Phycologia (1995) Volume 34 (5), 396-408

Small cells in the life history of dinoflagellates (Dinophyceae): a review

E.S. SILVA! AND M.A. FAUST2

llnstituto Nacional de Saude, Laboratorio de Microbiologia Experimental, A v. Padre Cruz, 1699 Lisboa, Portugal
2Department of Botany, National Museum of Natural History, Smithsonian Institution, Museum Support Center, Suitland,
M D 20746, USA

E.S. SILVA AND M.A. FAUST. 1995. Small cells in the life history of dinoflagellates (Dinophyceae): a review. Phycologia
34: 396-408.

Small cells are observed in dense populations of many dinoflagellate species, both in clonal cultures and in natural bloom
populations. They range in size from 0.5 to 0.16 of the normal cell volume and have reduced nucleus and cytoplasmic
components. Two possible origins are considered for such small cells: unequal cell division or budding-like division, and
successive 'depauperating' cell divisions. Small cells, which are reported here for 34 species, can proliferate actively in the
late stages of exponential growth, and may become the dominant form in cultures when nutrients are exhausted. When
new nutrients are added to the exhausted growth medium, small cells increase in size and structural components, and may
give rise to forms of a more typical size. Small cells may also serve as gametes. In the sexual cycle of the species studied
here, gametes were morphologically indistinguishable from vegetative small cells. Fusion was isogamous in Gymnodinium
splendens Lebour, Gyrodinium instriatum Freudenthal et Lee, Alexandrium lusitanicum Balech and Coolia monotis Meunier,
but anisogamy was also observed in Gyrodinium instriatum. The nuclear structure of several stages of the life history was
examined, including small cells. The results agree with earlier observations: the dinoflagellate nucleus is polyploid or
aneuploid and the chromosomes are polytenic. The existence of small forms may have implications for dinoflagellate
taxonomy.

INTRODUCTION found in the toxic species Gyrodinium cf. aureoleum Hulburt

Small cells of dinoflagellates showing the typical form of the
species often occur in clonal populations of typical vegetative
cells (Braarud 1957; Silva 1965, 1967; Partensky et al.
1988) and DNA analysis of Gymnodinium cf. nagasakiense
Takayama et Adachi by flow cytometry has demonstrated
that small and typical cells belong to a single genetically
homogeneous population (Partensky & Vaulot 1989). The
formation of small cells has been attributed to unequal asexual
cell division (amitosis) by budding (Apstein 1911; Silva 1971)
and/or successive depauperating cell divisions, i.e. divisions
giving rise to cells lower in mass and poorer in plastids and
pigments than normal cells (von Stosch 1973). The emergence
of small cells may occur after meiosis (gametogenesis) or
sexual conjugation between anisogamous gametes (Pfiester &
Anderson 1987).
The earliest observations of small cells were in species of
Ceratium and they were variously considered to be juvenile
forms (Hensen 1887, as reported by von Stosch 1964), seasonal
variants (Lohmann 1908), mutant cells (Kofoid 1909) or
gametes (Skoczylas 1958; Cao Vien 1967). Silva (1959-1989)
observed small cells in a number of dinoflagellates in clonal
cultures and bloom populations. Von Stosch (1964) described
the fusion of a small and a normal cell of Ceratium tripos
(O.F. Miiller) Nitzsch and considered the former to be a
micro-swarmer or male gamete. Later, when investigating
sexual reproduction in Gymnodinium pseudopalustre Schiller
and Woloszynskia apiculata von Stosch, von Stosch (1973)
also recognized fusing gametes, again smaller than typical
cells. For a review of small cells in dinoflagellates, see Pfiester
& Anderson (1987). More recently, dimorphic cells were
(partensky et al. 1988), Gymnodinium cf. nagasakiense (Parten­
sky & Vaulot 1989), Dinophysis acuta Ehrenberg and Dino­
physis cf. acuminata Claperede et Lachmann (MacKenzie
1992) and Dinophysis nor vegica Claparede et Lachmann
(Hansen 1993). This led to increased interest in comparing
the ecology, physiology and taxonomy of small and typical
forms.
For 30 years (1959-1989), the first author has been studying
the growth, cytology and nuclear cycle of dinoflagellates. The
purpose of the present publication is to summarize this work
and review the literature. We discuss the origin of small cells,
compare the cytology and size differentiation of small and
typical cells in culture and assess the role of small cells in the
life history.
MATERIALS AND METHODS
The species used were isolated from seawater samples collected
along the Portuguese coast by E.S. Silva (1962-1989). The
origin of ten clonal cultures producing small cells and the
growth media used are listed in Table 1. Clone 170, Coolia
monotis Meunier, was isolated by Dr M.A. Sampayo from a
bloom in a fish-pond at Setubal, Portugal. Axenic cultures
were obtained by using Provasoli's AM-9 antibiotic mixture.
They were maintained in Provasoli's ASPI medium at a
salinity of 36. 5%0, ASP2 at 24. 0%0 or ASP7 medium at 30. 8%0
(Provasoli 1963) at 20°C in a L: D regime of 14 : 10 h.
Provasoli's AM-9 antibiotic mixture was sometimes added
before the transfer of aged cells to new medium.
Light microscopy
Live or preserved clonal specimens were examined with a
Zeiss Photomicroscope III fitted with phase-contrast and

396 © 1995 International Phycological Society

 Sil va & Faust: Small cells in dinoflagellates 397

Table 1. Source of clones and growth media used

Species Clone Isolation Location! Medium2

Alexandrium lusitanicum Balech 18 June 1962 Obidos ASP7

Cochlodinium sp. 131 March 1975 St Andre ASPI
Coolia monotis Meunier 170 March 1977 Sado Estuary ASP l
Glenodinium foliaceum Stein 218 Nov. 1988 Melides ASP7
Goniodoma pseudogonyaulax Biecheler 20 1 Nov. 1981 Albufeira ASPI
Gonyaulax spinifera (Claparede & Lachmann) Diesing 226 Nov. 1982 St. Andre ASPI &2
Gymnodinium splendens Lebour 293 Oct. 1986 Mongedo ASP7
Gyrodinium instriatum Freudenthal et Lee 335 March 1988 Obidos ASPI &2
Gyrodinium resplendens Hulburt 93 Sept. 1973 St Andre ASPI &2
Scrippsiella trochoidea (Stein) Loeblich III 155 Nov. 1978 Albufeira ASP?

!Clones were collected from lagoons and from the Sado and Mondego estuaries on the Portuguese coast.
2See Provasoli ( 1963).

10 JJm

Figs 1-6. Gymnodinium splendens. Figs 1, 3-6 live cells, Fig. 2 fixed cell.
Fig. 1. Typical living cell.
Fig. 2. Unequal nuclear division (amitosis). Feulgen stained.
Fig. 3. Intermediate cell.
Fig. 4. Isogamic pair of fusing gametes.
Fig. 5. Budding: small cell still connected to the parent cell.
Fig. 6. Large planozygote.

dark-field optics. Cells were fixed in a mixture of acetic acid: Transmission electron microscopy
formaldehyde : ethanol (1: 1 : 1) or Bouin's fluid, followed by
Feulgen's staining procedure. Contrast was enhanced by For TEM, cultures were pre-fixed III 25% glutaraldehyde
staining with light green (Silva & Franca 1985). (GA) in culture medium (1: 10) for 15 min, followed by a

© 1995 International Phycological Society, Phycologia, 34, 396-408

398 Silva & Faust: Small cells in dinoflagellates

10 }Jm

7 8

11
10 pm
-

16
Figs 7-16. Gyrodinium instriatum, live cells, except in Figs 8, 10, 1 1.
Fig. 7. Typical cell.
Fig. 8. Unequal nuclear division. Feulgen stained.
Fig. 9. Typical cell with small cell in the hypocone.
Fig. 10. Dividing cell containing two large, darkly stained nuclei and a third smaller one. Feulgen stained.
Fig. 11. Small cell separating from the parent cell. Feulgen stained.
Fig. 12. Small cell.
Fig. 13. Dividing small cell.
Fig. 14. Two isogamous fusing gametes.
Fig. 15. Two anisogamous fusing gametes.
Fig. 16. Late planozygote with very large nucleus.

second fixation in 3% GA in cacodylate buffer or Karnov­
sky's fixative with 0.2 M sucrose for 1-2 h. Cells were
post-fixed in 1% OS04 in Ryter-Kellenberg buffer overnight,
stained en bloc with buffered 1% uranyl acetate for I h,
dehydrated in an ethanol series and embedded in Spurr's
resin. Specimens were thin-sectioned with a LKB ultramicro­
tome and sections contrasted with 50% uranyl acetate in
ethanol followed by Reynold's lead citrate. Thin sections
were observed with a Philips 30 1 transmission electron
microscope.

© 1995 International Phycological Society, Phycologia, 34, 396-408

 Sil va & Faust: Small cells in dinoflagellates 399

Table 2. Size comparison of typical and small cells in seven species Table 4. Occurrence of different cell forms in cultures of Glenodin­
of dinoflagellates ium Joliaceum

Cell size (lengthxwidth) /Lm Growth period (days)!

Species Typical cells Small cells 54 74 134

Alexandrium lusitanicum 29x2 1 1 7x 10 Cell forms % of population

Coolia monotis 52x 35 29x 18
Glenodinium Joliaceum 34x20 12x6 Typical cells 52.7 31.8 13.8
Gonyaulax spinifera 46x 32 28x20 Intermediate cells 19.6 21.7 20.5
Gymnodinium splendens 67x52 30x2 3 Small cells 26. 1 46.1 64.1
Gyrodinium instriatum 54x 32 26x10 Budding cells 0.2 0.1 0.4
Scrippsiella trochoidea 4 1x 32 22x 16 Pairs of small cells 0.3 0.1 0.4
Planozygotes 1. 1 0.1 0.8

!At least 100 cells were counted of each cell form.

RESULTS AND DISCUSSION

The difference in cell size between small and typical cells

Origin of small cells
Some ideas on the origin of small cells were mentioned in the
introduction: 'depauperating' divisions (von Stosch 1973) or
unequal nuclear fragmentation (amitosis) by budding (Borgert
19 10; Apstein 191 1; Silva 197 1, 1977). Small cells may form
within parent cells and separate after rupture of the parent
cell wall (Silva 197 1). They are capable of dividing vegetatively
(Silva 1971) and may serve as gametes or pre-gametic stages
(Braarud 1957; Pfiester 1989; Silva 1989).
Such small cells generally have the typical form of the
species (e.g. Figs 12, 26) but they contain a smaller nucleus,
reduced cytoplasmic components and a thin wall (Silva 1971).
can be approximately 0.5-0. 16 the cell volume, e.g. in Gymno­
dinium splendens Lebour (Figs 1, 3), Gyrodinium instriatum
Freudenthal et Lee (Figs 7, 8, 12), Glenodinium foliaceum
Stein (Figs 17, 20), Alexandrium lusitanicum Balech (Figs 24,
26), Gonyaulax spinifera (Claparede et Lachmann) Diesing
(Figs 29, 31) and Coolia monotis (Figs 34, 37). Cell sizes of
typical and small cells of seven dinoflagellate species are listed
in Table 2. Small cells serving as gametes are illustrated in
Gymnodinium splendens (Fig. 4), Gyrodinium instriatum
(Fig. 14) and Alexandrium lusitanicum (Fig. 27) but small cells
have been observed in many other species (Table 3).
Small cells may be abundant in the late exponential growth

Table 3. Dinoflagellates known to form small cells

Species Reference

Alexandrium lusitanicum Balech Balech 1985

Amphidinium carterae Hulburt Cao Vien 1967
Ceratium comutum (Ehrenberg) Claparede et Lachmann Skoczylas 1958
Ceratium tripos (O.F Muller) Nitzsch Apstein 19 1 1; von Stosch 1964
Cochlodinium heterolobatum Silva Silva 1962, 1967
Cochlodinium sp. Silva 1965, 1977
Coolia monotis Meunier Silva & Franca 1985
Dinophysis acuta Ehrenberg MacKenzie 1992; Hansen 1993
Dinophysis norvegica Claparede et Lachmann Hansen 1993
Dinophysis schuetti Murray et Whitting Taylor 1976
Dinophysis swezyae Kofoid et Skogsberg Taylor 1976
Goniodoma pseudogonyaulax Biecheler Silva 1965
Goniodoma sp. Silva 1965, 1969
Glenodinium Joliaceum Stein Silva 1962
Gonyaulax diacantha (Meunier) Schiller Silva 1961
Gonyaulax polyedra Stein Silva 1971
Gonyaulax spinifera (Claparede et Lachmann) Diesing Silva 1971
Gymnodinium aureolum Hulburt Partensky et al. 1988
Gymnodinium cf nagasakiense Takayama et Adachi Partenski & Vaulot 1989
Gymnodinium splendens Lebour Silva 1971, 1978
Gymnodinium pseudopalustre (Woloszynska) Schiller von Stosch 1973
Gyrodinium cohnii Schiller Kubai & Ris 1969
Gyrodinium instriatum Freudenthal et Lee Silva 1980
Gyrodinium resplendens Hulburt Silva 197 1
Gyrodinium uncatenum Hulburt Braarud 1957; Coats et al. 1984
Gyrodinium sp. Silva 1959
Omithocercus quadratus Schott Taylor 1973
Peridinium trochoideum (Stein) Lemmermann Braarud 1957
Peridinium cinctum (Muller) Ehrenberg Pfiester 1984
Peridinium volzii Lemmermann Pfiester &Anderson 1987
Prorocentrum micans (Ehrenberg) Dodge Silva 1961, 1965
Prorocentrum minimum (Pavillard) Schiller Graneli et al. 1985
Scrippsiella trochoidea (Stein) Loeblich III Silva 1980
Wolozynskia apiculata von Stosch von Stosch 1973

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400 Sil va & Faust: Small cells in dinoflagellates

phase in cultures of Gymnodinium splendens, Glenodinium
foliaceum, Alexandrium lusitanicum, Gonyaulax spinief ra and
Goniodoma pseudogonyaulax Biecheler and they remain viable
for different periods of time (Silva 1971). In nitrogen-depleted
medium small cells appear within days after the transfer of
populations in the mid-exponential growth phase. A rapid
increase in the number of small cells in a population suggests
that the frequency of division is rapid (Figs 13, 59). In cultures
of Gymnodinium cf. nagasakiense the generation time of small
cells reported by Partensky & Vaulot (1989) was 1 d, and of
large cells 1.7 d. The relative abundance of typical, intermedi­
ate and small cells in cultures varies during the growth period

17

10,um.
22

23

10 J.lm 10 ,um
4 -

25

Figs 17-23. Glenodinium foliaceum, live cells unless stated otherwise.

Fig. 17. A large typical and five small cells.
Fig. 18. Elongated nucleus during budding. Fixed cell, Feulgen stained.
Fig. 19. A large cell and a detached bud forming a small cell. Fixed cell, Feulgen stained.
Fig. 20. SIpall cell with long flagellum.
Fig. 21. Four small cells enclosed in a hyaline envelope.
Fig. 22. Four recently separated small daughter cells.
Fig. 23. Cell enclosed in a multilayered cyst wall.
Figs 24-28. Alexandrium lusitanicum, live cells, except in Fig. 25.
Fig. Typical cell.
24.
Fig. Two adj acent cells, a parent cell and a small cell. Feulgen stained.
25.
Fig. Two small cells of equal size.
26.
Fig. Fusing isogamous gametes.
27.
Fig. 28. Large planozygote discharging the ceIJ covering.

© 1995 International Phycological Society, Phycologia, 34, 396-408

 Silva & Faust: Small cells in dinoflagellates 40 1

(Table 4, Glenodinium foliaceum). The intermediate forms,
budding cells and pairs of small cells remain similar in
proportion, with less than 1% being planozygotes. Small cells
persist much longer in the above species than in any other
species grown in culture or studied in blooms.
Addition of fresh nutrient medium to the culture generally
causes an increase in size of the small cells, including enlarge­
ment of the nucleus and increase in chromosome density.
This phenomenon was also observed in cultures of Gymnodin­
ium cf. nagasakiense (Partensky & Vaulot 1989). Enlargement
of small cells of Goniodoma pseudogonyaulax to resemble
typical forms has also been seen in aged cultures without
nutrient addition (Silva 1965). The cultures behaved almost
as if fresh medium were added; the typical cells probably
developed by utilization of organic substances released after
cell lysis (Silva 1965) or in the presence of bacteria (Silva

10 ,..m

37 39

Figs 29-33. Gonyaulax spinifera, live cells.

Fig. 29. Typical cell.
Fig. 30. Unequal cell division.
Fig. 31. Small cell.
Fig. 32. Planozygote shedding the wall covering.
Fig. 33. A late planozygote.
Figs 34-39. Coolia monolis, live cells at the same magnification.
Fig. 34. Typical cell (slightly crushed).
Fig. 35. Budding cell.
Fig. 36. Two different-sized cells, small cell still attached to parent cell.
Fig. 37. Small cell.
Fig. 38. Isogamous gamete fusion.
Fig. 39. Zygote with a large nucleus.

© 1995 International Phycological Society, Phycologia, 34, 396-408

402 Sil va & Faust: Small cells in dinoflagellates

1978, 1982; Grandi et al. 1985). Cultures of some species
were maintained without addition of fresh medium for more
than a year (Silva 1962, 1965).
Development of small cells may occur within a hyaline­
enveloped pellicular cyst where 2, 4, 8 or occasionally 16 cells
are formed by amitotic cell divisions (Glenodinium foliaceum,
Figs 2 1, 22) (Silva 1962); small cysts were also found in
Gyrodinium sp. and in Goniodoma sp. (Silva 1959, 1969). Von
Stosch ( 1973) described identical hyaline-membraned cysts in
Woloszynskia apiculata which he interpreted as either division
cysts or zoosporangia.
Nuclear behaviour during the cell cycle
The dinoflagellate nucleus has a species-specific shape, struc­
ture, size and position in the cell (Figs 7, 12, 44). Nuclear
division is by mitosis or amitosis, but the distinction between
the two processes is not always clear. During mitosis, the
splitting of chromosomes may start at one end of the chromo-
some and proceed uniformly along the chromosome
(Alexandrium lusitanicum, Fig. 40; Cochlodinium sp., Fig. 5 1).
Irregular fragmentation of the chromosomes was often
observed, for example in Goniodoma pseudogonyaulax and
Scrippsiella trochoidea (Stein) Loeblich III (Figs 54, 55).
Before nuclear division by amitosis, an increase in the
number of chromosomes and the size of the nucleus occurs
(Silva 1965, 1977). The chromosomes divide transversely,
obliquely or in different planes, and occasionally form buds
(Cochlodinium sp., Fig. 51; Scrippsiella trochoidea, Fig. 55)
(Leadbeater & Dodge 1967; Silva 1971, 1977). Dense areas of
condensed chromatin are often present as nodules in the
chromosomes of Gyrodinium instriatum (Fig. 52), as more
elongate structures in Scrippsiella trochoidea (Fig. 53), or as
small units scattered in the nucleus of Gyrodinium resplendens
Hulburt (Fig. 58). The increase in number of small chromo­
somes before amitosis is a result of multiple fragmentation of
specific chromosomes. Small chromosomes may perhaps also

Figs 40-43. Nuclear features of dinoflagellates.

Fig. 40. Longitudinally dividing chromosome (arrowhead) in the nucleus of Alexandrium lusitanicum.
Fig. 41. Dividing nucleus in Scrippsiella trochoidea penetrated by channels (arrowheads).
Fig. 42. Ultrastructure of the nucleus in a small cell of Gonyaulax spinifera.
Fig. 43. Isolation of small nuclear fragment in the nucleus of Scrippsiella trochoidea (arrowheads).

© 1995 International Phycological Society, Phycologia, 34, 396-408


ongmate in the nucleolus (Goniodoma pseudogonyaulax,
Fig. 54) (Silva 1977).
Small chromosomes are illustrated here in Alexandrium
lusitanicum (Fig. 45), Coolia monotis (Fig. 4S) and Goniodoma
pseudogonyaulax (Fig. 54). The chromosomes are shorter and
more numerous compared to chromosomes at the start of
mitosis.
Grell (1952) described the nucleus of dinoflagellates as
polyploid or aneuploid and Grasse & Dragesco (1957) con­
sidered the chromosomes to be polytenic. This was confirmed
by Soyer & Haapala (1974). Polyteny and aneuploidy in
dinoflagellates may result from fragmentation of chromo­
somes followed by growth of the fragments due to intensive
DNA synthesis (Silva 1977). Polyteny and aneuploidy may
result in the development of single small cells containing a
small nucleus and a reduced number of chromosomes. This
occurs in populations of typical cells and was observed in
most species examined by us. Small cells no doubt have a
complete genome, but the nucleus has a reduced number of
Figs 44-47. Ultrastructure of the nucleus during the life cycle of Alexandrium lusitanicum.
Fig. 44. Morphology of the resting nucleus in typical cell.
Fig. 45. Possible unequal nuclear fragmentation (arrowheads).
Fig. 46. Small cell with very reduced nucleus.
Fig. 47. Constriction of the nuclear envelope (arrowheads) in an intermediate cell.
© 1995 International Phycological Society, Phycologia, 34, 396-408
Silva & Faust: Small cells in dinoflagellates 403
chromosomes (Figs 43, 46). The nucleus generally shows
dense nuclear material, suggesting that intensive DNA syn­
thesis has occurred. After transfer of small cells to fresh
medium, the cells increase in size, the nucleus enlarges, the
chromosome number increases and the cells develop into the
typical forms. Many typical cell populations were established
from single small cells.
Nuclear behaviour during budding
Budding is a rather common type of cell division during late
log-phase growth in Gymnodinium splendens (Figs 2, 5), Gyro­
dinium instriatum (Figs 10, II), Glenodinium foliaceum (Figs
IS, 19) and Coolia monotis (Figs 35, 36), both in culture and
in natural bloom populations. It occurs when a fragment of
the nucleus separates and is enclosed by its own cell mem­
brane. It may form within the mother cell (Gyrodinium instria­
tum, Fig. 9). The cell wall of the parent cell ruptures and the
small cell is released (Gyrodinium instriatum, Fig. 11; Alexan-
404 Silva & Faust: Small cells in dinoflagellates
drium lusitanicum, Fig. 25). This process has been observed
in Gyrodinium instriatum (Silva 197 1), Alexandrium lusitan­
icum, Goniodoma pseudogonyaulax and Goniodoma sp. (Silva
1965, 1969). The ultrastructure of Coolia monotis during
budding is shown in Figs 49 and 50.
Borgert (1910) and Apstein ( 19 11) were the first to notice
budding in dinoflagellates, which Borgert described as ami­
tosis giving rise to two different-sized cells. However, von
Stosch ( 1964) considered the pairs of different-sized cells in
Ceratium tripos to represent anisogamic fusion. Recently
Pfiester & Anderson (1987) described unequal amitosis in
Gyrodinium sp. in which the two daughter cells possessed
different numbers of chromosomes. Partensky & Vaulot ( 1989)
described the small form of Gymnodinium cf. nagasakiense as
the result of an atypical budding-like division. The nucleus
of the small cells appeared more dense but contained the
same amount of DNA as the large cells. In the small cells of
Gymnodinium cf. nagasakiense the nucleus was 3-6 times
smaller than in the typical form. Cells of different sizes
contained different numbers of chromosomes, some of which
were smaller than in typical cells.
During amitosis, part of the dividing nucleus may detach
from the two daughter nuclei and become isolated into a
small cell when the two daughter cells are freed. This was
observed in cells of Gyrodinium instriatum (Fig. 10) and
Cochlodinium heterolobatum (Silva 1967) and in thin sections
of Scrippsiella trochoidea (Figs 4 1, 42), Alexandrium lusitan­
icum (Figs 45, 47) and Coolia monotis (Fig. 48). During
mitosis, the nucleus was crossed by channels (Figs 4 1, 45), as
reported by Leadbeater & Dodge (1967) and Kubai & Ris
( 1969).
Figs 48-50. Ultrastructure of the nucleus in Coolia monotis at different stages of the cell cycle.
Fig. 48. Isolated small nuclear fragment (Nf ) and a large nucleus (N).
Figs 49, 50. Two different sections during budding-like cell division from the same cell with elongated nucleus (N), small nucleolus (nu) and
plastids (pI).
After budding, the small cells rapidly transform into inter­
mediate forms. Grell ( 1952) claimed that polyploidy or aneu­
ploidy can explain both the formation of the small cells with
a reduced nucleus and small-cell development into the typical,
larger form. Von Stosch ( 1964) observed a marked increase
in the density of nuclear material and lack of a nucleolus in
microswarmers of Ceratium tripos. The dense nuclear material
in small cells seen in the present study (Figs 12, 19, 25)
corresponds to compaction of chromosomes (Figs 43, 46, 57,
58). Similarly, condensed chromosomes were present in the
dense nucleoplasm in small cells of Gymnodinium cf. naga­
sakiense, whereas the chromosomes of typical cells were less
dense (Partensky & Vaulot 1989).
'Depauperating' divisions as a form of rejuvenation of small
cells
Von Stosch (1973) considered the development of small cells
as 'depauperating divisions which led to gamete differen­
tiation, but he found that each gamete could be cultured to
provide a clone. 'Depauperating' divisions may also occur
within a hyaline membrane (Silva 1959, 1969, 1971, 1977; von
Stosch 1964). In Peridinium volzii Lemmermann and Alexan­
drium tamarense (Lebour) Balech Pfiester & Anderson (1987)
referred to this phenomenon as parthenogenesis. Pfiester
(1984) observed cell division in small cells of Peridinium
cinctum (O.F. Muller) Ehrenberg in a nitrogen-depleted
medium. The daughter cells behaved as gametes. Partensky
& Vaulot (1989) referred to small cells in cultures of Gymnodin­
ium cf. nagasakiense as asexually dividing small cells, in
contrast to sexually formed gametes. They interpreted the
© 1995 International Phycological Society, Phyc% gia, 34, 396-408
 Silva & Faust: Small cells in dinoflagellates 405

small cell forms as potential duality and referred to the foliaceum, the intermediate cell form constituted 19.6% and
abundance of small cells in Gymnodinium cf. nagasakiense 21.7% of the cells after 54 or 74 days incubation respectively
cultures as rejuvenation of the cell population. It may lead to (Table 4). Intermediate cells result from cell divisions in which
prolongation of a bloom or to successive blooms (Silva 1989) cell sizes decrease or from the increase in size of vegetative
in which the small cells are responsible for the rapid increase small cells or they represent zygotes of small cells. In cultures
of the dinoflagellate populations. In red tides along the of Alexandrium lusitanicum, Gonyaulax spinifera and Gonyau-
Portuguese coast some small cell types were often dominant,
dividing rapidly. This was seen in blooms of Prorocentrum
micans Ehrenberg, Prorocentrum minimum (Pavillard) Schiller,
Gyrodinium instriatum, Glenodiniumfoliaceum and Scrippsiella
trochoidea (Silva 1980).
Many authors have mentioned that small dinoflagellate
cells may behave either as vegetative cells or as gametes
(Skoczylas 1958; Cao Vien 1967; Partensky & Vaulot 1989;
Pfiester 1989). Pairs of dividing small cells may be dis­
tinguished from fusing gametes by having the two cingula in
the same plane and nuclear division ends before cytokinesis,
as seen in Gymnodinium splendens and Gyrodinium instriatum
(Figs 5, 13); in pairs of fusing gametes the respective cingula
may be obliquely positioned (Gyrodinium instriatum and
Alexandrium lusitanicum, Figs 4, 14, 27; Coolia monotis (Faust
1992)) and the zygote bears two longitudinal flagella (see
Pfiester 1989). From observations on live or fixed populations,
dividing small cells appeared in higher numbers in young
densely growing populations, while fusion of gametes was
more frequent in older cultures. The ultrastructure of the
nucleus in Gonyaulax spinief ra is similar in a dividing small
cell (Fig. 59) and fusing gametes (Fig. 60).
During sexual reproduction fusing gametes may be equal
in size (isogamy) as in Gymnodinium splendens (Fig. 4), Gyrod­
inium instriatum (Fig. 14), Alexandrium lusitanicum (Fig. 27)
and Coolia monotis (Fig. 38) (Faust 1992). In other cases,
gametes are unequal in size (anisogamic) (also seen in Gyrod­
inium instriatum (Fig. 15)). Anisogamy may be mistaken for
budding but a small gamete has the specific dinoflagellate
morphology with furrows and flagella, while a bud is an
undifferentiated small entity (Glenodinium foliaceum, Fig. 19;
Coolia monotis, Figs 35, 36). After gamete fusion, the zygote
(planozygote) increases in size and rounds up (Faust 1992).
It forms a cyst which may enlarge during cyst maturation
(Faust 1992). A newly formed zygote may be larger than the
typical cell, as in Gymnodinium splendens (Fig. 6), Gyrodinium
instriatum (Fig. 16) and Coolia monotis (Faust 1992). The
planozygote of the latter species and of G. instriatum (Fig. 16)
has an unusually large nucleus, which occupies about half the
total cell volume. Paired and single chromosomes undergo a
circular, simultaneous movement, known as nuclear cyclosis,
within the nuclear envelope (Figs 16, 33, 39)(Biecheler 1952;
von Stosch 1973). Cyst maturation is accompanied by forma­
tion of starch grains and one or more red accumulation bodies
and a layered wall (Figs 23, 28). The planozygote form of
Gonyaulax spinifera (Fig. 32) represents an early resting stage
in the life history.

Appearance of intermediate cells

Figs 57, 58. Gyrodinium resplendens, thin sections of typical and

Intermediate-sized cells may be found in young dinoflagellate small cells.

Fig. 57. Typical cell.
populations. An intermediate cell form of Gymnodinium splen­ Fig. 58. Small cell showing small fragments of chromosomes that
dens is illustrated in Fig. 3 for comparison with a typical cell have become free in the nucleus (arrowheads); a large mitochond­
(Fig. 1) and a small cell (Fig. 4). In cultures of Glenodinium rion (mi) and a plastid (pi).

© 1995 International Phycological Society, Phycologia, 34, 396-408

406 Silva & Faust: Small cells in dinoflagellates

lax polyedra, intermediate forms resulted from decreasing cell mlUm instriatum, Glenodinium foliaceum (Silva 1978) and
divisions and preceded the appearance of small cells (Silva Coolia monotis, small cells often appeared in culture before
1971). However, in cultures of Gymnodinium splendens, Gyrod- the intermediate forms (Silva & Franca 1985).

Figs 51-56. Multiplication of chromosomes in dinoflagellates during different types of cell division.
Fig. 51. Splitting of chromosomes (arrowhead) and budding (double arrow) in the nucleus of Cochlodinium sp.
Fig. 52. Densely stained chromosome nodules (arrowheads) and budding chromosome segments (double arrows) in Gyrodinium instriatum.
Fig. 53. Striated chromosomes nodules (arrowheads) in Scrippsie//a trochoidea.
Fig. 54. Densely aggregated chromosomes in the nucleus (N), showing irregular chromosome fragmentation and a few chromosomes within
the nucleolus (nu) of Gonidiodoma pseudogonyaulax (arrowheads).
Figs 55, 56. Budding and irregular fragmentation of chromosomes (arrowheads) in Scrippsiel/a trochoidea.

© 1995 International Phycological Society, Phycologia, 34, 396-408


Figs 59, 60. Thin sections of Gonyaulax spinifera.
Fig. 59. Small dividing cell, the two daughter nuclei have separated.
Fig. 60. Ultrastructure of the nucleus in typical cell.
Taxonomic significance of small cells
When typical and small cells are found together in natural
populations, it may sometimes be difficult to ascertain whether
they belong to the same species. Dimorphic cells of Dinophysis
most likely represent different stages of the life cycle, probably
sexual. Hansen (1993) raised the question of whether some
species of Dinophysis may be variants of other species. He
considered cells of Dinophysis debilor Paulsen and D. crassior
Paulsen to be different morphotypes in the life history of D.
norvegica Claparede et Lachmann. Similarly, cells identified
as D. dens Pavillard may occur as a result of gametogenesis
in D. acuta Ehrenberg. MacKenzie (1992) observed pairs of
D. acuta and D. dens joined at their ventral margins. He
suggested that sexual conjugation was taking place, inferring
that D. acuta and D. dens are morphotypes of the same
species. Division stages leading to small cells have also been
observed in D. sacculus Stein (small cells resembling D. skagii
Paulsen) (Bardouil et al. 1991) and D. caudata Saville-Kent
(small cells resembling D. diegensis Kofoid) (Reguera et al.
1990).
Partensky & Vaulot (1988) compared the Japanese species
Gymnodinium cf. nagasakiense with the north European
species Gyrodinium cf. aureolum and found them morphologi­
cally indistinguishable. However, the European species was
able to form two subpopulations of vegetative cells, large and
small, whereas the Japanese species produced small forms
towards the end of a bloom. The two species possessed the
© 1995 International Phycological Society, Phycologia, 34, 396-408
Sil va & Faust: Small cells in dinoflagellates 407
60
same number of chromosomes, but the Japanese species
possessed about 40% more DNA than the European species.
CONCLUSIONS
Small cells may have two different functions: (1) they may
serve as vegetative cells which divide rapidly, thereby increas­
ing the population; (2) they may serve as gametes which after
fusion develop into a resting cyst.
Small cells may be formed by: (a) budding, unequal nuclear
fragmentation (amitosis), or by successive depauperating div­
isions; (b) gametogenesis or by sexual conjugation between
anisogamous gametes in nitrogen-depleted conditions.
Small cells may have the typical form of the species but
with a smaller nucleus, reduced cytoplasmic components and
a thin wall.
ACKNOWLEDGEMENTS
The first author wishes to thank Dr M.A. Sampayo for
the culture of Coolia monotis. Thanks are also due to Mrs
A. Rosa for technical assistance. This investigation was
partially supported by a grant from the Juncta National de
Investigacao Cientifica e Tecnologica, Lisbon. We convey very
special thanks to Dr G.M. Hallegraeff, Associate Editor and
unknown reviewers for their valuable comments.
408 Silva & Faust: Small cells in dinoflagellates
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Accepted 25 February 1995
© 1995 International Phycological Society, Phycologia, 34, 396-408