ISSN 1063-0740, Russian Journal of Marine Biology, 2008, Vol. 34, No. 5, pp. 325–328. © Pleiades Publishing, Ltd., 2008.
Original Russian Text © Yu.A. Ivanovskii, V.N. Kulepanov, 2008, published in Biologiya Morya.
The brown alga
has been culti-
vated for a long period in Southeastern Asia countries
and the Russian Far East. Biotechnological methods of
cultivation of this species have been created in detail
and described in the literature [1, 4, 5, 7]. The cited
papers also described the morphological peculiarities
of different stages of gametogenesis and the develop-
ment of male and female gametophytes, zygotae, and
early sporophyte stages. This caused the choice of Jap-
anese kelp as the object of our studies.
In macrophytic algae it is possible to ﬁnd or to
experimentally obtain a callus, a form of unorganized
growth. Several scientists have raised calluses from
somatic tissues in algae of different taxonomic groups,
cultivating it in agar-containing media [11–13]; the list
of such algae included kelps . After long-term cul-
tivation of kelp in callus cultures the development of
male and female gametophytes, as well as sporophytes,
has been recorded [8, 11].
As is known, both the generative and somatic cells
of algae are capable of forming multicellular structures.
In the members of the order Laminariales, if the tem-
perature is too high for development of gametangia,
gametophytes can live for more than 1 year and reach
more than 1 cm in diameter. If an oocyte, for any rea-
son, occurs unattached to the wall of oogonium, fertili-
zation still occurs; however the further development of
the sporophyte is abnormal. In the latter case, a spheri-
cal aggregation of cells arises, which dies in one month
. During the cultivation of
the maturation of gametes in gametophytes is retarded,
whereas their vegetative growth is induced. As a result,
both male and female gametocytes transform into non-
organized cell conglomerates. After being replanted on
nutritive medium, such a callus could survive indeﬁ-
In the available literature we failed to ﬁnd any refer-
ences to the role of dihaploidization of gametophytes in
the induction of unorganized cell growth and somatic
embryogenesis. However, this is a standard technique
cultivation of anthers in higher plants. The
most commonly used method is the treatment of cells
with colchicine. The other commonly used method is
stress stimulus with cold temperature, which switches
the developmental program of sporogenic cells in the
anthers from the gametophytic to sporophytic one .
From the above reasoning, the goal was set to obtain
a gametophytic callus and induce sporophytic morpho-
genesis in Japanese kelp by exposing its spores to fac-
tors that cause chromosome duplication. Such factors
include treatment with colchicine and
which we used in combination with cold stress.
Initiation of Gametophytic Callus and Somatic Embryogenesis
to Mutagenic Agents
Yu. A. Ivanovskii
and V. N. Kulepanov
Far Eastern State University, Vladivostok, 690950 Russia
TINRO-Center, Vladivostok, 690950 Russia
Accepted May 16, 2008
at the stage of zoospore germination ﬁxed to glass slides were irradiated
-rays in doses of 50, 100, or 250 Gy; or treated with colchicine at a concentration of 4
% for 5 days.
The cultivation was conducted in vessels with seawater at a temperature of 12
N and illumination of 4000 lux
for one month. Once a day, from day 22 to day 30, the temperature was reduced to 0
N for 12 h. As a result, in
experimental samples gametophytes appeared that did not form gametangia; these appeared by the third day of
cultivation, as plaques up to 2 cm in diameter (1–2 plaques per slide). In the same culture we found structures
(1–2 per slide) consisting of strictly radially arranged rows of somatic cells attached to the slides. Later, these
disks transformed into cones up to 0.5 cm in diameter. We recorded the development of a single-layered sporo-
arising from the center of such a cone.
callus, somatic embryogenesis,
, cell cultures.