|
USA
|
KF112417
|
KF112178
|
KF112691
|
Wu et al. 2016
|
R. griseus
|
MB03-079
|
USA
|
—
|
KT824030
|
KT823997
|
Vadthanarat et al. 2016
|
R. griseus
|
snBoth
|
USA
|
KF030308
|
KF030414
|
—
|
Binder and Bresinsky 2002
|
R. griseus
|
Halling10162
|
USA
|
MT010608
|
MT010618
|
—
|
Liu et al. 2020
|
R. huanggangensis
|
HFJAU10002
|
China
|
OL744444
|
OL963527
|
OL963530
|
This study
|
R. huanggangensis
|
HFJAU10003T
|
China
|
OL744445
|
OL963526
|
OL963528
|
This study
|
R. huanggangensis
|
HFJAU10004
|
China
|
OL744446
|
—
|
OL963529
|
This study
|
R. kauffmanii
|
HKAS68590
|
Yunnan, SW China
|
KP739283
|
—
|
—
|
Zeng et al. 2016
|
R. kauffmanii
|
HKAS 63584E
|
Yunnan, SW China
|
KT990634
|
KT990828
|
KT990465
|
Wu et al. 2016
|
R. kauffmanii
|
OR0278
|
Yunnan, SW China
|
—
|
MG212601
|
MG212643
|
Vadthanarat et al. 2018
|
R. kauffmanii
|
HKAS63584
|
SW China
|
KP739282
|
KP739301
|
KP739299
|
Zeng et al. 2016
|
R. nigerrimus
|
Tyni1
|
Japan
|
AF456832
|
—
|
—
|
Binder and Bresinsky 2002
|
R. nigrogriseus
|
BC0179
|
Thailand
|
—
|
MT085474
|
MT085478
|
Chuankid Boontiya et al. 2021
|
R. nigrogriseus
|
OR049
|
Thailand
|
—
|
KT824033
|
KT824000
|
Vadthanarat et al. 2016
|
R. nigrogriseus
|
FHMU2045
|
Southern Chin
|
MH367475
|
MH367487
|
—
|
Zeng et al. 2018
|
R. nigrogriseus
|
FHMU2800T
|
Southern Chin
|
MH367476
|
MH367488
|
—
|
Zeng et al. 2018
|
R. ornatipes
|
201/97 T *
|
USA
|
AF456815
|
—
|
—
|
Binder and Bresinsky 2002
|
R. ornatipes
|
Halling10163
|
USA
|
MT010617
|
MT010626
|
—
|
Liu et al. 2020
|
R. ornatipes
|
MBsn
|
USA
|
—
|
MT219516
|
MT219515
|
Chuankid Boontiya et al. 2021
|
R. ornatipes
|
93/97
|
USA
|
AF456825
|
—
|
—
|
Binder and Bresinsky 2002
|
R. pseudogriseus
|
Zeng647
|
Southern Chin
|
MT010613
|
MT010623
|
—
|
Liu et al. 2020
|
R. pseudogriseus
|
FHMU375
|
Southern Chin
|
MH367477
|
MH367489
|
—
|
Zeng et al. 2018
|
R. pseudogriseus
|
FHMU2205
|
Fujian, SE China
|
MH367478
|
MH367490
|
—
|
Zeng et al. 2018
|
R. pseudogriseus
|
HKAS 83950T
|
SE China
|
KP739285
|
—
|
—
|
Zeng et al. 2016
|
R. retipes
|
96/97
|
USA
|
AF456830
|
—
|
—
|
Binder and Bresinsky 2002
|
R. retipes
|
22/97
|
USA
|
AF456831
|
—
|
—
|
Binder and Bresinsky 2002
|
R. retipes
|
116/96
|
USA
|
AF456823
|
—
|
—
|
Binder and Bresinsky 2002
|
R. sinensis
|
HKAS83957
|
SE China
|
KP739291
|
KP739303
|
—
|
Zeng et al. 2016
|
R. sinensis
|
HKAS59832
|
SE China
|
KT990633
|
KT990827
|
KT990464
|
Wu et al. 2016
|
R. sinensis
|
HKAS83955
|
SE China
|
KP739289
|
KP739302
|
—
|
Zeng et al. 2016
|
R. sinogriseus
|
LJ258
|
NE China
|
MT010610
|
MT010620
|
—
|
Liu et al. 2020
|
R. sinogriseus
|
LJ260T
|
NE China
|
MT010609
|
MT010619
|
—
|
Liu et al. 2020
|
R. aff. kauffmanii
|
HY56
|
NE China
|
MT010616
|
—
|
—
|
Liu et al. 2020
|
R. vinaceipes
|
CFMR:DR-1035T
|
Dominican Republic
|
MN250180
|
—
|
—
|
Ortiz-Santana et al. 2007
|
R. vinaceipes
|
CFMR:BZ-2386
|
Belize
|
MN250190
|
—
|
—
|
Ortiz-Santana et al. 2007
|
R. zhangfeii
|
HKAS59699
|
SE China
|
JQ928627
|
JQ928582
|
JQ928603
|
Wu et al. 2016
|
R. zhangfeii
|
HKAS 83963
|
Fujian, SE China
|
KP739297
|
—
|
—
|
Zeng et al. 2016
|
R. zhangfeii
|
HKAS 83961
|
Fujian, SE China
|
KP739295
|
KP739305
|
—
|
Zeng et al. 2016
|
R. zhangfeii
|
HKAS53418
|
China
|
KT990630
|
KT990824
|
KT990462
|
Wu et al. 2016
|
Tylopilus sp.
|
204/97
|
USA
|
AF456813
|
—
|
—
|
Binder and Bresinsky 2002
|
Sequences obtained in this study are shown in bold. SW = southwestern; NE = northeastern; SE = southeastern.
T= holotype, E= epitype, T *= type species
Taxonomy
Retiboletus huanggangensis F. Zhou, Y. Gao, HJ. Hu, HY.Song & DM. Hu, sp. nov.
MycoBank No: 842302
Figures. 2 and 3
Etymology: “huanggangensis” referring to the collection site of this species (Huanggang Mountain).
Type: China. Jiangxi Province: Shangrao City, Yanshan County, Wuyishan Nature Reserve, Huanggang mountain, alt. 1600 m, August 23, 2021, Fan Zhou, WYS236 (holotype: HFJAU10003).
Description
Basidiomata small to medium-sized. Pileus 18.4–72.2 mm diam, hemispherical to applanate, surface dry, densely subtomentose, pale yellowish white (3A1) to grayish black (5E4) at the margin and black (30F1) towards center; context 4–15 mm in thickness in the center of the pileus, pale yellow (4A2), changing orange yellow (2B6) to rusty yellow (5B7) when injured. Hymenophore always adnate around the stipe; pores angular, tubes up to 11 mm long, white (29A2) when young and yellowish white (1A2) in age, becoming brownish yellow (30B4) when injured. Stipe 13–30 × 26–150 mm, central, clavate to flexuous, solid; surface dry, prominently and coarsely reticulate over the upper 3/4 or overall, pale yellow (2A2) in the upper part and black (1F3) downwards when young, white (3A1) on upper part and lower part black (3F4) in age, changed to pale yellow (5A2) when injured; basal mycelium white. Odor indistinct.
Basidia 30–36 × 10.5–13 μm, thin-walled, clavate, four-spored; sterigmata 2–5 μm long. Basidiospores [60/3/3] (7.0–)10–13.5 × 3.5–5.5 μm, Q = (1.75–)2.00–3.00(–3.33), Qm = 2.40 ± 0.34, amygdaloid to subfusiform to ellipsoid, grayish yellow to pale yellow to yellowish brown in KOH, smooth. Hymenophoral trama boletoid. Cheilo- and Pleurocystidia 27–46 × 8–10.5 μm, abundant, ventricose to fusiform or ellipsoid, thin-walled, hyaline or with golden yellow contents, without encrustations. Pileipellis a trichoderm about 130–220 μm thick, Composed of intertwined hyphae, brown to grayish brown in KOH, thin-walled hyphae 5–12 μm diam; terminal cells 22–74 × 4–12 μm, clavate or subterete, with obtuse apex. Pilea trama composed of thin- walled hyphae 3–11 μm diam. Stipitipellis hymeniform about 30–80 μm thick, composed of smooth, hyaline or grayish brown hyphae, narrowly or broadly clavate, ventricose or subfusiform; terminal cells 20–45 × 5–12.5 μm and occasionally with clavate, four-spored basidia 22–42 × 9–12 μm. Stipe trama composed of cylindrical, slightly thick-walled (up to 0.5 μm) parallel or interlaced hyphae 5–11 μm diam. Clamp connections absent in all tissues.
Habitat: Solitary or group on the ground in forests of Pinaceae and Fagaceae.
Distribution: Jiangxi Province, China.
Additional specimens examined: China. Jiangxi Province: Shangrao City, Yanshan County, Wuyishan Nature Reserve, Huanggangshan, alt. 1300 m, 31 July 2021 WYS083 and alt. 1650 m August 23, 2021, WYS277 (HFJAU10002; HFJAU10004).