1021-4437/04/5103- © 2004
Russian Journal of Plant Physiology, Vol. 51, No. 3, 2004, pp. 379–385. From Fiziologiya Rastenii, Vol. 51, No. 3, 2004, pp. 422–428.
Original English Text Copyright © 2004 by Shan, X. W. Li, D. Li, Shao, Liu.
In recent years, in order to understand the molecular
basis for organogenesis and somatic embryogenesis of
plant tissue culture, many researchers have gone
beyond the limitation of morphology and extended to
the ﬁelds of molecular biology. Among the studies, the
research on speciﬁc proteins during plant tissue culture
has become a very important direction in the ﬁeld.
Notable progress has been achieved with regard to the
molecular mechanism of somatic embryogenesis [1, 2],
but much less is known for molecular changes accom-
In the present study, the cotyledons of tomato seed-
lings were induced to produce calli and regenerated
plants via organogenesis. Based on a stable and optimal
regeneration system, the components and content of the
speciﬁc proteins occurring at the dedifferentiation
stage, redifferentiation stage, and plant regeneration
stage were analyzed. In addition, components and con-
tent of the proteins in embryogenic and nonembryo-
genic calli were evaluated. This work might have laid a
basis for further study in the genetic regulatory and
molecular basis for plant organogenesis.
MATERIALS AND METHODS
Establishment of Tissue Culture System
The cotyledons of a tomato (
L., cv. Jia Fen-10) were used as explants.
Tissue culture and plant regeneration.
sterilized conventionally, the cotyledons were inocu-
lated on the medium for callus induction and rediffer-
entiation (MS, 0.05 mg/l IAA, 1.0 mg/l BA, 3%
sucrose, and 7% agar, pH 5.8–6.2). The cotyledons ded-
ifferentiated into calli within 10 days in darkness.
When the cotyledons with calli were shifted to contin-
uous light conditions, they redifferentiated into many
shoot buds surrounding calli within 25 days. During the
entire culture process, the temperature was 25
When the buds grew up into 1.5-cm test-tube plantlets,
the plantlets could be cut off and transferred to the
medium for root induction (half-strength MS, 1.0 mg/l
IBA, 3% sucrose, and 7% agar, pH 5.8–6.2) in order to
obtain completely regenerated plants. When the root
system of the regenerated plants fully developed, they
were transplanted to soil. The plants could grow to
maturity and set fruits regularly.
Analysis of Expression Difference of Speciﬁc Proteins
during Tissue Culture
Various organs and structures were used
as experimental materials. These include cotyledons at
different dedifferentiation stages, calli at different red-
Differential Expression of Specific Proteins during
H. Y. Shan, X. W. Li, D. Li, S. Q. Shao, and B. Liu
Institute of Genetics and Cytology, Northeast Normal University, Changchun, 130024 China;
Received August 15, 2003
—The cotyledons of tomato (
L., cv. Jia Fen-10) seedlings were induced to
produce calli and regenerate plants via organogenesis. Utilizing this system, the composition and content of
stage-speciﬁc proteins associated with organogenesis were analyzed. Moreover, a comparison of the protein
composition and content between embryogenic and nonembryogenic calli was conducted. The SDS-PAGE
results and laser densitometric scanning maps showed that there were different speciﬁc proteins expressed at
different stages. Among them, six proteins (61, 54, 38, 37, 35, and 23 kD) were associated with the morpho-
genesis of organs, and two proteins (39 and 24 kD) were related to the morphogenesis of calli. Although no
distinctive difference in protein components of embryogenic calli was noted, there were different trends of
changes, both for the content of the proteins 39 and 24 kD, and for the content of the total proteins, at different
developmental stages of embryogenic calli. The results obtained from the embryogenic and nonembryogenic
calli indicated that these two materials were distinct in the protein components as well as in its content; for
example, the protein 54 kD was detected in nonembryogenic but not in embryogenic calli. The total protein con-
tent in nonembryogenic calli was lower than that in the embryogenic calli.
Key words: Lycopersicon esculentum - organogenesis - speciﬁc proteins - change in protein content
: BA—benzyladenine; IBA—indolylbutyric acid;
MS—Murashige and Skoog nutrient medium.
This article was submitted by the authors in English.