Agriculture Reference
In-Depth Information
wild-type showed bending by hook formation. However, as shown in Figure
1b the mutant
R3-1
germinated stranding upward without hook formation.
Enzymatic Analysis of the Wild-Type and the Mutant
R3-1
The resistance to strong sunlight will confer the mutant
R3-1
the capacity
to produce a high yielding. Under strong sunlight, chlorophyll will be excited
to generate triplet chlorophyll, which will result in the excitation of triplet
oxygen generated by the photolysis of H
2
O to
2H
2
O
→
3
O
2
+ 4H
+
+ 4e
-
to singlet oxygen (
1
O
2
). Getting energy exciting triplet oxygen to singlet
oxygen the process of energy transfer gave much more energy and the singlet
oxygen will move at a range of path length of 500 µm.
The soluble fraction and membrane fraction of leaves under strong
sunlight from the wild-type and the mutant
R3-1
were prepared as described in
Materials and Methods. By use of labeling method of histidine kinase by
incubating samples of soluble fraction and the membrane fraction with 10
-9
M
[
γ
-
32
P]ATP, high molecular weight histidine kinases, and low molecular
weight histidine kinases were labeled. As shown in Figure 2a, we could detect
the increase in the phosphorylation of 40, 48, 63, and 70 kDa histidine kinases
in
R3-1
in the soluble fraction, and 63 and 72 kDa histidine kinases in
membrane fraction compared with those in the wild-type by long exposure of
X-ray film.
As shown in Figure 2b obtained by short exposure of X-ray film, the
increase in the phosphorylation of these high molecular weight histidine
kinases were under the control of low molecular weight histidine kinases of
NDPK1 corresponding to 15 kDa, transit peptide NDPK2 corresponding to 19
kDa and leader sequence NDPK3 corresponding to 17 kDa. The increase in
the phosphorylation of transit peptide NDPK2 Ile12Leu Glu205Lys mutant
molecule of NDPK2 resulted in the faster mobility in the SDS gel
electrophoresis, which conferred the enhanced phosphorylation of large
molecular weight histidine kinases in soluble fraction and membrane fraction.
In our previous study (Haque et al., 2010), we reported that transit peptide
NDPK2 Ile12Leu Glu205Lys, the mutant molecule of NDPK2 with
glutathione
S
-transferase showed not only enhanced autophosphorylation but
