American Journal of Plant Sciences, 2012, 3, 1266-1271 Published Online September 2012 (
Understanding the Mechanism of Gamete Release in
Sargassum vulgare C. Agardh
Inderdeep Kaur1, Reeta Kumari2*
1SGTB Khalsa College, Department of Botany, University of Delhi, Delhi, India; 2Environmental Biology Laboratory, Department of
Botany, University of Delhi, Delhi, India.
Email:, *
Received June 25th, 2012; revised July 23rd, 2012; accepted August 5th, 2012
Sargassum vulgare C. Agardh shows androgynous receptacles, each bearing on an average 12 unisexual conceptacles
which open outside by ostiole, and wherein gametangia (antheridia or oogonia) lie interspersed with paraphyses. Since
out-put of eggs is extremely low, 4 - 6 per female conceptacle, Sargassum sp. ensures its survival under all eco-
physiological conditions. The released oogonium is “wrapped” in sulphated polysaccharide-rich wall layer known to
provide protection against desiccation. Oogonia after being “extruded” out of ostiole, are “incubated” on receptacle,
where they grow into eggs that are easily contacted by spermatozoids. Gamete release is synchronous and almost si-
multaneous ensuring high rates of fertilization. The release occurs on days falling near a full moon or new moon, during
low tides when conceptacles lie exposed. Gamete release occurs first from upper conceptacles, which “house” mature
gametangia while lower ones are still developing. This results in gamete release over an extended period of time. The
zygote dispersal and propagule recruitment also show adaptations selectively advantageous for the alga.
Keywords: Gamete Release; Mesochiton Stalk; Oogonium Incubation; Propagule Disp ersal; Sargassum vulgare
1. Introduction
Several reviews over the years have discussed various
factors affecting the developmental stages of algal
strands including gamete release, fertilization, dispersal
period, settlement, attachment, recruitment and subse-
quent growth [1-4]. Amongst marine algae, Fucales have
evoked a lot of interest in phenomena related to gamete
release, zygote formation and germling dispersal [5-8].
Significant progress has been made in understanding the
mechanisms that increase fertilization success of fucoids
inhabiting intertidal zones marked with periods of high
water velocities [9]. Biological and environmen tal factors
affect the intertidal populations, and amongst all, wave
action has received much attention over the last 20 years
[10,11]. Direct effects of wave forces on intertidal or-
ganisms include damage, detachment and displacement
[12]. Reproductive periodicity in fucoid algae, can be
correlated with lunar or tidal cycles as in Silvetia com-
pressa [13], Fucus distichus [14], Fucus vesiculosus [15]
and Sargassum vestitum [16]. High water motion can
inhibit gamete release in Fucus vesiculosus, F. distichous,
Pelvetia fastigiata and result in low fertilization success
as not all conceptacles expel eggs at the same time [17].
This mechanism allows cross fertilization and prevents
potential inbreeding that result in selfing. Repro- ductive
timing and synchronized reproduction itself may increase
fertilization success. This synchronous gamete release
(spawning) integrates various environmental signals [18,
19]. Inspite of detailed account of various aspects of
gamete release, the comparisons between the male and
female conceptacles is lacking. In this research paper
besides giving a detailed account of mucilage associated
with gamete release the authors bring out certain features
of differences in male and female conceptacles.
2. Material and Methods
The plants of Sargassum vulgare C. Agardh a brown
seaweed (Phaeophyceae, Fucales) were collected during
January (2005) from Port Okha (22˚28.528'N, 069˚
04.322'E), Gujarat (India). Plants were washed with sea-
water to remove debris and brought to laboratory in air
tight plastic bags. Selected portions were fixed in 10%
aqueous acrolein, post fixed in 1% mercuric chloride for
24 h to stabilize polyphenols, thereafter rinsed with dis-
tilled water and processed for light microscopic studies
[20]. The tissue was dehydrated at 4˚C with three succes-
sive changes in 2 methoxyethanol for 24 h each, fol-
lowed by 100% ethanol and prop anol for 24 h and fin ally
two successive changes were given with n-butanol for 24
*Corresponding a uthor.
Copyright © 2012 SciRes. AJPS
Understanding the Mechanism of Gamete Release in Sargassum vulgare C. Agardh 1267
h. The dehydrated material was infiltered and finally
embedded in glycol methacrylate. Sections (2 µ thick)
were cut with glass knives using a rotary microtome. The
sections were stained with Periodic Acid Schiff reagent
and 0.5% Toluidine Blue O at pH 4.4 [21]. Total proteins
were localized using Coomassie Brilliant Blue [22]. Al-
cian blue was used to stain sulphated polysaccharides
For scanning electron microscopy the plants were
fixed in 6% Glutaraldehyde, post fixed in OsO4. The por-
tions were then dehydrated in ascending acetone series,
ranging from 10 through 90% followed by two changes
in absolute acetone for 10 min. The material was later
passed through dry acetone (absolute acetone in CuSO4)
followed by critical point drying at 20˚C and 800 lb/
square inch with pressure for 30 min. This temperature
and pressure were raised to 35˚C and 1200 lb/square inch
respectively. These were then coated with gold (100 Å
thickness) under vacuum on sputter coating unit (Edward
coating unit) for 2 min to eliminate charging. The coated
material was examined and photographed with LEO 435
VP, variable pressure scanning electron microscope at an
accelerating voltage of 15 KV.
3. Results
Sargassum vulgare has a perennial existence in intertidal
pools where the plants undergo a period of active vegeta-
tive growth from July to October and reproductive growth
from October to February. When the temperature dips
and illumination is low during November, the plants en-
ter a period of sexual reproduction which culminates in
gamete release and zygote formation in the month of
February. The germlings are dispersed around February
and March. After an active reproductive phase, the
fronds shows senesce and die back leaving only peren-
nial holdfast which regenerates in the month of May
giving rise to a stipe and leaves.
3.1. Gametogenesis
During fertile phase, plants bear abundant androgynous
receptacles (Figure 1(A)). This unisexual conceptacles
are initiated in the cortical region of receptacles (Figure
Amongst several female conceptacles only 3 or 4 male
conceptacles are seen in a single receptacle (Figure
1(C)). The young conceptacle is oval to pear- shaped with
a narrow neck that opens outside through ostiole (Figure
1(D)). Inside the conceptacle, differentiation of oogonia,
(or antheridia) and also the associated paraphyses takes
place. With further development, ostiole narrows down
while the base widens to accommodate developing game-
tangia (Figures 1(E), (F)) and thus the mature concep-
tacle assumes a spherical shape. The cells of conceptacle
floor wall are the progenitors of gametangia and associ-
ated paraphyses. The oogonia arise only from the con-
ceptacles cells whereas, antheridia are produced on para-
physes from all over the conceptacle lining.
Male gametangium or antheridium is unilayered when
young but, at maturity acquires a second layer (Figure
1(G)). Later, a space develops between two layers which
Figure 1. (A) Scanning electron micrograph showing a re-
ceptacle with raised regions that correspond to conceptacles
whose ostioles (os) are seen as pores; (B, C) L.S. of recepta-
cles showing conceptacles (co) at various stages of develop-
ment (arrows). Number of female conceptacles (oo) is
greater than male (an) conceptacles in (C). TBO stained; (D)
Young female (oo) conceptacle with a narrow ostiole and
associated plug (pl) material. TBO stained; (E, F) CBB
stained sections showing paraphyses (arrows) in oogonial
(oo) conceptacle; In (F) three oogonia (oo) are at different
stages of development; (G, H) Male conceptacles showing
mature antheridia with two wall layers (arrows) and a sul-
phated-rich pad like structure. Ostiole remains closed by
compactly arranged cells in (H). TBO stained.
Copyright © 2012 SciRes. AJPS
Understanding the Mechanism of Gamete Release in Sargassum vulgare C. Agardh
gets filled with sulphated polysaccharides. The cyto-
plasm encloses 64 nuclei that correspond to the number
of spermatozoids. The ostiole remains closed due to com-
pactly arranged meristoderm cells, until antheridia ma-
ture (Figure 1(H)). In S. vulgare approximately, 64
spermatozoids are produced per antheridium and many
antheridia per conceptacles, while only one egg per oo-
gonium and 4 or 5 oogonia per conceptacle are pr oduced
(Figure 1(F)).
The young oogonium has an alginate-rich single lay-
ered wall known as exochiton (Figure 2(A)). Dur ing fur-
ther development two more layers namely mesochiton
rich in sulphated polysaccharides and endochiton, a mix
of sulphated and carboxylated polysaccharides are formed.
Period of mesochiton formation is the most active stage
when oogonial nucleus is prominent and nuclear mem-
brane is irregular in outlin e (Figure 2(B)). Nuclear blebs
are formed which establish a “cross talk” between cyto-
plasm and nucleus. The perinuclear region is full of phy-
sodes (Figure 2(C)). The conceptacle opening remains
occluded with a polysaccharide-rich plug Figure 2(D)).
3.2. Gamete Release and Role of Mucilage
Sargassum vulgare releases its gametes profusely in
packets on a full moon and new moon day during low
tides. The oogonia are released while they are “wrapped”
in mucilage (Figure 2(E)). The mesochiton, which acts
as a protective covering at this stage shows a granular
texture (Figures 2(F), (G)). A maximum of two oogonia
may be released through the ostiole at a given time. This
is because the released oogonia are held back to the
conceptacle floor by a mesochiton stalk and the narrow
ostiole which can dilate only a little cannot accommodate
more than two stalks (Fi gure 2(H)).
On the contrary, the spermatozoids are released “en
masse” through ostiole where the meristoderm loosens
and the gametes “wrapp ed” in mucilage are released. Th e
conceptacle cavity shows scanty mucilage (Figures 3(A),
(B)). The antheridial contents take up a tadpole-like
shape which facilitates th eir exit through the na rrow osti-
ole (Figure 3(C) ).
There is copious amount of mucilage in the concept-
tacle cavity which houses oogonia in contrast to male
conceptacle (Figure 3(B)). The paraphyses which co-
exist with oogonia (Figure 3(D)) possess mucilage with
staining properties similar to plug material (Figures 3(E),
(F)) and are therefore act as a source of polysaccharides.
3.3. Propagule Settlement
Mesochiton stalk ho lds the zygote and initial d ivisions in
germling formation occur while it is retained on recepta-
cle (Figures 3(G), (H)). This “parental care” is extend-
edto the germlings that are shed with a well defined rhi-
Figure 2. (A) Section through young female conceptacle
showing oogonium (oo) with only one layered (exochiton)
wall (arrow). TBO stained; (B, C) the nucleus (n) is pro-
minent, with undulated (arrow) envelope and a prominent
nucleolus. Nuclear blebs (double arrow) are seen in (C); (D)
Plug (pl) material from female conceptacles stains well with
PAS and Alcian blue (inset) indicating its complex nature;
(E) Scanning electron micrograph showing released oogonia
with a mesochition stalk (st) and mucilage (mu); (F, G) Oo-
gonia at the time of release showing ruptured exochition
(arrows), thick granular, mesochition (ms) and physode
rich cytoplasm. TBO stained; (H) Two oogonia released
from a conceptacle with lightly stained mesochiton stalks
(arrows). TBO stained.
zoidal end (Figure 3(G)).
4. Discussion
Sargassum is one of the most widely investigated alga
with various events in reproductive phase having been
elucidated [24,25]. Great emphasis has been laid down
Copyright © 2012 SciRes. AJPS
Understanding the Mechanism of Gamete Release in Sargassum vulgare C. Agardh 1269
Figure 3. (A)-(C) Sections through male conceptacles where
ostiole (os) “gapes” apart in (A); conceptacle has reduced
polysaccharides in the cavity (Alcian Blue stained) in (B)
and spermatozoids “wrapped” in a mucilage showing their
release in (C). (A) and (C) are TBO stained; (D)-(F) Para-
physes (pa) from oogonial conceptacles; (D) phase contrast
micrograph; (E) stained with Alcian blue; (F) TBO staine d;
(G, H) Incubated germlings with a smaller vacuolated
rhizoidal (rh) and a larger physode (ph) rich cell. The ar-
rows show walls laid down in the germlings.
on gamete release and post fertilization recruitment sta-
ges [2,26,27]. Reproductive events are governed by a
number of environmental cues which evolve as a mecha-
nism for populations to increase the probability of fer-
tilization by releasing gametes at the same time. Gamete
release at low tide period is stated to be a consequence of
mechanism selected to permit successful fertilization.
Sexual reproduction is enhanced in habitats or at the
times where water motion is low and water rela- tively
calm [26].
According to Pearson and Brawley [14] ability of Fu-
cus distichous to synchronise gamete release during pe-
riods of low water motion is an extremely valuable ad-
aptation for an organism inhabiting intertidal zone. Low
tides provide ideal conditions for making large number
of gametes available which increase chances of gamete
collision. According to Suto [28], the period of anthero-
zoid motility is short and the adaptations are therefore
towards quick encounter with eggs. Another feature
which ensures a high success rate of fertilization and
establishment is the mucilage around th e female gametes
and propagules [29 ].
The present study highlights for the first time, that
architecture of male and female conceptacles is fine-
tuned for sexual act. The male conceptacle in Sargassum
heterophyllum is reported to possess an ostiolar plug [29]
which plays an important role in determining the time of
gamete release. In the present study, male conceptacle in
S. vulgare lacks ostiolar plug and remains closed by
tightly fitted ostiolar (meristoderm) cells. On the other
hand, female conceptacle has a plug. The present study
indicates that these plug helps in isolating the developing
structures from outer eco-physical pressures and also
provides gametangia insulated environment. This regu-
lates oogonial release by not allowing female to exit till
spermatozoids are made available. Removal of such a
plug from female conceptacle might take a little longer
because it is not only copious in amount but also a mix-
ture of polysaccharides. Thus it needs to be dissolved in
order to make way for oogonia being released. Since
plug is absent from antheridial conceptacle, the release of
spermatozoids is easier and it may have a bearing on
temporal difference in gamete release. In such a situation,
it is quite likely that gamete release in this androgynous
species is protandrous.
One feature reported for the first time is the difference
in number of paraphyses in male and female concepta-
cles. Paraphyses are known to be associated with mu-
cilage secretion that fills the cavity, bathes oogonia and
occludes ostiole. The role of paraphysial secretions in
early oogonial development is to check desiccation. At
maturity, the secretions help in making ostiole slippery
hence facilitating gamete release [8]. Whether the ostio-
lar closure in Sargassum conceptacles has any evolution-
ary bearing on the callose plugs during micro and mega
sporogenesis of higher plants, needs further investiga-
5. Acknowledgements
Inderdeep Kaur gratefully acknowledges the financial
support provided by the Department of Science and
Technology under their Women Scientist Scheme (WOS-
Copyright © 2012 SciRes. AJPS
Understanding the Mechanism of Gamete Release in Sargassum vulgare C. Agardh
A). Reeta Kumari is thankful to the University Grants
Commission, New Delhi (India) for award of Rajiv
Gandhi National Fellowship for the period 2006-07.
[1] A. R. O. Chapman, “Demography,” In: M. Littler, D. S.
Littler, Eds., Handbook of Phycological Methods, Eco-
logical Field Methods: Macroalgae, Cambridge Univer-
sity Press, Cambridge, 1985, pp. 253-268.
[2] B. Santelices, “Patterns of Reproduction, Dispersal and
Recruitment in Seaweeds,” Oceanography Marine Biol-
ogy Annual Review, Vol. 28, 1990, pp. 177-276.
[3] M. N. Clayton, “Propagules of Marine Macro Algae:
Structure and Development,” British Journal of Phycol-
ogy, Vol. 27, No. 3, 1992, pp. 219-232.
[4] R. L. Vadas, S. Johnson and T. A. Norton, “Recruitment
and Mortality of Early Post-Settlement Stages of Benthic
Algae,” British Journal of Phycology, Vol. 27, No. 3,
1992, pp. 331-351.
[5] T. A. Norton, “Gamete Expulsion and Release in Sar-
gassum muticum,” Botanica Marina, Vol. 24, No. 8, 1981,
pp. 465-470. doi:10.1515/botm.1981.24.8.465
[6] P. A. Mooney and V. Staden, “Lunar Periodicity of the
Levels of Endogenous Cytokinins in Sargassum hetero-
phyllum (Phaeophyceae),” Botanica Marina, Vol. 27, No.
10, 1984, pp. 467-472. doi:10.1515/botm.1984.27.10.467
[7] I. Kaur and M. R. Vijayaraghavan, “Oogonial Develop-
ment, Maturation and Release in Sargassum vulgare C.
Agardh and S. johnstonii Setchell & Gardner,” Aquatic
Botany, Vol. 42, No. 2, 1992, pp. 173-185.
[8] I. Kaur and M. R. Vijayaraghavan, “Histochemical Stud-
ies on the Mesochiton-Stalk, Egg and Zygote of Sargas-
sum vulgare C. Agardh (Phaeophyceae, Fucales),” Japa-
nese Journal of Phycology, Vol. 40, 1994, pp. 431-436.
[9] S. H. Brawley and L. E. Johnson, “Gametogenesis, Gam-
etes and Zygotes: An Ecological Perspective on Sexual
Reproduction in the Algae,” British Journal of Phycology,
Vol. 27, No. 3, 1992, pp. 233-252.
[10] C. L. Hurd, “Water Motion, Marine Macroalgal Physiol-
ogy and Production,” Journal of Phycology, Vol. 36, No.
3, 2000, pp. 453-472.
[11] D. I. Taylor and D. R. Schiel, “Wave-Related Mortality in
Zygotes of Habitat-Forming Algae from Different Expo-
sures in Southern New Zealand: The Importance of
‘Stickability’,” Journal of Experimental Marine Biology
and Ecology, Vol. 290, No, 2, 2003, pp. 229-245.
[12] M. W. Denny, “Predicting Physical Disturbance: Mecha-
nistic Approaches to the Study of Survivorship on Wave-
Swept Shores,” Ecological Monographs, Vol. 65, No. 4,
1995, pp. 371-418. doi:10.2307/2963496
[13] G. A. Pearson, E. A. Serrao, M. J. Dring and R. Schmid,
“Blue and Green Light Signals for Gamete Release in the
Brown Alga, Silvetia compressa,” Oceanologia, Vol. 138,
No. 2, 2004, pp. 193-201.
[14] G. A. Pearson and S. H. Brawley, “Reproductive Ecology
of Fucus distichus (Phaeophyceae): An Intertidal Alga
with Successful External Fertilization,” Marine Ecology
Programme Series, Vol. 143, 1996, pp. 211-223.
[15] G. A. Pearson, E. A. Serrao and S. H. Brawley, “Control
of Gamete Release in Fucoid Algae: Sensing Hydrody-
namic Conditions via Carbon Acquisition,” Ecology, Vol.
79, No. 5, 1998, pp. 1725-1739.
[16] D. I. May and M. N. Clayt on, “Oogenesis, the Formation
of Oogonial Stalks and Fertilization in Sargassum vesti-
tum (Fucales, Phaeophyta) from Southern Australia,”
Phycologia, Vol. 30, No. 3, 1991, pp. 243-256.
[17] S. H. Brawley, L. E. Johnson, G. A. Pearson, V. Sper-
ansky, R. Li and E. Serrao, “Gamete Release at Low Tide
in Fucoid Algae: Maladaptive or Advantageous?” Ameri-
can Zoologist, Vol. 39, 1999, pp. 218-229.
[18] G. A. Pearson and E. A. Serrao, “Revisiting Synchronous
Gamete Release by Fucoid Algae in the Intertidal Zone:
Fertilization Success and Beyond?” Integrative and Com-
parative Biology, Vol. 46, No. 5, 2006, pp. 587-597.
[19] E. A. Serrao, G. A. Pearson, L. Kautsky and S. H. Braw-
ley, “Successful External Fertilization in Turbulent Envi-
ronments,” Proceedings of National Academy of Science,
USA, Vol. 93, No. 11, 1996, pp. 5286-5290.
[20] N. Feder a nd T . P. O’ Br ie n, “Pl ant Mi cr ote c hni que : Some
Principles and New Methods,” American Journal of Bot-
any, Vol. 55, No. 1, 1968, pp. 123-142.
[21] M. E. McCully, “Histological Studies on the Genus Fucus
I. Light Microscopy of the Mature Vegetative Plant,”
Protoplasma, Vol. 62, No. 4, 1968, pp. 287-305.
[22] K. Weber and M. Osborn, “Proteins and Sodium Dodecyl
Sulphate: Molecular Weight Determination on Poly-
acrylamide Gels and Related Procedures,” In: H. Neurath
and R. L. Hill, Eds., The Protein Vol. 1, 3rd Edition, Aca-
demic Press, New York, 1975, pp. 179-223.
[23] B. C. Parker and A. G. Diboll, “Alcian Stains for Histo-
chemical Localization of Acid and Sulphated Polysaccha-
rides in Algae,” Phycologia, Vol. 6, No. 1, 1966, pp. 37-
46. doi:10.2216/i0031-8884-6-1-37.1
[24] N. Chikako, M. Taizo and T. Terunober, “Degeneration
and Extrusion of Nuclei during Oogenesis in Silvetia
babingtonii, Cystoseira hakodatensis and Sargassum con-
fusum (Fucales, Phaeophyceae),” Phycologia, Vol. 40, No.
5, 2001, pp. 411-420. doi:10.2216/i0031-8884-40-5-411.1
[25] D. Zou, G. Kunshan and C. Weizhou, “Photosynthetic
Carbon Acquisition in Sargassum henslowianum (Fucales,
Phaeophyta), with Special Reference to the Comparison
between the Vegetative and Reproductive Tissues,” Pho-
tosynthetic Research, Vol. 107, No. 2, 2011, pp. 159-168.
Copyright © 2012 SciRes. AJPS
Understanding the Mechanism of Gamete Release in Sargassum vulgare C. Agardh
Copyright © 2012 SciRes. AJPS
[26] C. Monteiro, H. E. Aschwin, A. S. Ester and S. Rui,
“Habitat Differences in the Timing of Reproduction of the
Invasive Alga Sargassum muticum (Phaeophyta, Sargas-
saceae) over Tidal and Lunar Cycles,” Journal of Phy-
cology, Vol. 45, No. 1, 2009, pp. 1-7.
[27] B. Santelices, “Recent Advances in Fertilization Ecology
of Macro Algae,” Journal of Phycology, Vol. 38, No. 1,
2002, pp. 4-10.
[28] S. Suto, “Studies on the Shedding, Swimming and Fixing
of Spores of Seaweeds,” Bulletin of the Japanese Society
of Scientific Fisheries, Vol. 16, No. 1, 1950, pp. 1-9.
[29] A. T. Critchley, V. M. Peddemors and R. N. Pienaar,
“Reproduction and Establishment of Sargassum hetero-
phyllum (Turner) C. Ag. (Phaeophyceae, Fucales),” Euro-
pean Journal of Phycology, Vol. 26, 1991, pp. 303-314.