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Genomics and Applied Biology 2014, Vol. 5, No. 2, 1-6
http://gab.biopublisher.ca
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2.3 Expression Analysis of arginase genes under
NaCl treatment during seed germination in
Arabidopsis thaliana
The results of the analysis of the arginase genes
expression show that the arginase genes expressed
abundantly during seed germination. To further
investigate the change of the arginase genes expression
under the NaCl treatment, this research conducted
SqRT-PCR and QRT-PCR methods to do the
experiments to study it.
2.4 Expression Analysis of arginase genes under
the NaCl treatment in different concentrations
To choose the suitable NaCl density, we read
numerous academic papers, finally choose 0, 50, 100,
150, 200 mM as 5 concentration gradients to be
treatment conditions.
The result shows that the expression levels of
ARGAH1
and
ARGAH2
genes had risen with the
increasing of NaCl concentration. Under 200 mM
NaCl treatment, the expression levels of
ARGAH1
and
ARGAH2
reached the highest.
Figure 3 Expression analysis of
ARGAHs
genes under NaCl
treatment by SqRT-PCR
2.5 Expression analysis of arginase genes during
different germination stages under NaCl treatment
We can see that the expression levels of
ARGAH1
and
ARGAH2
were increased with increasing NaCl
concentration. In order to detect the expression levels
of
ARGAH1
and
ARGAH2
under NaCl treatment, we
chose 100 mM as the experimental concentration. The
seed sterilized was sown in 1/2 MS medium and 1/2
MS medium adding 100 mM NaCl. The seed was
vernalized at 4
℃
for two days and then transferred to
the culture chamber to culture. RNA was extracted
from the germinating seed after different culture time
(0, 12, 24, 36, 48) h, and then was reverse transcripted
to cDNA, the expression levels of
ARGAH1
and
ARGAH2
was detected by Real-time quantitative PCR.
Experimental results are shown in Figure 4, both of
ARGAH1
and
ARGAH2
were largely expressed in the
seed; the expression level of
ARGAH1
gene is induced
by 100 mM NaCl treatment compared with the control
group with no NaCl added. The expression level of
ARGAH1
is a little higher after 0 h culture. The
speculated reason is that the material was taken out
from low temperature (4
℃
), and it is reported that low
temperature induce the expression of
ARGAHs
. In this
study, the expression model of
ARGAH2
is similar to
ARGAH1
, except for the time after culture 36 h and 48
h at which time expression of
ARGAH2
gene under
NaCl treatment was a bit lower than control. It is not
clear that the reason causing this situation, so further
research is needed to find it out.
Figure 4 Real-time quantitative expression analysis of
ARGAHs
gene under NaCl treatment during seed germination. A
represented the expression change of
ARGAH1
gene treated
with 100mM NaCl during seed germination; B represented the
expression change of
ARGAH2
gene treated with 100mM NaCl
during seed germination
2.6 The enzyme activity assays of arginase
In this study, we analyzed the arginase activity
under100 mM NaCl treatment during seed
germination. We extracted the whole protein from
fresh materials and detected the arginase activity. We
can see that the arginase activity in seed was the
highest, and the arginase activity is higher under NaCl
treatment compared with control.There is an increase
in 36 h, while the possible reason needs further
analysis to find out.