ISSN 1021-4437, Russian Journal of Plant Physiology, 2008, Vol. 55, No. 3, pp. 385–389. © Pleiades Publishing, Ltd., 2008.
Original Russian Text © M.F. Song, Y Z. Han, 2008, published in Fiziologiya Rastenii, 2008, Vol. 55, No. 3, pp. 426–430.
Phosphoinositide-speciﬁc phospholipase C (PI-PLC)
plays an important role in cell signal transduction. It
can hydrolyze phosphatidyl inositol 4,5-bisphosphate
) into two second messengers, inositol 1,4,5-tri-
) and diacylglycerol (DAG). IP
the release of Ca
from the intracellular stores and gen-
oscillation, which regulates the Ca
dent enzymes and channels .
Previous studies of the PLC-IP
signal pathway in
plants were mainly focused on their response to the
extracellular stimuli, such as drought [2, 3], NaCl ,
cold stress , IAA , ABA [2, 4, 5], GA , and
pathogen attack .
In plants, the ﬁrst cDNA encoding functional PI-
PLC was cloned from
after, plant cDNAs encoding PI-PLC proteins have
been obtained from
, and rice . The structure
of PI-PLCs is composed of an exchange factor hand
(EF-hand) motif, an X domain and a Y domain
(together forming the catalytic domain), and a C2
domain, but the protein lacks pleckstrin homology (PH)
domain . This structure is very similar to animal PI-
: BA—benzyladenine; DAG—diacylglycerol;
GSP—gene-speciﬁc primer; IP
NGSP—nest gene-speciﬁc primer; PIP
4,5-bisphosphate; PI-PLC—phosphoinositide-speciﬁc phospho-
lipase C; RACE—rapid ampliﬁcation of cDNA ends.
This text was submitted by the authors in English.
Different PI-PLC isoforms have different biological
functions. The functions of most of PI-PLC isoforms
are still obscure.
an important ornamental plant from the tropical zone.
Here, we report a novel PI-PLC gene cloned from this
plant species. Our data may help to better understand
the role of PI-PLC in plants.
MATERIALS AND METHODS
Plant materials and treatments.
were grown in the greenhouse at a 16-h pho-
toperiod. Roots, stems, leaves, ﬂowers, stamens, pistils,
siliques, and seeds were collected. For investigation of
responses to stresses, plants were treated with NaCl
(250 mM or 300 mM, for 4 h)or subjected to dehydra-
tion (on paper towel, for 4 h). For investigation of
responses to hormones, plants were sprayed with IAA
M), BA (10
M), or ABA (50
4 h later, selected materials were collected, frozen in
, and stored until required.
3'-RACE of the
Total RNA was
using Trizol Extraction Kit
(Promega, United States). According to the manufac-
turer’s instructions, mRNA was converted into
5'-RACE-Ready cDNA and 3'-RACE-Ready cDNA
RACE cDNA Ampliﬁcation Kit
(Clontech, United States). For 3'-RACE, a 3'-RACE
gene-speciﬁc primer (GSP) 5'-GGDGCKCARATGRT-
NGCNTTYAAYAT-3' and a 3'-RACE nest gene-spe-
ciﬁc primer (NGSP) 5'-AATGGHGGNTGYGGNTW-
YRTBAARAARC-3' were designed according to the
Molecular Cloning and Characterization of a Phosphoinositide-
Specific Phospholipase C from
M. F. Song and Y. Z. Han
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences,
China Agriculture University, Beijing, 10094 China;
fax:+86-10-6273-3491; e-mail: firstname.lastname@example.org
Received January 23, 2007
—A full-length gene encoding phosphoinositide-speciﬁc phospholipase C (PI-PLC), named
was cloned from
by RT-PCR and RACE, using the designed degenerate primers. The
gene is a 2087-bp-long sequence including one open reading frame encoding a polypeptide of 601 amino acids.
Northern blot analysis showed that the
gene was expressed predominantly in leaves and stems and at
very low levels in other tissues. The expression of
was strongly induced under the conditions of high
salt or ABA treatment.
Key words: Torenia fournieri - hormone - phospholipase C - stress