Diatom Research, 2019

https://doi.org/10.1080/0269249X.2018.1551246

NOTE

Molecular phylogeny suggests the affinity of the planktonic diatoms Climacodium and
Bellerochea (Lithodesmiales, Mediophyceae)
1, ∗ 2,3,4 2,4
FERNANDO GÓMEZ † , LU WANG & SENJIE LIN
1 Carmen Campos 3, E-11500 Puerto de Santa María, Spain
2 Marine Biodiversity and Global Change Research Center and State Key Laboratory of Marine Environmental Science,
Xiamen University, Xiamen, People’s Republic of China
3 Institute of Oceanography, Minjiang University, Fuzhou 350108, People’s Republic of China
4 Department of Marine Sciences, University of Connecticut, Groton, CT, USA

Climacodium frauenfeldianum is a planktonic chain-forming diatom able to proliferate in severely oligotrophic waters of tropical seas.
In this study, a strain of C. frauenfeldianum, the type of Climacodium, was established in culture from the tropical southwestern Atlantic
Ocean off Brazil. SEM observations showed weakly silicified cells, without visible surface sculpturing. In the small subunit (SSU) rRNA
and ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene phylogeny, C. frauenfeldianum clustered with sequences
of Bellerochea malleus (the type of Bellerochea) and B. horologicalis. Contrary to previous classifications that placed Climacodium in
the Hemiaulaceae, Climacodium and Bellerochea should be placed in the same family, Bellerocheaceae. The species Bellerochea yucata-
nensis needs further research because sequences of the single strain available clustered with the genus Lithodesmioides. In adaptation to
the oligotrophic open ocean environment, C. frauenfeldianum seems to have evolved an appearance closer to Hemiaulus, i.e. very lightly
silicified flat cells with long elevations, and with diazotrophic cyanobionts.
Keywords: Bacillariophyceae, Bacillariophyta, DNA barcoding, molecular phylogenetics, phylogeny

The usually nutrient depleted stratified surface waters of
tropical oceans are unfavourable environments for most
diatoms that are usually well adapted to highly turbulent,
nutrient-rich waters (Margalef 1978). Several species of
Rhizosolenia Brightwell and Hemiaulus Heiberg are able
to proliferate in oligotrophic waters when living in mutu-
alistic symbioses with diazotrophic filamentous cyanobac-
teria (Carpenter et al. 1999). Semina & Levashova (1993)
remarked on the unusual distribution of Hemiaulus hauckii
Grunow ex Van Heurck and Climacodium frauenfeldianum
Grunow that were only found under low phosphate and
silicate concentrations at the surface of tropical or subtrop-
ical seas. Carpenter & Janson (2000) examined cells of
C. frauenfeldianum by epifluorescence microscopy, find-
ing that the cells contained 20–30 globular endosymbionts
with cyanobacterial-type fluorescence. Molecular analyses
revealed that the endosymbionts were closely related to
the diazotrophic cyanobacterium, Cyanothece spp. (Falcón
et al. 2002). Coccoid cyanobacteria have also been reported
in Neostreptotheca subindica von Stosch (Hallegraeff &
Jeffrey 1984).
Grunow (1868) described the genus Climacodium
Grunow, with C. frauenfeldianum as the type species, from
samples collected from waters surrounding the Nicobar
Islands, Indian Ocean. Taxonomic treatment of taxa around
Climacodium has been unstable. Climacodium jacobii
Cleve, described from the warm North Atlantic Ocean
(Cleve 1897), and Climacodium atlanticum Mangin (Man-
gin 1910) are considered synonyms of C. frauenfeldianum
(Ostenfeld 1903, Sournia 1968). Climacodium japonicum
Schröder (Schröder 1906) was later synonymized with
Hemiaulus membranaceus Cleve (Sournia 1968) and some
records of Climacodium biconcavum Cleve (later trans-
ferred to Eucampia Ehrenberg) are considered misidentifi-
cations of H. membranaceus or Eucampia zodiacus Ehren-
berg (Rivera et al. 2003). Hasle & Syvertsen (1997) main-
tained that Climacodium comprises C. frauenfeldianum
and C. biconcavum, distinguishing them by the apertures
(spaces between valves of adjacent cells in chains), which
are barely oval in C. frauenfeldianum, elliptic-lanceolate in
C. biconcavum (Hasle & Syvertsen 1997). Records of C.
biconcavum are scarce and questionable while C. frauen-
feldianum is a distinct and widespread species in stratified
waters of tropical seas (Semina & Levashova 1993).
Climacodium has been classified within the family
Eucampiaceae, order Biddulphiales (Cupp 1943), while

*Corresponding author. Email: fernando.gomez@fitoplancton.com

† F. Gómez and L. Wang contributed equally to this paper.

Associate Editor: Teofil Nakov

(Received 15 July 2018; accepted 4 October 2018)

© 2019 The International Society for Diatom Research

Published online 14 Jan 2019

2 Gómez et al.

Sournia (1968) placed it in the Biddulphiaceae, with
Biddulphia S.F. Gray and many other genera, such
as Bellerochea Van Heurck, Cerataulina H. Peragallo
ex F. Schütt, Ditylum J.W. Bailey ex L.W. Bailey,
Eucampia, Hemiaulus, Isthmia C. Agardh, Lithodesmium
Ehrenberg, Helicotheca Ricard ( = Streptotheca Shrub-
sole) and Triceratium Ehrenberg. Round et al. (1990)
and Hasle & Syvertsen (1997) placed Climacodium in
the Hemiaulaceae, together with Hemiaulus, Eucampia
and Cerataulina. In their description of Climacodium,

Figs 1–7. Light and scanning electron microscopic pictures of Climacodium frauenfeldianum. Figs 1–2. Wild cells. Cells with epibiontic
Pseudobodo sp. Arrow (Fig. 2) indicates the central area, which is devoid of plastids. Figs 3–7. Cells of strain FG22. Fig. 3. Pigmentation is
concentrated at the centre of the cells (arrow in the boxed area). Fig. 4. Glutaraldehyde fixed cell. Figs 5–7. Scanning electron micrographs.
Figs 4–5. Note the variability in the length of the apical projections. Figs 6–7. The arrows indicate the potential bilabiate portula. Fig. 6.
The inset shows the end of the apical projection. Fig. 7. The inset shows the external opening of the potential bilabiate portula surrounded
by smoother valve surface. Scale bars = 20 μm (Figs 1–5) or = 2 μm (Figs 6–7).
 Affinity of the planktonic diatoms Climacodium and Bellerochea 3

Figs 8. SSU rDNA phylogenetic tree of the Lithodesmiales and other mediophycean diatoms with Coscinodiscus radiatus as the out-
group. The inset focuses on the Lithodesmiales, illustrating support values at all the nodes. The newly sequenced species is shown in bold
type. Numbers after each taxon name are GenBank accession numbers. The bootstrap values ≥ 70 from Maximum Likelihood analysis
(right) and posterior probabilities ≥ 0.8 (left) from Bayesian Inference are indicated in the internodes. Scale bar = 0.02 substitutions per
site.
4 Gómez et al.

Figs 9. RuBisCO large subunit (rbcL) gene phylogenetic tree of the Lithodesmiales and other mediophycean diatoms with Coscinodis-
cus radiatus as the outgroup. The newly sequenced species is shown in bold type. Numbers after each taxon name are GenBank accession
numbers. The bootstrap values ≥ 70 from Maximum Likelihood analysis (right) and posterior probabilities ≥ 0.8 (left) from Bayesian
Inference are indicated in the internodes. Scale bar = 0.02 substitutions per site.
 Affinity of the planktonic diatoms Climacodium and Bellerochea
Round et al. (1990, p. 264) reported that ‘The valve sur-
face appears structureless in the SEM, except for a small
rimmed pore midway between the two processes. This
pore may not be present on all the valves and varies in
its appearance’. In this study, we provide the first molec-
ular data for Climacodium based on a culture of the type
species, C. frauenfeldianum.
The strain C. frauenfeldianum FG22 was established
from cells isolated off Ubatuba, São Paulo State, Brazil
(23° 31 27.80 S, 45° 04 59.48 W). Materials and meth-
ods are reported in the Supplementary Information. The
cultured cells were H-shaped, joined by the extended valve
apices, elevations with obtuse ends, to form chains that
were somewhat twisted about the pervalvar axis. The size
of the pervalvar and apical axes were 11–23 and 60–
110 μm, respectively. The apertures between adjacent cells
were almost rectangular, usually wider than the cell was
deep. The ends of the elevations were obtuse (Figs 1–4)
and the length of the elevations was variable (Figs 4–5).
Under scanning electron microscopy, the very lightly sili-
cified valve surface was unstructured (Figs 5–7) although
the frustule apices were wrinkled (Fig. 6). There was a cir-
cular smooth area in the centre of the valve, with a central
pore bordered by two semicircular ridges, which may be a
bilabiate portula (Fig. 7). Observations of the internal valve
surface, necessary to confirm whether this structure is a
bilabiate process, were not possible because acid cleaning
dissolved the frustules.
In the SSU rRNA gene phylogeny, sequences of Dity-
lum, Helicotheca, Lithodesmioides von Stosch, Mediopyxis
Medlin & S. Kühn, Bellerochea and Climacodium clus-
tered together with high support (Bootstrap Proportion,
BP, 100; Posterior Probability, PP, 1). This Lithodesmi-
ales clade was divided into four groups: (1) Ditylum sol
(Grunow) De Toni and D. brightwellii (T. West) Grunow
in Van Heurck, (2) Lithodesmium undulatum Ehren-
berg, L. intricatum Ehrenberg and two strains retrieved
from GenBank as Helicotheca tamesis (Shrubsole) Ricard;
and (3) Lithodesmioides polymorpha Stosch and Belle-
rochea yucatanensis von Stosch, and other two sequences
retrieved from GenBank as B. malleus (Brightwell) Van
Heurck and D. brightwellii (Fig. 8). The fourth highly
supported clade (BP, 95; PP, 0.99) contained C. frauenfel-
dianum, five sequences identified as B. malleus, B. horo-
logicalis von Stosch, Mediopyxis helysia S. Kühn, Harg-
reaves & Halliger, Helicotheca tamesis and Helicotheca sp.
(Fig. 8).
For rbcL, there were fewer available sequences, partly
because some of the data appear to originate from misiden-
tified or contaminated cultures. For example, of the three
sequences for B. malleus, only one clustered within the
lithodesmioid diatoms (Fig. 9). Sequences with accession
numbers DQ514763 and KM010149 (retrieved from Gen-
Bank as B. malleus) are not related to the lithodesmioid
diatoms but clustered with the Plagiogrammaceae and
5
Naviculales, respectively. Sequences of Bellerochea, Cli-
macodium, Ditylum, Helicotheca, Lithodesmioides, Lith-
odesmium and Mediopyxis clustered together with high
support (BP, 100; PP, 1) (Fig. 9). The sequence from C.
frauenfeldianum was sister to sequences of B. malleus iso-
late Har-1Bellmall (KC309557) and B. horologicalis strain
ECT3829 (HQ912536), with moderate support (BP, 88; PP,
1). As observed in the SSU rDNA phylogeny, Bellerochea
was not monophyletic. The rbcL sequence KJ577881 from
B. yucatanensis did not cluster together with B. malleus/B.
horologicalis (Fig. 9). Further information on the species
and sequence data of Bellerochea, including its classi-
fication and the misidentification of cultured strains, is
provided in the Supplementary Information.
In summary, our phylogenetic analysis of the SSU
rRNA and rbcL genes revealed that Climacodium is more
closely related to Bellerochea and other members of the
Lithodesmiales (Figs 8–9) than to members of the Hemi-
aulales, where it is currently placed. We therefore recom-
mend that Climacodium be moved to the Bellerocheaceae,
with B. malleus and B. horologicalis. Our analyses also
provide further evidence that some genera within the Lith-
odesmiales are not monophyletic, and several species (e.g.
B. yucatanensis) need further research for more appropriate
generic placement.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This work was supported by the Brazilian Conselho Nacional de
Desenvolvimento Cientifico e Tecnologico [grant number BJT
370646/2013–14 to F.G]; MEL Senior Fellowship at Xiamen
University, China [grant number MELRS1711 to F.G.]; Chinese
Visiting Scholarship Council and the National Natural Science
Foundation of China [grant number #41330959 to L.W. and S.L.].
This is contribution #2018303 of MEL, Xiamen University.
Supplemental data
Supplemental data for this article can be accessed http://dx.doi.org/
10.1080/0269249X.2018.1551246.
ORCID
Fernando Gómez http://orcid.org/0000-0002-5886-3488
Lu Wang http://orcid.org/0000-0003-4559-3607
Senjie Lin http://orcid.org/0000-0001-8831-6111
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