ISSN 1021-4437, Russian Journal of Plant Physiology, 2009, Vol. 56, No. 5, pp. 654–662. © Pleiades Publishing, Ltd., 2009.
Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 5, pp. 726–735.
In ﬂowering plants, microspores develop into the
male gametophytes with one vegetative cell and two
sperm cells. One sperm cell fuses with the egg, and the
other fuses with the central cell in the female gameto-
phyte, and these events result in the formation of the
zygote and primary endosperm cell, respectively. It
drew an interest to switch the gametophytic develop-
ment of microspores into embryogenesis, especially in
the agronomically important crops, such as wheat ,
tobacco [2, 3], rice , barley [5, 6], maize , and
Microspore embryos have been generated in anther
culture [10–13], and these switching can be induced by
stress treatment, such as starvation and/or heat shock
[3, 8, 14, 15]. Even though detail mechanisms and
involved factors are not elucidated, it has been reported
that stress-related proteins, such as heat shock proteins,
are involved in the microspore embryo induction in
rape [15, 16], pepper , tobacco , and other plants.
The expression of the
subfamily of napin
genes, seed storage protein coding genes, is also known
This text was submitted by the authors in English.
to coincide with the induction of oilseed rape
microspore embryogenesis .
Embryo-speciﬁc genes temporally expressed during
the early stages of pollen embryogenesis have also been
identiﬁed in several plants, such as wheat [19, 20] and
oilseed rape ; and these genes seem to be develop-
mentally regulated, suggesting the involvement of these
genes in the development of embryos. Even though
there is no report showing the overall gene expression
patterns during the pepper anther development until
now, anthers at stage 2 with microspores, covering the
period from just before microspore mitosis to mitosis,
are responsive to the induction signals, such as external
stresses for embryogenesis [10, 22]. This implies that
unknown gene(s) triggering embryo induction may be
either more or less transcribed at stage 2 in response to
the induction signals.
We report here eighty one differentially expressed
genes (DEGs) identiﬁed by GeneFishing PCR using
120 random primers during pepper anther develop-
ment, and the sequence analysis of seven representing
DEGs that are either up- or down-regulated at stage 2.
We also documented the changes in mRNA expression
patterns of DEG13, a novel gene that seems to be
related to stresses, upon heat-treating microspores as
well as during anther development.
A Novel Stress-Related Gene in Developing Pepper Anthers
, K. Chae
, J. Ha
, J. S. Lee
, I. C. Jang
, S. Jeong
, M. Kim
, and M. Yoon
Department of Technology Marketing, Mokwon University, 800 Doan-dong Seo-gu, Taejon, 302-729, Korea
Department of Life Sciences, Mokwon University, 800 Doan-dong Seo-gu, Taejon, 302-729, Korea;
fax: 8242-829-7580; e-mail: email@example.com
Received April 2, 2008
—To study gene expression patterns and to ﬁnd genes related with microspore embryogenesis during
L.) anther development, mRNA expression patterns were investigated at four devel-
opmental stages distinguished according to the size of ﬂower bud, the color of anthers, and the cytological fea-
ture of microspores. Through GeneFishing using 120 random primers, 81 genes were found to be differentially
expressed as anthers develop. We directly sequenced seven of them, which were either up- or down-regulated
at stage 2, since microspores at stage 2 are known to be responsive to the induction signals for microspore
embryogenesis. Nucleotide sequence analysis of the isolated differentially expressed genes (DEGs) and the
comparison of these sequences with the GenBank data indicate that DEG13 is a novel gene, which is highly
homologous to a stress-related gene of potato,
91%) and to alcohol dehydrogenase gene of
70%), whose expression is also tightly related to stresses. In vitro data also showed that DEG13 was
more abundantly expressed in heat-treated microspores than in untreated microspores. Here, we report devel-
opmental stage-speciﬁc gene expression patterns during anther development and a novel stress-related gene,
DEG13, which may be involved in microspore embryogenesis in response to heat treatment.
Key words: Capsicum annuum - anther development - DEGs - microspore embryogenesis - stress
: ACP—annealing control primer; DAPI—4',6'-diami-
dino-2-phenylindol; DEG—differentially expressed gene.