Results

Prelimilary screening killing effect and impact on

cruciferous plants of crude extracts of Capsicum
annuum L. fruits, Momordica cochinchinensis L. seeds
and Melia azedarach leaves
The synergistic effect of those three crude extracts from
CA fruits, MC seeds and MA leaves on both the insect and the
vegetable together have been investigated. Crude extracts were
prepared with distilled water and sprayed on potted Brassica
juncea (L.) Czerm and Coss plants containing 4
th
instars of Pieris
rapae.
















The mortality rates of the insect tested on the plants were
not as high as on the petri dishes in previous report (Nguyn
Ngc Ho et al., 2011) (figure 1). However the killing effect of
those extracts were still quite high. After 72 h the dead rate of the
insect sprayed with mixture of MC and CA was the highest ( 80,4
6,94%). Inevitably, there were synergistic insect killing effect in
all three paired mixtures since the their killing activities were
higher than the ones of individual extract with double
concentration. Unexpectedly, it was observed that all Brassica
juncea plants sprayed with MA leave extract both individually and
in mixture were gradually blighted and died (data not show). So it
could be inferred that Melia azedarach aqueous leave extract
inhibit the growth of the vegetables. This result raised our
awareness of using MA leave extract from our further
development of a combined herbal pesticide.



Introduction
Pieris rapae, Spodoptera litura and Plutella xylostella are the most common serious worms that damage the main cruciferous plants in the world and they can cause 90-100% crop loss
(Eltayeb et al., 2010). There have been several investigations by Vietnamese scientists, aiming at plant extracts to control these insects
(Nguyn Th Minh Tm, 1992; Phm Th Trn Chu et al., 2002), however no strongly effective formulations has been invented sofar. In-Lab and in-greenhouse experiments, we found three
extracts from from fruits of hot chilli (Capsicum annuum L.) leaves of Melia azedarach L. and seeds of Momordica cochinchinensis L. have very strong pesticidal and antifeedant activities.
Moreover, the combination of these three extracts, XGO, resulted a synergistic effect that caused 100% insects death in some hours. However the detail modes of action of individual plant
extract and of the mixture on the insect pests, and the exact candidate insecticidal secondary metabolites or proteins in the extracts have not been known yet. Further more, no field trial of
XGO has been done to evaluate its practical lethal dose to each insect, durability of protective effects and the agronomical characteristics of the cruciferous plants which would be sprayed
with this mixture of plant extracts
Conclusion
- Mixed plant extracts of C. annuum fruit , M. cochinchinesis seed, and M. azedarach exhibited synergistic insecticidal activity towards Pieris rapae.
- Most of Pieris rapae digestive proteases are belong to the serine group, and largely inhibited by PPIs from seeds of M. cochinchinesis and Sophora japonica.
- The ethanol extract from C. annuum fruit yielded the most pesticidal activity and less color than the water extract
- All preparations containing extract of M. azedarach inhibit the growth, reduced the nutritious quality of cabbage, and should not be used as a herbal pesticides.
- 10 time diluted mixed extract of C. annuum fruit and M. cochinchinesis seed was the most potential preparation for the use as a novel biopestide. Its pesticidal efficacy is equal to the commercial biopesticide
References
- -Barbero, G.F., Palma, M., Barroso, C.G. (2006) Pressurized liquid extraction of capsaicinoids from peppers. J .Agric. Food. Chem. 54 (9):32316
-Phm Th Trn Chu, Phan Th H,
Mai Ngc Ton, Trnh Hng Thi, Trn Quang Tn, Hong Th Vit, Nguyn u Ton, Phm Th Hnh(2000). Tc dng tr su hi rau ca ch phm momosertatin tch t ht gc (momordica cochinchinensis). TC Khoa hc: Khoa hc t nhin (i hcquc gia H
Ni).- 2000.- no.1.-Tp16. - tr. 1-11.
-Nguyn Ngc Ha, inh Th Phng, Nguyn Vn Du, Lu Th Phng, Nguyn Th Cm Chu, Nguyn Vn Giang, Nguyn Th PhngTho, ng Xun Nghim (2011): Investigation of Killing and Antifeedant Effects on Pieris rapae Larvae of Extracts from
Some Potential Plant Species. Journal of Sciences and Development (Vietnamese Iss.) 4: 535-542
-
Figure 7. Weekly average number of
Pieris rapae larvae per cabbage plant.
Development of a novel bioinsecticide based on XGO extract
from plants for the control of the cabbageworm (Pieris rapae)
and other major pests of cruciferous vegetables
Hi Qunh, Bi Th Thanh, Nguyn Vn Thng, Nguyn Khc Hi, Nguyn c Thnh, ng Xun Nghim
Faculty of Biotechnology, Hanoi University of Agriculture,
Corresponding authors email: quynhdohai@gmail.com
.












The results indicated that capsaicinoids were extracted most with
methanol and least with water at 90
o
C.Our result is similar to other
reports in which methanol (Kirschbaum-Titze et al.,2002, Karnka et al.,
2002), ethanol (Barbero et al., 2006) are solvents that are normally
employed for the extraction of capsaicinoids in various extraction
techniques, such as Soxhlet extraction, maceration, and extraction by
magnetic stirring. Water is not a good solvent for extracting
capsaicinoids,
0
10
20
30
40
50
60
70
80
90
100
MC+CA MC+MA MA+CA MA MC CA H2O
K
i
l
l
i
n
g

a
c
t
i
v
i
t
y

(
%
)

Extracts
12h
24 h
36 h
48 h
60 h
72 h
Figure 1. Killing activities of different extractions. The concentration of
each extract in the mixtures is one tenth, and in the unmixed extracts
is one fifth of the original crude extracts.
Table 1. Antifeedant activites of different
extracts of CAfruits and MC seeds
Extracts Components
Antifeedant
activities (%)
CT1 CA+ Hexane 79,1 1,0
CT2 CA+ Acetone 67,5 2,4
CT3 CA+ Methanol 91,6 1,6
CT4 CA+ Ethanol 85,4 1,2
CT5
CA+ 90
o
C +
H2O
63,9 1,5
CT6
CA+50
o
C +
H2O
69,2 2,2
CT7 MC + H2O 93,0 1,0
CT8
MC + 90
o
C
+H2O
93,3 1,6
CT9 H2O 0
Extraction of Capsaicinoids from CA fruits
Extractions of Capsaicinoids were performed by applying
different solvents or water at different temperature on dry powder
of CA fruits (8:1 v/w) The clear extracts were collected after
centrifugation at 9000 rpm (figure 2).
The efficiency of different extractions were compared by applying
the ten-time -diluted solutions on the cabbage leaves or/and
larvae for antifeedant and killing activities (Table 1, figure 3)

Evaluation of pesticidal afficacy of preparations against
Pieris rapae in the field conditions
Seven preparations of plant extracts were used in this
field evaluation. The negative control were sprayed with water,
commercial biopesticide Kuang Hwa Bao WP 1600IU/mg were
used as positive control Since the concentration of each
compositional extract in the mixtures is one half of the individual
one. Figure 5 showed that the synergistic effect against Pieris
rapae were observed in all mixed extracts, and the mixture of
CA and MC extracts showed the highest protective property of
all including the commercial product (table 2 and figure 7).
Table 2: Nine preparations of plant extracts for field trials
Plant
extracts
Components (1:10 diluted)
Formula1 C. annuum
Formula 2 M. cochinchinesis
Formula 3 M. azedarach
Formula 4 C. annuum+ M. cochinchinesis+ M. azedarach
Formula 5 C. annuum+ M. cochinchinesis
Formula 6 C. annuum+ M. azedarach
Formula 7 M. cochinchinesis+ M. azedarach
Formula 8 water
Formula 9 Kuang Hwa Bao WP 16000 IU/mg (ctr commercial
one)
0
0.5
1
1.5
2
2.5
3
3.5
L
a
r v
a
e
a
n
d
p
u
p
a
e
p
e
r
p
l a
n
t
Plant extracts
Figure 3. A. Killing activities of different extracts
(see table1 for components of extracts)
Figure 5. Killing and antifeedant activity
of diluted extracts from seeds of MC
LD50 of Capsaicinoids from CA fruits and protease inhibitors form
MC seeds
After 48 h, more than 76% of the larvae were dead when the plants
were sprayed with 8-time diluted heated extract from seeds of MC
(Figure 3). In other experiments more than 98% of the insects were
killed with 8-time diluted methanol extract from fruits of CA but this rate
significantly decreased at the next dilution as only 62.5% of them dead
with the 16-time diluted extract. The mortality rates of 8-time diluted
extracts from MC and CA are equal to or even higher than the efficacy
of some other biopesticides (Nguyn Th Minh Tm, 1992; Nguyn Vn
Sn, 2001), so they were chosen for the field tests.
0
0.5
1
1.5
2
2.5
3
Formula 1 formula 2 Formula 3 Formula 4 Formula 5 Formula 6 Formula 7 Formula 8 Formula 9
P
r
o
d
u
c
t
i
v
i
t
y

(
k
g
/
p
l
a
n
t
)
Mixed Plant extracts
0
0.5
1
1.5
2
2.5
3
3.5
4
S
u
g
a
r
c
o
n
t
e
n
t
(
w
/ w
%
)
Mixed plant extracts
0
0.005
0.01
0.015
0.02
0.025
0.03
0.035
V
i
t
a
m
i
n
C
c
o
n
t
e
n
t
(
w
/
w
%
)
Mixed plant extracts
Figure 10. Vitamin C contents of tested cabbage
MA+MC MC
MC+CA
CA
A
B C
Figure 6. Killing and antifeedant
activity of diluted methanol extracts
from fruits of CA
Figure 9: Average reduced sugar contents of tested cabbage
Figure 8:A.Cabbage productivity: average marketable weight per plant
Extraction protease inhibitor from MC
seeds
Extractions of PI were performed by
applying water at different temperature on
dry powder of MC seeds (8:1 v/w). The result
were shown in table 1 and figure 3.
There are no different in MC
extractions with or without heat treat. PI of
MC seeds were reported as thermostable
protein (Tsoi et al., 2004). This result were
confirm with SDS-PAGE (Figure 4), in which,
heat-treat extraction still contain under 10
kdal PI. This was a good result since
treatment at high temperature could reduce
protein content and vicosity of MC seed
extracts and facilitate the use of spraying
machine on field
Figure 4. Major proteins in
seeds of Momordica
cochinchinensis (1-untreated;
1-90
o
C for an hour)
Figure 11. Field trial of mixed plant extracts with cabbage (Brassica oleracea) and
Pieris rapae Larva
Figure 2. Extracts of CA fruits in different solvents