Lactifluus (Russulaceae) diversity in Central America and the Caribbean: melting pot between realms

Abstract

Species of the ectomycorrhizal genus Lactifluus , and often entire sections, are typically unique to a single continent. Given these biogeographic patterns, an interesting region to study their diversity is Central America and the Caribbean, since the region is closely connected to and often considered a part of the North American continent, but biogeographically belong to the Neotropical realm, and comprises several regions with different geologic histories. Based on a multi-gene phylogeny and morphological study, this study shows that Central America, Mexico and the Caribbean harbour at least 35 Lactifluus species, of which 33 were never reported outside of this region. It was found that species from the Caribbean generally show affinities to South American taxa, while species from the Central American mainland generally show affinities to Northern hemispheric taxa. We hypothesise that host specificity and/or climate play a crucial role in these different origins of diversity. Because of these different affinities, Caribbean islands harbour a completely different Lactifluus diversity than the Central American mainland. The majority of species occurring on the islands can be considered endemic to certain islands or island groups. In this paper, detailed morphological descriptions are given, with a focus on the unique diversity of the islands, and identification keys to all hitherto described Lactifluus species occurring in Central America and the Caribbean are provided. One new section, Lactifluus sect. Nebulosi , and three new species, Lactifluus guadeloupensis , Lactifluus lepus and Lactifluus marmoratus are described.

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Persoonia 44, 2020: 278–300 ISSN (Online) 1878-9080
www.ingentaconnect.com/content/nhn/pimj https://doi.org/10.3767/persoonia.2020.44.10
RESEARCH ARTICLE
INTRODUCTION
The Russulaceae family represents a dominant group of ecto-
mycorrhizal fungi in almost all forest ecosystems worldwide. In
2008, molecular research showed that the traditionally recog-
nised genera Russula and Lactarius were not monophyletic,
resulting in a rearrangement of the generic landscape into four
separate genera; Lactarius sensu novo, Russula, Multifurca
and Lactifluus (Buyck et al. 2008, 2010). The genus Lactifluus
comprises around 200 described species worldwide, although
its total diversity is estimated to be much higher, and has its
highest species richness in the tropics (De Crop 2016). Lacti-
fluus species, and often sections, are usually unique to a single
continent, so there is little to no overlap in species diversity
between continents.
Central America, part of Mexico, and the Caribbean present a
geographically interesting area to study, since they are often
considered part of the North American continent and are closely
connected to it, but biogeographically belong to the Neotropical
realm together with the South American continent. Given the
biogeographic patterns in Lactifluus, the question arises whether
species occurring in Central America and the Caribbean are
most closely related to North or to South American species.
Most of Central America and the Caribbean is situated on the
Caribbean plate. Cuba is an exception as it originated on the
Caribbean plate but became stuck on the North American
plate. The northern part of Hispaniola is a piece of Cuba that
was sheared off as the Caribbean plate moved eastward. The
Central American mainland is located on the western edge of
this plate and consists of seven countries: Belize, Costa Rica,
El Salvador, Guatemala, Honduras, Nicaragua and Panama. It
is a tectonically active region and the landscape of the mainland
of Central America is strongly determined by mountain ranges.
The region contains diverse forest types: tropical moist broad-
leaf forests, tropical dry broadleaf forests, tropical coniferous
forests and montane forests. The tropical lowlands of Central
America are dominated by plants of South American origin,
examples of ectomycorrhizal hosts are Coccoloba, Guapira,
Neea and Pisonia. This lowland vegetation strongly differs from
the montane vegetation, which includes mainly species with
temperate North American origins, such as Alnus, Pinus and
Quercus. In addition, the southern part of Mexico also belongs
to the Neotropical realm, and consists of habitats similar to
those present in Central America.
Central America is bordered in the east by the Caribbean
Sea. The Caribbean islands comprise two main geographical
groups: the Greater Antilles and the Lesser Antilles, which have
independent origins. The Antilles are mostly volcanic in origin,
arisen due to the subduction of the North American plate under
the Caribbean plate. The Greater Antilles are geologically the
oldest and lie along the Northern edge of the Caribbean plate.
Lactifluus (Russulaceae) diversity in Central America
and the Caribbean: melting pot between realms
L. Delgat1,*, R. Courtecuisse2, E. De Crop1, F. Hampe3, T.A. Hofmann4,
C. Manz5, M. Piepenbring5, M. Roy6, A. Verbeken1
Key words
Antilles
Basidiomycota
ectomycorrhizal fungi
French West Indies
integrative taxonomy
Latin America
Middle America
new taxa
Russulales
1 Ghent University, Department of Biology, Research group Mycology, K.L.
Ledeganckstraat 35, 9000 Ghent, Belgium;
corresponding author e-mail: lynn.delgat@ugent.be.
2 Université Lille Nord France, Faculté des Sciences Pharmaceutiques &
Biologiques, F-59006 Lille, France.
3 Straße des Friedens 11, 99330 Gräfenroda, Germany.
4 Vicerrectoría de Investigación y Posgrado, Centro de Investigaciones
Micológicas (CIMi), Herbario UCH, Universidad Autónoma de Chiriquí,
0427 David, Panamá.
5 Goethe University, Department of Biological Sciences, Research group
Mycology, Max-von-Laue-Str. 13, 60438 Frankfurt am Main, Germany.
6 Université Toulouse 3 Paul Sabatier, CNRS, ENFA, UMR5174 EDB (Labo-
ratoire Évolution & Diversité Biologique), 118 route de Narbonne, 31062
Toulouse, France.
Abstract Species of the ectomycorrhizal genus Lactifluus, and often entire sections, are typically unique to a single
continent. Given these biogeographic patterns, an interesting region to study their diversity is Central America and
the Caribbean, since the region is closely connected to and often considered a part of the North American conti-
nent, but biogeographically belong to the Neotropical realm, and comprises several regions with different geologic
histories. Based on a multi-gene phylogeny and morphological study, this study shows that Central America, Mexico
and the Caribbean harbour at least 35 Lactifluus species, of which 33 were never reported outside of this region.
It was found that species from the Caribbean generally show affinities to South American taxa, while species
from the Central American mainland generally show affinities to Northern hemispheric taxa. We hypothesise that
host specificity and /or climate play a crucial role in these different origins of diversity. Because of these different
affinities, Caribbean islands harbour a completely different Lactifluus diversity than the Central American mainland.
The majority of species occurring on the islands can be considered endemic to certain islands or island groups. In
this paper, detailed morphological descriptions are given, with a focus on the unique diversity of the islands, and
identification keys to all hitherto described Lactifluus species occurring in Central America and the Caribbean are
provided. One new section, Lactifluus sect. Nebulosi, and three new species, Lactifluus guadeloupensis, Lactifluus
lepus and Lactifluus marmoratus are described.
Article info Received: 4 July 2019; Accepted: 9 March 2020; Published: 12 June 2020.

279
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
They comprise the islands of Cuba, Cayman Islands, Jamaica,
Hispaniola (i.e., Dominican Republic and Haiti), Puerto Rico
and the Virgin Islands. The largest Greater Antillean islands are
Cuba, Hispaniola and Jamaica. Several different forest types
occur on the Greater Antilles, such as lowland forests (e.g.,
rainforests with Carapa guianensis; semi-deciduous forests with
Coccoloba diversifolia, Guapira fragrans; seasonal evergreen
forests with Ceiba pentandra), submontane forests (e.g., rain-
forests with Cecropia peltata, Dacryodes excelsa, Mora abbottii,
Oxandra laurifolia, Sloanea berteriana; pine forests with Pinus
caribaea, Pinus cubensis; sclerophyllous forest with Coccoloba
retusa, Quercus oleoides subsp. sagraeana) and montane
forests (e.g., rainforests with Alchornea latifolia, Magnolia spp.,
Ocotea ekmanii, Solanum acropterum; (mixed) pine forests
with Pinus cubensis, Pinus occidentalis) (Panagopoulos 1999).
The Lesser Antilles on the other hand are a series of small
islands on the eastern edge of the Caribbean plate. The major
part of the Lesser Antilles is volcanic in origin, though a small
group of islands (i.e., Antigua and Barbuda, Grande-Terre of
Guadeloupe, Marie-Galante and Barbados) is of coral (calcar-
eous) origin, building an arc dorsally contiguous to the main
volcanic one. This yields a special interest to Guadeloupe,
which comprises both ecologies (i.e., volcanic for Basse-Terre
and calcareous for Grande-Terre). In contrast, Trinidad and
Tobago are continental islands that were cut off by sea level
rise from the South American mainland while the other islands
emerged from the ocean floor. Trinidad, Guadeloupe and Mar-
tinique are the largest Lesser Antillean islands. Vegetation of
the Lesser Antilles has been studied in detail, and according
to the literature the forests can be classified in four main types:
1. altitudinal, montane or cloud forest;
2. hygrophytic or submontane rain forest;
3. mesophytic or (semi-)evergreen seasonal forest; and
4. xerophytic or tropical semi-deciduous forest.
Montane forests occupy small areas near the highest moun-
tains of Martinique and the vegetation consists of shrubs, most
typically Clusia plukenetii. In hygrophytic forests, the annual
rainfall is over 2 700 mm/year and a pronounced dry season is
absent. In these forests, epiphytes are abundant and tree spe-
cies are virtually all evergreen. Dominant tree species include
Dacryodes excelsa, and the endemic Licania ternatensis, Mag-
nolia dodecapetala and Sloanea massonii. Mesophytic forests
have a lower annual rainfall and are characterised by species
of Inga, Nectandra, Simarauba and Tabebuia, but patches of
several ectomycorrhizal Coccoloba spp. are also present. In
xerophytic forests annual rainfall is lower than 1 250 mm/year
and there is a distinct dry season. Approximately half of the tree
species are deciduous and the canopy commonly consists of
species of Bursera, Ceiba, Coccoloba, Guapira, Lonchocarpus,
and Pisonia (Stehle 1947, Beard 1948, Pegler & Fiard 1983,
Panagopoulos 1999, Joseph 2013). Coccoloba, Guapira and
Pisonia are known ectomycorrhizal hosts (e.g., Tedersoo et
al. 2010, Alvarez-Manjarrez et al. 2018) and it is in these xero-
phytic forests that ectomycorrhizal fungi predominate (Pegler
& Fiard 1983).
Extensive collections of agaricoid Basidiomycota were made on
the Lesser Antilles, making it the best studied region in Central
America and the Caribbean, and a total of ten Lactarius species
were recorded in this area (Pegler & Fiard 1979, 1983). Re-
cently, most of these species have been combined in Lactifluus
(Montoya et al. 2012, Verbeken et al. 2012). However, these
species are in need of a thorough molecular study. Other areas
in Central America and the Caribbean have been studied less
intensively, yielding only three species of Lactifluus described
from the Greater Antilles, one Lactifluus species from Central
America and four species from southern Mexico (Singer 1973,
Montoya et al. 1996, 2012, Miller et al. 2000, Montoya & Ban-
dala 2004, Crous et al. 2019, Delgat et al. 2019).
In this paper, a phylogeny based on multiple loci, detailed
morphological information and identification keys for Lactifluus
species occurring in the Caribbean, and Central America and
Mexico are provided. Three new species are described here:
Lactifluus guadeloupensis, *L. lepus and L. marmoratus. A
new section, L. sect. Nebulosi is described, L. castaneibadius
and L. murinipes are synonymised, and Lactarius coccolobae,
Lactarius fuscomarginatus, Lactarius pegleri and Lactarius sect.
Panuoidei are combined in Lactifluus.
(* L. = Lactifluus)
MATERIAL AND METHODS
Sampling
Collections for the Lesser Antilles were made during regular
fieldtrips to the islands Martinique and Guadeloupe between
2003 and 2015, organised within the program ‘Les champig-
nons des Petites Antilles: diversité, écologie et protection’
initiated and conducted by R. Courtecuisse and permitted since
2006 by grants and funding from different organizations (see
Acknowledgements). Field work focused on different habitats
such as xero-, meso- and hygrophytic forests. The collections
from Western Panama were obtained during a field trip to
Chiriquí province in 2018, sampling mostly montane forests,
but to a lesser extent also lowlands. Herbarium vouchers were
gathered for other regions.
Morphological analysis
Macroscopic characters were observed from fresh material,
with colour codes referring to Kornerup & Wanscher (1978).
Microscopic characters were observed from dried material.
Basidiospores were mounted in Melzer’s reagent and hyme-
nium, pileipellis and stipitipellis were studied in Congo red in
L4. The basidiospore measurements (i.e., length, width and
Q = quotient of length and width) are given as [Ava-2xSDa]–
Ava–Avb –[Avb+2xSDb], in which Ava = lowest mean value
for the measured specimens, Avb = greatest mean value,
SDa/ b = standard deviation of the lowest and greatest mean
value, respectively (number of spores measured per speci-
men = 15– 20). If there was only one specimen, measurements
are given as [Av-2xSD]–Av–[Av+2xSD]. Basidiospores were
measured in side view excluding ornamentation. Measurements
of basidia, cystidia and pileipellis terminal elements are given
as [Av-2xSD]–Av– [Av+2xSD], based on minimum 10 measure-
ments per structure and species. Measurements of basidia do
not include sterigmata. Line drawings were made with the aid
of a drawing tube at following magnifications: 6000× for spores
(Zeiss Axio Scope 2 microscope), 1500× for other hymenial
elements and sections (Olympus cx31 microscope).
Molecular analysis
DNA was extracted from fresh material preserved in CTAB
(Cetyl trimethylammonium bromide) using the CTAB extraction
method described in Nuytinck & Verbeken (2003). The protocol
described by Dentinger et al. (2010) was used for dried collec-
tions from Kew herbarium, a modified CTAB protocol (Tel-Zur
et al. 1999; modified by Agentschap Plantentuin Meise) was
used for other dried collections. PCR amplification protocols
follow Le et al. (2007). Four nuclear markers previously shown
as informative within this genus (Stubbe et al. 2010, Van de
Putte et al. 2010, Montoya et al. 2012) were used:
1. the internal transcribed spacer region of ribosomal DNA
(ITS), comprising the ITS1 and ITS2 spacer regions and
the ribosomal gene 5.8S, using primers ITS-1F and ITS4
(White et al. 1990, Gardes & Bruns 1993);

280 Persoonia – Volume 44, 2020
Lactarius fuliginosus MTB 97-24 GENT JQ446111 JQ446180 JQ446240 KR364392
Lactarius hatsudake FH 12-052 GENT KR364085 KR364215 KR364285 KR364411
Lactarius miniatescens AV 11-177 GENT KR364059 KR364187 KR364315 KR364443
Lactarius olympianus ED 08-018 GENT KR364089 KR364220 KR364320 KR364448
Lactarius scrobiculatus JN 2001-058 GENT KF432968 – KR364344 KR364474
Lactarius tenellus ADK 3598 GENT KF133280 KF133313 KF133345 KR364482
Lactifluus acicularis KVP 08-002 GENT HQ318226 HQ318132 HQ328869 JN389131
Lactifluus acrissimus EDC 11-112 GENT KR364041 KR364168 KR364254 KR364366
Lactifluus albomembranaceus EDC 12-046 GENT KR364064 KR364193 KR364257 KR364369
Lactifluus allardii JN 2004-008 GENT KF220016 KF220125 KF220217 KR364370
Lactifluus ambicystidiatus HKAS J7008 HKAS KR364108 KR364239 KR364309 KR364437
Lactifluus annulatoangustifolius BB 00-1518 GENT, PC AY606981 KR364253 – –
Lactifluus annulifer TH 9014 BRG, DUKE KC155376 KC155376 – –
G4819 PC KM073086 – – –
Lactifluus armeniacus EDC 14-501 MFLU, GENT KR364127 – – –
Lactifluus aurantiifolius AV 94-063 GENT KR364017 KR364144 – –
Lactifluus aureifolius AV 11-074 GENT KR364056 KR364183 KR364259 KR364371
Lactifluus auriculiformis AV 12-050 GENT KR364086 KR364216 KR364260 KR364372
Lactifluus bertillonii JN 2012-016 GENT KR364087 KR364217 KR364261 KR364373
Lactifluus bhandaryi TENN 051830 TENN KR364111 KR364140 – –
Lactifluus bicapillus EDC 12-176 GENT KR364070 KR364199 KR364300 KR364428
Lactifluus bicolor DS 06-247 GENT JN388955 JN388987 JN375590 JN389186
Lactifluus brachystegiae AV 99-002 GENT KR364018 KR364145 KR364262 KR364374
Lactifluus brunellus TH 9130 BRG, DUKE JN168728 – – –
Lactifluus brunneocarpus AB 305 GENT KR364035 KR364162 KR364343 KR364473
Lactifluus brunneoviolascens AV 13-038 GENT KR364123 KR364246 KR364264 KR364376
Lactifluus brunnescens AV 05-083 GENT KR364019 KR364146 KR364263 KR364375
Lactifluus caribaeus CL/MART06.014 LIP MK046841 MK046791 – –
J.P. Fiard 818A K MK046852 – – –
J.P. Fiard 827B K MK046853 – – –
Lactifluus carmineus AV 99-099 GENT KR364131 KR364251 KR364265 –
Lactifluus castaneibadius J.P. Fiard 837A K MK046854 – – –
Lactifluus chamaeleontinus JD 946 MEISE KR364079 KR364208 KR364267 KR364377
Lactifluus chiapanensis Bandala 4374a GENT GU258297 GU265580 GU258316 KR364378
Lactifluus chrysocarpus LE 253907 LE JX442761 JX442761 – –
Lactifluus clarkeae MN 2004002 L KR364011 HQ318205 KR364268 KR364379
Lactifluus coccolobae OKM-27240 CFMR MK774805 – – –
Lactifluus cocosmus ADK 4462 GENT KR364013 KR364141 KR364269 KR364380
Lactifluus conchatulus LTH 457 GENT GU258296 GU265659 GU258399 KR364381
Lactifluus corrugis AV 05-392 GENT JQ753822 KR364143 JQ348127 –
SA A12 L2 GENT KR364088 KR364218 KR364361 KR364491
Lactifluus crocatus KVP 08-034 GENT HQ318243 HQ318151 HQ328888 JN389145
Lactifluus cyanovirescens JD 988 GENT KR364082 KR364211 KR364270 KR364382
Lactifluus denigricans EDC 11-218 GENT KR364051 KR364178 KR364272 KR364384
Lactifluus densifolius AV 11-111 GENT KR364057 KR364184 KR364273 KR364385
Lactifluus dissitus AV-KD-KVP 09-134 GENT JN388978 JN389026 JN375628 JN389172
Lactifluus distantifolius LTH 288 GENT HQ318274 HQ318193 KR364274 JN389155
Lactifluus domingensis ANGE1035 JBSD MK931340 MN128988 MK937131 –
ANGE542 JBSD MK931339 MN128987 MK937130 –
ANGE837 JBSD MK931342 MN128989 MK937133 MN128629
ANGE838 JBSD MK931341 MN128990 MK937132 –
Lactifluus dwaliensis Asia LTH 55 GENT KF220111 KF220204 KF220278 KR364386
Lactifluus edulis FN 05-628 GENT KR364020 KR364147 KR364275 KR364387
Lactifluus flammans JD 941 MEISE KR364078 KR364207 KR364303 KR364431
Lactifluus flocktonae JET1006 MEL JX266621 JX266637 – –
Lactifluus foetens ADK 2840 MEISE KR364023 KR364150 KR364279 KR364391
Lactifluus fuscomarginatus LM4719 XAL HQ168368 – – –
LM 4379 XAL HQ168367 HQ168367 – –
Lactifluus genevievae GG-DK 17-02-05 GENT GU258294 GU265657 GU258397 KR364401
Lactifluus gerardii AV 05-375 GENT GU258254 GU265616 GU258353 KR364403
JN 2007-029 GENT GU258224 GU265582 GU258318 –
Lactifluus glaucescens LGAM 2010-0132 LGAM-AUA KR364105 KR364236 KR364280 KR364407
Lactifluus guadeloupensis RC/Guad11-023 LIP KP691412 KP691421 KP691430 –
Lactifluus guanensis GUA-104 CFMR MK046851 – – –
Lactifluus gymnocarpus EDC 12-047 GENT KR364065 KR364194 KR364282 KR364408
Lactifluus hallingii R. E. Halling 4977 NY MK931343 – – –
R. E. Halling 7993 NY MK931333 – MK937128 –
NVE 520 KF937338 – – –
R. E. Halling 7938 NY MK931327 MN128984 MK937127 –
FH 18-077 GENT MK931338 MN128991 MK937129 MN128628
Lactifluus hygrophoroides AV 05-251 GENT HQ318285 HQ318208 HQ328936 KR364413
GenBank accession numbers
Species Herbariumnumber Herbarium ITS LSU RPB2 RPB1
Table 1 Specimens and GenBank accession numbers of DNA sequences used in the molecular analyses.

281
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
Lactifluus igniculus LE 262983 LE JX442759 JX442759 – –
Lactifluus kigomaensis AV 11-006 GENT KR364052 KR364179 KR364288 KR364415
Lactifluus kivuensis JR Z 310 GENT KR364027 KR364154 – –
Lactifluus lamprocystidiatus EH 72-195 GENT KR364015 – – –
Lactifluus leae FH 12-013 GENT KF432957 KR364213 KR364292 KR364419
Lactifluus leonardii GG 07-02-04 GU258308 GU265668 GU258408 KR364495
Lactifluus leoninus DS 07-454 GENT KF220055 JN388989 JN375592 JN389188
Lactifluus leptomerus AV-KD-KVP 09-131 GENT JN388972 JN389023 JN375625 JN389169
Lactifluus lepus RC/Guad 08-042 LIP KP691414 KP691423 KP752179 –
RC/Guad 05-029 LIP MK046811 MK046774 – –
Lactifluus leucophaeus LTH 182 GENT KF220059 KF220157 KF220243 KR364420
Lactifluus longipilus LTH 184 GENT HQ318256 HQ318169 HQ328905 JN389152
Lactifluus luteolus AV 05-253 GENT KR364016 KR364142 KJ210067 KR364440
Lactifluus luteopus EDC 11-087 GENT KR364049 KR364176 KR364312 KR364441
Lactifluus madagascariensis BB 99-409 PC AY606977 DQ421975 DQ421914 –
Lactifluus marmoratus LD 15-066 GENT MK046805 MK046768 MK063907 MK089574
Lactifluus melleus AV 11-183 GENT KR364060 KR364189 KR364277 KR364389
Lactifluus multiceps TH 9154A BRG, DUKE JN168731 – – –
Lactifluus murinipes LD 15-018 GENT MK046814 – – –
LD 15-057 GENT MK046815 – – –
LD 15-054 GENT MK046828 MK046778 MK063911 MK089578
LD 15-015 GENT MK046832 MK046782 MK063915 MK089580
LD 15-020 GENT MK046836 MK046785 MK063918 MK089581
LD 15-032 GENT MK046838 MK046787 MK063920 MK089579
CL/MART06.019 LIP KP691417 KP691426 – –
F.1890 LIP KP691418 – – –
J.P. Fiard 849A K MK046855 – – –
Lactifluus nebulosus LD 15-059 GENT MK046804 MK046766 MK063905 MK089572
LD 15-061 GENT MK046833 MK046783 MK063916 MK089573
PAM-Mart12-90 LIP KP691415 KP691424 KP691432 KR364396
PAM/Mart 05-091 LIP MK046850 MK046799 – –
J.P. Fiard 828A K MK046856 – – –
Lactifluus neotropicus R.W.G. Dennis 97 K MN102680 – – –
Lactifluus nonpiscis AV 11-137 GENT KR364058 KR364185 KR364317 KR364445
Lactifluus ochrogalactus AV/KD/KVP 09-120 GENT KR364130 KR364248 KR364318 KR364446
Lactifluus oedematopus KVP 12-001 GENT KR364100 KR364232 KR364319 KR364447
Lactifluus pallidilamellatus Montoya 4716 GENT JQ753824 JQ348268 – MN128632
AV 17-012 GENT MN102682 – – –
AV 17-015 GENT MN102684 – – –
Lactifluus panuoides D.N. Pegler 3133 K MK046858 – – –
RC/Guy10_024 LIP KJ786647 KJ786551 KP691428 –
RC/Mart 03-077 LIP MK046807 MK046770 MK063908 MN128627
CL/GUAD06.045 LIP MK046843 – – –
R.W.G. Dennis 109A K MK046859 – – –
Lactifluus pegleri LD 15-042 GENT MK046831 MK046781 MK063914 MK089582
LD 15-014 GENT MK046835 MK046784 MK063917 MK089583
LD 15-049 GENT MK046837 MK046786 MK063919 –
PAM-Mart12-91 LIP KP691416 KP691425 KP691433 KR364397
PAM/Mart 05-088 LIP MK046808 MK046771 – –
PAM-Mart12-54 LIP MK046809 MK046772 – –
J.P. Fiard 821B K MK046860 – – –
Lactifluus pelliculatus JD 956 MEISE KR364080 KR364209 KR364321 KR364449
Lactifluus persicinus EDC 12-001 GENT KR364061 KR364190 KR364298 KR364426
Lactifluus petersenii AV 05-300 GENT GU258281 GU265642 GU258382 KR364450
Lactifluus phlebonemus EDC 12-023 GENT KR364062 KR364191 KR364322 KR364451
Lactifluus phlebophyllus BB 00-1388 PC AY606974 DQ421979 DQ421918 –
Lactifluus pilosus LTH 205 GENT KR364006 KR364134 KR364323 KR364452
Lactifluus pinguis AV-RW 04-023=LTH117 GENT HQ318211 HG318111 HQ328858 JN389126
Lactifluus piperatus 2001 08 19 68 GENT KF220119 KF241840 KF241842 KR364453
Lactifluus pulchellus KW 304/FH 12-037 GENT KR364092 KR364223 KR364306 KR364434
Lactifluus pumilus EDC 12-066 GENT KR364067 KR364196 KR364332 KR364462
Lactifluus putidus LD 15-006 GENT MK046818 – – –
LD 15-017 GENT MK046819 – – –
LD 15-002 GENT MK046820 – – –
LD 15-040 GENT MK046821 – – –
LD 15-041 GENT MK046822 – – –
LD 15-065 GENT MK046823 – – –
LD 15-011 GENT MK046824 – – –
LD 15-033 GENT MK046825 – – –
LD 15-034 GENT MK046826 MK046777 MK063910 MK089575
LD 15-062 GENT MK046827 – – –
LD 15-004 GENT MK046829 MK046779 MK063912 MK089577
LD 15-039 GENT MK046830 MK046780 MK063913 MK089576
LD 15-030 GENT MK046834 – – –
PAM/Mart 11-013 LIP KP691413 KP691422 KP691431 –
PAM-Mart12-88 LIP MK046806 MK046769 – –
RC/Mart 05-110 LIP MK046810 MK046773 – –
GenBank accession numbers
Species Herbariumnumber Herbarium ITS LSU RPB2 RPB1
Table 1 (cont.)

282 Persoonia – Volume 44, 2020
Lactifluus putidus (cont.) RC/Guad 06-005 LIP MK046813 MK046776 – –
PAM/Mart 05-030 LIP MK046840 MK046790 – –
PAM-Mart12-103 LIP MK046845 MK046795 – –
RC/Mart 03-001 LIP MK046849 MK046798 – –
PAM/Mart 05-085 LIP MK046839 MK046788 – –
RC/Mart10 011 LIP MK046812 MK046775 MK063909 –
Lactifluus ramipilosus EDC 14-503 MFLU, GENT KR364128 – – –
Lactifluus reticulatovenosus EH 6472 GENT GU258286 GU265649 GU258389 –
Lactifluus robustus JN 2011-074 GENT KR364047 KR364173 KR364358 KR364488
Lactifluus roseolus AV 99-160 GENT KR364032 KR364159 KR364333 KR364463
Lactifluus roseophyllus JN 2011-076 GENT KF220107 KF220202 KF220276 KR364464
Lactifluus rubiginosus JD 959 MEISE KR364081 KR364210 KR364304 KR364432
Lactifluus rubrobrunnescens EH 7194 GENT KR364115 – – –
Lactifluus rubroviolascens EDC 12-051 GENT KR364066 KR364195 KR364334 KR364465
Lactifluus rufomarginatus ADK 3358 MEISE KR364033 KR364160 KR364335 KR364466
Lactifluus rugatus EP 1212/7 LGAM-AUA KR364104 KR364235 KR364337 KR364467
Lactifluus russulisporus REH 9398 NY KR364097 KR364229 KR364307 KR364435
Lactifluus ruvubuensis JD 303 MEISE KR364009 KR364137 KR364310 KR364438
Lactifluus sainii PUN 7046 PUN KM658971 – – –
Lactifluus sepiaceus PL 40509 GU258287 GU265650 GU258390 KR364475
PL 34204 GU258288 GU265651 GU258391 –
Lactifluus sp. AV 05-325 GENT MK931329 MN128985 – –
TH7460 BRG, DUKE KT339233 KT339233 – –
TH7880 BRG, DUKE KT339212 KT339212 – –
F1068593 F MN102703 – – –
MAN_MAN919 FLOR KY936896 – – –
AV 04-195 GENT KF220045 KF220146 KF220232 KR364404
AV 05-374 GENT KF220049 KF220150 KF220236 KR364405
AV 07-056 GENT KR364008 KR364136 KR364293 KR364421
AV 99-012 GENT KR364021 KR364148 KR364276 KR364388
AV 05-249 GENT MK931325 – MK937125 –
AV-KD-KVP 09-121 GENT JN388979 JN389014 JN375616 JN389160
DS 06-003 GENT GU258231 GU265588 GU258325 JN389185
AV 04-181 GENT MK931328 – DQ421935 –
EDC 12-068 GENT KR364068 KR364197 KR364299 KR364427
EDC 12-195 GENT KR364071 KR364200 KR364301 KR364429
JD 907 GENT KR364076 KR364205 KR364302 KR364430
JN 2011-014 GENT KF220104 KF220199 KF220273 KR364406
JN 2011-036 GENT KF220105 KF220200 KF220274 KR364454
JN 2011-071 GENT KR364043 KR364169 KR364255 KR364367
JN 2011-072 GENT KF220106 KF220201 KF220275 KR364455
JN 2011-077 GENT KR364044 KR364170 KR364256 KR364368
AV 05-283 GENT GU258259 GU265621 GU258358 –
KVP 08-011 GENT HQ318232 HQ318139 HQ328876 JN389135
JN 2011-012 GENT KR364045 KR364171 KR364294 KR364422
KVP 08-031 GENT HQ318240 HQ318148 HQ328885 JN389142
AV 05-330 GENT JQ753828 HQ318129 JQ348140 –
AV 05-275 GENT MK931336 – – –
LTH 270 GENT EF560685 GU265598 GU258335 KR364402
LTH 274 GENT KR364107 KR364238 KR364325 KR364457
AV 05-227 GENT JQ753832 JQ348284 JQ348150 –
AV 05-293 GENT JQ753834 JQ348287 JQ348153 –
AV 17-011 GENT MN102681 – – –
AV 17-014 GENT MN102683 – – –
AV 17-016 GENT MN102685 – – –
FH 18-054 GENT MN102686 MN101703 MN120442 MN120459
FH 18-060 GENT MN102687 MN101704 MN120454 MN120471
FH 18-061 GENT MN102688 MN101705 MN120456 MN120467
FH 18-062 GENT MN102689 MN101706 MN120450 MN120472
FH 18-063 GENT MN102690 MN101707 MN120458 MN120469
FH 18-067 GENT MN102691 MN101708 MN120443 MN120460
FH 18-068 GENT MN102692 MN101709 MN120447 MN120464
FH 18-069 GENT MN102693 MN101710 MN120452 MN120474
FH 18-124 GENT MN102694 MN101711 MN120451 MN120473
FH 18-128 GENT MN102695 MN101712 MN120457 MN120468
FH 18-129 GENT MN102696 MN101713 MN120455 MN120470
FH 18-131 GENT MN102697 MN101714 MN120445 MN120461
FH 18-132 GENT MN102698 MN101715 MN120448 MN120465
FH 18-133 GENT MN102699 MN101716 MN120446 MN120462
FH 18-134 GENT MN102700 MN101717 MN120449 MN120466
FH 18-135 GENT MN102701 MN101718 MN120444 MN120463
FH 18-141 GENT MN102702 MN101719 MN120453 MN120475
MCA 3937 GENT KR364109 KR364240 KR364350 –
RC/Guy 09-004bis LIP KJ786643 KP691419 KP691427 –
RC/Guy 09-036 LIP KJ786645 KJ786550 KP752178 –
RC/Guy 12-007 LIP KJ786668 MK046789 – –
Halling 8262 NY GU258235 GU265593 GU258330 –
MR/Guy 13-145 PC KJ786691 KJ786595 MK063906 –
G3185 PC KJ786694 KJ786603 KP691434 KR364399
MR/Guy 13-032 PC KJ786686 KJ786581 – –
G3264 PC KJ786706 KJ786620 KP691435 KR364400
MR-GUY-14-195 PC KM073084 – – –
GenBank accession numbers
Species Herbariumnumber Herbarium ITS LSU RPB2 RPB1
Table 1 (cont.)

283
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
Lactifluus sp. (cont.) G4727 PC KM073087 – – –
G4836 PC KM073083 – – –
G4797 KM073085 – – –
JLC 060310-01 KJ786714 – – –
TENN 064342 TENN KR364103 KR364234 KR364324 KR364456
TENN 065854 TENN KR364101 MN128986 KR364271 KR364383
TENN 065929 TENN KR364102 KR364233 KR364308 KR364436
LM4640 XAL HQ168369 – – –
LM-UNAH 0072 XAL HM639277 – – –
LM-UNAH 0073 XAL HM639278 – – –
PGK13-130 KP691436 KR605507 – –
ASM 12,075 MN102679 – – –
Lactifluus subclarkeae REH 9231 NY KR364095 KR364227 KR364346 KR364477
Lactifluus subgerardii AV 05-269 GENT GU258263 GU265625 GU258362 KR364478
Lactifluus subiculatus SLM 10114 BRG, RMS JQ405654 – – –
Lactifluus subkigomaensis EDC 11-159 GENT KR364050 KR364177 KR364295 KR364423
Lactifluus subpruinosus JN 2011-061 GENT KR364046 KR364172 KR364357 KR364487
Lactifluus subvellereus AV 05-210 GENT KR364010 KR364138 KR364347 KR364479
Lactifluus subvolemus KVP 08-048 GENT JQ753927 – KR364356 KR364486
Lactifluus sudanicus AV 11-174 GENT HG426469 KR364186 KR364348 KR364480
Lactifluus tanzanicus AV 11-017 GENT KR364053 KR364180 KR364296 KR364424
Lactifluus tenuicystidiatus JN 2011-080 GENT KR364048 KR364174 KR364359 KR364489
Lactifluus uapacae AV 07-048 GENT KR364007 KR364135 KR364352 KR364483
Lactifluus urens EDC 14-032 GENT KR364124 KR364247 KR364353 –
Lactifluus vellereus ATHU-M 8077 ATHU-M KR364106 KR364237 KR364354 KR364484
Lactifluus velutissimus JD 886 MEISE KR364075 KR364204 KR364355 KR364485
Lactifluus venezuelanus RC/Mart 03-120 LIP MK046842 MK046792 – –
RC/Guad 03-048 LIP MK046847 MK046797 – –
RC/Guad11-017 LIP KP691411 KP691420 KP691429 –
PAM/Guad 10-036 LIP MK046844 MK046794 MK063922 MN128630
PAM/Guad 10-037 LIP MK046846 MK046796 MK063923 MN128631
F.1980 MK046848 – – –
Lactifluus veraecrucis M 8025 ENCB KR364112 KR364241 – –
Lactifluus versiformis AV-KD-KVP 09-045 GENT JN388967 JN389031 JN375632 JN389177
Lactifluus vitellinus KVP 08-024 GENT HQ318236 HQ318144 HQ328881 JN389138
Lactifluus volemoides TS 0705 GENT KR364038 KR364165 – –
Lactifluus volemus KVP 11-002 GENT JQ753948 KR364175 KR364360 KR364490
REH 9320 NY KR364096 KR364228 KR364362 KR364492
Lactifluus wirrabara GG 24-01-04 GU258307 GU265667 GU258407 KR364494
Lactifluus xerampelinus MH 201176 GENT KR364099 KR364231 KR364364 KR364496
GenBank accession numbers
Species Herbariumnumber Herbarium ITS LSU RPB2 RPB1
Table 1 (cont.)
2. a part of the ribosomal large subunit 28S region (LSU),
using primers LR0R and LR5 (Moncalvo et al. 2000);
3. the region between the conserved domains 6 and 7 of the
second largest subunit of the RNA polymerase II (RPB2),
using primers bRPB2-6F and fRPB2-7cR (Liu et al. 1999,
Matheny 2005); and
4. the region between domains A and C of nuclear gene en-
coding the largest subunit of RNA polymerase II (RPB1),
using primers RPB1-Ac and RPB1-Cr (Stiller & Hall 1997,
Matheny et al. 2002).
PCR products were sequenced using an automated ABI 3730
XL capillary sequencer (Life Technology) at Macrogen. Forward
and reverse sequences were assembled into contigs and edited
with Sequencher v. 5.0 (Gene Codes Corporation, Ann Arbor,
MI, USA) or BioloMICS (BioAware SA NV). The dataset was
supplemented with closely related sequences from GenBank
and worldwide reference sequences from De Crop et al. (2017)
(Table 1). Sequences were aligned online in the multiple se-
quence alignment program MAFFT v. 7 (Katoh & Toh 2008),
using the E-INS-I strategy. Trailing ends were trimmed and
the alignment was manually edited where needed in Mega 6
(Tamura et al. 2013). The alignment is deposited in TreeBASE
(Submission ID 24693). First, the ITS+LSU alignment was
partitioned into partial 18S, ITS1, 5.8S, ITS2 and partial 28S.
Both RPB1 and RPB2 alignments were partitioned into the
intron(s) and the first, second and third codon positions of
the exon. PartitionFinder was used to find the appropriate
partitioning scheme (Lanfear et al. 2017). Maximum likelihood
(ML) analyses, using RAxML v. 8.0.24 (Stamatakis 2014), were
combined with the Rapid Bootstrapping algorithm with 1 000
replicates under the GTRCAT option (Stamatakis et al. 2008).
There was no supported conflict between the separate gene
trees, so they were concatenated. ML analyses were conducted
on the CIPRES Science Gateway (Miller et al. 2010).
Distribution of closest relative(s)
To investigate whether species occurring in Central America
and the Caribbean are more closely related to North or to
South American species, the following steps were undertaken.
For each largest possible clade consisting only of species oc-
curring in Central America and/or the Caribbean (i.e., a total
of 22 clades), the closest relative(s) was/were determined by
considering the closest node with a bootstrap support value of
at least 70. If there was only one most closely related clade, or
if those most closely related clades originated from the same
continent, a value of one was added towards the count for that
continent. If the closest relatives of a clade were originating
from multiple continents, a value of one was added for each
of these continents. For each Central American or Caribbean
clade, hosts and altitude were noted when possible, and counts
of the closest relatives of different plant families and altitudes
were performed in the same way. These counts were visualised
in a bar plot using R (R Core Team 2018).

284 Persoonia – Volume 44, 2020
Fig. 1 Maximum Likelihood tree based on concatenated ITS, LSU, RPB2 and RPB1 sequence data. Maximum Likelihood bootstrap values > 70 are shown.
LD 15−061 Lactifluus nebulosus Martinique
J.P. Fiard 828A Lactifluus nebulosus holotype Martinique
LD 15−059 Lactifluus nebulosus Martinique
GUA−104 Lactifluus guanensis holotype Virgin Islands
RC/Guad11−023 Lactifluus guadeloupensis sp. nov. holotype Guadeloupe
LD 15−036 Lactifluus nebulosus Martinique
LD 15−031 Lactifluus nebulosus Martinique
PAM/Mart 05−091 Lactifluus nebulosus Martinique
LD 15−066 Lactifluus marmoratus sp. nov. holotype Martinique
PAM−Mart12−103 Lactifluus putidus Martinique
RC/Mart10 011 Lactifluus putidus Martinique
OKM−27240 Lactifluus coccolobae holotype Puerto Rico
LD 15−039 Lactifluus putidus Martinique
LD 15−004 Lactifluus putidus Martinique
PAM/Mart 11−013 Lactifluus putidus Martinique
LD 15−057 Lactifluus murinipes Martinique
LD 15−018 Lactifluus murinipes Martinique
F.1890 Lactifluus murinipes Martini que
LD 15−020 Lactifluus murinipes Martinique
LD 15−015 Lactifluus murinipes Martinique
CL/MART06.019 Lactifluus murinipes Martinique
J.P. Fiard 837A Lactifluus castaneibadius holotype Martinique
J.P. Fiard 849A Lactifluus murinipes holotype Martinique
J.P. Fiard 827B Lactifluus caribaeus paratype Martinique
Bandala 4374a Lactifluus chiapanensis Mexico
TH 9130 Lactifluus brunellus Guyana
G3185 Lactifluus sp. French Guiana
JLC 06031001 Lactifluus sp. French Guiana
RC/Guad 05−029 Lactifluus lepus sp. nov. holotype Guadeloupe
D.N. Pegler 3133 Lactifluus panuoides Guadeloupe
R.W.G. Dennis 109A Lactifluus panuoides paratype Trinidad And Tobago
TH7460 Lactifluus sp. Guyana
RC/Guy10 024 Lactifluus panuoides French Guiana
REH 9231 Lactifluus subclarkeae Australia
JET1006 Lactifluus flocktonae Australia
MN 2004002 Lactifluus clarkeae Australia
CL/GUAD06.045 Lactifluus panuoides Guadeloupe
PGK13−130 Lactifluus sp. New Caledonia
EDC 12−023 Lactifluus phlebonemus Cameroon
JD 941 Lactifluus flammans Congo
EDC 12−046 Lactifluus albomembranaceus holotype Cameroon
EDC 12−047 Lactifluus gymnocarpus Cameroon
AV 11−017 Lactifluus tanzanicus Tanzania
AV 11−137 Lactifluus nonpiscis Togo
AV 05−253 Lactifluus luteolus United States
EH 7194 Lactifluus rubrobrunnescens holotype Indonesia
AV 13−038 Lactifluus brunneoviolascens Italy
REH 9398 Lactifluus russulisporus holotype Australia
ADK 2840 Lactifluus foetens holotype Benin
AV 05−083 Lactifluus brunnescens Malawi
HKAS J7008 Lactifluus ambicystidiatus China
LM−UNAH 0073 Lactifluus sp. Honduras
LTH 55 Lactifluus dwaliensis Asia Thai land
ASM 12 075 Lactifluus sp. United States
AV 05−374 Lactifluus sp. United States
JN 2011−014 Lactifluus sp. Viet Nam
FH 18−063 Lactifluus sp. Panama
LTH 182 Lactifluus leucophaeus Thailand
TENN 064342 Lactifluus sp. United States
JN 2011−072 Lactifluus sp. Viet Nam
JN 2011−036 Lactifluus sp. Viet Nam
2001 08 19 68 Lactifluus piperatus France
KVP 12−001 Lactifluus oedematopus Germany
KVP 08−011 Lactifluus sp. Thailand
AV 17−015 Lactifluus pallidilamellatus Mexico
LTH 288 Lactifluus distantifolius Thailand
KVP 08−031 Lactifluus sp. Thailand
LTH 184 Lactifluus longipilus Thailand
REH 9320 Lactifluus volemus Australia
AV−KD−KVP 09−121 Lactifluus sp. India
KVP 08−034 Lactifluus crocatus Thailand
AV−KD−KVP 09−131 Lactifluus leptomerus holotype India
KVP 08−024 Lactifluus vitellinus holotype Thailand
AV−KD−KVP 09−134 Lactifluus dissitus Indi a
AV−KD−KVP 09−045 Lactifluus versiformis holotype India
KVP 08−002 Lactifluus acicularis Thailand
AV 17−014 Lactifluus sp. Mexico
KVP 08−048 Lactifluus subvolemus Slovenia
FH 18−134 Lactifluus sp. Panama
AV 05−227 Lactifluus sp. United States
KVP 11−002 Lactifluus volemus Belgium
FH 18−067 Lactifluus sp. Panama
JN 2011−061 Lactifluus subpruinosus Viet Nam
JN 2011−074 Lactifluus robustus Viet Nam
EH 72−195 Lactifluus lamprocystidiatus holotype Papua New Guinea
JN 2011−080 Lactifluus tenuicystidiatus Viet Na m
TENN 051830 Lactifluus bhandaryi holotype Nepal
AV 12−050 Lactifluus auriculiformis holotype Thailand
LTH 457 Lactifluus conchatulus isotype Thail and
GG−DK 17−02−05 Lactifluus genevievae holotype Australia
KW 304/FH 12−037 Lactifluus pulchellus holotype Thailand
LE 262983 Lactifluus igniculus holotype Viet Nam
Halling 8262 Lactifluus sp. Costa Rica
LM 4379 Lactifluus fuscomarginatus holotype Mexico
LM4640 Lactifluus sp. Mexico
AV 05−269 Lactifluus subgerardii United States
EH 6472 Lactifluus reticulatovenosus holotype Indonesia
LTH 270 Lactifluus sp. Thailand
FH 12−013 Lactifluus leae Thailand
AV 05−375 Lactifluus gerardii United States
F1068593 Lactifluus sp. Costa Rica
AV 17−016 Lactifluus sp. Mexico
AV 05−283 Lactifluus sp. United States
JN 2007−029 Lactifluus cf. gerardii var. fagicola Canada
DS 06−003 Lactifluus sp. Malaysia
PL 40509 Lactifluus sepiaceus New Zealand
GG 24−01−04 Lactifluus wirrabara Australia
LTH 274 Lactifluus sp. Thailand
AV 05−300 Lactifluus petersenii United States
AV/KD/KVP 09−120 Lactifluus ochrogalactus India
86
100
90
88
84
100
87
94 PAM−Mart12−90 Lactifluus nebulosus Martinique
71
PUN 7046 Lactifluus sainii India
RC/Mart 05−110 Lactifluus putidus Martinique
PAM/Mart 05−030 Lactifluus putidus Martinique
PAM−Mart12−88 Lactifluus putidus Martinique
RC/Mart 03−001 Lactifluus putidus Martinique
LD 15−033 Lactifluus putidus Martinique
LD 15−006 Lactifluus putidus Martinique
LD 15−011 Lactifluus putidus Martinique
PAM/Mart 05−085 Lactifluus putidus Martinique
LD 15−062 Lactifluus putidus Martinique
LD 15−030 Lactifluus putidus Mart inique
LD 15−002 Lactifluus putidus Mart inique
80 LD 15−065 Lactifluus putidus Marti nique
LD 15−041 Lactifluus putidus Mart inique
LD 15−040 Lactifluus putidus Mart inique
LD 15−017 Lactifluus putidus Martinique
RC/Guad 06−005 Lactifluus putidus Guadeloupe
100 LD 15−034 Lacti fluus putidus Martinique
100 LD 15−032 Lactifluus murinipes Martinique
LD 15−054 Lactifluus murinipes Martinique
CL/MART06.014 Lactifluus caribaeus Martinique
97
98 J.P. Fiard 818A Lactifluus caribaeus holotype Martinique
81
100 RC/Guad 08−042 Lactifluus lepus sp. nov. Guadeloupe
100 RC/Mart 03−077 Lactifluus panuoides Martini que
81
92
100
70
100
98
98
100
100
100
98
100
90
100 100
99
94
100
100
100
89
100
88 LGAM 2010−0132 Lactifluus glaucescens Greece
AV 04−195 Lactifluus sp. United States
100
99100
JN 2011−076 Lactifluus roseophyllus Viet Nam
FH 18−061 Lactifluus sp. Panama
100 FH 18−128 Lactifluus sp. Panama
84
100
100
8279
AV 05−293 Lactifluus sp. United States
91 AV 05−330 Lactifluus sp. United States
94
73
93
95
98
100 AV 17−011 Lactifluus sp. Mexico
AV−RW 04−023 LTH117 Lactifluus pinguis holotype Thailand
FH 18−132 Lactifluus sp. Panama
100 FH 18−068 Lactifluus sp. Panama
83
100
100 100 FH 18−054 Lacti fluus sp.Panama
SA A12 L2 Lactifluus corrugis United States
100 AV 05−392 Lactifluus corrugis United States
AV 17−012 Lactifluus pallidilamellatus Mexico
100 Montoya 4716 Lactifluus pallidilamellatus Mexico
100 FH 18−135 Lactifluus sp. Panama
FH 18−131 Lactifluus sp. Panama
99 FH 18−133 Lactifluus sp. Panama
98
100
100
98
99
100
100
100
95 LM4719 Lactifluus fuscomarginatus Mexico
FH 18−062 Lactifluus sp. Panama
99 FH 18−124 Lactifluus sp. Panama
95
100
97
100
97
98
73
DS 06−247 Lactifluus bicolor Malaysia
GG 07−02−04 Lactifluus leonardii Australia
100 PL 34204 Lactifluus sepiaceus New Zealand
92
85
99
FH 18−069 Lactifluus sp. Panama
100 FH 18−141 Lactifluus sp. Panama
Central America
Caribbean
L. sect.Lacti fluus
L. sect. Tenuicystidiati
L. sect. Gerardii
L. sect. Piperati
JN 2004−008 Lactifluus allardii United States L. sect. Allardii
L. sect. Ambicystidiati
L. subg. Lactifluus
L. sect.Nebulosi
L. sect. Panuoidei
L. sect. Tomentosi
L. sect. Luteoli
L. sect. Gymnocarpi
L. sect.Phlebonemi
L. sect. Gymnocarpi
Unnamed clade 1

285
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
ANGE1035 Lactifluus domingensis Dominican Republic
TENN 065854 Lactifluus sp. United States
JN 2011−071 Lactifluus sp. Viet Nam
AV 05−249 Lactifluus sp. United States
R. E. Halling 7993 Lactifluus hallingii Costa Rica
FH 18−077 Lactifluus hallingii holotype Panama
AV 05−275 Lactifluus sp. United States
NVE 520 Lactifluus hallingii Colombia
R. E. Halling 4977 Lactifluus hallingii Colombia
FH 18−129 Lactifluus sp. Panama
ATHU−M 8077 Lactifluus vellereus Greece
LM−UNAH 0072 Lactifluus sp. Honduras
AV 05−210 Lactifluus subvellereus United States
LTH 205 Lactifluus pilosus holotype Thailand
JN 2012−016 Lactifluus bertillonii Germany
AV 11−074 Lactifluus aureifolius Tanzania
AV 99−160 Lactifluus roseolus Zimbabwe
EDC 12−068 Lactifluus sp. Cameroon
FN 05−628 Lactifluus edulis Malawi
BB 00−1388 Lactifluus phlebophyllus Madagascar
EDC 14−032 Lactifluus urens Zambia
AB 305 Lactifluus brunneocarpus Guinea
JD 303 Lactifluus ruvubuensis Gabon
JD 988 Lactifluus cyanovirescens Congo
DS 07−454 Lactifluus leoninus Thailand
EDC 12−195 Lactifluus sp. Cameroon
ADK 3358 Lactifluus rufomarginatus Benin
G4836 Lactifluus sp. French Guiana
PAM/Guad 10−036 Lactifluus venezuelanus Guadeloupe
RC/Mart 03−120 Lactifluus venezuelanus Martinique
MAN MAN919 Lactifluus sp. Brazil
RC/Guy 12−007 Lactifluus sp. French Guiana
G4797 Lactifluus sp. French Guiana
TH7880 Lactifluus sp. Guyana
TH 9014 Lactifluus aff. annulifer Guyana
MR/Guy 13−032 Lactifluus sp. French Guiana
SLM 10114 Lactifluus subiculatus paratype Guyana
G4727 Lactifluus sp. French Guiana
RC/Guy 09−004bis Lactifluus sp. French Guiana
G3264 Lactifluus sp. French Guiana
TH 9154A Lactifluus multiceps Guyana
EDC 11−112 Lactifluus acrissimus Tanzania
LE 253907 Lactifluus chrysocarpus holotype Viet Nam
EDC 14−503 Lactifluus ramipilosus holotype Thailand
BB 99−409 Lactifluus madagascariensis Madagascar
AV 99−012 Lactifluus sp. Zimbabwe
JD 886 Lactifluus velutissimus Congo
AV 07−048 Lactifluus uapacae holotype Cameroon
BB 00−1518 Lactifluus annulatoangustifolius Madagascar
JD 946 Lactifluus chamaeleontinus Congo
JD 956 Lactifluus pelliculatus Congo
ADK 4462 Lactifluus cocosmus holotype Togo
AV 07−056 Lactifluus sp. Cameroon
AV 11−111 Lactifluus densifolius Tanzania
AV 11−183 Lactifluus melleus isotype Togo
AV 99−002 Lactifluus brachystegiae holotype Zimbabwe
JD 907 Lactifluus sp. Congo
RC/Guy 09−036 Lactifluus sp. French Guiana
MCA 3937 Lactifluus sp. Guyana
MR/Guy 13−145 Lactifluus sp. French Guiana
LD 15−014 Lactifluus pegleri Martinique
PAM−Mart12−91 Lactifluus pegleri Martinique
LD 15−049 Lactifluus pegleri Marti nique
M 8025 Lactifluus veraecrucis holotype Mexico
J.P. Fiard 821B Lactifluus pegleri paratype Martinique
PAM/Mart 05−088 Lactifluus pegleri Martinique
PAM−Mart12−54 Lactifluus pegleri Martinique
EDC 12−051 Lactifluus rubroviolascens Cameroon
EDC 11−218 Lactifluus denigricans Tanzania
EDC 11−159 Lactifluus subkigomaensis paratype Tanzania
AV 11−006 Lactifluus kigomaensis holotype Tanzania
AV 99−099 Lactifluus carmineus holotype Zimbabwe
AV 94−063 Lactifluus aurantiifolius holotype Burundi
TS 0705 Lactifluus volemoides holotype Tanzania
EDC 14−501 Lactifluus armeniacus holotype Thailand
JN 2011−012 Lactifluus sp. Viet Nam
FH 18−060 Lactifluus sp. Panama
TENN 065929 Lactifluus sp. United States
MH 201176 Lactifluus xerampelinus Mozambique
EDC 12−001 Lactifluus persicinus holotype Cameroon
JR Z 310 Lactifluus kivuensis holotype Congo
EDC 12−176 Lactifluus bicapillus paratype Cameroon
EDC 12−066 Lactifluus pumilus Cameroon
JD 959 Lactifluus rubiginosus Congo
AV 11−174 Lactifluus sudanicus isotype Togo
EDC 11−087 Lactifluus luteopus Tanzania
EP 1212/7 Lactifluus rugatus Greece
AV 05−251 Lactifluus hygrophoroides United States
JN 2001−058 Lactarius scrobiculatus Slovakia
FH 12−052 Lactarius hatsudake Thailand
ED 08−018 Lactarius olympianus United States
MTB 97−24 Lactarius fuliginosus Sweden
ADK 3598 Lactarius tenellus Benin
AV 11−177 Lactarius miniatescens Togo
76
83
100
100
JN 2011−077 Lactifluus sp. Viet Nam
97 AV 04−181 Lactifluus sp. United States
100
76
94
ANGE542 Lactifluus domingensis Dominican Republic
100 ANGE837 Lactifluus domingensis Dominican Republic
ANGE838 Lactifluus domingensis holotype Dominican Republic
100 AV 05−325 Lactifluus sp. United States
82
100
72 R. E. Halling 7938 Lactifluus hallingii Costa Rica
78
99
95
100
77
100
100
100
78
95
95
100
100
94
100
100
100
99
91
100
100
100
98 MR−GUY−14−195 Lactifluus sp. French Guiana
PAM/Guad 10−037 Lactifluus venezuelanus Guadeloupe
F.1980 Lactifluus venezuelanus Martinique
RC/Guad 03−048 Lactifluus venezuelanus Guadeloupe
73 RC/Guad11−017 Lactifluus venezuelanus Guadeloupe
7571
70
86
100
88
81 G4819 Lactifluus cf. annulifer French Guiana
95 R.W.G. Dennis 97 Lactifluus neotropicus holotype Trinidad And Tobago
100
81
71
100
100
95
100
100
100
97
100
95
100
100 LD 15−042 Lactifluus pegleri Martinique
97
100
100
100
72
100
99
93
100
100
100
100
100
100
100
100
74
96
100
0.1
L. sect. Neotropicus
L. sect. Lactariopsis
L. sect. Russulopsidei
L. sect. Edules
L. sect. Albati
L. subg. Lactariopsis
L. sect. Pseudogymnocarpi
L. sect. Xerampelini
L. sect. Aurantiifolii
L. sect. Rubroviolascentini
L. sect. Polyspaerophori
Outgroup
100
L. subg. Pseudogymnocarpi
RESULTS
Taxonomy
Lactifluus subg. Lactifluus
Lactifluus sect. Lactifluus
Lactifluus pallidilamellatus (Montoya & Bandala) Van de
Putte, Mycotaxon 120: 444. 2012
≡ Lactarius pallidilamellatus Montoya & Bandala, Cryptog. Mycol. 25 (1):
16. 2004.
Ecology — Found in mesophytic (subtropical) forest, with Car-
pinus caroliniana.
Distribution — Known from Veracruz, Mexico.
Notes — Lactifluus pallidilamellatus is characterised by
its slender habit, orange pileus and stipe, crowded, cream to
yellowish lamellae, brown discolouring context, copious white
latex, prominent lamprocystidia, basidiospores measuring
(6.4–)7.2–8.8 × (5.6–)6.4 –7.5(– 8) µm, with a heavy reticulum
and relatively short terminal elements in the pileipellis, measur-
ing 16–40.8 × 2.4–12.8 µm (Montoya & Bandala 2004). The
most closely related described species is L. oedematopus from
Europe (Fig. 1), which is morphologically very similar, and has
also been reported from forests with Carpinus (Van de Putte
et al. 2016), but their different distribution easily distinguishes
them from each other.
No other species in L. sect. Lactifluus have been described from
Central America, nor the Caribbean or South America. However,
our phylogeny reveals four undescribed species from this area
in this section, three from Panama and one from Mexico (Fig. 1).
Since this section represents a difficult species complex (Van
de Putte 2012), in which morphological differences are often
very subtle, a detailed study on this complex will be needed to
further unravel these species.
Fig. 1 (cont.)

286 Persoonia – Volume 44, 2020
Lactifluus sect. Gerardii (A.H. Sm. & Hesler) Stubbe, Per-
soonia 38: 76. 2016
Lactifluus fuscomarginatus (Montoya, Bandala & I. Haug)
Delgat, comb. nov. — MycoBank MB831635
≡ Lactarius fuscomarginatus Montoya, Bandala & I. Haug, Mycologia 104
(1): 176. 2012.
Ecology — Found in Fagus grandifolia var. mexicana forest.
Distribution — Known from Veracruz, Mexico.
Notes — Lactifluus fuscomarginatus is characterised by
the dark pileus and stipe, whitish lamellae with blackish brown
lamellae edges, pigmented, projecting and subcylindrical cheilo-
leptocystidia and basidiospores measuring (8–)9 –10–10.2 – 11
× (7–)8 –9 – 9.2 –10.5(–11) µm (Q = 1.08–1.14), with reticulate
ornamentation, with rounded to somewhat sharp ridges up to
0.5 µm high. Lactifluus fuscomarginatus is situated in a clade
together with three undescribed species from Mexico and Cen-
tral America, and the closest described relatives to this clade
are L. reticulatovenosus from Indonesia and L. subgerardii
from North America (Fig. 1). Lactifluus fuscomarginatus thus
belongs to L. sect. Gerardii (Fig. 1), and not to Lactarius subg.
Plinthogalus, and is here combined in Lactifluus.
Lactifluus subg. Gymnocarpi (R. Heim ex Verbeken)
De Crop, Persoonia 38: 75. 2016
Lactifluus sect. Nebulosi Delgat, sect. nov. — MycoBank
MB828339
Typus. Lactarius nebulosus Pegler, Kew Bull. 33: 610. 1979 (≡ Lactifluus
nebulosus (Pegler) De Crop).
Pileus medium to large sized, firm, convex with central de-
pression to infundibuliform, with inflexed margin when young;
surface slightly velutinous, often slightly wrinkled, sometimes
smooth, irregular or strongly wrinkled, white-grey with brown
spots to brown. Stipe central, cylindrical to tapering downwards,
firm to stuffed, whitish, staining brown. Lamellae subdecurrent
to deeply decurrent, often rather distant, white to cream; edge
concolourous. Context white, in most cases turning brown.
Odour in most cases unpleasant, fishy. Latex watery white,
in most cases staining brown and taste mild, in the case of
L. murinipes unchanging and acrid.
Basidiospores subglobose to ellipsoid; ornamentation com-
posed of isolated, rounded warts up to 1 µm high. True pleuro-
macrocystidia present in most species, up to 140–185 µm long,
cylindrical to subfusiform with rounded, mucronate, rostrate or
slightly moniliform apex. Pileipellis a trichoderm to palisade;
terminal elements in most cases thin-walled.
Notes — Lactifluus sect. Nebulosi corresponds to clade 9 in
De Crop et al. (2017) and belongs in L. subg. Gymnocarpi. In
contrast to the other described sections in this subgenus that
completely lack true pleurocystidia, most species of L. sect.
Nebulosi have conspicuous pleuromacrocystidia. The section
contains only Neotropical collections and is characterised by
dull fruiting body colours (a brown-grey pileus and a white-grey
Fig. 2 Basidiocarps. a. Lactifluus putidus, LD 15-002 (L. Delgat ); b. Lactifluus nebulosus, LD 15-061, inset LD 15-059 (L. Delgat); c. Lactifluus guadeloupensis,
RC/Guad11-023 (R. Courtecuisse); d. Lactifluus marmoratus, LD 15-066 (L. Delgat); e. Lactifluus murinipes, LD 15-020 (L. Delgat); f. Lactifluus lepus, RC/
Guad 05-029 (R. Courtecuisse); g. Lactifluus panuoides, RC/Mart 03-077 (R. Courtecuisse); h. Lactifluus venezuelanus, PAM/Guad 10-036 (P.-A. Moreau);
i. Lactifluus pegleri, LD 15-014 (L. Delgat).

287
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
stipe), an unpleasant fishy odour in all species except L. murini-
pes, and broadly ellipsoid spores with isolated, rounded warts
up to 1 µm high.
Lactifluus putidus (Pegler) Verbeken, Mycotaxon 120: 446,
2012 — Fig. 2a, 3
≡ Lactarius putidus Pegler, Kew Bull. 33 (4): 620. 1979.
Pileus 14–66 mm diam, plano-convex with central depression
and inflexed margin when young, infundibuliform with straight to
deflexed margin when adult; surface somewhat to very wrinkled,
brown (6E3–4, 6F3–5) with yellowish light brown areas, often
in the centre (4A3–5). Stipe 8–30 × 4 –14 mm, cylindrical, white
to yellowish white (4A1–4), with brown stains (5D5, 6E3–5).
Lamellae decurrent, medium spaced to distant, sometimes
slightly intervenose, white to cream (4A2–3), staining brown
(6E5, 6F4–7); edge entire and concolourous. Context white,
firm, turning dark brown, often first turning violaceous brown,
dark green blue with guaiac, greyish blue-green with FeSO4.
Odour very unpleasant, like rotten fish or urine. Taste mild. Latex
very abundant, watery white, staining brown, mild.
Basidiospores broadly ellipsoid to ellipsoid, 6.5–7.7– 9.3 –10.6
× 5.3–6.2–6.9 –7.7 µm (Q = 1.04 –1.23 –1.36 –1.53); orna-
mentation amyloid, composed of isolated rounded warts up
to 1 µm high, rarely connected by fine connective lines; plage
inamyloid. Basidia 51–67.5 – 84.5(– 88) × 9–11–13.5 µm, sub-
clavate, 4-spored. Pleuromacrocystidia 72.5–121.5–170.5
× 5.5–8–10.5 µm, cylindrical to subfusiform with rounded,
mucronate or tapering apex, rarely branching, thin-walled. Pleu-
ropseudocystidia scarce, 4 µm diam, not or slightly emergent.
Lamellar edge fertile. Hymenophoral trama mixed, with hyphae,
lactifers and sphaerocytes. Pileipellis a palisade; elements of
the suprapellis 29–41.5–53.5(–55) × 2.5– 3.5– 4 µm, cylindrical
to capitate up to 10.5 µm; subpellis composed of isodiametric
cells. Stipitipellis resembling pileipellis.
Ecology — Found in xerophytic and mesophytic forests,
with Bursera simaruba, Coccoloba diversifolia, Coccoloba pu-
bescens, Coccoloba swartzii, Guapira fragrans, Inga laurina,
Lonchocarpus roseus and Pisonia subcordata.
Distribution — Known from the islands of Martinique and
Guadeloupe.
Specimens examined. Martinique, Réserve naturelle La Caravelle, Com-
mune de Trinité, Bois de Pointe rouge, soil with Guapira fragrans, Coccoloba
pubescens, 03 Nov. 2015, Lynn Delgat, LD 15-004 (GENT); ibid., soil with
Guapira fragrans, Coccoloba pubescens, Coccoloba swarzii, 03 Nov. 2015,
Lynn Delgat, LD 15-011 (GENT); Réserve naturelle La Caravelle, Trinité,
Tartane, vicinity of ruins of Chateau Dubuc (14.769397°, -60.889991°, alt.
30.7 m), soil with Guapira fragrans, Coccoloba swartzii, 08 Nov. 2015, Lynn
Delgat, LD 15-030 (GENT); Réserve naturelle La Caravelle, Commune de
Trinité, Bois de Pointe rouge (14.757943°, -60.932939°, alt. 47.8 m), soil
with Coccoloba pubescens, Guapira fragrans, 10 Nov. 2015, Lynn Delgat,
LD 15-039 (GENT); Commune du Prêcheur, Anse Lévrier (14.844459°,
-61.218130°, alt. 64.1 m), soil with Guapira fragrans, 14 Nov. 2015, Lynn
Delgat, LD 15-062 (GENT); ibid., (14.844562°, -61.217664°, alt. 78.3 m),
soil with Guapira fragrans, 14 Nov. 2015, Lynn Delgat, LD 15-065 (GENT).
Notes — Lactifluus putidus is a commonly encountered
species and is probably the most frequent Lactifluus species
in the area (numerous specimens from various collectors have
been accumulated through the years and are mainly deposited
in K and LIP). It can easily be recognised in the field by the
strongly wrinkled cap in mature specimens, the strong, un-
pleasant odour and the very abundant latex. When the fruiting
body is cut, the latex often first turns violaceous brown before
turning dark brown. Microscopically, the presence of capitate
terminal elements in the pileipellis and stipitipellis is a distinctive
character, which was not observed in any of the other species
from the Antilles. Molecularly, L. putidus is closely related to
L. coccolobae (Fig. 1), but can be distinguished from it by the
strongly wrinkled pileus, the capitate elements in the pileipellis
and the absence of gelatinised hyphae in the pileipellis, in addi-
tion to L. coccolobae being associated with Coccoloba uvifera
in dune habitats in the Greater Antilles.
Lactifluus coccolobae (O.K. Mill. & Lodge) Delgat, comb.
nov. — MycoBank MB828348
≡ Lactarius coccolobae O.K. Mill. & Lodge, Mycologia 92 (3): 564. 2000.
Ecology — Found in sand on dunes under Coccoloba uvi-
fera.
Distribution — Known from Puerto Rico and Guana Island,
British Virgin Isles.
Notes — Based on the original description this species fits
morphologically in L. sect. Nebulosi, notably because of the
dull basidiocarp colours, the brown staining of the latex and
context, the strong fishy odour and the spore ornamentation with
isolated warts (Miller et al. 2000). The placement in this section
is confirmed molecularly by the position of the type sequence
in the phylogeny (Fig. 1). A similar species is L. guanensis,
which also occurs in the Greater Antilles with Coccoloba uvifera,
but L. coccolobae can be distinguished from this species by
its narrower basidia (8–9.5 µm wide), slightly shorter spores
(7.2–9(–10.8) µm long), lower spore ornamentation (up to 0.3
µm) and a gelatinised pileipellis (Miller et al. 2000). Lactifluus
coccolobae is closely related to the Lesser Antillean L. putidus
(Fig. 1), but can easily be distinguished from it by the absence
of a strongly wrinkled pileus, the absence of capitate elements
in the pileipellis and the presence of gelatinised hyphae in the
pileipellis.
Fig. 3 Lactifluus putidus (LD 15-004, LD 15-011, LD 15-030, LD15-065).
a. Basidiospores; b. basidia; c. macrocystidia; d. pseudocystidium; e. pileipel-
lis terminal elements; f. section through the pileipellis. — Scale bars = 10 µm.
c
b
d
a
ef

288 Persoonia – Volume 44, 2020
Lactifluus guadeloupensis Delgat & Courtec., sp. nov. —
Myco Bank MB828343; Fig. 2c, 4
Diagnosis: Differs from L. nebulosus and L. marmoratus by the irregular pileus
surface, the pruinose lamellae and the presence of many 2-spored basidia.
Typus. Guadeloupe, Grande-Terre, Commune de Saint François, Baie
Olive (16.249905°, -61.283412°, alt. 2.9 m), with Guapira and Fabaceae,
06 Aug. 2011, R. Courtecuisse, RC/ Guad11-023 (LIP).
Etymology. Refers to the island where the species was found.
Pileus up to 40 mm diam, convex with slight central depression;
young basidiocarps sometimes umbilicate; surface irregularly
bumpy, finely pubescent to velvety, marmorate greyish brown
mixed with yellowish to yellowish ochraceous, becoming dull
rusty brown. Stipe 20 × 6–10 mm, short, slightly tapering
downwards; surface pruinose, with rather variable colours,
dirty cream when young to yellowish brown or marmorate when
adult. Lamellae adnate to subdecurrent, distant, thick, more or
less intervenose through irregular ridges or bumps, pruinose,
yellowish cream, turning rusty brownish. Context cream, turning
brown, slightly stuffed. Odour unpleasant, reminding of fish or
rotting material, then becoming a more classical lactarioid odour
while drying or reminding the odour of L. volemus (i.e., agree-
able shellfish-like, Jerusalem artichokes). Taste not observed.
Latex watery white, colour change and taste not observed.
Basidiospores mostly broadly ellipsoid, some subglobose or
ellipsoid; size very variable, 7.1–9.4–9.6–11.8(–11.9) × 5.8–
7.5–7.9–9.4 (–9.5) (Q = 1.08–1.23 –1.26 –1.41); ornamentation
amyloid, composed of rounded warts, up to 1 µm high, isolated;
plage inamyloid. Basidia (54–) 56.5–75.5 –94.5 × 8–10.5–
13(–14) µm, cylindrical to subclavate, majority 4-spored but
many 2-spored and some 1- and 3-spored, sometimes with
wall locally thickened. Pleuromacrocystidia 53–104.5–155.5
× 6.5–8–10 µm, abundant, cylindrical with moniliform apex,
thin-walled. Pleuropseudocystidia absent. Lamellar edge fertile.
Hymenophoral trama mixed, with hyphae, lactifers and sphaero-
cytes. Pileipellis a palisade to trichopalisade; elements of the
suprapellis 29–60–91.5 × 2.5–4.5 –6 µm, cylindrical, some
septate, some branching; subpellis composed of isodiametric
cells and more elongated cells. Stipitipellis resembling pilei-
pellis, terminal elements slightly shorter, 29.5– 42.5–56 × 3–5–
6.5(–7) µm.
Ecology — Found in xerophytic forests with Guapira and Fa-
baceae.
Distribution — Only known from type locality.
Notes — Lactifluus guadeloupensis is morphologically simi-
lar to L. nebulosus and L. marmoratus. However, it can be macro-
scopically distinguished from these two species by an irregular
pileus surface and pruinose lamellae. Microscopically it can
be distinguished from L. nebulosus by longer basidia, longer
terminal elements in the pileipellis and potentially by the pre-
sence of many 2-spored and some 1- and 3-spored basidia.
For microscopic differences with L. marmoratus, see the notes
on that species. The relationships of L. guadeloupensis are not
completely resolved, and it is closely related to L. coccolobae,
L. guanensis, L. marmoratus, L. nebulosus and L. putidus
(Fig. 1). However, Lactifluus guadeloupensis is recorded from
Grande-Terre of Guadeloupe and it is the only hitherto known
Lesser Antillean Lactifluus species found on calcareous soil of
coral origin, in contrast to the volcanic origin of the localities of
the other Lactifluus species.
Lactifluus marmoratus Delgat, sp. nov. — MycoBank
MB828344; Fig. 2d, 5
Diagnosis. Differs from L. guadeloupensis and L. nebulosus by the slightly
larger spores, the larger and especially wider basidia, the presence of
branching pleuromacrocystidia and the large isodiametric cells in the
pileipellis and stipitipellis.
Typus. Martinique, Commune du Prêcheur, Anse Lévrier (14.845595°,
-61.216511°, alt. 56 m), soil with Guapira fragrans, 14 Nov. 2015, Lynn Delgat,
LD 15-066 (GENT).
Etymology. Refers to the marbled colour of the pileus.
Pileus 39 mm diam, convex with slight central depression and
deflexed margin; surface chamois-leather like, grey (8E2), with
some white, and greyish orange (5B4) and light brown (6D4)
spots. Stipe 27 × 6 mm, cylindrical, somewhat tapering down-
wards; surface chamois-leather like, greyish brown (6E2) to light
brownish grey (6C1, 6D2), marbled with light brown (6D5) spots.
Lamellae decurrent, rather distant, with some bifurcations near
the margin, white, staining brown; edge even and concolourous.
Context white, slightly stuffed, unchanging, dark green blue with
guaiac, slowly dark greenish grey with FeSO4. Odour fishy, like
fresh fish, shrimps or crayfish. Taste insignificant. Latex rather
scarce, watery white, staining brown; taste mild.
Basidiospores broadly ellipsoid to ellipsoid, rarely subglobose,
8.9–10.5–12.1 × 7.3–8.3–9.3 µm (Q = (1.11–)1.13–1.26–
1.40); ornamentation amyloid, composed of rounded warts,
up to 1 µm high, mostly isolated, sometimes connected by
fine connective lines; plage inamyloid. Basidia 68– 85 –102 ×
12.5–14–16 µm, subclavate, 4-spored. Pleuromacrocystidia
84–134–184 × 7–9–11 µm, medium abundant, cylindrical with
rounded, mucronate or slightly moniliform apex, some branch-
ing in two, thin-walled. Pleuropseudocystidia 4.5–10 µm diam,
not emergent; content oleiferic. Lamellar edge fertile. Hymeno-
phoral trama mixed, with hyphae, lactifers and sphaerocytes.
Pileipellis a palisade; elements of the suprapellis 22–50–78 ×
3–5–8 µm, cylindrical to lageniform, with brown intracellular
Fig. 4 Lactifluus guadeloupensis (RC/Guad11-023). a. Basidiospores;
b. basidia; c. macrocystidia; d. pileipellis terminal elements; e. section through
the pileipellis. — Scale bars = 10 µm.
c
b
d
a
e

289
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
pigmentation, often septate; subpellis composed of very large
isodiametric cells, 15–45 µm diam. Stipitipellis resembling
pileipellis, but without brown intracellular pigment in the ter-
minal cells.
Ecology — Found in mesophytic forests with Guapira fra-
grans.
Distribution — Only known from type locality.
Notes — Lactifluus marmoratus macroscopically resem-
bles L. guadeloupensis and L. nebulosus. However, there are
some subtle macroscopic differences: L. nebulosus differs by
the lack of brown tinges in the pileus and the white stipe, and
L. guadeloupensis differs by the irregular pileus surface and the
pruinose lamellae. In contrast to the other species, the context
of L. marmoratus does not change colour when cut, although
this could be caused by the scarcity of the latex in the studied
specimen. Microscopically, L. marmoratus differs from both
species by the slightly larger spores, the larger and especially
wider basidia, the presence of branching pleuromacrocystidia
and the large isodiametric cells in the pileipellis and stipitipellis.
Molecularly, it is not clear what the most closely related species
is to L. marmoratus, but it is closely related to L. coccolobae,
L. guanensis, L. nebulosus and L. putidus (Fig. 1).
Lactifluus nebulosus (Pegler) De Crop, Persoonia 38: 76.
2016 — Fig. 2b, 6
≡ Lactarius nebulosus Pegler, Kew Bull. 33 (4): 610. 1979.
Pileus 7–83 mm diam, hemispherical with depressed centre
and inflexed margin when young, infundibuliform with straight
to deflexed margin when adult; surface chamois-leather like,
often slightly wrinkled, grey (4C2, 5B-C2, 5D3, 6B1) and white,
with some yellow (4A2–4) areas, and brown spots (5D4). Stipe
5–30 × 3–15 mm, cylindrical; surface irregular, white, with
brown (6D4, 6E5) stains. Lamellae decurrent to deeply decur-
rent, distant, often bifurcating near the margin, white to cream
(4A2), staining brown (5C4, 6E5); edge even and concolourous.
Context white, stuffed, turning brown when cut (7D3), with
the middle part of the stipe remaining unchanged in adult
specimens, dark green-blue with guaiac, dark greenish grey
with FeSO4. Odour like Russula xerampelina; shellfish. Taste
insignificant, mild. Latex abundant to very abundant in young
specimens, watery white, staining brown; taste mild.
Basidiospores broadly ellipsoid to ellipsoid, 7.3–9–9.5 –11.3
(–12.8) × 5.9 – 6.9 –7.2 – 8.4(–9.7) µm (Q = 1.09–1.30–1.32–
1.46); ornamentation amyloid, composed of rounded warts, up
to 1 µm high, isolated, rarely connected by fine connective lines;
plage inamyloid. Basidia 47.5– 59.5–72 × 10–11.5–12.5(–13)
µm, subclavate, 4-spored. Pleuromacrocystidia (54–)59.5 –
112.5–166 × 6–9.5 –13 µm, very abundant, cylindrical with
rounded, mucronate or slightly moniliform apex, thin-walled.
Pleuropseudocystidia very scarce, not emergent, content olei-
feric. Lamellar edge fertile. Hymenophoral trama mixed, with
hyphae, abundant lactifers and abundant sphaerocytes. Pilei-
pellis a palisade; elements of the suprapellis 18–43–68 (–72)
× 2.5–4–5 µm, cylindrical to lageniform, often septate; sub-
pellis composed of isodiametric cells. Stipitipellis resembling
pileipellis.
Ecology — Found in xerophytic forests with Coccoloba pu-
bescens, Coccoloba swartzii and Guapira fragrans.
Distribution — Only known from the island of Martinique.
Fig. 5 Lactifluus marmoratus (LD 15-066). a. Basidiospores; b. basidia;
c. macrocystidia; d. pseudocystidia; e. pileipellis terminal elements; f. section
through the pileipellis. — Scale bars = 10 µm.
Fig. 6 Lactifluus nebulosus (LD 15-036, LD 15-059, LD 15-061). a. Basi-
diospores; b. pseudocystidium; c. ma crocystidia; d. basidia; e. pileipellis
terminal elements; f. section through the pileipellis. — Scale bars = 10 µm.
c
b
d
a
ef
c
b
d
a
e f

290 Persoonia – Volume 44, 2020
Specimens examined. Martinique, Réserve naturelle La Caravelle, Com-
mune de Trinité, Tartane, vicinity of ruins of Chateau Dubuc (14.769464°,
-60.890001°, alt. 31.2 m), soil with Guapira fragrans, Coccoloba pubes-
cens, 08 Nov. 2015, Lynn Delgat, LD 15-031 (GENT); ibid., (14.769232°,
-60.887195°, alt. 24.3 m), soil with Coccoloba pubescens, 08 Nov. 2015, Lynn
Delgat, LD 15-036 (GENT); Réserve naturelle La Caravelle, Commune de
Trinité, Tartane, close to bar ‘Le Phare’, above the mangrove, Anse Four à
Chaux (14.765185°, -60.897090°, alt. 20.1 m), soil with Coccoloba swartzii,
Guapira fragrans, 12 Nov. 2015, Lynn Delgat, LD 15-059 (GENT); ibid.,
(14.764985°, -60.897280°, alt. 18.8 m), soil with Coccoloba pubescens,
Guapira fragrans, 12 Nov. 2015, Lynn Delgat, LD 15-061 (GENT).
Notes — Lactifluus nebulosus is sometimes confused with
L. caribaeus, only differing by the absence of grey tinges in
the pileus of L. caribaeus and its stronger unpleasant odour.
However, microscopic differences are clear: L. nebulosus has
larger spores, a palisade structure of the pileipellis, and con-
spicuous macrocystidia, while L. caribaeus has a trichoderm
and no macrocystidia. Concordant with these clear microscopic
differences, L. nebulosus and L. caribaeus are molecularly
not sister species. In contrast, L. nebulosus is probably most
closely related (although unsupported: bootstrap = 69) to the
recently described L. guanensis from the British Virgin Isles
(Fig. 1). However, this species can easily be distinguished
from L. nebulosus by the presence of an amyloid plage, the
absence of pleurocystidia and the lamprotrichoderm structure
of the pileipellis (Crous et al. 2019). Lactifluus nebulosus is
also macro- and microscopically similar to L. guadeloupensis
sp. nov. and L. marmoratus sp. nov.; for comparison see the
notes on those respective species.
Lactifluus guanensis Delgat & Lodge, Persoonia 43: 375.
2019
Ecology — Found on sandy soil under Coccoloba uvifera.
Distribution — Only known from Guana Island, British Virgin
Isles.
Notes — This recently described species fits morphologically
in L. sect. Nebulosi, notably because of the dull basidiocarp
colours, the brown staining of the latex and context, the unplea-
sant odour and the spore ornamentation with isolated warts. The
placement in this section is confirmed molecularly by the posi-
tion of the type sequence in the phylogeny (Fig. 1). Compared
to L. coccolobae, which also occurs on Guana Island, L. guan-
ensis has broader basidia (9.5–12–14(–14.5) µm), slightly
longer spores ((7.3–)7.5–9.5–11.4(–11.7) µm), higher spore
ornamentation (up to 1 µm high) and non-gelatinised pileipellis
(Crous et al. 2019). Compared to Lactifluus species from the
Lesser Antilles, L. guanensis is easily distinguishable, notably
due to the often amyloid plage, the absence of macrocystidia
and the lamprotrichoderm structure of the pileipellis consisting
of thick-walled elements. In addition, only L. coccolobae was
also found in association with Coccoloba uvifera.
Lactifluus murinipes (Pegler) De Crop, Persoonia 38: 76.
2016 — Fig. 2e, 7
≡ Lactarius murinipes Pegler, Kew Bull. 33 (4): 623. 1979.
= Lactifluus castaneibadius (Pegler) De Crop, Persoonia 38: 76. 2016.
≡ Lactarius castaneibadius Pegler, Kew Bull. 33 (4): 622. 1979.
Pileus 8–75 mm diam, plano-convex with inflexed margin when
young, infundibuliform with straight to deflexed margin when
adult; surface chamois-leather like, sometimes slightly wrinkled,
dark brown (6F4–6, 7F4, 8F4–5), sometimes with some lighter
areas (5B4, 5D4, 6D4). Stipe 11–30 × 5–14 mm, cylindrical to
slightly tapering downwards, white to yellowish white (4A2),
turning light brown (5B3, 5C3, 5D4–5, 6D4), sometimes with
orange stains (6C6). Lamellae subdecurrent to decurrent,
medium to distant, often bifurcating near the margin, slightly
transvenose, white to cream (4A2), staining brown (5B3–4,
6C6); edge crenulate and concolourous. Context white, firm,
slowly turning brown, dark green blue with guaiac, no reaction
with FeSO4. Odour weakly sweet. Taste first mild, then acrid
to very acrid, sometimes mild when latex is scarce or absent.
Latex scarce to abundant, watery white, not changing colour;
taste acrid to very acrid or first mild, then acrid to very acrid.
Basidiospores subglobose to broadly ellipsoid, some slight-
ly ellipsoid, 6.1–8–8.2 – 9.8(–10.1) × 5.5– 6.5 – 6.8–8 µm
(Q = 1.04–1.19–1.23–1.36), in some collections rare large
spores are present (11.3–14.5 × 8.6–12.9 µm); ornamentation
amyloid, composed of rounded warts, up to 0.5–1 µm high,
mostly isolated, sometimes connected by fine connective lines;
plage inamyloid. Basidia 50.5–65.5–81 × (8–)8.5 –10.5 –12
µm, subclavate, mostly 4-spored, sometimes 1- or 2-spored.
Pleuromacrocystidia 62–102–142 × 7–9–11(–11.5) µm, very
abundant, cylindrical with rounded, mucronate or rostrate apex,
thin-walled. Pleuropseudocystidia very abundant, 4–7 µm diam,
not or slightly emergent; positive reaction with sulphovanillin.
Lamellar edge fertile. Hymenophoral trama filamentous, with
hyphae and abundant lactifers. Pileipellis a trichoderm; ele-
ments of the suprapellis 16–61.5–106.5(–150) × (2–) 2.5– 4–5
µm, cylindrical, some septate; subpellis composed of hyphae;
with emerging dermatocystidia 41–55.5–70 × 4–5.5–7 µm and
dermatopseudocystidia 5.5–7 µm diam. Stipitipellis resembling
pileipellis, terminal elements slightly shorter than in the pileipel-
lis, 22–45.5–68.5 × 3–4– 5 µm.
Ecology — Found in xerophytic forests with Coccoloba diver-
sifolia, Coccoloba pubescens, Coccoloba swartzii, Guapira
fragrans, Lonchocarpus sp. and Tabebuia sp.
Distribution — Only known from the island of Martinique.
Fig. 7 Lactifluus murinipes (LD 15-015, LD15-018, LD15-020, LD 15-054,
LD 15-057). a. Basidia; b. basidiospores; c. macrocystidia; d. pseudocystidia;
e. dermatocystidia; f. pileipellis terminal elements; g. section through the
pileipellis. — Scale bars = 10 µm.
cb
d
a
efg

291
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
Specimens examined. Martinique, Réserve naturelle La Caravelle, Com-
mune de Trinité, Tartane, close to bar ‘Le Phare’, above the mangrove, Anse
Four à Chaux (14.761724°, -60.925873°, alt. 16.9 m), soil with Guapira fra-
grans, 04 Nov. 2015, Lynn Delgat, LD 15-015 (GENT); ibid., (14.767778°,
-60.888451°, alt. 13.3 m), soil with Guapira fragrans, 04 Nov. 2015, Lynn
Delgat, LD 15-018 (GENT); ibid., (14.769612°, -60.890165°, alt. 33.7 m),
soil with Guapira fragrans, Coccoloba pubescens, 04 Nov. 2015, Lynn Delgat,
LD 15-020 (GENT); ibid., (14.765021°, -60.897273°, alt. 18.9 m), soil with
Coccoloba swartzii, Guapira fragrans, 12 Nov. 2015, Lynn Delgat, LD 15-054
(GENT); ibid., (14.764985°, -60.897280°, alt. 18.8 m), soil with Coccoloba
swartzii, Guapira fragrans, 12 Nov. 2015, Lynn Delgat, LD 15-057 (GENT);
Réserve naturelle La Caravelle, Commune de Trinité, Tartane, vicinity of ruins
of Chateau Dubuc (14.768835°, -60.889188°, alt. 23.9 m), soil with Coccoloba
swartzii, 08 Nov. 2015, Lynn Delgat, LD 15-032 (GENT).
Notes — Lactifluus murinipes is easily distinguishable from
the other Antillean species in L. sect. Nebulosi. Macroscopically,
it differs by the dark brown pileus, by the latex that tastes acrid
and does not turn brown, by the absence of an unpleasant
odour and by the absence of a reaction with FeSO4.
Microscopically, the trichodermial pileipellis with abundant
conspicuous dermatomyositis helps distinguish L. murinipes
from most other similar species.
It was previously already suspected that L. castaneibadius
and L. murinipes were synonyms (De Crop et al. 2017). Our
phylogeny (Fig. 1) confirms the synonymy of these species
molecularly, since sequences of the holotypes of both species
are part of the same well-supported clade (p-distance = 0.002).
They were previously morphologically distinguished from each
other by the colour of the stipe, the sulphovanillin reaction of the
pseudocystidia and the spore size and shape. However, stipe
colour appears quite variable because of the colour-changing
context. Additionally, it was observed that all recent collections
belonging to this clade have a positive sulphovanillin reaction,
but the reaction time differs between collections, which could
explain the perceived absence of the reaction in collections that
react slower. The different spore sizes mentioned in the original
description (7.5–8–9 × 5.5–7– 8 µm in L. castaneibadius and
9–9.5–10.5 × 6.5 –7.5 – 8.5 µm in L. murinipes) can be con-
sidered intraspecific variation. The difference in spore shape
(Q = 1.16 in L. castaneibadius and Q = 1.28 in L. murinipes)
could be explained by the difference in spore size, since the
measurements of the recent collections of this species show
that larger spores generally have slightly bigger Q values. In
conclusion, morphological differences between these species
were subtle and are now known to be intraspecific variation.
Lactifluus caribaeus (Pegler) Verbeken, Mycotaxon 120: 446.
2012 — Fig. 8
≡ Lactarius caribaeus Pegler, Kew Bull. 33 (4): 617. 1979.
Pileus 40–75 mm diam, plano-convex or depressed to almost in-
fundibuliform, with involute margin when young; surface slightly
velutinous, slightly wrinkled, ivory yellow, bruising with cinnamon
to clay spots. Stipe 20–40 × 10–25 mm, tapering downwards,
very thick and robust, white, turning cinnamon. Lamellae deeply
decurrent, slightly distant, thick, occasionally anastomosing,
cream; edge entire and concolourous. Context whitish, firm,
turning light cinnamon, light green blue with guaiac, rapidly dark
green with FeSO4. Odour strong, unpleasant, recalling urine.
Taste mild. Latex white, staining cinnamon.
Basidiospores subglobose to broadly ellipsoid, rarely slightly el-
lipsoid, 6.6–7.6–8.5 × 5.8–6.3– 6.8 µm (Q = 1.06–1.20–1.35);
ornamentation amyloid, composed of rounded warts, up to 0.5
µm high, isolated, sometimes connected by fine connective lines;
plage inamyloid. Basidia 47.5–59–71 × 8–9–10.5 µm, cylindric
to subclavate, 4-spored. Pleuromacrocystidia absent. Pleuro-
pseudocystidia 3–9 µm diam, not emergent to emergent; apex
rounded or mucronate; content oleiferic. Lamellar edge fertile.
Hymenophoral trama mixed, with hyphae, lactifers and sphaero-
cytes. Pileipellis a trichoderm; elements of the suprapellis
26.5–49–72 × 2.5–3.5– 4.5 µm, cylindrical, some septate; sub-
pellis composed of hyphae. Stipitipellis resembling pileipellis.
Ecology — Found in xerophytic forests with Coccoloba pu-
bescens.
Distribution — Only known from the island of Martinique.
Specimens examined. Martinique, Sainte-Luce, Trois Rivières, Ravine
Saint-Pierre, towards Ravin Gros Vaisseaux (north of the nationale 5)
(14.467552°, -60.926485°, alt. 22.1 m), soil, 28 Nov. 2006, J.P. Fiard, CL/
MART06.014 (LIP); Réserve naturelle La Caravelle (alt. 5 m), soil with Coc-
coloba pubescens, Myrica coriacea and Zanthoxylon caribaeum, 26 Oct.
1976, J.P. Fiard, J.P. Fiard 818A, holotype (K).
Notes — Only one recent collection of this species was
found, which lacks a field description and picture, so the macro-
scopic part of the description is based on the original description
and picture (Pegler & Fiard 1979, 1983). Lactifluus caribaeus
can be recognised macroscopically by the combination of the
ivory yellow pileus and the strong, unpleasant odour. Molecu-
larly, L. caribaeus is most closely related to L. chiapanensis
from Mexico (Fig. 1), and microscopically, they share the
trichodermial structure of the pileipellis. The only other Lesser
Antillean species with this type of pileipellis structure is L. mu-
rinipes. However, L. murinipes is macroscopically clearly distinct
from L. caribaeus because of the dark brown pileus, the acrid
unchanging latex and the absence of an unpleasant odour.
In contrast, L. nebulosus does look macroscopically similar
to L. caribaeus and is sometimes confused with this species,
as already discussed in the notes on L. nebulosus. Another
morphologically similar species is L. coccolobae, known from
Fig. 8 Lactifluus caribaeus (CL/ MART06.014). a. Basidiospores; b. basidia;
c. pseudocystidia; d. pileipellis terminal elements; e. section through the pilei-
pellis. — Scale bars = 10 µm.
c
b
d
a
e

292 Persoonia – Volume 44, 2020
Puerto Rico and Guana Island (BVI). This species differs from
L. caribaeus, notably by the gelatinised pileipellis, the adnate
and crowded lamellae and its dune habitat in association with
Coccoloba uvifera.
Lactifluus chiapanensis (Montoya, Bandala & Guzmán) De
Crop, Persoonia 38: 76. 2016
≡ Lactarius chiapanensis Montoya, Bandala & Guzmán, Mycotaxon 57:
412. 1996.
Ecology — Found in tropical forest with Gymnopodium anti-
gonoides.
Distribution — Known from Chiapas, Mexico.
Notes — Lactifluus chiapanensis fits morphologically in
L. sect. Nebulosi, notably because of the brown staining of the
latex and context, and the spore ornamentation with isolated
warts. It is most closely related to L. caribaeus (Fig. 1), from
which it can be distinguished by the dark to greyish brown
pileus colours and the presence of macrocystidia, measuring
92–110.4 × 10.4–13.6 µm (Montoya et al. 1996).
Lactifluus unnamed clade 1
Lactifluus lepus Delgat & Courtec., sp. nov. — MycoBank
MB828346; Fig. 2f, 9
Diagnosis. Differs from species of L. sect. Nebulosi by the striate pectinate
margin, the widely spaced lamellae, the considerably smaller spores, the
thick-walled terminal elements of the pileipellis and the presence of many
1- and 2-spored basidia.
Typus. Guadeloupe, Petit-Bourg, Tambour, valley of the Tambour river
(16.189618°, -61.595192°, alt. 22.7 m), 03 Sept. 2005, R. Courtecuisse et
F. Lurel, RC/Guad 05-029 (LIP).
Etymology. Referring to the many 2-spored basidia with rather long
sterigmata, reminding of the silhouette of a hair (lepus).
Pileus 35 mm diam, convex with slight central depression, mar-
gin pectinate-striate, slightly bumpy on the striae but not strongly
lumpy; surface clearly rugose to wrinkled, more or less radially
in the middle and more concentrically towards the margin in
some places, pale yellow with some patches more brownish
or rusty, in the centre a bit darker (reminding of the colours of
Russula fellea). Stipe 20 × 7 mm, cylindrical, slightly curved,
surface finely pruinose, smooth or slightly wrinkled, cream with
pale orange hue. Lamellae decurrent, very distant, somewhat
thick, pale cream, edge entire and concolourous. Context very
pale cream, staining brown. Odour fruity. Taste not observed.
Latex white; taste extremely acrid.
Basidiospores subglobose to broadly ellipsoid, rarely glo-
bose, 5.6–6.1–6.6 × 4.8–5.4–5.9(– 6) µm (Q= 1.01–1.14 –
1.27(–1.28)); ornamentation amyloid, composed of isolated
rounded warts, up to 1 µm high; plage inamyloid. Basidia
53–65.5–77.5(–79) × 7.5–8–9 µm, subclavate, many 1-, 2-
and 4-spored basidia, rarely 3-spored. Pleuromacrocystidia
49.5–85.5–121.5(–136) × 5–7–8.5 µm, abundant, cylindrical to
slightly fusiform, sometimes branching, thin-walled, arising from
deep in the hymenium. Pleuropseudocystidia very abundant,
3–4 µm diam, sometimes branching, not or slightly emergent.
Lamellar edge sterile; marginal cells 20–40 × 2.5–4 µm. Sub-
hymenium cellular. Hymenophoral trama filamentous, with
hyphae and abundant lactifers. Pileipellis a lampropalisade;
elements of the suprapellis 28–78.5–129.5 × 4–5–6 µm,
cylindrical, often with subcapitate apex, thick-walled; subpellis
composed of isodiametric cells. Stipitipellis a trichoderm; ter-
minal elements as in pileipellis.
Ecology — Found in hygrophytic forests.
Distribution — Only known from the island of Guadeloupe.
Notes — In our phylogeny, as well as in the worldwide mo-
lecular analysis of Lactifluus by De Crop et al. (2017), L. lepus
is a rather isolated species in L. subg. Gymnocarpi. So far only
one closely related species is known: an undescribed species
from French Guiana (Fig. 1). Lactifluus lepus can easily be
distinguished from the species of L. sect. Nebulosi by the striate
pectinate margin, the widely spaced lamellae, the considerably
smaller spores and the thick-walled terminal elements of the
pileipellis.
Lactifluus sect. Panuoidei (Singer) Delgat, comb. nov. — Myco-
Bank MB831636
≡ Lactarius sect. Panuoidei Singer, Kew Bull. 7 (3): 301. 1952. — Typus.
Lactifluus panuoides (Singer) De Crop, Persoonia 38: 76. 2016.
≡ Lactarius panuoides Singer, Kew Bull. 7 (3): 300. 1952.
≡ Pleurogala panuoides (Singer) Redhead & Norvell, Mycotaxon 48:
377. 1993.
Lactifluus panuoides (Singer) De Crop, Persoonia 38: 76.
2016 — Fig. 2g, 10
≡ Lactarius panuoides Singer, Kew Bull. 7 (3): 300. 1952.
≡ Pleurogala panuoides (Singer) Redhead & Norvell, Mycotaxon 48: 377.
1993.
Basidiocarp pleurotoid. Pileus 10–23 × 8–24 mm diam, spatuli-
form, irregularly lobed; surface finely pubescent, slightly furfura-
ceous, wrinkled at the insertion, yellowish cream, staining pale
rosy rusty brown, sometimes almost completely purplish brown
when old. Stipe excentric and reduced. Lamellae converging
Fig. 9 Lactifluus lepus (RC/ Guad 05-029). a. Macrocystidia; b. basidio-
spores; c. basidia; d. pseudocystidia; e. pileipellis terminal elements; f. section
through the pileipellis. — Scale bars = 10 µm.
c
b
d
a
ef

293
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
to the stipe, not crowded, cream, staining pale rusty brown.
Context thin. Odour not remarkable. Taste mild. Latex watery
white, changing to rosy-brownish. Subiculum tomentose, white
to cinnamon.
Basidiospores subglobose to broadly ellipsoid, 5.7–6.8–7.9
(–8.7) × 5–5.8 –6.7(–6.9) µm (Q = 1.05 –1.17–1.29), orna-
mentation amyloid, composed of rounded to irregular warts,
up to 1 µm high, connected by thick connectives, subreticu-
late, plage completely amyloid. Basidia 27–37.5–47.5(– 48)
× 8–9–10 µm, subclavate, 4-spored. Pleuromacrocystidia
absent. Pleuropseudocystidia 7.5–9.5 µm diam, thick-walled,
content granular and very refringent, slightly emergent. Lamellar
edge fertile. Hymenophoral trama filamentous, with hyphae
and abundant lactifers. Pileipellis a palisade; elements of the
suprapellis 18.5–88–158 × 4.5–5–6 µm, cylindrical with ob-
tuse apex, thick-walled, often septate; subpellis composed of
isodiametric cells.
Ecology — Found on wood in hygrophytic forests.
Distribution — Known from the Lesser Antilles and French
Guiana.
Specimens examined. French Guiana, Commune de Regina, Savane-
Roche Virginie trail, PK 122,7 (4.309524°, -52.133775°, alt. 2.8 m), slope
forest, 26 Apr. 2010, R. Courtecuisse, RC/Guy 10-024 (LIP). – Guadeloupe,
Basse-Terre, Sofaia trail, on fallen twigs and debris in hygrophytic forest,
25 Oct. 1977, D.N. Pegler, Pegler 3133 (K). – Martinique, Fort-de-France,
Les Nuages, la Médaille, Rivière Blanche forest, on trunk, 04 Sept. 2003,
R. Courtecuisse, RC/Mart 03-077 (LIP). – trinidad, Naranja, 02 Oct. 1949,
Dennis 109 (K), holotype.
Notes — Lactifluus panuoides is the only pleurotoid Lacti-
fluus species known to occur in the Lesser Antilles. This species
has some striking characters: the thick-walled pseudocystidia
and the completely amyloid plage, which is common in Russula,
but rare in Lactifluus. Other Neotropical pleurotoid species are
L. brunellus and L. multiceps, both described from Guyana.
Lactifluus panuoides can be easily distinguished from them.
Lactifluus brunellus also belongs in L. subg. Gymnocarpi,
but it differs from L. panuoides by the brown basidiomes with
white margin, the shaggy fasciculate subiculum, the slightly
larger basidia (43–53 × 9–10.5 µm) and spores (6.8– 8.4 ×
(5.6–)6 –7.2 µm), the inamyloid plage, the rarely connected
echinulate-spinuose basidiospore ornamentation and the
pileipellis hairs which frequently have swollen bases. Lactifluus
multiceps on the other hand belongs to L. subg. Lactariopsis.
It can be distinguished from L. panuoides by the golden yellow
to brownish orange basidiomes, the well-developed stipe, the
strong acrid taste, the larger spores (7.2–9.2 × 6.4– 8 µm), the
plage without a large amyloid spot and the longer pileipellis
hairs (40–220 × 4– 7 µm) which frequently have swollen bases.
Additionally, both L. brunellus and L. multiceps are found in
forests dominated by Dicymbe corymbosa (Miller et al. 2002).
Molecularly, L. panuoides represents an isolated clade (Fig. 1;
De Crop et al. 2017). The collection from Guyana represents
a second species in this clade (Delgat, unpubl. results). Given
that L. panuoides is the type of Lactarius sect. Panuoidei, this
clade represents L. sect. Panuoidei, which is recombined in
this paper.
Lactifluus subg. Lactariopsis (Henn.) Verbeken
Lactifluus sect. Neotropicus J. Duque, Delgat, Verbeken,
M.A. Neves & A.A. Carvalho, Syst. Biodivers. 18: in press.
2020.
Lactifluus venezuelanus (Dennis) De Crop, Persoonia 38:
77. 2016 — Fig. 2h, 11
≡ Lactarius venezuelanus Dennis, Kew Bull. Add. Ser. 3: 467. 1970.
Pileus 28–80 mm diam, convex with deep central depres-
sion, striate for a 1/3 to a 1/2 of the radius; surface smooth to
slightly bumpy, pale apricot orange to bright brownish orange,
hygrophanous. Stipe 15– 35 × 4–10 mm, tapering downwards,
fistulose; surface apically with small ridges which are continuing
from the decurrent teeth of the gills, pale apricot, rustier towards
the base. Lamellae subdecurrent to decurrent, not crowded,
slightly pruinose, yellowish cream to pale rusty ochre; edge
entire and concolourous. Context white to very pale apricot.
Odour weak, more or less fruity or like rubber. Taste insignificant
or slightly acrid, very astringent in young specimens. Latex
white, quickly disappearing, hence absent in most specimens,
not changing colour.
Basidiospores subglobose to broadly ellipsoid, 5.9–6.6– 6.8 –
7.7 × 5.2–5.6–5.8– 6.5 µm (Q = 1.06–1.14 –1.17–1.34(–1.39));
ornamentation amyloid, composed of rounded to irregular warts,
up to 1 µm high, often connected by lower connective lines,
forming an incomplete reticulum; plage inamyloid. Basidia 34.5–
43–52 × 8–10–11.5(–12) µm, subclavate, mostly 4-spored,
some 1- and 2-spored present. Pleurocystidia absent. Pleuro-
pseudocystidia abundant, 6–10 µm diam, not or slightly
emergent. Lamellar edge sterile; marginal cells 15.5–25–34.5
× 2.5–3.5–4.5 µm, narrowly cylindric to subfusiform, often
tapering near apex. Hymenophoral trama mixed, with hyphae,
lactifers and sphaerocytes. Pileipellis a palisade; elements of
the suprapellis 8–65–122.5 × 2.5–3–3.5(– 4) µm, cylindrical
with obtuse apex, sometimes septate; subpellis composed of
isodiametric cells. Stipitipellis resembling pileipellis.
Ecology — Found in hygrophytic or mesophytic forests with
Guapira fragrans.
Distribution — Known from the islands of Martinique and
Guadeloupe.
Fig. 10 Lactifluus panuoides (Pegler 3133). a. Pseudocystidia; b. basidia;
c. basidiospores; d. section through the pileipellis. — Scale bars = 10 µm.
c
b
d
a

294 Persoonia – Volume 44, 2020
Specimens examined. Guadeloupe, Basse-Terre, commune de Saint-
Claude, Beausoleil, Trace du plateau Dimba (16.026758°, -61.702051°,
alt. 440.6 m), 09 Sept. 2003, R. Courtecuisse, RC/Guad 03-048 (LIP);
Basse-Terre, Petit-Bourg, Route de la Traversée, near the national park
of Guadeloupe, 14 Aug. 2010, P.-A. Moreau, PAM/GUAD 10.037 (LIP);
Basse-Terre, Goyave, Chutes de Moreau, 05 Aug. 2011, R. Courtecuisse,
RC/Guad11-017 (LIP). – Martinique, Fort-de-France, Plateau Perdrix, 05
Sept. 2003, R. Courtecuisse, RC/Mart 03-120 (LIP).
Notes — Lactifluus venezuelanus is easily distinguishable
from other Antillean Lactifluus species due to the orange striate
pileus, the apical ridges on the stipe and the absence of latex
in most fruiting bodies. This species is placed in L. subg. Lac-
tariopsis, which is confirmed morphologically by the absence
of true pleurocystidia and the absence of a colour change of
the context and/or latex as in most species of this subgenus.
The most closely related described species is L. subiculatus
from Guyana (Fig. 1), which shares the striate margin of the
pileus and the absent or scarce latex. This species can hardly
be confused with L. venezuelanus because of the smaller
fruiting bodies (9–30 mm), the presence of a well-developed
subiculum, the larger and much wider basidia (45–60 × 15–18
µm wide) and the larger spores (7.6–9.2(– 9.6) × 6.4 –7.6 µm)
(Miller et al. 2012). The most closely related Antillean species is
L. neotropicus (Fig. 1), which can easily be distinguished from
L. venezuelanus because of the presence of an annulus, the
absence of a striate margin and the presence of thick-walled
terminal elements in the pileipellis.
Lactifluus neotropicus (Singer) Nuytinck, Mycotaxon 118:
450, 2011 — Fig. 12
≡ Lactarius neotropicus Singer, Kew Bull. 7 (3): 299. 1952.
Pileus 30–40 mm diam, convex with central depression; sur-
face smooth at the centre, slightly velutinous, with indistinct
squamules towards the margin, brown to umber, with a paler
ochraceous zone at the margin. Stipe 10–15 × 7–8 mm, cy-
lindrical or tapering downwards, solid, pale buff, with apical
annulus just below lamellae insertion; annulus narrow, mem-
branous with a fibrillose margin, concolourous with the stipe
surface. Lamellae adnexed to subdecurrent, crowded, thin,
pale buff; edge concolourous. Context pale ochraceous buff
to pale vinaceous cinnamon, unchanging. Odour none. Taste
very acrid. Latex watery white, scarce.
Basidiospores subglobose to broadly ellipsoid, 6.2–7–7.9(–8)
× 5.3–6.1–6.9 (–7) µm (Q = 1.03–1.16 –1.30(–1.36)); orna-
mentation amyloid, composed of rounded warts, up to 0.5 µm
high, often connected by lower connective lines, forming an
incomplete reticulum; plage inamyloid. Basidia 30–37–43.5 ×
8.5–9.5–11 µm, subclavate, 4-spored. Pleurocystidia absent.
Pleuropseudocystidia not observed. Hymenophoral trama
mixed, with hyphae, lactifers and sphaerocytes. Pileipellis a pali-
sade; elements of the suprapellis 18–54.5–117.5 × 3.5– 6–8.5,
cylindrical with obtuse apex, sometimes tapering near the top,
thick-walled; subpellis composed of isodiametric cells.
Ecology — Found in hygrophytic forests.
Distribution — Only known from type locality.
Fig. 11 Lactifluus venezuelanus (RC/Guad 03-048, RC /Guad11-017, RC/
Mart 03-120). a. Basidiospores; b. basidia; c. marginal cells; d. pseudocysti-
dia; e. pileipellis terminal elements; f. section through the pileipellis. — Scale
bars = 10 µm.
c
b
d
a
ef
Fig. 12 Lactifluus neotropicus (R.W.G. Dennis 97). a. Basidiospores; b. ba-
sidia; c. pileipellis terminal elements; d. section through the pileipellis.
— Scale bars = 10 µm.
c
b
d
a

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L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
Specimens examined. trinidad, Diego Martin, between Maqueripe trail
and North Post, 30 Sept. 1949, R.W.G. Dennis, R.W.G. Dennis 97 (K),
holotype.
Notes — The holotype of L. neotropicus clusters in the phylo-
geny together with a collection from French Guiana (Fig. 1), but
since only ITS1 is available for the holotype and ITS variability
is relatively low in L. sect. Neotropicus, further research is ne-
ces sary to determine whether this collection could represent
L. neotropicus. Since no certain recent collections were found of
L. neotropicus, the macroscopic part of the description is based
on the original description (Singer 1952). Besides the holotype,
two other collections identified as L. neotropicus exist, collected
in 1977 by Fiard. However, one of them was not available for
loan due to the scarcity of the material. Microscopical study of
the second collection, i.e., J.P. Fiard 1043A from Guadeloupe,
showed that, compared to the holotype, this collection has much
longer basidia (55–64.5–80 × 7–8.5–9.5 µm) and subcapitate
pileipellis hairs, which suggest it could represent a new species.
Aside from this putative new species, L. neotropicus is the only
species in the Antilles with velum. This velum, together with the
absence of a striate margin and the presence of thick-walled
terminal elements in the pileipellis, makes L. neotropicus easily
distinguishable from L. venezuelanus. Both species belong
in L. subg. Lactariopsis, which is the only subgenus in which
secondary velum is observed (Montoya et al. 2012). Another
Neotropical Lactifluus species which has velum is L. annulifer,
which is described from the Brazilian Amazon. Lactifluus annu-
lifer is quite similar to L. neotropicus but differs notably by the
mild taste of the context, the much larger spores (8.8–13 × 8–10
µm) and the presence of macrocystidia (Singer et al. 1983).
Lactifluus sect. Albati (Bataille) Verbeken, Mycotaxon 118:
451. 2011
Lactifluus domingensis Delgat & Angelini, IMA Fungus 10
(14): 7. 2019
Ecology — Found in montane forests with Pinus occidentalis.
Distribution — Only known from Dominican Republic, on the
island of Hispaniola.
Notes — This recently described species fits morphologically
in L. sect. Albati, because of the large white basidiomes, the
acrid taste and the velutinous cap. The placement in this section
is supported molecularly by the position of the type sequence
in the phylogeny (Fig. 1). Compared to L. hallingii, which
also belongs in L. sect. Albati, but occurs in Central America,
L. domingensis has slightly higher spore ornamentation (up
to 1.7 µm), somewhat shorter basidia ((42–)43 –53 –63.5 ×
9.5–12.5–15.5 µm), somewhat differently shaped macrocys-
tidia (i.e., more often a mucronate or moniliform apex, more
rarely with a rounded or tapering apex) and a loosely arranged
cutis to trichoderm as a pileipellis. In addition, L. hallingii is
associated with Quercus, while L. domingensis is associated
with Pinus occidentalis (Delgat et al. 2019).
Lactifluus hallingii Delgat & De Wilde, IMA Fungus 10 (14):
10. 2019
Ecology — Found in montane forests with Quercus hum-
boldtii, Quercus seemanii, Quercus copeyensis, Quercus sp.
Distribution — Known from Costa Rica, Panama and Co-
lombia.
Notes — This recently described species fits morphologically
in L. sect. Albati, because of the large white basidiomes, the
acrid taste and the velutinous cap. The placement in this section
is confirmed molecularly by the position of the type sequence in
the phylogeny (Fig. 1). For a comparison with L. domingensis,
see the notes on L. domingensis.
Lactifluus subg. Pseudogymnocarpi (Verbeken) De Crop,
Persoonia 38: 75. 2016
Lactifluus sect. Polysphaerophori (Singer) Verbeken, Myco-
taxon 120: 445. 2012
Lactifluus pegleri (Pacioni & Lalli) Delgat, comb. nov. — Myco-
Bank MB826723; Fig. 2i, 13
≡ Lactarius pegleri Pacioni & Lalli, Mycotaxon 44 (1): 182. 1992.
Pileus 13–90 mm diam, hemispherical with involute margin and
central depression when young, infundibuliform with straight
margin when adult; surface slightly velutinous, wrinkled near
the margin, burgundy red (9D7–9E4–10D4) with some paler,
more orange areas (5A4–6, 5B5, 6B3, 7D3), especially near
the margin. Stipe 11–50 × 6–20 mm, cylindrical; surface slightly
velutinous, reddish yellow-orange (4A4, 5A3–4, 6A2, 6B3,
7C3, 8C4, 8D5). Lamellae subdecurrent to decurrent, rather
distant, some intervenose, some bifurcating near the margin,
cream (3A2, 4A2); edge even and concolourous. Context white,
firm, slowly dark green blue with guaiac, slowly darkening with
FeSO4. Odour insignificant. Taste mild. Latex scarce to abun-
dant, watery white, not changing colour, taste mild.
Basidiospores broadly ellipsoid, some subglobose or ellipsoid,
6.8–7.4–8.2 – 9.3 × 5.5– 6.1– 6.7–7.5 µm (Q = 1.06–1.22–
1.38); ornamentation amyloid, composed of rounded warts,
Fig. 13 Lactifluus pegleri (LD 15-042, LD15-049). a. Pseudocystidia; b. basi-
diospores; c. basidia; d. pileipellis terminal elements; e. section through the
pileipellis. — Scale bars = 10 µm.
c
b
d
a
e

296 Persoonia – Volume 44, 2020
up to 0.2–0.5 µm high, mostly isolated, sometimes connected
by fine connective lines; plage inamyloid. Basidia (51–)53–
68.5–84.5 × 7–8.5–10(–10.5) µm, cylindrical to subclavate,
slightly refringent, 4-spored, often arising deep in the hymenium.
Pleuro cystidia absent. Pleuropseudocystidia 6–7.5 µm diam,
not emergent. Lamellar edge fertile. Hymenophoral trama
cellular, with sphaerocytes and broad lactifers. Pileipellis a
palisade; elements of the suprapellis 16.5–52– 87 × 3.5– 4–4.5,
cylindrical with obtuse apex, slightly thick-walled, some sep-
tate; subpellis composed of rather small, slightly thick-walled
isodiametric cells. Stipitipellis resembling pileipellis.
Ecology — Found in xerophytic forests with Coccoloba pu-
bescens and Guapira fragrans.
Distribution — Only known from the island of Martinique.
Specimens examined. Martin ique, Réserve naturelle La Caravelle,
Commune de Trinité, Tartane, close to bar ‘Le Phare’, above the mangrove,
Anse Four à Chaux (14.761724°, -60.925873°, alt. 16.9 m), soil with Guapira
fragrans, 04 Nov. 2015, Lynn Delgat, LD 15-014 (GENT); ibid., (14.762887°,
-60.897124°, alt. 10.6 m), soil with Coccoloba pubescens, Guapira fragrans,
12 Nov. 2015, Lynn Delgat, LD 15-049 (GENT); Réserve naturelle La Cara-
velle, Commune de Trinité, Bois de Pointe Rouge, soil with Coccoloba pubes-
cens, Guapira fragrans, 10 Nov. 2015, Lynn Delgat, LD 15-042 (GENT).
Notes — Lactifluus pegleri is an easily recognizable spe-
cies because of its fleshy habitus and notable burgundy red
to orange pileus colour. It is known from several localities
in Martinique and additional specimens collected during the
2003–2015 field trips (coll. R. Courtecuisse) are deposited in
LIP. This species was first identified by Pegler as L. hygropho-
roides, and only later proposed as a new species (Lalli & Pacioni
1992). Indeed, macroscopically, L. pegleri strongly reminds
of L. hygrophoroides. However microscopically, L. hygro-
phoroides has more elongated spores (Q = 1.35) and a more re-
ticulate spore ornamentation (Lalli & Pacioni 1992). Additionally,
the ecology of these two species is very different, with L. hygro-
phoroides occurring with temperate broad-leaved species (e.g.,
oaks). An important distinguishing feature reported in the origi-
nal description, was the trichodermial structure of the pileipellis
(Lalli & Pacioni 1992), however, our microscopical study of the
collections shows that L. pegleri has a lampropalisade structure
as a pileipellis, similar to L. hygrophoroides. Despite the similar-
ity between these two species, the phylogeny confirms that they
are different species and shows that they are not even closely
related, although they both belong to L. subg. Pseudogymno-
carpi (Fig. 1). Lactifluus pegleri is the only Antillean species in
this subgenus and like most species in this subgenus, L. pegleri
has a lampropalisade, unchanging latex and an orange to
reddish brown pileus. The most closely related species is
L. veraecrucis from Mexico (Fig. 1), which strongly resembles
L. pegleri according to the original description (Singer 1973)
and a microscopic study of the isotype. Recent collections of
L. veraecrucis are needed to find characters, besides their dif-
ferent distribution, to distinguish them. Given that L. veraecrucis
has been designated as the type of L. sect. Polysphaerophori,
L. pegleri belongs to this section as well. Next to these two
Central American species, this section also contains at least
three unnamed South American species. Species previously
classified in this section based on morphology, such as L. vene-
zuelanus and L. nebulosus, do not belong in this section and
the section is in need of an updated morphological description.
Lactifluus veraecrucis (Singer) Verbeken, Mycotaxon 120:
445. 2012
≡ Lactarius verae-crucis Singer, Beih. Sydowia 7: 104. 1973.
≡ Lactarius veraecrucis Singer. 1973.
Ecology — Found in tropical rain forest.
Distribution — Known from Chiapas and Veracruz, Mexico.
Notes — Lactifluus veraecrucis represents the type species
of L. sect. Polysphaerophori. Its lamellar trama is completely
cellular, and the presence of sphaerocytes in the trama was
one of the original characters used to define this section, which
caused numerous species to be included in this section. Cur-
rently only two described species are confirmed to belong in this
section: L. veraecrucis and L. pegleri. Lactifluus veraecrucis is
closely related to L. pegleri (Fig. 1) from the Lesser Antilles, and
morphologically closely resembles it. The spores of L. verae-
crucis are slightly more subglobose (7–8.3 × 6.5–7 µm) than
those of L. pegleri, but the easiest way to distinguish them is
their different distribution. Recent collections of L. veraecrucis
could help to find additional characters to distinguish between
these species.
KEY TO CARIBBEAN SPECIES
1. Context and latex not changing colour .............2
1. Context, pileus, lamellae, stipe and /or latex changing to
brown.......................................4
2. Basidiospore ornamentation mostly composed of isolated
warts; basidia 51–84.5 µm long; pileus colour burgundy
red with orange; context mild...............L. pegleri
2. Basidiospore ornamentation with many connectives be-
tween the warts; basidia shorter, 30– 52 µm long; pileus
colour orange to brown; context slightly to very acrid (subg.
Lacta riopsis) .................................3
3. Annulus or velar remnants present on the stipe; pileus not
striate; terminal elements of the pileipellis thick-walled ..
.................................L. neotropicus
3. Annulus or velar remnants absent; pileus rather thin and
striate; terminal elements of the pileipellis thin-walled...
................................L. venezuelanus
4. Basidiocarp large and white; pileus velutinous; context
acrid; associated with Pinus occidentalis L. domingensis
4. Not this combination of characters (subg. Gymnocarpi) 5
5. Habitus pleurotoid ....................L. panuoides
5. Habitus agaricoid; basidiospores with isolated verrucae 6
6. Pileipellis structure a trichoderm; pleurocystidia absent 7
6. Pileipellis structure a trichopalisade to palisade; pleuro-
macro cystidia 49.5–184 × 5 –13 µm ............... 9
7. Terminal elements of the pileipellis thick-walled ........
...................................L. guanensis
7. Terminal elements of the pileipellis thin-walled .......8
8. Pileipellis gelatinised, found in dunes with Coccoloba uvi-
fera...............................L. coccolobae
8. Pileipellis not gelatinised, found in forests with other Cocco-
loba species......................... L. caribaeus
9. Pileus surface very irregular with a striate pectinate mar-
gin ....................................L. lepus
9. Pileus surface rather smooth or wrinkled, margin not stri-
ate ........................................10
10. Taste of latex and context acrid; latex not changing colour;
no reaction with FeSO4; odour not distinctly unpleasant .
................................... L. murinipes
10. Taste of latex and context mild; latex changing colour to
brown; context reacting green-grey with FeSO4; odour
distinctly unpleasant .......................... 11
11. Pileus colour brown, often light brown in the centre; pileus
surface often strongly wrinkled; odour characteristically
very strongly unpleasant; terminal elements up to 55 µm
..................................... L. putidus
11. Pileus colour marbled; pileus surface at most slightly
wrinkled, surface chamois-leather like; terminal elements
up to 68–91.5 µm ............................12

297
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
12. Brown tinges present in pileus; stipe colour variable (grey-
dirty cream); basidiospores broadly ellipsoid (average
Q = 1.24–1.26) ..............................13
12. No brown tinges in pileus; stipe white; basidiospores el-
lipsoid (average Q = 1.30–1.32) .........L. nebulosus
13. Basidia rather broad (average width 14 µm); large sphaero-
cytes (15–45 µm) in the pileipellis ..... L. marmoratus
13. Basidia rather slender (average width 10.5 µm); small sphae-
rocytes (up to 15 µm) in the pileipellis ...............
.............................. L. guadeloupensis
KEY TO CENTRAL AMERICAN AND MEXICAN SPECIES
1. Basidiocarps large and white; pileus velutinous; taste acrid
...................................... L. hallingii
1. Basidiocarps not white; pileus not velutinous .........2
2. Pileus colour orange ............................ 3
2. Pileus colour greyish brown to blackish brown ........ 4
3. Latex unchanging; pleurocystidia absent; spore ornamen-
tation up to 0.5 µm high, not forming a reticulum........
...................................L. veraecrucis
3. Latex staining brown; conspicuous pleurolamprocystidia;
spore ornamentation up to 1–2 µm high, forming a complete
reticulum ....................... L. pallidilamellatus
4. Latex unchanging; pleuromacrocystidia present; spore orna-
mentation not forming a reticulum ...... L. chiapanensis
4. Latex staining brown; pleurocystidia absent; spore ornamen-
tation reticulate ..................L. fuscomarginatus
Distribution of closest relative(s)
Figure 14 shows the distribution of the closest relative(s):
i. for taxa originating from Central America or the Caribbean;
ii. according to host association of Central American and
Caribbean taxa; and
iii. by altitude of those taxa.
The majority of Caribbean clades have a South American closest
relative, while Central American clades show a more mixed
result. Clades found asso ciated with Fabaceae, Polygonaceae
or Nyctaginaceae have closest relatives in South America,
while clades found associated with Betulaceae, Fagaceae or
Pinaceae have closest relatives from North America, Asia and/
or Europe. Clades reported from altitudes lower than 500 m
were found to have South American relatives, while clades
reported from higher altitudes, mainly over 1 000 m, were found
to have North American, Asian and/or European relatives.
DISCUSSION
Diversity of Lactifluus spp. in the Antilles
Studying Lactifluus diversity in the Antilles has not only revealed
new species, but also a new section. The newly described
section Lactifluus sect. Nebulosi contains six Lesser Antillean
species, as well as L. chiapanensis from Mexico, and L. coc-
colobae and L. guanensis from the Greater Antilles (Fig. 1).
The section belongs in L. subg. Gymnocarpi. In contrast to
the other described sections in this subgenus that completely
lack true pleurocystidia, most species of L. sect. Nebulosi have
conspicuous pleuromacrocystidia. The section contains only
Neotropical collections and is characterised by dull fruiting
body colours (a brown-grey pileus and a white-grey stipe),
an unpleasant fishy odour in all species except L. murinipes,
and broadly ellipsoid spores with isolated, rounded warts up
to 1 µm high. This type of isolated spore ornamentation is
relatively rare in the genus and is reminiscent of the African
L. sect. Phlebonemi in its traditional morphological delineation,
which contains the species L. angustus, L. arsenei, L. nonpiscis,
L. phlebonemus and L. pisciodorus. It is noteworthy that all
these species also exhibit an unpleasant and often fishy odour
(Verbeken & Walleyn 2010). Interestingly, two other Antillean
species, L. lepus and L. pegleri, have isolated spore ornamen-
tation. Both species do not belong in L. sect. Nebulosi, but in
different sections in L. subg. Gymnocarpi and L. subg. Pseudo-
gymnocarpi, respectively. The high proportion of this type of
ornamentation in the Lesser Antilles, which occurs in eight out
of eleven species with three independent origins, combined
with the relatively rare occurrence globally, raises the question
whether this type of spore ornamentation could represent an
evolutionary advantage in the habitats of the Lesser Antilles
and if it could be correlated to certain environmental variables.
Besides the six species of L. sect. Nebulosi, five other species
were recorded, bringing the total known diversity of the Lesser
Antilles to eleven Lactifluus species, although some species
Caribbean
Central.America
Europe
Asia
North America
South America
Own distribution
0
5
10
15
20
25
Polygonaceae
Nyctaginaceae
Fabaceae
Betulaceae
Pinaceae
Fagaceae
Europe
Asia
North America
South America
Associated host(s)
0
5
10
15
0−500
500−1000
1000+
Europe
Asia
North America
South America
Altitude
0
2
4
6
8
10
12
Distribution of closest relative(s) compared to
Fig. 14 Number of taxa with closest relative(s) from Europe, Asia, North America or South America compared to: a. Central American or Caribbean distribu-
tion; b. different host families of Central American and Caribbean taxa; c. different altitudes of Central American and Caribbean taxa.
c
b
a

298 Persoonia – Volume 44, 2020
remain rare (i.e., known from only one or two collections).
Even though intraspecific variation is poorly known or unknown
for these rare species, we are convinced it is appropriate to
describe them, given the extensive sampling and the clear
morphological and molecular differences. Of the eleven species
occurring in the Lesser Antilles, eight or nine have not been
reported from other areas and can be classified as endemic
to the Lesser Antilles. Only L. panuoides and L. venezuelanus
have been reported from the mainland, from French Guiana
(Fig. 1) and Venezuela (Dennis 1970), respectively. Lactifluus
neotropicus might also occur in French Guiana (Fig. 1), but
more research is needed to confirm this record. Lactifluus nebu-
losus was previously reported from the British Virgin Islands
(Miller et al. 2000), but this collection (GUA-104) was shown to
represent a different species: L. guanensis (Crous et al. 2019),
so no overlap in Lactifluus species between the Greater Antilles
and the Lesser Antilles has been found thus far. The Greater
Antilles are considerably less diverse in Lactifluus species than
the Lesser Antilles, with only three known species, despite the
much larger surface area. Considering the small surface area
of the Lesser Antilles, less than 15 000 km2, these islands can
be considered highly diverse in Lactifluus species. This result
contrasts with an earlier observed pattern of ectomycorrhizal
communities being less diverse in smaller areas (Peay et al.
2007). However, the distribution of these species across the
different islands of the Lesser Antilles is still poorly studied.
Given the small distances between the islands, it could be
expected that most species occur on multiple islands, although
it could be possible that some species are endemic to a single
island. To investigate this, more sampling is needed across
the archipelago.
The case of the Lesser Antilles demonstrates that sampling dif-
ferent forest types is important, even though in tropical climate
ectomycorrhizal fungi are considered to be more abundant in
xerophytic forests (Pegler & Fiard 1983). Our results confirm
the predominance of Lactifluus at the xerophytic forests of the
Lesser Antilles, with about 75 % of the collections originating
from this type of forests. Species occurring in these forests are
occasionally also found in hygrophytic or mesophytic forests,
often in association with Guapira fragrans, a common host
tree in xerophytic forests that can be present in mesophytic
or hygrophytic forests as well. Despite their association with a
host tree that is abundant in xerophytic forests, some species,
such as L. venezuelanus, show a clear preference for meso-
phytic or hygrophytic forests. Other species, such as L. lepus,
L. marmoratus and L. panuoides, are also exclusively known
from these forest types. Because of the presence of these
unique species, mesophytic and hygrophytic forests should not
be overlooked when studying the diversity of ectomycorrhizal
fungi, even though collections can be less abundant and/or
less conspicuous, and therefore harder to find.
Diversity of Lactifluus spp. in Central America and Mexico
In Central America and Mexico, our phylogeny revealed the
presence of at least 21 species of Lactifluus, of which only five
species have been described. Many of these species occur in
species complexes, such as L. sect. Gerardii, L. sect. Lactifluus
and L. sect. Piperati. These sections belong to L. subg. Lacti-
fluus, and no Neotropical representatives other than L. fusco-
marginatus were reported for this subgenus before (De Crop
et al. 2017). Therefore, it seems likely that these 16 unnamed
clades represent new undescribed species. However, given
the difficulty of delimiting species in complexes such as these,
with morphological differences often being very subtle, these
complexes will need detailed study to allow describing these
species. Despite the limited number of described species, our
phylogeny shows that there is a high diversity of Lactifluus spp.
in Central America and Mexico. In addition, many species are
known from only one or two collections and are originating
from a limited number of regions, so it can be expected that
the number of species will rise even more.
Contrasting diversity patterns of Lactifluus spp. between
Central America/ Mexico and the Antilles
No overlap has been found between species from Central
America and Mexico and species from the Caribbean, contrary
to what was detected for several other ectomycorrhizal fungi
associated with Coccoloba uvifera or Pinus for example (Ortiz-
Santana et al. 2007, Põlme et al. 2017). Moreover, some con-
trasting biogeographical patterns between the two regions
can be observed, such as how the species are distributed. In
the Antilles, 14 species were found, distributed across seven
clades, while in Central America and Mexico, 20 species were
found, distributed across 17 clades. This shows that there were
a relatively limited number of colonisations in the Antilles, after
which some clades radiated, especially L. sect. Nebulosi, while
in Central America there were many repeated colonisations,
with only limited radiation. In addition, the majority of Central
American species occur in L. subg. Lactifluus, and the ma-
jority of Caribbean species in L. subg. Gymnocarpi. Another
contrasting pattern is their different affinities, with Antillean
taxa mainly showing South American affinities, and Central
American and Mexican taxa mainly showing North American,
Asian and Euro pean affinities (Fig. 14). Previously, the Lesser
Antilles were suspected to contain Lactifluus species of both
North and South American origin (Pegler & Fiard 1983, Hackel
2014). However, the Antillean L. pegleri turns out to be unrelated
to the North American L. hygrophoroides. In fact, all Lesser An-
tillean Lactifluus species are embedded within South American
clades in the phylogeny (Fig. 1), so they all appear to have
more recent common ancestors with South American species.
Lesser Antillean species were mainly found associated with
Coccoloba (Polygonaceae) and Guapira (Nyctaginaceae), both
important ectomycorrhizal hosts in South America, showing that
host specificity could be responsible for the South American
origin of these taxa. In contrast, the three species from the
Greater Antilles have mixed origins, with L. coccolobae and
L. guanensis belonging to L. sect. Nebulosi, which consists
mainly of Lesser Antillean species and has South American
affinities, and L. domingensis having North American affinities
(Fig. 1). Lactifluus coccolobae and L. guanensis were found with
Coccoloba uvifera (Polygonaceae), while L. domingensis was
found in montane forests with Pinus occidentalis (Pinaceae).
In Central America and Mexico on the other hand, the majority
of the species were found to have North American, Asian or
European affinities, and only few species were found to have
South American affinities, despite being connected to this conti-
nent. Species with South American affinities are L. veraecrucis,
of which the host relationship is unknown, and L. chiapanensis,
which was found associated with Gymnopodium floribundum,
a species found at low altitudes in Mexico, Guatemala and
Belize. The genus Gymnopodium is only known from Central
America, but belongs to the Polygonaceae family, to which also
the ectomycorrhizal Coccoloba belongs. Coccoloba is distribu-
ted across Mexico, Central America, the Caribbean and South
America, and was reported as a host for most other species in
the same section as L. chiapanensis. In contrast, other Central
American species were found with Carpinus (Betulaceae),
Fagus or Quercus (Fagaceae), important ectomycorrhizal
host genera in North America, Europe and Asia. These plant
families can, however, also be found in the western mountain
ranges in South America, and L. hallingii for example, which is
associated with Quercus, was also reported from localities at
high altitudes in Colombia.

299
L. Delgat et al.: Lactiuus diversity in Central America and the Caribbean
Affinities of Lactifluus spp. with North and South American
taxa in the light of tectonics
The land masses of Central America arose as a result of the sub-
duction of the Cocos plate under the Caribbean plate, with
Costa Rica and Panama forming a separate block neighbouring
the Nasca plate in the south and the South American plate in
the southeast (Kellogg et al. 1995). The land masses emerged
first in the northern part of Central America during Oligocene
and Miocene, and finally the isthmus was closed probably in
the area of Panama approximately until 4 million years ago
(e.g., Haug et al. 2001). This process allowed plants and their
fungi to move southwards from North America and to colonise
Central America before the isthmus was closed, while north-
wards dispersal from South America was at first impeded by a
broad water passage and later only possible by island hopping
over an island archipelago that later fused into the land bridge
of Panama.
The Antilles are located at the northern and eastern boarder
of the Caribbean plate and are mostly the result of complex
subduction events of the Atlantic plate underneath the Carib-
bean plate. The Caribbean plate originated from the Pacific on
the western side of Northern South America and moved first
northwards and later eastwards resulting in shear zones with
the South American plate until it attained its modern aspect by
the end of the Middle Eocene (Freeland & Dietz 1971). There-
fore, there were possibly more opportunities for dispersal from
South America onto the Antilles than from North America. In the
middle Oligocene connections between North temperate flora
and at least Puerto Rico existed, as pollen records showed that
for example Fagus occurred on the island (Graham & Jarzen
1969). However, suitable habitats for the temperate flora were
located at high elevation, and due to the intense erosion on
Caribbean islands, those habitats have disappeared. The lower
elevation of islands today could act as an additional barrier to
the dispersion of North American taxa or their Central American
relatives.
CONCLUSIONS
In summary, we hypothesise that host specificity, possibly at
host family level, is a crucial factor causing the observed bio-
geographical patterns. Taxa associated with Fabaceae, Polygo-
naceae and Nyctaginaceae show South American affinities
and taxa associated with Betulaceae, Fagaceae and Pinaceae
show North American, Asian and European affinities (Fig. 14).
Sampling ectomycorrhizae may help to further determine to
which degree host specificity shapes Lactifluus diversity. How-
ever, since different plant taxa exhibit different distributions,
the effects will also be strongly associated with abiotic factors,
such as climate and altitude. We found that species occurring
at low altitudes show affinities to South American taxa, and
species occurring at high altitudes, in montane forests, show
affinities to North American, Asian and European taxa (Fig. 14).
These high altitudes have a similar climate and vegetation to
temperate regions, while at low altitudes climate and vegetation
are similar to tropical regions. Therefore, it is not possible to
separate the effects of climate and host association, and both
could be considered fundamental explanatory factors of the
ability of a taxon to colonise a certain region. In addition, the
different geological histories of Central America/Mexico and the
Antilles may also have contributed to the contrasting diversity
patterns between the regions.
These results suggest that host specificity and climate could
shape phylogenetic patterns in ectomycorrhizal fungi, and
encourage studying the effects of host specificity and climate
on Lactifluus diversity patterns on a global scale.
Acknowledgements The first author was funded by a doctoral scholarship of
the Special Research Fund (BOF, grant BOF-DOC-2015-007001). E. De Crop
is supported by the ‘Special Research Fund Ghent University’ (BOF, grant
BOF-PDO-2017-001201). The collecting trips to the Lesser Antilles (2006–
2015) were financially supported by various organizations and structures
such as ONF (French Forestry Office), DIREN, DEAL (Regional counterparts
of the French Ministry for Ecology in Guadeloupe and Martinique), Regional
administrations (Martinique in 2015) and Regional Parks (PNRM – Parc
naturel régional de Martinique). We thank the Environmental Ministry of
Panama (MiAmbiente) for issuing collection and export permits (SE/ PH-4-18,
SEX/H-2-18). O. Cáceres, M. Cuevas and J. Rodríguez are thanked for
logistic support during fieldwork in Panama. Part of the molecular work was
funded by the Laboratoire d’Excellence CEBA (ANR-10-LABX-25-01) and the
Bibliothèque du vivant sequencing project. The authors thank L. Martinez-Suz
and B. Dentinger for the DNA extractions of the dried collections from Kew
herbarium. The authors would also like to thank P.-A. Moreau, for his collec-
tions and the picture of L. venezuelanus. A. Verbeken wants to thank J. Perez
Moreno for organizing the fieldtrip in Mexico at the occasion of IWEMM9.
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