ArticlePDF Available

Russula vinosoflavescens sp. nov., from deciduous forests of Northern Alsace, France

ISSN (print) 0093-4666 (online) 2154-8889 Mycotaxon, Ltd. ©2017
October–December 2017—Volume 132, pp. 707–721
Russula vinosoavescens sp. nov.,
from deciduous forests of Northern Alsace, France
Jean Michel Trendel1, Felix Hampe2 & Annemieke Verbeken2*
1 7 rue des Coquilles, 67500 Haguenau, France
2 Department of Biology, Ghent University,
K. L. Ledeganckstraat 35, 9000 Ghent, Belgium
Correspondence to: *
Abstract—Based on morphological, molecular, and ecological data, a new species of
Russula sect. Russula, found on several occasions under deciduous trees in Northern Alsace
is described and illustrated as: Russula vinosoavescens, belonging to R. subsect. Sardoninae.
Key wordsBasidiomycota, Russulaceae, ITS, Russula persicina, phylogeny
Russula Pers. (Russulales, Basidiomycota) is an ectomycorrhizal genus with
a world-wide distribution that is particularly well studied in temperate regions,
notably in Europe (Romagnesi 1967; Sarnari 1998, 2005). e ectomycorrhizal
associations, which oen involve a selective fungus–plant partnership, were
sometimes advantageously used to delimit infrageneric taxa. Such has been
the case for R. subsect. Sardoninae Singer (Sarnari 1998, emend.), which is
divided into series—provisionally dened by Sarnari (1998)—comprising, on
one hand, species strictly associated with conifers (R. ser. Sardonia and R. ser.
Sanguinea, both essentially corresponding to R. subsect. Sardoninae as
conceived by Romagnesi 1967, 1987) and, on the other hand, species associated
with deciduous trees, sometimes in a host-specic relationship (R. ser.
Exalbicans = R. subsect. Exalbicantinae Singer as retained by Romagnesi, with
species found only under birches) and sometimes with a broader host-range
(R. ser. Persicina = R. subsect. Persicinae Romagn., species that can be found
under oaks, hornbeams, beeches, birches, chestnuts, poplars, willows).
708 ... Trendel, Hampe & Verbeken
Table 1. Russula, Gymnomyces, and uncultured specimens with GenBank
and UNITE sequence accession numbers used in the molecular analyses.
Species Specimen Country Sequence
G. gilkeyae JT 2572 USA AY239346
G. monosporus OSC 117360 USA EU669222
root sample (uncultured) France JQ890299
— — JN197654
— USA EF434062
— USA GU997950
— USA GU997948
R. americana F 18871 USA HQ604839
R. aquosa TU 101831 Estonia UDB015988
TU 101708 Estonia UDB011290
R. atrorubens TU 101691 Estonia UDB011358
TU 101851 Estonia UDB016006
R. bresadolae
(“R. atropurpurea”) HUE 178 Germany UDB000313
PRM 858109 Czech Republic HG423574
R. cavipes RT 9149 Italy JF908681
HUE 163 Germany AF418623
Russula cf. ammodica F 18874 USA HQ604840
R. citrinochlora TU 101905 Norway UDB016045
20024 Sweden UDB002550
R. consobrina TU 118108 Estonia UDB011223
TU 106965 Estonia UDB011207
R. depallens TU 101829 Estonia UDB015986
TU 101838 Estonia UDB015994
R. emetica F 14309 AY228360
DG 18 United Kingdom JQ888196
DG 44 United Kingdom UDB001628
R. exalbicans 2-1117IS76 — AY061674
R. fageticola (“R. mairei”) FH 12262 Germany KT934013
R. fageticola (“R. nobilis”) HUE 054 Germany UDB000346
R. fellea HUE 218 Germany UDB000345
HUE 177 Germany UDB000314
R. fragilis UE 2006-11-08-22 Italy UDB018436
UE 2004-20-09-04 Sweden UDB018434
R. gracillima TU 101725 Estonia UDB011361
TU 106437 Estonia UDB011176
R. luteotacta TU 106379 Estonia UDB011166
FH 12-187 Germany KT933991
R. ochroleuca TU 118113 Estonia UDB011228
LW 115 Germany UDB000295
R. persicina TU 101826 Estonia UDB015984
TU 106950 Estonia UDB011196
R. pumila F 133 Finland UDB009582
L 3X87 FM993279
R. queletii FH 12-237 Germany KT934007
RT 5140 Ital y JF908668
Russula vinosoavescens sp. nov. (France) ... 709
R. renidens TU 101809 Estonia UDB015975
JR 1758F IT Finland UDB011117
R. rhodomelanea FH 2011-BT307 Germany UDB018429
IB 92/451 HT Italy UDB018435
R. sanguinea TU 106710 Estonia UDB019728
FH 12240 Germany KT934008
R. sardonia TU 106951 Estonia UDB011197
FH 12215 Germany KT933999
R. silvestris JK 11080504 Sweden UDB018431
JK 11080802 Sweden UDB018430
R. thindii HT BSHC KD11-095 India KM386693
R. torulosa TU 106444 Estonia UDB011177
TU 101626 Italy UDB016261
R. vinosoavescens JMT 05081006 France UDB031189
JMT 08081512 France UDB031188
JMT 14090502 HT France UDB031187
JMT 05081007 France UDB031190
JMT 10080701 France UDB024104
JMT 13090715 France UDB024105
R. puellula (“R. puellaris”) MC 01-502 Denmark UDB000010
R. cuprea FH 12250 Germany KT934010
R. olivobrunnea TU 101883 Finland UDB016034
HT holotype; IT isotype.
e aim of this paper is to introduce a new Russula species occurring in
deciduous forests of Northern Alsace (NE France) and belonging to R. subsect.
Sardoninae. Our initial morphological concept of the species, based on several
collections from various localities, is fully supported by molecular data. ITS
sequence analysis shows that the dierent collections form a well-supported
clade in the Russula phylogenetic tree.
Materials & methods
All collections of the new taxon were made by Jean Michel Trendel and are deposited
in GENT Herbarium. ese collections are referred to with an eight-digit code
YYMMDDXX (year; month; day; collection number). All collecting sites are located
in Northern Alsace (NE France); the code of the municipality is that assigned by the
National Institute for statistics and economic studies (INSEE); names (in quotation
marks) of forest or place are those indicated on the topographic maps (1:25000 scale)
from the Institut national de l’information géographique et forestière (IGN); map
coordinates of the collections refer to the Universal Transverse Mercator (UTM)
32N kilometer grid system; geological data (underlying rock) were mainly obtained
710 ... Trendel, Hampe & Verbeken
from geological maps (1:50000) edited by the Bureau de Recherches Géologiques et
Minières (BRGM).
Morphological analysis
Specimens were photographed in situ. e pictures are available at
Microscopic observations were carried out
on fresh material. Pileipellis elements were studied on radial sections made by hand
midway from the cap margin, mounted either in SDS Congo red (Clémençon 1999)
or in distilled water. e search for acido-resistant encrustations was performed by
staining with Ziehl carbol fuchsin followed by a rapid dierentiation using a 1M HCl
solution. Pileocystidia content was revealed using sulfovanillin (SV) freshly prepared
with a 50% or 80% H2SO4 aqueous solution. Hymenial elements were studied in the
above-mentioned observation media. In some cases, additional controls were carried
out on very ne cap sections made from dried material and further rehydrated in a
moisture-saturated closed chamber for 24 h before observation in water. Basidiospores
(from spore deposits) were examined in Melzer’s reagent and spore measurements
(ornamentation excluded) were recorded randomly for 300 spores (6 collections) in
side view. Measurements are given as (MINa) [AVa – 2*SD]–AvaAVb–[AVb + 2*SD]
(MAXb), with AVa = lowest mean value for the measured collections and MINa the
minimum value corresponding with this mean value, AVb = greatest mean value and
MAXb the maximum value corresponding with this mean value, and SD = standard
deviation calculated for the measurements of one collection (minimum and maximum
value are only given if not in the 2*SD-interval). Q stands for spore “length/width ratio
and is given as (MINQa)–QaQb–(MAXQb) with Qa and Qb being the lowest, and the
highest respectively, mean ratio for the measured specimen. e colour of the spore
deposit, referring to Romagnesi’s (1967) scale, was assessed against a personal chart
(JMT) and the Dagrons chart (unpublished). Macrochemical reactions were determined
using FeSO4 in crystalline form, a 4% phenol solution, and a strong Guaiac solution
made of permanently soaked Guaiac wood in 95% ethanol.
Molecular analysis
Total genomic DNA was extracted from dried material according to Nuytinck &
Verbeken (2003), with modications described in Van de Putte et al. (2010). e ITS
region was amplied using the primers ITS1F–ITS4 (White et al. 1990, Gardes & Bruns
1993) and with polymerase PerfectTAQ (5 PRIME, Hilden, Germany) in accordance
with the manufacturer’s recommendation. PCR amplication followed Eberhardt
(2012), and the PCR products were puried using the Qiaquick PCR Purication
Kit (Qiagen, Hilden, Germany) and directly sequenced with BigDye 3.1 technology
(Applied Biosystems, now ermo Fisher Scientic, Wilmington, USA). Specimens
JMT-05081006, JMT-05081007, and JMT-05081512 were extracted and sequenced in
Muséum National d’Histoire Naturelle de Paris (Marc-André Selosse) following Séne
et al. (2015).
Raw sequences were edited in the BioEdit Sequence Alignment Editor version 7.2.5
(Hall, 2013) or Sequencher version 4.8 (Gene Codes Corporation). Edited sequences
Russula vinosoavescens sp. nov. (France) ... 711
were aligned by MAFFT version 7 using the strategy E-INS-i (Katoh & Standley 2013).
Maximum-likelihood searches for tree building were carried out locally with 100
replicates with the GTR+GAMMA model, selecting the best solution of all replicates
analysis in RaXML 8.1.12 (Stamatakis 2014). Fast bootstrap searches were done locally
or through the CIPRES Science Gateway (Miller et al. 2010) with 10 000 replicates.
e nal alignment included a total number of 67 ITS sequences, with 21 sequences
corresponding to the current concept of R. subsect. Sardoninae (Sarnari 1998). Except
for the R. vinosoavescens collections cited as studied material all sequences were
retrieved from GenBank or UNITE (Table 1). e tree with the highest log likelihood
(–4523.1303) is shown. Initial tree(s) for the heuristic search were obtained automatically
by applying the Maximum Parsimony method.
Russula vinosoavescens Trendel & F. Hampe, sp. nov. Figures 1, 2
MycoBank MB 819428
Diers from Russula persicina by its colour range, its mild or only slightly acrid taste,
its reticulate spore ornamentation, and the presence of granular pigments in cuticular
Type: France, Alsace, Morsbronn-les-Bains (67303), “Niederwald”, UTM 32N:
0406318/5419158, 213 m alt., on argillaceous soil with eumull humus (underlying
rock: upper Keuper red marls), under Quercus robur and Carpinus betulus with some
Fagus sylvatica nearby, in an area characterized by calcicolous mycoora with numerous
Phlegmacium (Cortinarius) species, 5 Sept. 2014, JMT-14090502 (Holotype, GENT;
UNITE UDB031187).
Etymology: Referring to the purplish-red wine-coloured (vinoso) cap and the
tendency of the cap to become brown-yellowish or ochraceous (avescens) as well as
when handling the stipe.
Pileus 4.0–6.5 cm diam., more or less eshy, rather rm to very rm but with
margin occasionally slightly elastic, oen irregular, at times knotty, convex-
attened with the margin sometimes incurved or even inrolled, then broadly
but shallowly depressed; margin not or shortly sulcate, oen exuose-undulate
or lobed, in some cases showing a very clear, whitish border extending to the
edge of the gills (giving a “festooned” appearance); surface rugulose, more or
less radially veined, frequently uneven-bumpy in the centre, a little shiny in wet
conditions, but soon dry (except sometimes in the centre which remains a little
greasy) and then more mat, purple-red, carmine-red, vinaceous-red, somewhat
violet, rather vivid, but frequently paler, livid pinkish-vinaceous, with mauve
(lilac) greyish shades that can entirely replace the reddish tinges, even more
clearly greyish purple at the margin, becoming yellowish cream-ochraceous
from the centre or showing locally ochre-bistre areas merging in the reddish
and greyish mauve tints, sometimes becoming almost totally beige-brown,
712 ... Trendel, Hampe & Verbeken
ochreous-brown, with livid purple-violet glints mostly near the margin and a
residual purple-brown at centre. Lamellae adnate or attenuated-subdecurrent,
sometimes sub-emarginate, with a variable number of lamellulae, crowded to
rather spaced, interveined, anastomosing and more or less forked at every
level, in some cases more particularly near the stem (exceptionally regular,
without any forking or anastomosing), (sub)acute at the cap margin (rarely
subobtuse), narrow (0.3–0.5 cm, exceptionally broader 0.7 cm), whitish or
very pale cream; edge entire but oen somewhat irregularly wavy. Stipe 2–6 ×
0.9–1.5(–2) cm, cylindrical or slightly enlarged downwards, usually rounded
at the base (in a few cases tapering), sometimes eccentric, rm to almost hard,
with a rm medulla (exceptionally hollow-stulous); surface nearly smooth,
nely wrinkled, white with occasionally a purple tinge (in one case remarkably
grey-purplish, somewhat brownish), distinctly yellowing (dirty, oen with
greyish-brown shades) when handled or bruised. Context white, in the pileus
almost unchanging, in medulla of stipe slightly yellowing, sometimes a little
purple-violet beneath the cuticle in the most coloured forms; taste mild (or
almost mild, occasionally with an unpleasant aertaste, somewhat bitter),
mild to slightly (and sometimes volatile) acrid in the gills, exceptionally more
strongly acrid; smell not distinctive.
Macrochemical reactions FeSO4: rose-orange; Phenol: brown; Guaiac:
in most cases reacting on the stem surface almost immediately (<2 s, in one
case more slowly), but of little intensity at the beginning, developing (10–15 s)
in blue of medium intensity, exceptionally with a stronger reaction, (dark blue);
on gills, the reaction developing slowly and weakly (blue-green to light blue).
Spore print cream, II(a)b–c.
Basidiospores subglobose to broadly ellipsoid, 6.4–7.28.1–9.0 × 5.9–6.5
6.9–7.7 µm (n = 300), Q = 1.06–1.111.20–1.30; ornamentation amyloid,
oen with incompletely amyloid warts; warts conical-obtuse, or more acute,
sometimes somewhat truncated, generally around 0.4–0.8 µm high, but
also frequently reaching 1.0 µm or higher, sometimes locally catenulate,
interconnected by crests or more frequently by (oen very ne) connectives,
forming a well-developed and oen nearly complete (with numerous meshes)
reticulum, which may appear confused in some cases; suprahilar plage
moderately amyloid, more or less bordered by small warts or low crests.
Basidia 4-spored, 42–57 × 9.5–12.5 µm, including sterigmata (about 5 µm
long). Pleuromacrocystidia 60–90 × 10.5–14.5 µm, numerous, fusiform
or more or less clavate, most commonly with a pointed appendage, which
can show constrictions, more rarely ending in a short obtuse protuberance,
Russula vinosoavescens sp. nov. (France) ... 713
Fig. 1. Russula vinosoavescens: colour forms observed. A, B. holotype, collection JMT-14090502;
C. collection JMT-05082042, with marked reddish pigmentation; D. collection JMT-05081006,
with marked reddish pigmentation; E. collection JMT-08081512, with predominating ochraceous
colour; F. collection JMT-10080701, with mauve–lilac colour reminiscent of a Griseinae.
714 ... Trendel, Hampe & Verbeken
strongly staining with SV. Pileipellis composed of hyphae containing reddish-
violaceous granular pigments; epicuticular elements (2.0–)2.5–4.0 µm diam.,
sometimes branched, exuose, sometimes inated up to 5.5 µm; terminal cell
obtuse, or slightly thickened with a short appendage, also subcapitulate, or
(exceptionally) pear-shaped 7.0 µm diam., but also conversely more or less
attenuated (sometimes abruptly). Pileocystidia of two types; type 1 usually
unicellular (occasionally with 1 septum), rarely bid, clavate–fusiform, less
frequently cylindric, 4.0–9.5 µm diam., oen appendiculate–capitulate or
showing an obtuse apical protuberance, sometimes slender, (at least 120 µm
long, narrowed at the base to 2.5–3.0 µm diam.), with a content reacting variably
depending on the collection—but also on a cuticular section—when treated
with SV, staining (rather) strongly (grey-blackish) or weakly (grey-pinkish)
or even remaining inert (pinkish); type 2 lactiferoid (some clearly termini of
ascending lactifers), pluriseptate, more or less regularly cylindrical, 6.0–10.5
µm diam., with variably shaped terminal elements that sometimes taper (with
possible constrictions), sometimes are somewhat thickened or subclavate, or
with type 1 type appendages, originating in the deeper pellis, distinctly reacting
with SV (except in one specimen where the granular content of pileocystidia
type 2 is inert or reacts only very weakly in SV); without encrusted elements
aer treatment with Ziehl fuchsin.
Ecology & distribution—Under deciduous trees, associated with
Carpinus betulus, or Quercus (Q. robur or Q. petraea), or both (possibly also
with Fagus sylvatica), in a rather wet environment (possibly periodically drier),
on more or less argillaceous soils, neutral to supercially slightly acidic, usually
nutrient-rich with a rather high base saturation level (calcium-rich), with
mesotrophic to eutrophic mull humus. Phenology: late July to early September
(summer fruiting). Known only from six sites (9 collections) in Northern
Alsace (France).
Additional specimens examinedFRANCE, Alsace. Bas-Rhin, Mattstall,
commune associated with Lembach (67263), ‘Sauerhald’, UTM 32N: 0409918/5426543,
alt. 203 m, underlying rock: Upper Muschelkalk marly calcareous formation, under
Quercus, Carpinus, Fagus (forest locally rich in orchids, e.g., Epipactis microphylla),
JMT-13090715 (GENT, UNITE: UDB024105); Dauendorf (67087), ‘Herrenwald’,
0402753/5410066, 168 m, Loess from the Quaternary (and more ancient alluvial
deposits?), in a wet environment, nitrogen-rich and with a high base saturation level,
under Carpinus, Quercus, Alnus glutinosa, Fraxinus excelsior, Ulmus sp., and with a
herbaceous layer comprising Allium ursinum, Carex sylvatica, Circaea lutetiana, Geum
urbanum, Glechoma hederacea, Paris quadrifolia, Stachys sylvatica, JMT-10080701
(GENT, UDB024104); 0402762/5409965, 171 m, Loess from the Quaternary (and
more ancient alluvial deposits?), under Carpinus, Quercus, JMT-05082042 (GENT);
Russula vinosoavescens sp. nov. (France) ... 715
Fig. 2. Russula vinosoavescens (holotype, collection JMT-14090502). A. fruitbody in sectional
view; B. pileus epicutis: pileocystidia (with content shown schematically) and hyphal terminal
elements; C. hymenial cystidia on gill sides; D. spores as observed in Melzer’s reagent. Scale bars:
A = 1 cm; B = 10 µm; C = 20 µm; D = 5 µm.
716 ... Trendel, Hampe & Verbeken
Bettwiller (67036), ‘Buchwald, 0365394/5416535, 347 m, Upper Muschelkalk marly
calcareous formation, under Quercus, Carpinus, with some Fagus and Prunus avium,
JMT-05081006 (GENT, UDB031189) and JMT-05081007 (GENT, UDB031190), most
likely belonging to the same mycelium; Forstheim (67141), ‘der Wald’, 0405631/5415509,
211 m, Pliocene sands and clays, wet environment, under Quercus, Carpinus, Populus
tremula with Russula lutensis Romagn. and R. rutila Romagn., JMT-08081512 (GENT,
UDB031188); Drusenheim (67106), ‘Barrwald’, approximately 0420900/5401000, 124 m,
Quaternary alluvial deposits, wet environment, on bare blackish ground, with Carpinus,
Quercus, Alnus glutinosa, Fraxinus excelsior, 25/07/1981, JMT-81072501 (GENT, spore
print only available).
Molecular analysis
e six ITS sequences of the newly described R. vinosoavescens form
an independent clade, which receives high bootstrap support (Fig. 3). e
clade corresponding to R. vinosoavescens is nested within a large, supported
(bootstrap value 75) clade that includes the typical European representatives of
R. subsect. Sardoninae (R. sardonia Fr., R. queletii Fr., R. torulosa Bres.). Beside
these, it contains a number of species (e.g., R. cavipes Britzelm., R. exalbicans
(Pers.) Melzer & Zvára, R. gracillima Jul. Schä., R. luteotacta Rea, R. persicina
Krombh., R. renidens Ruots. et al., R. sanguinea Fr.) included by Sarnari (1998)
in his emended concept of R. subsect. Sardoninae but attributed to dierent
(although closely related) infrageneric taxa by previous authors (Romagnesi
1967, Singer 1986, Bon 1988). In addition to the abovementioned species,
which fairly well represent R. subsect. Sardoninae sensu Sarnari, the clade
includes extra-European Sardoninae species such as R. thindii K. Das & S.L.
Miller and some poorly known species with an uncertain systematic position
(e.g., R. americana Singer, R. citrinochlora Singer).
e deeper phylogenetic relationships within the Sardoninae clade as
well as its external relationships with R. fellea (Fr.) Fr., R. consobrina (Fr.) Fr.,
R. ochroleuca Fr., and the large clade representing R. subsect. Russula
s.l. (including a sequence of the generic type R. emetica (Schae.) Pers.)
remain poorly resolved in the ITS analysis. Within the Sardoninae clade,
R. vinosoavescens occupies a rather isolated position in a weakly supported
subclade containing R. renidens and samples identied as R. citrinochlora.
Morphological analysis
Despite its mild taste (in most cases only weakly acrid in gills),
R. vinosoavescens shows a combination of characters—a more or less eshy
rm pileus, cuticle oen rugulose with polychromic colours, cream spore
Russula vinosoavescens sp. nov. (France) ... 717
Fig. 3. Molecular phylogenetic analysis by Maximum Likelihood method-based tree with the
highest log likelihood (–4523.1303), based on ITS sequences.
718 ... Trendel, Hampe & Verbeken
print, well-characterised and non-encrusted pileocystidia—that is consistent
with its position in R. subsect. Sardoninae sensu Sarnari. Moreover, by its
habitus, somewhat decurrent narrow gills, and clear yellowing of its stipe when
bruised, this species is very reminiscent of certain forms of R. persicina. Russula
vinosoavescens diers, however, unambiguously from the latter by its colours,
which are never ‘purered. Indeed, even in its reddest forms, the cap always
displays some purple-vinaceous tinges, presumably reecting the existence of
one or several blue pigments, occurring in sucient amount to signicantly
shade the red ones. e presence of these blue pigments becomes even more
apparent when grey-mauve (lilac) tints replace—sometimes completely—the
reddish colouring. In addition, this russula appears to fade readily to yellowish
ochre, or even brown-beige, and therefore colour forms devoid of any reddish
or bluish shades might be expected.
Other striking features are the oen rugulose surface of the cuticle and the
strong tendency of the lamellae to become anastomosed-forked, though it
should be kept in mind that these characters are variable. e same is true for
the variability of the speed and intensity of the Guaiac reaction, which is mostly
in an average position on the reactivity scale.
One of the most striking microscopical features is the aggregation of
pigments into granules (sometimes distinctly combining red and blue
pigments) that are generally easily observed in the cap cuticular hyphae. e
cuticular structure of specimens for which we were unable to examine in fresh
condition was re-examined in dried material. Concerning the least pigmented
collection JMT-08081512 (Fig. 1-E), cuticular sections from the central part
of the cap—apparently the richest in residual red and blue pigments—revealed
the presence of pigmented grains. Our search was much less conclusive for the
vividly coloured specimens JMT-05081006 (Fig. 1-D), and even if we were able
to show the presence of sparse coloured granules, it appears that a signicant
amount of pigment is either vacuolar (soluble) or at least not distinctly granular.
In addition, examination of the cuticle of collection JMT-05082042 (Fig. 1-C),
not studied in a fresh state, clearly revealed the presence of pigmented granules.
As reported by Romagnesi (1967) and Sarnari (1998), granular pigments are
highly characteristic of R. subg. Heterophyllidia but are also found occasionally
in some species belonging to other groups. In this respect, one should especially
mention R. renidens, a Nordic russula growing under birches, which shows
abundant granular red-purplish pigments, the latter probably co-occurring
in a soluble form accounting for the intensity of its colours (Sarnari 1998).
A similar situation may also exist for some of our vividly coloured collections
Russula vinosoavescens sp. nov. (France) ... 719
(notably JMT-05081006 and JMT-05081007). Indeed, it seems likely that
pigment aggregates contribute less eciently to global colouring compared to
those in solution, and that a strong predominance of granular pigments over
their soluble counterparts may explain the relative paleness of some of our
e morphology of the terminal hyphal cells of the epicuticular elements
appears too variable to be usable for species recognition. On the other hand,
pileocystidial shape seems informative at species level. It is also signicant that
this characteristic can be found in almost all members of R. subsect. Sardoninae
as dened by Sarnari, where the new species belongs.
Another important diagnostic feature is the strongly reticulate spore
ornamentation that distinguishes R. vinosoavescens from similar species
sharing the same cream spore print and also growing under deciduous trees,
such as R. persicina or the birch associates, R. exalbicans and R. renidens.
Within Russula subsect. Sardoninae, partitioned by Sarnari (1998) into
ecological series based on Russula/tree partnerships, R. vinosoavescens ts in
R. ser. Persicina, devoted to species associated with broadleaf trees (but not
strictly with birches); the other series cover species linked strictly to either
conifers (R. ser. Sardonia and R. ser. Sanguinea) or birches (R. ser. Exalbicans).
Indeed, R. vinosoavescens appears associated with oak (Quercus robur or
Q. petraea) or hornbeam (Carpinus betulus) or both, the only trees present
on all collecting sites. A wider host association might include beech (Fagus
sylvatica, generally well represented in the forests of interest but sometimes
rather far away from the fruiting bodies) or, less likely, aspen (Populus tremula,
which can occur in more complex environments). Birch (Betula spp.), absent
from all the prospected sites, should not be considered a potential associate.
However, regarding Romagnesis (1967) initial concept of Persicinae, which
refers to “intermediate forms between Emeticinae and Sardoninae [both sensu
Romagnesi!], with less reticulate spores and gills more decurrent or weeping
than the former, and more purely red in colour than the latter” [a concept at
least partly shared by Sarnari (1998)—“pileus pure red”], it becomes necessary
to restrict Sarnari’s R. ser. Persicina denition exclusively to ecology, as in
R. vinosoavescens colouration is not restricted to pure red and its spores are
rather strongly reticulate. In his classication, Bon (1988) does not retain
Romagnesi’s Persicinae group but treats the dierent taxa of R. persicina in his
emended R. subsect. Exalbicantinae. In this context, R. vinosoavescens would
take its place beside R. exalbicans, which also oen displays washed-out greyish
720 ... Trendel, Hampe & Verbeken
colours, and R. renidens, genetically a very closely related species sharing some
outstanding morphological features such as pigments that are at least partly
granular or lamellae that are more or less forked.
We are extremely thankful to Josie Lambourdière and Prof. Marc-André Selosse
(Muséum national d’Histoire Naturelle, Paris) for some additional molecular analyses,
and to Dr. Philippe Schaeer for helpful discussions. Dr. Ursula Eberhardt and Georey
Kibby are acknowledged for useful comments and reviewing the paper.
Literature cited
Bon M. 1988. Clé monographique des russules d’Europe. Doc. Mycol. 18(70–71): 1–120.
Clémençon H. 1999. Vom Umgang mit Kongorot. Schweiz. Z. Pilzk. 77(5): 247–250.
Eberhardt U. 2012. Methods for DNA barcoding of fungi. 183–205, in: WJ Kress, DL Erickson
(eds). DNA Barcodes: Methods and Protocols, Methods in Molecular Biology, vol. 858. New
York, Humana Press.
Gardes M, Bruns TD. 1993. ITS primers with enhanced specicity for basidiomycetes –
application to the identication of mycorrhizae and rusts. Mol. Ecol. 2(2): 113–118.
Hall T. 2013. BioEdit, Biological sequence alignment editor for Windows 5/98/NT/2000/XP/7,
version 7.2.5. Ibis Biosciences, Carlsbad.
(accessed 15 March 2016).
Katoh K, Standley DM. 2013. MAFFT multiple sequence alignment soware version 7:
improvements in performance and usability. Mol. Biol. Evol. 30(4): 772–780.
Miller MA, Pfeier W, Schwartz T. 2010. Creating the CIPRES Science Gateway for inference of large
phylogenetic trees. 1–8, in: Proceedings of the Gateway Computing Environments Workshop
(GCE), 14 Nov. 2010, New Orleans, Louisiana.
Nuytinck J, Verbeken A. 2003. Lactarius sanguiuus versus Lactarius vinosus – molecular and
morphological analyses. Mycol. Progress 2(3): 227–234.
Romagnesi H. 1967. Les Russules d’Europe et d’Afrique du Nord. Paris, Bordas.
Romagnesi H. 1987. Statuts et noms nouveaux pour les taxa infragénériques dans le genre Russula.
Doc. Mycol. 18(69): 39–40.
Sarnari M. 1998. Monograa illustrata del Genere Russula in Europa, vol 1. Vicenza, Associazione
mycologica Bresadola (AMB), Fondazione Centro studi micologici.
Sarnari M. 2005. Monograa illustrata del Genere Russula in Europa, vol 2. Vicenza, Associazione
mycologica Bresadola (AMB), Fondazione Centro studi micologici.
Séne S, Avril R, Chaintreuil C, Georoy A, Ndiaye C, Diédhiou AG, Sadio O, Courtecuisse R, Sylla
SN, Selosse MA, Bâ A. 2015. Ectomycorrhizal fungal communities of Coccoloba uvifera (L.) L.
mature trees and seedlings in the neotropical coastal forests of Guadeloupe (Lesser Antilles).
Mycorrhiza 25: 547–559.
Singer R. 1986. e Agaricales in modern taxonomy. Königstein, Koeltz Scientic Books.
Stamatakis A. 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large
phylogenies. Bioinformatics 30(9): 1312–313.
Russula vinosoavescens sp. nov. (France) ... 721
Van de Putte K, Nuytinck J, Stubbe D, Le HT, Verbeken A. 2010. Lactarius volemus sensu lato
(Russulales) from northern ailand: morphological and phylogenetic species concepts
explored. Fungal Diversity 45(1): 99–130.
White TJ, Bruns T, Lee S, Taylor J. 1990. Amplication and direct sequencing of fungal
ribosomal RNA genes for phylogenetics. 315–322, in: MA Innis et al. (eds). PCR Protocols:
a guide to methods and applications. San Diego, Academic Press.
... monospo rusʼ), which probably belongs to subsect. Sardoninae(Trendel et al. 2017). ...
Full-text available
A comprehensive morphological and genetic study of type material and new collections of sequestrate Russulales species formerly belonging to the genera Arcangeliella, Elasmomyces, Gymnomyces, Hydnangium, Hymenogaster, Macowanites, Martellia, Secotium and Zelleromyces is here undertaken, for the purpose of providing a complete taxonomical revision of sequestrate Russulaceae species in the Mediterranean and temperate regions of Europe. As a result, seven distinct taxa in the genus Lactarius and 18 in the genus Russula are identified. Six of them are new species: L. populicola, L. subgiennensis, R. bavarica, R. candidissima, R. hobartiae and R. medi­terraneensis, and seven represent new combinations: L. josserandii (≡ Zelleromyces josserandii), L. soehneri (≡ Hydnangium soehneri), R. candida (≡ Hydnangium candidum), R. cerea (≡ Hydnangium cereum), R. messapica var. messapicoides (≡ Macowanites messapicoides), R. meridionalis (≡ Zelleromyces meridionalis) and R. neuhoffii (≡ Hydnangium neuhoffii). Twenty-two of the 25 taxa are illustrated, while descriptions, microscopy images, as well as extensive information on the ecology, chorology and phylogeny for all taxa are provided. A key is further included to facilitate their identification.
Full-text available
Two new species of Russulaceae from China are herein described and illustrated based on their morphologies and phylogenies. A hypogeous gasteroid species, Lactarius sulphosmus sp. nov. and an agaricoid species, Russula vinosobrunneola sp. nov. are introduced. The latter is morphologically distinguished from R. sichuanensis, although the ITS-based phylogeny was unable to distinguish them. Therefore, a multi-gene phylogenetic analysis of the nLSU, ITS, mtSSU, and tef-1α gene sequences of Russula subsection Laricinae was carried out, which supports the assertion that they are different species.
Full-text available
We studied belowground and aboveground diversity and distribution of ectomycorrhizal (EM) fungal species colonizing Coccoloba uvifera (L.) L. (seagrape) mature trees and seedlings naturally regenerating in four littoral forests of the Guadeloupe island (Lesser Antilles). We collected 546 sporocarps, 49 sclerotia, and morphotyped 26,722 root tips from mature trees and seedlings. Seven EM fungal species only were recovered among sporocarps (Cantharellus cinnabarinus, Amanita arenicola, Russula cremeolilacina, Inocybe littoralis, Inocybe xerophytica, Melanogaster sp., and Scleroderma bermudense) and one EM fungal species from sclerotia (Cenococcum geophilum). After internal transcribed spacer (ITS) sequencing, the EM root tips fell into 15 EM fungal taxa including 14 basidiomycetes and 1 ascomycete identified. Sporocarp survey only weakly reflected belowground assessment of the EM fungal community, although 5 fruiting species were found on roots. Seagrape seedlings and mature trees had very similar communities of EM fungi, dominated by S. bermudense, R. cremeolilacina, and two Thelephoraceae: shared species represented 93 % of the taxonomic EM fungal diversity and 74 % of the sampled EM root tips. Furthermore, some significant differences were observed between the frequencies of EM fungal taxa on mature trees and seedlings. The EM fungal community composition also varied between the four investigated sites. We discuss the reasons for such a species-poor community and the possible role of common mycorrhizal networks linking seagrape seedlings and mature trees in regeneration of coastal forests.
Full-text available
Phylogenies are increasingly used in all fields of medical and biological research. Moreover, because of the next generation sequencing revolution, datasets used for conducting phylogenetic analyses grow at an unprecedented pace. RAxML (Randomized Axelerated Maximum Likelihood) is a popular program for phylogenetic analyses of large datasets under maximum likelihood. Since the last RAxML paper in 2006, it has been continuously maintained and extended to accommodate the increasingly growing input datasets and to serve the needs of the user community. I present some of the most notable new features and extensions of RAxML, such as, a substantial extension of substitution models and supported data types, the introduction of SSE3, AVX, and AVX2 vector intrinsics, techniques for reducing the memory requirements of the code and a plethora of operations for conducting post-analyses on sets of trees. In addition, an up-to-date, 50 page user manual covering all new RAxML options is available. The code is available under GNU GPL at
Full-text available
We report a major update of the MAFFT multiple sequence alignment program. This version has several new features, including options for adding unaligned sequences into an existing alignment, adjustment of direction in nucleotide alignment, constrained alignment and parallel processing, which were implemented after the previous major update. This report shows actual examples to explain how these features work, alone and in combination. Some examples incorrectly aligned by MAFFT are also shown to clarify its limitations. We discuss how to avoid misalignments, and our ongoing efforts to overcome such limitations.
Full-text available
Lactarius volemus (Fr.: Fr.) Fr. is a well known and morphologically easily recognizable milkcap of the Northern hemisphere, forming ectomycorrhiza with both deciduous and coniferous trees. It was originally described from Europe, but is also reported in other continents. Although it is characterized by several unique macro- and micromorphological features, substantial variation in colour, lamellae spacing and changing and staining of the latex has been recorded and it is therefore considered as a putatively unresolved species complex. This study explores the concordance between morphological and phylogenetic species concepts within L. volemus sensu lato of northern Thailand, combining a critical morphological scrutiny with a multiple gene genealogy based on LSU, ITS and rpb2 nuclear sequences. Twelve strongly supported monophyletic clades and six terminal branches are discernable in all phylogenetic trees and represent 18 phylogenetic species. Six of the monophyletic clades can be morphologically distinguished and are described as new species: L. acicularis, L. crocatus, L. distantifolius, L. longipilus, L. pinguis and L. vitellinus. Five other clades also show some morphological differences, but these are too subtle and do not allow for a clear-cut species delimitation without the corroboration of molecular data. Lactarius volemus sensu lato of northern Thailand is therefore still considered as a partially cryptic species complex. Pleurolamprocystidia, pileipellis hairs and to a lesser degree also pileus colour are important diagnostic characteristics. Spore morphology, latex discoloration and pileus surface texture are less useful as diagnostic features. Whether this rich diversity is the result of in situ Pleistocene survival or post-glacial expansion and subsequent radiation, has yet to be revealed. Keywords Russulaceae -Cryptic species-Species complex-Morphology-Multiple gene phylogeny
We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a ‘universal’ primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the ‘universal’ ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.
Lactarius vinosus is often considered as a variety of L. sanguifluus. Morphological (especially macroscopical characters and spore-ornamentation) and molecular arguments (based on ITS-sequencing) are given here to treat it as a separate species. Its relationship to closely related species of Lactarius section Deliciosi is discussed.
This chapter describes methods currently used for DNA barcoding of fungi, including some comments on the barcoding of aged herbarium material. The collecting procedures are focussed on macro-fungi. The laboratory methods are for medium-throughput DNA barcoding, targeted at the 96-well format, but without the assistance of robotics. In the absence of an approved and standardized DNA barcoding locus for fungi, the chapter outlines the amplification and sequencing of nuclear ribosomal genes, ITS, and LSU D1/D2 which are most widely used for the identification of fungi from diverse environments.