Eur. J. Phycol.

(2015), 1–14

A new definition of Adenoides eludens, an unusual marine
sand-dwelling dinoflagellate without cingulum, and
Pseudadenoides kofoidii gen. & comb. nov. for the species
formerly known as Adenoides eludens

FERNANDO GÓMEZ1,3, RYO ONUMA2, LUIS F. ARTIGAS3 AND TAKEO HORIGUCHI4

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1

Laboratory of Plankton Systems, sala 100, Oceanographic Institute, University of São Paulo, Praça do Oceanográfico 191,
Cidade Universitária, Butantã, São Paulo 05508-900, Brazil
2
Department of Natural History Sciences, Graduate School of Science, Hokkaido University, North 10, West 8, 060-0810
Sapporo, Japan
3
Laboratoire d’Océanologie et Géosciences, CNRS UMR 8187, Université du Littoral Côte d’Opale, MREN ULCO, 32 av.
Foch, 62930 Wimereux, France
4
Department of Natural History Sciences, Faculty of Science, Hokkaido University, North 10, West 8, 060-0810 Sapporo,
Japan
(Received 9 Jul 2014; revised 24 Sep 2014; accepted 8 Nov 2014)
The species Amphidinium eludens, as described by Herdman (1922; Proc. Trans. Liverpool Biol. Soc. 36: 18) based on her
drawing in fig. 1, has been investigated for the first time by scanning electron microscopy and phylogenetic analysis. The
morphological and molecular data reveal that this species is distantly related to other known dinoflagellates. Balech [1956, Rev.
Algol., n. ser. 2(1–2): 30] cited Amphidinium eludens Herdman (1922, fig. 1) as the basionym of the type of Adenoides, while he
described and illustrated Amphidinium kofoidii Herdman (1922, fig. 2) as Adenoides eludens. As the nomenclatural rules do not
allow the change of basionym, we have re-defined the genus Adenoides based on the characteristics of Amphidinium eludens
Herdman (1922, fig. 1). The thecal plate formula of Adenoides eludens is Po, 5′, 6″, 0c, 3+s, 5′″, 3p, 1″″. This species lacks a
cingulum. The apical pore complex resembles that of peridinioid dinoflagellates, while the absence of a cingular groove is
reminiscent of desmokont prorocentroids. We also propose Pseudadenoides kofoidii gen. & comb. nov. based on Herdman’s 1922
fig. 2 of Amphidinium kofoidii which was described by Balech in 1956 and re-named Adenoides eludens.
Key words: Adenoides kofoidii, Dinophyta, microphytobenthos, molecular phylogeny, new genus, psammophilic Dinophyceae

INTRODUCTION
Dinoflagellates are unicellular organisms with two
dimorphic flagella. Based on the arrangement of the
flagella, the dinokonts (= two flagella in grooves) are
dinoflagellate cells in which two flagella are inserted
ventrally; one flagellum is transverse, wrapped around
the cell, and housed in a groove called the cingulum or
girdle (equatorial or transverse) and the other one is a
trailing longitudinal flagellum and housed in a sulcus
(longitudinal). The other type, desmokonts (= two
anterior flagella) are cells in which two dissimilar
flagella emerge from the anterior part of the cell.
They have two leading flagella inserted apically,
rather than ventrally. One flagellum extends forward
and the other circles its base, and there are no flagellar
grooves (cingulum or sulcus). The best known
Correspondence to: Fernando Gómez. (e‑mail:
fernando.gomez@fitoplancton.com)

example of desmokonts is the genus Prorocentrum
C.G. Ehrenberg. However, not all dinoflagellates fit
into this division. The descriptions of new benthic
species have largely increased in recent years
(Gómez, 2012). The presence of an incomplete cingulum is reported in some sand-dwelling dinokonts
(Amphidiniopsis J. Woloszynska, Cabra Sh. Murray &
D.J. Patterson, Herdmania J.D. Dodge, Rhinodinium
Sh. Murray, Hoppenrath, S. Yoshimatsu, S. Toriumi &
J. Larsen) (Murray & Patterson, 2004; Yamaguchi
et al., 2011). Planktonic dinoflagellates such as
Podolampas F. Stein lack a groove (cingulum) to
harbour a transversal flagellum that encircles the cell
(Gómez et al., 2010). However, the absence of a cingulum has never been reported in benthic dinokonts.
In one of the earlier studies of sand-dwelling dinoflagellates, Herdman (1922) described the species
Amphidinium eludens and A. kofoidii. In her fig. 1
she described Amphidinium eludens as oval to ovoid

ISSN 0967-0262 (print)/ISSN 1469-4433 (online)/15/000001-14 © 2015 British Phycological Society
http://dx.doi.org/10.1080/09670262.2015.1009174

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F. Gómez et al.
in shape, has a barely visible episome and a hump in
the sulcal area (Herdman, 1922, p. 22, fig. 1). In her
fig. 2 she described Amphidinium kofoidii as round to
squarish in shape, with a more visible button-like
episome (Herdman, 1922, p. 26, fig. 2). Balech
(1956) proposed the new thecate genus Adenoides
based on Herdman’s fig. 2 of Amphidinium kofoidii.
However, Balech (1956) proposed as basionym
Amphidinium eludens Herdman (1922, fig. 1). Later,
Dodge (1982) proposed Adenoides kofoidii for
Amphidinium kofoidii Herdman 1922, fig. 2. The taxonomic and nomenclatural history of Adenoides eludens is reported in Hoppenrath et al. (2003).
Adenoides eludens based on Balech’s description and
as described by Herdman’s fig. 2 of Amphidinium
kofoidii was investigated again by Dodge & Lewis
(1986) and Hoppenrath et al. (2003), and molecular
data have been available since Saldarriaga et al.
(2001). To date, there are 45 sequences of different
molecular markers of Adenoides eludens and another
39 sequences labelled as two uncultured Adenoides.
The sequences of Adenoides eludens available in
GenBank are from the cultures CCCM 683 and
CCMP 1891 isolated from the Pacific coasts of
Canada or field material collected from the same location. Another culture CCMP 2081 was isolated from
Europe. However, there are no molecular data for
Adenoides eludens from the European Atlantic coast
where the species was described. While Adenoides
eludens (= Amphidinium kofoidii) has been subjected
to morphological and molecular studies, little is known
about the species described as Amphidinium eludens
Herdman (1922, fig. 1). This organism is one of the
first described sand-dwelling dinoflagellates and
according to Herdman (1922) is responsible for
discolourations in the sands of the European Atlantic
coasts. However, the detailed morphology of this
species remains unknown.
In this study, we provide light microscopy pictures,
the first scanning electron microscopy pictures and the
first molecular data for Amphidinium eludens
Herdman (1922, fig. 1), and the first molecular data
for Amphidinium kofoidii Herdman (1922, fig. 2) from
the French coasts of the English Channel, where
Balech (1956) described the genus Adenoides. The
morphological and molecular data support the conclusion that the two species described in figs 1 and 2 by
Herdman (1922) belong to independent genera.
Amphidinium eludens is an unusual dinoflagellate,
characterized by the lack of a cingular groove similar
to desmokont dinoflagellates.

MATERIALS AND METHODS
Source, isolation and microscopy observations
This study was undertaken in the soft sandy sediments of the
shore of Wimereux, France (50º46′12″N, 1º36′42″E)

2
collected during low tide in June, 2012. Two sites on the
beach in front of the LOG laboratory (MREN ULCO and
Marine Station of Wimereux UL1; Gómez & Artigas, 2014)
were sampled: the moist sands around the border of a large
pool (~50 m diameter, ~1 m depth), and several smaller pools
and moist sands showing a faint brownish discolouration.
The upper centimetre of sand was collected with a spoon
and deposited into a bottle containing seawater collected at
the same location. Then, the sand with seawater was stirred
vigorously and the suspension settled in a composite settling
chamber. The settled material was examined with an inverted
microscope (Nikon Eclipse TE2000-S, Tokyo) and photographed with a Nikon Digital Sight DS-2M camera. The cell
size, described as length (apical to antapical axis) and depth,
i.e. the length along the lateral sides (ventral to dorsal
distance), was measured in 25 specimens. The width
(transdiameter) was measured in five specimens.
For scanning electron microscopy, the sand samples with
seawater were stirred vigorously, and the suspension was
fixed with glutaraldehyde (5%) and filtered onto a 0.8 µm
pore size Whatman Nuclepore® membrane filter, washed
with distilled water, fixed with osmium tetroxide, dehydrated
with a graded series of ethanol and critical point dried with
CO2. Filters were mounted on stubs, sputter-coated with gold
and viewed using a Hitachi S4800 scanning electron microscope. Images were presented on a black background using
Adobe Photoshop CS3.

PCR amplification and DNA sequencing
For molecular analysis, each specimen of Amphidinium eludens and A. kofoidii was micropipetted individually with a
fine capillary into a clean chamber and washed several times
in serial drops of 0.2 μm filtered and sterilized seawater.
Finally, 1–5 specimens of each species were deposited in a
0.2 ml Eppendorf tube filled with several drops of absolute
ethanol. The sample was kept at room temperature and in
darkness until the molecular analysis could be performed.
Prior to DNA extraction, the 0.2 ml Eppendorf tubes were
centrifuged for 10 min at 14462 × g in a TOMY MX-201
centrifuge (Tokyo, Japan). Ethanol was then evaporated in a
vacuum desiccator. Cells were resuspended in 10 μl of
QuickExtract DNA extraction solution (Epicenter,
Madison) and incubated at 56ºC for 1 h and 98ºC for 2 min
in a thermal cycler (GeneAmp PCR System 9700, Applied
Biosystems, Foster City). The product was used as DNA
template for the following polymerase chain reaction
(PCR). In the first round of PCR, to obtain almost complete
SSU rDNA and partial LSU rDNA sequences, two sets of
primers (SR1-SR12b for SSU rDNA and D1RF1 28-1483R
for LSU rDNA, respectively) were simultaneously applied.
In the second round of PCR, 0.5 µl of the first PCR products
was used as DNA template, and three pairs of primers (SR1SR5, SR4-SR9p and SR8-SR12b; see Yamaguchi
et al., 2006) were used for SSU rDNA amplification and
two pairs of primers (D1RF-25R1 and D3A-28-1483R; see
Daugbjerg et al., 2000) were applied for LSU rDNA amplification. PCR conditions for both rounds of amplification
consisted of one initial cycle of denaturation at 94ºC for 5
min, followed by 35 cycles of denaturation at 94ºC for 30 s,
annealing at 55°C for 30 s, and extension at 72ºC for 30 s.
The PCR process was completed by a final extension cycle

Pseudadenoides gen. & comb. nov. and Adenoides
at 72ºC for 7 min. The PCR products were directly
sequenced using the ABI PRISM BigDye Terminator
Cycle Sequencing Kit (Applied Biosystems) and a DNA
autosequencer ABI PRISM3100 Genetic Analyzer
(Applied Biosystems). Both forward and reverse strands
were sequenced.

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Phylogenetic analyses
Both SSU rDNA and LSU rDNA sequences were aligned
based on the secondary structure of the rDNA molecule
(database no longer available), and the alignments were
refined manually. We used sequences from GenBank of
Amoebophrya spp. as outgroups for SSU rDNA and a ciliate
Tetrahymena pyriformis (C.G. Ehrenberg) Lwoff and an apicomplexa Eimeria tenella (Raillet & Lucet) Fantham were
used for outgroups of LSU rDNA analyses, respectively. The
accession numbers of sequences included in the alignment
are indicated in each tree. The aligned sequences were examined using maximum likelihood (ML) analyses with PAUP
version 4.0b10 (Swofford, 2003), and Bayesian analysis with
MrBayes 3.2.1 (Huelsenbeck & Ronquist, 2001). The program Modeltest version 3.04 (Posada & Crandall, 1998),
which employs the hierarchical likelihood ratio test (hLRT)
was used to explore the best ML sequence evolution model
for the dataset. The hRLT model selected for ML analysis of
the dataset was TrN+I+G. In the ML analysis, a heuristic
search was performed with a TBR branch-swapping algorithm, and the starting tree was obtained by the neighborjoining (NJ) method. The parameters in this analysis were:
assumed nucleotide frequencies A = 0.2756, C = 0.1898, G =
0.2392 and T = 0.2954; substitution rate matrix with A<->C
= 1.0000, A<->G = 3.5126, A<->T = 1.0000, C<->G =
1.0000, C<->T = 7.7026 and G<->T = 1.0000; proportion
of sites assumed to follow a gamma distribution with shape
parameter = 0.5633; and number of rate categories = 4.
Bootstrap analysis for ML was calculated for 100 pseudoreplicates. For LSU rDNA analysis, the hLRT model selected
for ML analysis of the dataset was TrN+I+G. In the ML
analysis, a heuristic search was performed with a TBR
branch-swapping algorithm, and the starting tree was
obtained by the NJ method. The parameters in these analyses
were: assumed nucleotide frequencies A = 0.2865, C =
0.1806, G = 0.2701 and T = 0.2628; substitution rate matrix
with A<->C = 1.0000, A<->G = 2.8299, A<->T = 1.0000,
C<->G = 1.0000, C<->T = 6.6145 and G<->T = 1.0000;
proportion of sites assumed to follow a gamma distribution
with shape parameter = 0.6675; and number of rate categories
= 4. Bootstrap analysis for ML was calculated for 100
pseudo-replicates. For Bayesian analysis, GTR+I+G model
was selected by MrModeltest 2.2 (Nylander et al., 2004) as a
suitable evolutionary model. Markov chain Monte Carlo
iterations were carried out until 3 000 000 generations
were attained for SSU rDNA phylogeny, while 5 500
000 generations were required for LSU rDNA phylogeny,
when the average standard deviations of split frequencies
fell below 0.01, indicating a convergence of
the iterations. Our sequences were deposited in DDBJ/
EMBL/GenBank under accession numbers LC002839LC002848.

3
RESULTS
Based on the morphological and molecular data (see
below) the species described as Amphidinium eludens
Herdman (1922, fig. 1) and Amphidinium kofoidii
Herdman (1922, fig. 2) belong to separate genera. If
we had proposed a new genus name based on
Herdman (1922, fig. 1), it would have been illegitimate as a superfluous later homotypic synonym of
Adenoides [International Code of Nomenclature for
algae, fungi, and plants (I.C.N.), art. 52.1, McNeill
et al., 2012]. As defined by Balech (1956), the basionym of the type of Adenoides, Adenoides eludens, is
Amphidinium eludens Herdman (1922, fig. 1).
According to article 7.3 of I.C.N.: ‘A new combination or a name at new rank (Art. 6.10) is typified by the
type of the basionym even though it may have been
applied erroneously to a taxon now considered not to
include the type (but see Art. 48.1)’. The characteristics of the genus Adenoides are defined by
Amphidinium eludens Herdman (1922, fig. 1), independently of the fact that Balech (1956) provided for
Adenoides eludens the description and illustrations of
the species described as Amphidinium kofoidii by
Herdman (1922, fig. 2). Hereafter, we redefined the
genus Adenoides based on the morphology of
Amphidinium eludens Herdman (1922, fig. 1).
Taxonomic descriptions
Adenoides Balech emended F. Gómez, R. Onuma,
Artigas & Horiguchi
DIAGNOSIS: Armoured cell laterally compressed,
lacking the cingulum and flagella inserted ventrally.
Thecal plate formula Po, 5′, 6″, 0c, 3+s, 5′″, 3p, 1″″.
An alternative interpretation is Po, cp, 4′, 7″, 1c, 1+s,
5′″, x, 2p, 1″″.
TYPE SPECIES: Adenoides eludens (Herdman) Balech.
BASIONYM: Amphidinium eludens Herdman 1922,
p. 22, fig. 1.
SYNONYM: Adenoides kofoidii sensu Dodge 1982.
Adenoides eludens F. Gómez, R. Onuma, Artigas &
Horiguchi epitype
Given the lack of type material, we have designated
fig. 1 in Herdman (1922) as the lectotype for this
species and the cell shown in Fig. 23 of this study as
the epitype under Article 9.7 of the I.C.N. (McNeill
et al., 2012). A detailed description of the epitype is
presented below.
EPITYPE: Fig. 23. A SEM stub was deposited in the
herbarium of the Faculty of Science, Hokkaido
University as SAP114711.

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F. Gómez et al.
The slightly laterally flattened cells are ellipsoidal
in lateral view. The sulcal area is depressed and lies on
the anterior third of the cell, neither extending onto the
epitheca nor reaching the antapex. In lateral view, the
ventral contour of the cell showed a hump (Figs 2, 6).
The cells were 30 ± 2 (27–37) µm long, 23 ± 2 (18–27)
µm deep, and 13–17 µm wide (Figs 1–15). There is an
intergradation of the size with large and small specimens co-existing in the same sample (Fig. 6). The
epitheca is directly connected to the hypotheca, without any cingulum. We presume that the vestigial cingular groove was placed in the suture between the preand postcingular plates that is at the level of the
flagellar insertion.
The height of the epitheca is about one-third the
length of the cell. Some specimens showed a large
pusule in the anterior dorsal part of the cell (Figs 1–4).
The transversal flagellum encircled the cell (Fig. 5).
The cell possesses yellow-brown plastids. The species
Amphidinium kofoidii sensu Herdman is more
pigmented and brownish when compared with
Adenoides eludens (Fig. 7). Some specimens of
Adenoides were found devoid of pigments and with
numerous granules (Figs 8–10). The cell showed two
pyrenoids with a starch sheath, each pyrenoid placed
in the sides of the mid anterior hypotheca (Figs 6, 11).
The nucleus is oval and situated in the posterior
region, although hardly visible because it is hidden
by plastids. Some specimens experienced ecdysis or
exuviation, releasing empty thecae (Figs 11–12).
The empty theca was usually divided into two parts,
which are considered the epitheca and hypotheca
(Figs 13–15). One plate that is interpreted as a sulcal
anterior plate is attached to the epitheca (Fig. 13).
The tabulation is illustrated in detail in Figs 16–46.
The thecal plate pattern is an apical pore plate (Po),
five apical plates (5′), six precingular plates (6″), no
cingular plate (0c), at least three sulcal plates (3+s),
five postcingular plates (5′″), three posterior intercalary plates (3p) and one antapical plate (1″″). The cell
surface is smooth with round pores (approximately
0.15 µm). The pores are evenly distributed in the
plates, with a trend to form rows near the sutures
(Fig. 19). There is an accumulation of pores in the
posterior end of the first and second posterior intercalary plates (1p, 2p) and in the dorsal end of the
antapical plate (Figs 16, 40–41). The pores are absent
in the sulcal plates (Figs 22, 27–30, 34). The boundary
of the plates shows a thick (~1 µm wide) smooth and
depressed margin. These intercalary bands were more
conspicuous in the hypotheca (Figs 19, 21).
The apical pore plate showed a pentagonal to round
shape being bordered by five apical plates
(Figs 24, 37). The junction of the apical pore plate
and the apical series formed a ridge, except in the first
apical plate suture which is shorter than in the other
apical plates (Figs 24, 25, 37). The apical pore plate
contains a row of marginal pores (10–13 pores) and a

4
round central cover plate (Figs 26, 37). From the
central cover plate emerged a rim that protruded and
extended towards the anterior part of the first apical
plate (Figs 24, 26, 37).
There are five apical plates (Figs 20, 23, 37). Plate
3′ is the largest and is located in the dorsal side of the
epitheca. Plates 2′ and 5′ are intermediate in size.
Plates 4′ and 1′ are the smallest of the apical series.
The apical plates 2′ to 5′ showed a curved ridge in the
suture with the apical pore plate. Plate 1′ did not show
a ridge in the junction with the apical pore plate. Plate
1′ is a six-sided irregular smooth-surfaced polygon,
with very few pores when compared with the other
apical plates (Figs 24–26). An alternative interpretation of the apical tabulation is to consider plate 1′ as a
wide canal plate (cp). Plates 2′ and 4′ are pentagonal,
especially plate 4′ which is a quasi-regular pentagon.
Plates 3′ and 5′ are six-sided. Plate 5′ shows a distinctive undulating flange in the sutures, except in the
anterior end in the suture with plates 5″ and 6″
(Figs 25–26, 37).
At the level of the precingular plate series, there are
six precingular plates and a large smooth-surfaced and
concave plate. This plate is joined to the precingular
plates when the epitheca is separated from the
hypotheca (Fig. 13). Based on the proximity to the
flagellar pores, the differences in the shape (concave)
and the absence of ornamentation (smooth-surfaced
lacking pores and intercalary bands) as in the other
sulcal plates, we have considered it as the right anterior
sulcal plate (S.d.a.) (Figs 21–22, 34). An alternative
interpretation is to consider this large plate as plate 1″.
Following the interpretation as an anterior sulcal plate,
the first precingular plate (1″) is displaced to the left
side in the ventral view. Plates 1″ and 5″ are the
biggest of the epitheca, plates 2″, 3″ and 4″ are intermediate in size and plate 6″ is the smallest of the
precingular series (Figs 16–20, 35–36). Plate 1″ is
polygonal and curved in the suture with the anterior
sulcal plate (Figs 21–23). Plates 2″, 4″ and 5″ are
six-sided and plates 3″ and 6″ are quadrangular
(Figs 16–20). Plate 3″ is nearly square-shaped
(Fig. 20) and plate 6″ is an elongated rectangle oriented
along the antero-posterior axis (Figs 28, 34–35).
The cell does not have a cingulum. However,
when the theca is empty, it tended to separate
along the suture between the precingular and postcingular plate series (Figs 13–15). This suggests that
this thick suture could be occupying the space of the
cingulum of a hypothetical ancestor. The observations of live specimens revealed that the transversal
flagellum encircles this area (Fig. 5). The cell shows
a depression in the sulcal area (Figs 21–22, 32–35).
The left anterior sulcal (S.s.a.) plate is smooth
and located between the right anterior sulcal plate
(S.d.a.) and a large plate 1′″. This S.s.a. plate is
elongated and concave and it could be alternatively
interpreted as a first cingular plate (Figs 21–23, 34).

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Pseudadenoides gen. & comb. nov. and Adenoides

Figs 1–15. Light microscopy pictures of live specimens and empty thecae of the redefined Adenoides eludens (Fig. 7 also includes
Amphidinium kofoidii). Figs 1–4. Different views of the same specimen. Fig. 5. Note the transversal flagellum that encircles the cell.
Fig. 6. Note the different sizes. Fig. 7. Note the different pigmentation between Adenoides eludens (yellowish, on the left) and
Amphidinium kofoidii (brownish, on the right). Figs 8–10. Specimens lacking pigmentation. Figs 11–12. Ecdysis or exuviation from
the theca. Figs 13–15. Empty theca with separated epitheca and hypotheca. Fig. 13. Note that the sulcal right (= dexter) anterior plate
(S.d.a.) is attached to the epitheca. LF = longitudinal flagellum. TF = transversal flagellum. Scale bar = 10 µm.

There are two flagellar pores in the depressed area of
the sulcus. The longitudinal flagellum is inserted in
a pore located in the right side of the sulcus,
adjacent to the suture of plates 6″ and 5′″ (Figs 22,
27–30, 34). The right half of this pore is surrounded
by a prominent ridge (Figs 28–29). The pore of the
transversal flagellum is located in the middle of the
sulcus. It is bordered by a rectangular structure

(Figs 28–30). Below the pores, a large sulcal posterior plate with a rounded posterior end is subdivided
into three plates connected by wedge-shaped contours (Figs 29–30). The right anterior sulcal (S.d.a.)
plate is in contact with the flagellar pores (Fig. 22).
This plate shows an anterior-posterior oriented line,
which suggests the presence of a subdivision of the
plate (Fig. 30).

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F. Gómez et al.

Figs 16–30. Scanning electron micrographs of the re-defined Adenoides eludens. Figs 16–17. Right lateral view. Fig. 18. Ventral view.
Fig. 19. Left lateral view. Fig. 20. Dorsal view. Figs 21–23. Ventral lateral view. Figs 24–25. Detail of the apical plates. Fig. 26. Detail of
the apical pore plate. Figs 27–30. Detail of the sulcal plates. Fig. 30. The arrowheads point out tentative sutures in the sulcal plates. S.d.a. =
Sulcal dexter (= right) anterior plate. S.s.a. = Sulcal sinister (= left) anterior plate. S.p. = Sulcal posterior plate. LFP = longitudinal flagellar
pore. TFP = transversal flagellar pore. Scale bar = 10 µm, except Figs 22, 24–30 where scale bar = 1 µm.

There are five postcingular plates (Figs 16, 19).
Plate 1′″ is the longest plate of the cell and extended
for 2/3 the height of the hypotheca. The anterior part

of this five-sided plate is curved and overhangs at
the anterior end (Figs 18, 22, 34). Plates 2′″ and 4′″
are large and broad, plate 3′″ is intermediate in size

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Pseudadenoides gen. & comb. nov. and Adenoides

7

Figs 31–41. Scanning electron micrographs of a single specimen of Adenoides eludens. Fig. 31. Ventral view. Figs 32–33. Right
lateral view. Fig. 34. Detail of the sulcal area. Fig. 35. Latero-ventral view. Figs 36–37. Detail of the epitheca. Figs 38–39. Dorsal
view. Figs 40–41. Antapical view. Fig. 40. The inset shows the pores of the plates 2p (up) and 1″″ (down). S.d.a. = Sulcal dexter
(= right) anterior plate. S.s.a. = Sulcal sinister (= left) anterior plate. S.p. = Sulcal posterior plate. Scale bar = 20 µm, except Figs 34
and 37 where scale bar = 1 µm.

and plate 5′″ is the smallest of the postcingular
series. Plate 2′″ is four sided, plate 3′″ is a regular
pentagon and plate 4′″ is five-sided and elongated
posteriorly. Plate 5′″ is smaller and more anteriorly
placed than the other plates of the postcingular series (Figs 34–35). An alternative interpretation is that
plate 5′″ is equivalent to plate x reported in some
sand-dwelling dinoflagellates with incomplete
cingulum.
Four large plates that do not belong to the sulcal and
postcingular series are considered posterior intercalary and antapical plates. Three plates are considered

posterior intercalary plates and the plate situated at the
bottom of the cell is considered the antapical plate 1″″
(Figs 16, 19). The antapical plate is bordered by plate
1′″ and three posterior intercalary plates (1p, 2p, 3p).
The first and second posterior intercalary plates
(1p, 2p) are large and with the shape of an oblique
irregular pentagon (Figs 16, 19). Plates 1p and 2p
have an accumulation of pores in the posterior end,
at the contact with the antapical plate (Figs 39–41).
Plate 3p is an elongated pentagon oriented in the
longitudinal axis (Figs 32–33). The antapical plate is
hexagonal and it showed, at the dorsal end, the special

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F. Gómez et al.

8

Figs 42–46. Line drawings of the re-defined Adenoides eludens with the Kofoid system of tabulation. Fig. 42. Right lateral. Fig. 43.
Left lateral. Fig. 44. Apical. Fig. 45. Antapical. Fig. 46. Ventral.

Figs 47–56. Light microscopy pictures of live specimens (47–50) and empty thecae (51–56) of Pseudadenoides kofoidii gen. &
comb. nov. Fig. 47. The arrows point the transversal flagella encircling the cingulum. Fig. 50. Megacytic cell. Figs 51–56. Empty
thecae. Scale bar = 10 µm.

pore field found in plates 1p and 2p (Fig. 41). An
alternative interpretation of the tabulation of the
hypotheca may appear when plate 5′″ is considered
to be an x plate (Fig. 48), then plate 3p is considered to
be 5′″ plate.
A new genus name for Amphidinium kofoidii Herdman
(1922, fig. 2)

We had to redefine the genus Adenoides due to the
designation of Amphidinium eludens Herdman (1922,
fig. 1) as basionym of the type of Adenoides.
Morphological and molecular data reveal that the
species Amphidinium kofoidii Herdman (1922, fig. 2)
does not belong to the new definition of the genus
Adenoides or to the other known dinoflagellate genus.
Therefore, a new genus name is proposed here. The

Pseudadenoides gen. & comb. nov. and Adenoides

9

description of the genus and its type species is here
simplified because the morphological characteristics
have been already reported from the English Channel
and North Sea (Balech, 1956; Dodge & Lewis, 1986;
Hoppenrath et al., 2003). For that reason, we have
summarized the description which is similar to that
found, with more detail, in Hoppenrath et al. (2003).

central cover plate. From the central cover plate
emerged a narrow rim that protruded and extended
ventrally towards the anterior part of the first apical
plate (Fig. 62). The apical plates 1′ and 4′ are in
contact with the sulcus (Fig. 62). Plate 2′ is very
narrow and 4′ is relatively large, covering nearly the
entire right half of the epitheca. There are no precingular plates. The shallow cingulum consists of six
plates and there are four small sulcal plates surrounding the flagellar pore (s.a., s.s., s.p., s.d.) (Figs 59–60).
The hypotheca consists of 11 plates, comprised of five
postcingular, five posterior intercalary plates and one
antapical plate. Plates 1′″ and 2′″ lie at the left lateral
cell side in the upper fourth of the hypotheca. The
small, pentagonal plate 3′″ lies dorsally, and the pentagonal, posteriorly pointed plate 4′″ is relatively
large, lying at the right lateral side. The six-sided
plate 5′″ is in contact with the sulcus. All five posterior
intercalary plates are large and cover most of the
hypotheca (Figs 57–59). The plates 1p and 5p are in
contact with the sulcus and bordered its posterior
margin. Both are in contact with each other and form
a ‘ventral suture’ (Figs 59–60).

Pseudadenoides F. Gómez, R. Onuma, Artigas &
Horiguchi gen. nov.

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DIAGNOSIS: Armoured cell laterally compressed, with a
minute epitheca. The plate formula is Po, 4′, 0″, 6c, 4s,
5′″, 5p, 1″″ (or alternatively it can be interpreted as Po
4′, 0″, 6c, 5s, 5′″, 3p, 2″″).
SYNONYM: Adenoides sensu Balech 1956.
ETYMOLOGY: Pseudo-, pseud- (before vowels),
Ancient Greek ψευδής (pseudes): false, not genuine,
fake. The type species has been confused with
Adenoides eludens. The gender is feminine (art. 62.4
of I.C.N.).
TYPE SPECIES: Pseudadenoides kofoidii (Herdman) F.
Gómez, R. Onuma, Artigas & Horiguchi comb. nov.
BASIONYM: Amphidinium kofoidii Herdman (1922,
p. 26, fig. 2)
SYNONYM: Adenoides eludens sensu Balech 1956
EPITYPE: Fig. 60
Pseudadenoides kofoidii (Herdman) F. Gómez, R.
Onuma, Artigas & Horiguchi comb. nov.
Cells are asymmetrical, round to squarish, slightly flattened laterally, 28–37 µm long and 21–29 µm deep. The
minute epitheca is cup-shaped, depressed and scarcely
visible (Figs 47–50). The cingulum lies almost at the
anterior end of the cell, completely encircling the
epitheca and meeting without displacement. The sulcal
area lies on the anterior third of the cell, neither extending onto the epitheca nor reaching the antapex, and is
slightly depressed. The transverse flagellum completely
encircles the cell at the cingulum level (Fig. 47). Most of
the cells show one large pusule in the anterior
hypotheca. The cell is full of brown plastids. There are
two conspicuous pyrenoids. The nucleus is oval and
situated in the posterior region, although hardly visible
because it is hidden by plastids (Fig. 47–50).
The thecal plate pattern is an apical pore plate (Po),
four apical plates (4′), without precingular plates (0″),
six cingular plates (6c), four sulcal plates (4s),
five postcingular plates (5′″), five posterior intercalary
plates (5p) and one antapical plate (1″″) (Figs 51–62).
The apical pore plate is an angular square-shape
bordered by four apical plates which form a ridge
running around it (Fig. 60). The apical pore plate
contains few marginal pores (~7 pores) and a round

Molecular phylogeny
We have sequenced specimens of the species
Amphidinium eludens Herdman (1922, p. 22, fig. 1,
now Adenoides eludens emended), and the species
Amphidinium kofoidii Herdman (1922, p. 26, fig. 2,
now Pseudadenoides kofoidii gen. & comb. nov.). The
SSU- and LSU rDNA sequences of Amphidinium
kofoidii Herdman from specimens of the Pacific
Ocean are available in GenBank (retrieved as
Adenoides eludens). We have also provided the first
SSU- and LSU rDNA sequences from specimens of
the European Atlantic coasts, where Amphidinium
kofoidii was described. The aligned length for SSUrDNA was 1725 bp and 954 bp for LSU rDNA (D1–
D3 region excluding the variable region). The
sequences of the Atlantic and Pacific specimens of
Amphidinium kofoidii were identical (Figs 63, 64).
We provided the first SSU- and LSU rDNA sequences
of Amphidinium eludens. All the sequences obtained
from different samples were identical (Figs 63, 64).
The species Amphidinium eludens and Amphidinium
kofoidii described in Herdman (1922) are distantly
related in the SSU rDNA (Fig. 63) and LSU rDNA
trees (Fig. 64). Our data strongly support the splitting
of these two species into two distinct genera based on
the considerable evolutionary distance of their respective SSU- and LSU rDNA sequences. Because of the
low resolution of basal positions of the phylogenetic
trees, it is also difficult to establish the accurate relationships between these genera with other dinoflagellates (Figs 63, 64).

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F. Gómez et al.

10

Figs 57–62. Scanning electron micrographs of Pseudadenoides kofoidii gen. & comb. nov. Figs 57–58. Right lateral view. Fig. 59.
Ventral view. Fig. 60. Apical view. Fig. 61. Dorsal view. Fig. 62. Detail of the pore and apical plates. Scale bar = 20 µm, except
Fig. 62 where scale bar = 1 µm.

DISCUSSION
In pioneering studies of sand-dwelling dinoflagellates, Herdman (1922) described in her fig. 1
Amphidinium eludens as forming discolourations in
a beach of the British Isles. Despite the tradition of
taxonomic studies in the European Atlantic coasts
and the relative abundance of A. eludens, to date that
species has remained under investigation. Confusion
has been present since the original description.
Herdman (1922) illustrated Amphidinium eludens
as slightly larger than A. kofoidii in line drawings
lacking any reference to the size and reported a
length of 70 µm and 50–80 µm for A. eludens and
A. kofoidii, respectively. However, in a note at the
end of a subsequent publication (Herdman, 1923, p.
63) she explained that the size measurements were
overestimated and should be 30 µm and 25–40 µm
for Amphidinium eludens and A. kofoidii, respectively. Consequently, the cells of Amphidinium kofoidii tend to be larger than those of A. eludens. This

contrasts with the relative sizes of Herdman’s original figures. In our observations, we found an average length of 30 µm for Amphidinium eludens,
which is usually slightly less than for A. kofoidii.
The error in the relative size of the original description could have contributed to further confusion in
the identification of both species. When Balech
(1956) proposed Adenoides eludens, he considered
that Herdman’s figure 1 and 2 corresponded to the
same species. Dodge & Lewis (1986, p. 224) maintained this view when they reported ‘the cell contents of the two species is very similar it may be that
they are only variants’. As a consequence, the species Amphidinium eludens has received little attention, and has been cited as Adenoides eludens sensu
Balech (Paulmier, 1992) a situation that has deprived
us from knowing the unusual morphology of
Amphidinium eludens until now.
Our specimens studied from a sandy beach in the
English Channel correspond to Herdman’s fig. 1

11

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Pseudadenoides gen. & comb. nov. and Adenoides

Fig. 63. Maximum likelihood phylogenetic tree of Adenoides eludens and Pseudadenoides kofoidii with other dinoflagellates
inferred from SSU rDNA sequences. The species newly sequenced in this study are bold. The numbers at each node represent
bootstrap value (maximum likelihood, ML) and posterior probability (PP). Only values of > 70% (bootstrap) and > 0.9 (PP) are
indicated. Closed circle denotes 100%/1.0 = bootstrap/PP.

12

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F. Gómez et al.

Fig. 64. Maximum likelihood phylogenetic tree of Adenoides eludens and Pseudadenoides kofoidii with other dinoflagellates
inferred from partial LSU rDNA sequences. The species newly sequenced in this study are bold. The numbers at each node represent
bootstrap value (maximum likelihood, ML) and posterior probability (PP). Only values of > 70% (bootstrap) and > 0.9 (PP) are
indicated. Closed circle denotes 100%/1.0 = bootstrap/PP.

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Pseudadenoides gen. & comb. nov. and Adenoides
reported as Amphidinium eludens. The shared characteristics with the original description including (1)
general cell contour and size; (2) lack of apparent
raised epitheca viewed from the lateral side; (3) the
presence of a hump in the ventral contour of the cell
where flagella arise; and (4) the position and shape of
the nucleus and pyrenoids (Figs 1–6). The main characteristic of the redefined Adenoides eludens is the
absence of the cingular groove. With the exception
of Podolampas and allied genera, the absence of a
cingular groove is a rare feature in planktonic species
with a globular shape. Although there is no groove,
the transversal flagellum nonetheless encircles the
cell. As a general trend, the cell flattening of the
benthic species makes it difficult to interpret the tabulation. This is even more difficult in some sand-dwelling dinoflagellates that have an incomplete cingulum.
Adenoides is an extreme case, where the cingulum is
absent or the sulcal left anterior plate could be interpreted as a vestigial first cingular plate. We can speculate an evolution towards the loss of the cingular
groove as an adaptation to benthic habitats. In fact,
dinoflagellates that lack the cingular groove such as
Prorocentrum are widespread and diverse in benthic
habitats. Then, we can speculate that the loss of the
cingular groove may be an advantage in benthic habitats, and tentatively propose that the rotational movement of the transversal flagellum is less useful in
interstitial space between the sand grains than in pelagic habitats.
With the molecular phylogeny we can find a
relationship between the planktonic and benthic
dinoflagellates that is useful to interpret their tabulation. For example, as suggested by Saldarriaga et al.
(2003), the sand-dwelling genus Roscoffia Balech is
related to Podolampas (Gómez et al., 2010). The genera
Amphidiniopsis or Herdmania are related to planktonic
Protoperidinium-like cells such as Archaeperidinium
minutum (Kofoid) E. Jørgensen (Gómez et al., 2011;
Yamaguchi et al., 2011). The morphology of the apical
pore of Adenoides is reminiscent of peridinioid genera
such as Heterocapsa F. Stein or Azadinium Elbrächter &
Tillmann (Tillmann et al., 2009). The tabulation of the
epitheca of Adenoides is 5′ and 6″, but it can also be
interpreted as 4′ and 6″ or 5′ and 7″. These are common
tabulation features of dinoflagellates from different phylogenetic origins.
Taxonomic sampling in the phylogenetic trees
remains incomplete, and for the moment there are
no dinoflagellate sequences close to Adenoides or
Pseudadenoides. Numerous sand-dwelling dinoflagellates also remain as monotypic genera, lacking
any known relative with which they can be classified, at least at the family level. The lack of the
cingular groove in Adenoides or the absence of
precingular plates in Pseudadenoides makes these
genera particularly interesting for the evolution of
dinoflagellates.

13
ACKNOWLEDGEMENTS
We thank Pierre Compère and Gerry Moore for their
useful advice on nomenclature.

FUNDING
F.G. was supported by a UL1 post-doctoral grant and a
CNRS convention of research, and an invited lecturer
grant from Université Littoral-Côte Opale. F.G. is
currently supported by the Brazilian Conselho Nacional
de Desenvolvimento Científico e Tecnológico (grant number BJT 370646/2013-14).

AUTHOR CONTRIBUTIONS
F. Gómez: collection, isolation, light and electron microscopy, drafting and editing manuscript; R. Onuma: molecular analysis; L.F. Artigas: collection and editing
manuscript; T. Horiguchi: phylogenetic analysis and
editing manuscript.

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