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Molecular Entomology 2013, Vol.4, No.3, 13-21
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Figure 5 The effect of wheat and triticale seed extractions on
the protease activity of the carob moth gut
Note: Each point is average of the three replications
At the lowest concentration used (0.106 mg
protein/mL) the enzyme activity was inhibited 2.6%
and 15.0% for triticale and wheat seed extracts,
respectively. However, at the highest concentration of
the triticale and wheat seed extracts (1.7 mg
protein/mL) percentage of inhibition was 56.0% and
61.0%, respectively.
2.4 In gel amylase and protease assays
When amylase was assayed in the gel, two bands were
seen in all three parts. However, the middle part of the
midgut bands were clearly bigger than those of the
others indicating that the enzyme isoforms are more
active in the middle parts (Figure 2B). Relative
mobility of A1 band was 0.25 and A2 band was 0.43.
In the protease gel, two protease bands were seen in
the mid, anterior and posterior midgut and similar to
the amylase bands, protease bands of the mid part
were sharper than the anterior and posterior midgut
showing the higher activity of these proteases in the
middle parts (Figure 3B). Relative mobility of the P1
and P2 were 0.52 and 0.6, respectively.
2.5 The effect of wheat and triticale seed extracts
on α-amylase in gel assays
The effect of wheat and triticale seed extract on
α-amylase activity in the gel was similar to the effect of
these plants extracts on spectrophotometric enzyme
assays i.e. with increasing the seed extraction the
amount of the enzyme inhibition increased, too (Figure
6). As shown in figure 5 in first lane two bands seen but
in the other lanes where triticale seed extracts were
used second amylase band (A2) was eliminated and the
first amylase band (A1) intensity was decreased. The
same pattern of inhibition was seen when wheat seed
extract was used in enzyme assay (Figure 7). In control
(a) no inhibition was observed but in the second lane (b)
which the greatest amount of the wheat seed extract
was used (1.7 mg protein/mL) the least intensity of the
band is seen when compared with last lane (f) which
the least amount of the wheat seed extracts (0.16 mg
protein/mL) was tested.
Figure 6 The effect of the triticale seed extracts on the protease
activity of the carob moth gut
Note: a: control without triticale seed extract; b: 1.7 mg
protein/mL seed extract; c: 0.85 mg protein/mL: d: 0.425 mg
protein/mL; e: 0.212 mg protein/mL; f: 0.106 mg protein/mL
Figure 7 The effect of the wheat seed extracts on the α-amylase
activity of the carob moth gut
Note: a: control without triticale seed extract; b: 1.7 mg
protein/mL; c: 0.85 mg protein/mL; d: 0.425 mg protein/mL; e:
0.212 mg protein/mL; f: 0.106 mg protein/mL
2.6 The effect of wheat and triticale seed extracts
on protease activity in gel assay
Effect of the wheat seed extract on protease activity of
the gut was dose dependant (Figure 8). When high
dose of the extract (1.7 mg protein/mL) was used
almost all the two protease bands were eliminated and
as the wheat seed extract dose reduced the intensity of