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Frontiers in Crop Improvement
Vol 9 : 3012-3019 (Special Issue-VII) December 2021 Print ISSN : 2393-8234 Online ISSN : 2454-6011
Astha Foundation, Meerut (U.P.) India

Behavioral Response of Ladybird Beetles, Coccinella septempunctata Linn. and

Coccinella transversalis Fab. to Synthetic Herbivore-Induced Plant Volatiles (HIPV‘s) in
a Y-tube Olfactometer
Rakesh Kumar1*, Suresh M. Nebapure1, R.K. Sharma1, S.R. Sinha1, Rekha Kumawat2
1
Division of Entomology, ICAR-IARI New Delhi-110012
2
Agricultural Research Station (Agriculture University, Jodhpur), Mandor, Rajasthan
*Email : rakeshcsa8328@gmail.com

Abstract
Five synthetic herbivore-induced plant volatiles (HIPV’s) and their mixtures were tested for their ability to attract two coccinellid
species viz. Coccinella septempunctata Linn. and Coccinella transversalis Fab. in a Y-tube olfactometer under laboratory
conditions. The both sexes of C. septempunctata Linn. were attracted to Methyl salicylate (MeSA) at (5 ppm and 10 ppm) and
Cis-3-hexenyl acetate (5 ppm). Whereas, females of C. septempunctata Linn. also attracted to Methyl salicylate (1 ppm),
Cis-hexanol (5 ppm) and b-caryophyllene (5 ppm). On the contrary, male and female of C. transversalis Fab. were attracted to
Methyl salicylate at 10 ppm and 5 ppm respectively. Similarly, Cis-hexanol at 10 ppm concentration also attracted both sexes of
C. transversalis Fab. Among the HIPV’s mixtures, significantly higher responses of both sexes of C. septempunctata Linn. and
C. transversalis Fab. were recorded from Methyl salicylate (MeSA) + b-caryophyllene + Cis-hexanol (1:1:1) at almost all
concentrations tested (viz., 1, 5, 10 and 15 ppm). Whereas, combination of b-caryophyllene + Cis-hexanol (1:1) failed to attract
any sex of both species except 10 ppm concentration, that attracted male of C. septempunctata Linn. Results of present study
clearly indicated that the mixture of herbivore-induced plant volatiles gave higher responses of C. transversalis Fab. and C.
septempunctata Linn. adults than compounds tested singly. This is the first report of C. transversalis Fab. attraction to Methyl
salicylate in a Y-tube olfactometer.

Key words : HIPV’s, Coccinella septempunctata Linn., Coccinella transversalis Fab., Methyl salicylate (MeSA), Cis-hexanol

predation by predators in the field conditions (Zhu and

Introduction
The ladybird beetles (Coleoptera: Coccinellidae) are
potential predators of soft-bodied insects i.e., aphids,
scale insects, mealy bugs, phytophagous mites,
whiteflies, nymphs of pentatomid bugs, etc., in
agro-ecosystem. Thus, coccinellids have great economic
importance as natural enemies to contain insect pests
problems in agriculture crops. Coccinellids widely
distributed in India with two hundred sixty one predaceous
coccinellids species belonging to fifty seven genera
(Omkar and Pervez 2004). Herbivore damage to the
plants results in production of plant volatiles and these
volatile compounds refer to as “herbivore-induced plant
volatiles”. Herbivore-induced plant volatiles mediate
relationships between plants and insects through the
attraction of natural enemies and the repulsion of
herbivores (Takabayashi and Dicke 1996; Drukker et al.,
2000; Kessler and Baldwin, 2001; de Boer and Dicke,
2004; Uefune et al., 2012; Aartsma et al., 2017; Ray et al.,
2019). It is anticipated that site specific application of
herbivore-induced plant volatiles can be used to increase
natural enemy’s populations within an area of interest,
which will consecutively result in reduced insect pest
population. In recent years, studies have shown that
synthetic HIPVs attract bene?cial insects and increase
park, 2005; Rodriguez-Saona et al., 2011; Kessler and
Baldwin, 2001; James and Price, 2004; James, 2005;
Salamanca et al., 2017). In the present study, behavioral
response of Ladybird beetle, Coccinella septempunctata
Linn. and Coccinella transversalis Fab. to synthetic HIPVs
was investigated in laboratory conditions.
Materials and Methods
Behavioural responses of both Lady Bird beetle species
viz., Coccinella septempunctata Linn. and Coccinella
transversalis Fab. were recorded in a Y-tube olfactometer,
made up of transparent Plexiglas (3 cm ID; stem 10 cm,
arms 8 cm; stem-arms angle 130o) with each arm
connected to a glass tube holding the odor source.
Filtered air at the rate of 1.0 liters/min was drawn through
the olfactometer by a pump during the experimentation.
Adult male and females of Coccinella sp. were directly
collected from experimental field at I.A.R.I. New Delhi and
starved for 24 hours before bioassay. Synthetic herbivory
induced plant volatile’s were selected to find out their
possible effects on Coccinella septempunctata Linn. and
Coccinella transversalis Fab., on the basis of literature
accounts suggesting that each might have some attractive
properties to some natural enemies in different
agro-ecosystems. Test compounds were obtained from
 Rakesh Kumar et al.,
commercial sources (Table-1). Selected synthetic
compounds, singly, and as different mixtures were
dissolved in a solvent (hexane) at different concentrations
viz. 1 ppm, 5ppm, 10 ppm and 15 ppm. For the odor
source whatman filter paper (2×2 cm) impregnated with
10 µl of test solution was used and before introducing it
into one arm of Y-tube olfactometer, left to dry for 10s.
Whereas, untreated whatman filter paper (control) of the
same size was introduced into another arm of
olfactometer. Directional bias excluded by rotating the
apparatus 180o after five runs. Time to respond for
Coccinella septempunctata Linn. and Coccinella
transversalis Fab. standardized to record maximum
activity for the test compound. Male and female were
allowed for 15 minutes to respond to the test compound
and each insect was used only once. When insects
traveled at least 4 cm along arm connected to test
compound responses were considered positive. The
olfactometer disconnected from the glass tube and
thoroughly washed with water, rinsed in 70% ethanol, and
dried in an oven at 120oC after every three runs. All
experiments were conducted in day light at28±2oC and
60% relative humidity.
Results and Discussion
The responses of Coccinella septempunctata Linn., male
and female to synthetic HIPVs presented in Table 2. C.
septempunctata Linn., male attracted significantly higher
to methyl salicylate (MeSA) at 5 ppm (N= 20, P = 0.02)
and 10 ppm (N = 20, P = 0.004), respectively over the
control. Whereas, Cis-3-hexenyl acetate at 5 ppm elicited
significantly higher responses (N = 20, P = 0.004) among
all concentrations tested. On the contrary, the other
volatile compounds at all concentration tested failed to
elicit significant responses in male. Similarly, methyl
salicylate (MeSA) at 1 ppm (N= 20, P = 0.02) 5 ppm (N =
20, P = 0.001) and 10 ppm (N = 20, P = 0.004) elicited
significantly higher responses in C. septempunctata Linn.,
female over the control. C. septempunctata Linn., female
also responded to Cis-3-hexenyl acetate at 5 ppm (N= 20,
P = 0.02), Cis-hexanol at 5 ppm (N= 20, P = 0.02) and
b-caryophyllene at 5 ppm (N= 20, P = 0.02), respectively,
while the other concentrations of these volatile
compounds and all concentration of limonene tested,
failed to elicit significant responses (Table-2, fig.-1).
The responses of C. septempunctata Linn., male
and female to different combinations of synthetic HIPVs
are presented in Table 3. Significantly higher responses
of C. septempunctata Linn., male were recorded from
methyl salicylate (MeSA)+ b-caryophyllene + Cis-hexanol
(1:1:1) at 1 ppm (N = 20, P = 0.004), 5 ppm (N = 20, P ?
0.001), 10 ppm (N = 20, P = 0.0004), b-caryophyllene +
Cis-hexanol (1:1) at 10 ppm (N = 20, P = 0.02), methyl
3013
salicylate + Cis-hexanol (1:1) at 5 ppm (N = 20, P = 0.02),
15 ppm (N = 20, P = 0.004) and methyl salicylate (MeSA)
+ b-caryophyllene (1:1) at 10 ppm (N= 20, P= 0.02),
respectively. In contrast, b-caryophyllene + Cis-hexanol
(1:1) at 1 ppm (N = 20, P = 0.186), 5 ppm (N = 20, P =
0.384), 15 ppm (N = 20, P = 0.07), methyl salicylate +
Cis-hexanol (1:1) at 1 ppm (N = 20, P = 0.186), 10 ppm
(N = 20, P = 0.186), methyl salicylate (MeSA) +
b-caryophyllene (1:1) at 1 ppm (N = 20, P = 0.384), 5 ppm
(N = 20, P = 0.186), 15 ppm (N = 20, P = 1.00) and methyl
salicylate (MeSA)+ b-caryophyllene + Cis-hexanol (1:1:1)
at 15 ppm (N = 20, P= 0.07) failed to elicit significant
responses (Table-3, Fig.-2). Similarly, among the
different combinations of HIPVs tested, methyl salicylate
+ Cis-hexanol (1:1) at 5 ppm (N = 20, P = 0.02), 10 ppm
(N = 20, P = 0.004) and methyl salicylate (MeSA)+
b-caryophyllene + Cis-hexanol (1:1:1) at 1 ppm (N = 20, P
= 0.02), 5 ppm (N = 20, P =0.0004), 10 ppm (N = 20, P =
0.0004), 15 ppm (n= 20 P = 0.02) elicited significantly
higher responses over the control in C. septempunctata
Linn., female. On the contrary, â-caryophyllene +
Cis-hexanol (1:1) at 1 ppm (N = 20, P = 1.00), 5 ppm (N =
20, P = 0.384), 10 ppm (N = 20, P = 0.07), 15 ppm (N =
20, P = 0.07), methyl salicylate + Cis-hexanol (1:1) at 1
ppm (N = 20, P = 0.666), 15 ppm (N = 20, P = 0.07),
methyl salicylate (MeSA) + b-caryophyllene (1:1) at 1
ppm (N = 20, P = 0.666), 5 ppm (N = 20, P = 0.186), 10
ppm (N = 20, P = 0.384) 15 ppm (N = 20, P = 1.00) failed
to elicit significant responses (Table 3, Fig 2).
Our findings revealed that, C. septempunctata
Linn., male attracted significantly higher to methyl
salicylate (MeSA) at 5 ppm and 10 ppm over the control.
Whereas, Cis-3-hexenyl acetate at 5 ppm elicited
significantly higher responses over the control among all
concentrations tested. On the contrary, the other volatile
compounds at all concentration tested failed to elicit
significant responses. Similarly, methyl salicylate (MeSA)
at 1, 5 and 10 ppm elicited significantly higher responses
in C. septempunctata Linn., female over the control. C.
septempunctata Linn., female also responded to Cis-3-
hexenyl acetate at 5 ppm, Cis-hexanol at 5 ppm and b-
caryophyllene at 5 ppm respectively, while the other
concentrations of these volatile compounds and all
concentration of limonene tested, failed to elicit
significant responses (fig.-2). Whereas, significantly
higher responses of C. septempunctata Linn., male over
the control were recorded from methyl salicylate
(MeSA)+ b-caryophyllene + Cis-hexanol (1:1:1) at 1 ppm,
5 ppm, 10 ppm, b-caryophyllene + Cis-hexanol (1:1) at 10
ppm, methyl salicylate + Cis-hexanol (1:1) at 5, 15 ppm,
and methyl salicylate (MeSA) + b-caryophyllene (1:1) at
10 ppm, respectively. In contrast, b-caryophyllene +
Cis-hexanol (1:1) at 1 ppm, 5 ppm, 15 ppm, methyl
3014 Frontiers in Crop Improvement, Volume 9 (Special Issue-VII) December 2021

Table-1 : Details of synthetic HIPV’s used in the present study.

S. No. Compound Purity Supplier
1. Methyl salicylate > 99% Sigma-Aldrich
2. Cis-hexanol > 98% Sigma-Aldrich
3. b-caryophyllene > 80% Sigma-Aldrich
4. D-Limonene 97% Sigma-Aldrich
5. Cis-3-Hexenyl acetate =98% Sigma-Aldrich
Table-2 : Behavioral responses of Coccinella septempunctata Linn. in a Y-tube olfactometer to synthetic HIPVs.
Synthetic Concen- Male response (mean ±SE) Female response (mean ±SE)
HIPVs tration N Source Control P value N Source Control P value
Methyl salicylate 1ppm 20 13±0.11a 7±0.11a 0.186 20 15±0.10a 5±0.10b 0.02
5ppm 20 15±0.10a 5±0.10b 0.02 20 18±0.07a 2±0.07b <0.001
10ppm 20 16±0.00a 4±0.09b 0.004 20 16±0.09a 4±0.09b 0.004
15ppm 20 14±0.1a 6±0.1a 0.07 20 14±0.1a
6±0.1a 0.07
Limonene 1ppm 20 12±0.1a 8±0.1a 0.384 20 11±0.09a 9±0.09a 0.666
5ppm 20 13±0.11a 7±0.11a 0.186 20 12±0.1a 8±0.1a 0.384
10ppm 20 6±0.1a 14±0.1a 0.07 20 9±0.09a 11±0.09a 0.666
15ppm 20 9±0.09a 11±0.09a 0.666 20 7±0.11a 13±0.11a 0.186
Cis-3-Hexenyl 1ppm 20 13±0.11a 7±0.11a 0.186 20 12±0.1a 8±0.1a 0.384
acetate 5ppm 16±0.09a 4±0.09b 0.004 20 15±0.10a 5±0.10b 0.02
20
a a a
10ppm 20 13±0.11 7±0.11 0.186 20 12±0.1 8±0.1a 0.384
15ppm 20 8±0.11a 12±0.11a 0.384 20 9±0.09a 11±0.09a 0.666
a a a
Cis-hexanol 1ppm 20 9±0.09 11±0.09 0.666 20 13±0.11 7±0.11a 0.186
5ppm 20 13±0.11a 7±0.11a 0.186 20 15±0.10a 5±0.10b 0.02
10ppm 20 12±0.1a 8±0.1a 0.384 20 14±0.1a 6±0.1a 0.07
15ppm 20 9±0.09a 11±0.09a 0.666 20 8±0.11a 12±0.11a 0.384
b- caryophyllene 1ppm 20 13±0.11a 7±0.11a 0.186 20 13±0.11a 7±0.11a 0.186
5ppm 20 14±0.1a 6±0.1a 0.07 20 15±0.10a 5±0.10b 0.02
10ppm 20 8±0.11a 12±0.11a 0.384 20 9±0.09a 11±0.09a 0.666
15ppm 20 6±0.1a 14±0.1a 0.07 20 7±0.11a 13±0.11a 0.186
*For each odour source, different letters indicate significant differences from control (Paired t test).

Figure-1 : Behavioral responses (mean per cent response) of Coccinella septempunctata Linn (a) male and (b) female to synthetic
HIPVs in Y-tube olfactometer. Asterisks indicate significant differences over the control (Paired t test: N = 20, *P = 0.05, **P=
0.01, ***P= 0.001, ****P =0.0001).
 Rakesh Kumar et al., 3015

Table-3 : Behavioral responses of Coccinella septempunctata Linn. in Y-tube olfactometer to different combinations of
synthetic HIPVs.
Synthetic HIPVs Concen- Male (mean % ± SE response) Female (mean % ± SE response)
tration N Source Control P value N Source Control P value
b-caryophyllene + 1ppm 20 7±0.11a 13±0.11a 0.186 20 10±0.11a 10±0.11a 1.00
Cis-hexanol (1:1) 5ppm 0.384 20 0.384
20 12±0.1a 8±0.1a 12±0.1a
8±0.1a

10ppm 20 15±0.10 a
5±0.10 b 0.02 20 14±0.1a 6±0.1a 0.07
15ppm 20 14±0.1 a
6±0.1a 0.07 20 14±0.1 a
6±0.1a 0.07
Methyl salicylate+ 1ppm 20 13±0.11 a
7±0.11 a 0.186 20 11±0.11 a
09±0.11 a 0.666
Cis-hexanol (1:1) 5ppm a b 0.02 20 a b 0.02
20 15±0.10 5±0.10 15±0.10 5±0.10
10ppm 20 13±0.11 a
7±0.11 a 0.186 20 16±0.09 a
4±0.09 b 0.004
15ppm 20 16±0.09 a
4±0.09 b 0.004 20 14±0.1 a
6±0.1a 0.07
Methyl salicylate+ 1ppm 20 8±0.11 a
12±0.11 a 0.384 20 9±0.11 a
11±0.11 a 0.666
b-caryophyllene
5ppm 20 13±0.11 a
7±0.11 a 0.186 20 13±0.11 a
7±0.11 a 0.186
(1:1)
10ppm 20 15±0.10 a
5±0.10 b 0.02 20 12±0.1 a
8±0.1a 0.384
15ppm 20 10±0.11 a
10±0.11 a 1.00 20 10±0.11 a
10±0.11 a 1.00
b-caryophyllene + 1ppm 20 16±0.09 a
4±0.09 b 0.004 20 15±0.10 a
5±0.10 b 0.02
Cis-hexanol +
Methyl salicylate 5ppm 20 18±0.07 a
2±0.07 b <0.001 20 17±0.8 a
3±0.8b 0.0004
(1:1:1) 10ppm 20 17±0.8a 3±0.8b 0.0004 20 17±0.8a 3±0.8b 0.0004
15ppm 20 14±0.1 a
6±0.1a 0.07 20 15±0.10 a
5±0.10 b 0.02
* For each odour source, different letters indicate significant differences from control (Paired t test).

Figure-2 : Behavioral responses (mean per cent response) of Coccinella septempunctata Linn. (a) male and (b) female to
different combinations of synthetic HIPVs in Y-tube olfactometer. Code for treatments: CA+HE, â- caryophyllene +
Cis-hexanol (1:1); MeSA+HE, Methyl salicylate+ Cis-hexanol (1:1); MeSA+CA, Methyl salicylate+ â- caryophyllene
(1:1); CA+HE +MeSA, â- caryophyllene + Cis-hexanol + Methyl salicylate (1:1:1). Asterisks indicate significant
differences over the control (Paired t test: N = 20, *P ? 0.05, **P= 0.01, ***P= 0.001, ****P =0.0001).
3016 Frontiers in Crop Improvement, Volume 9 (Special Issue-VII) December 2021

Table-4 : Behavioral responses of Coccinella transversalis Fab. in Y-tube olfactometer to synthetic HIPVs.
Synthetic Concen- Male response (mean ±SE) Female response (mean ±SE)
HIPVs tration N Source Control P value N Source Control P value
Methyl salicylate 1ppm 20 10±0.11a 10±0.11a 1.00 20 9±0.09a 11±0.09a 0.666
5ppm 20 14±0.1a 6±0.1a 0.07 20 15±0.10a 5±0.10b 0.02
10ppm 20 16±0.09 a
4±0.09 b 0.004 20 13±0.11 a
7±0.11 a 0.186
15ppm 20 11±0.09a 9±0.09a 0.666 20 12±0.1a 8±0.1a 0.384
Limonene 1ppm 20 8±0.11 a
12±0.11 a 0.384 20 11±0.09 a
9±0.09 a 0.666
5ppm 20 11±0.09a 9±0.09a 0.666 20 11±0.09a 9±0.09a 0.666
10ppm 20 10±0.11 a
10±0.11 a 1.00 20 8±0.11 a
12±0.11 a 0.384
15ppm 20 8±0.11 a
12±0.11 a 0.384 20 7±0.11 a
13±0.11 a 0.186
Cis-3-Hexenyl 1ppm 20 11±0.09 a
9±0.09 a 0.666 20 10±0.11 a
10±0.11 a 1.00
acetate 5ppm 20 a b 0.02 20 a a 0.384
15±0.10 5±0.10 12±0.1 8±0.1
10ppm 20 12±0.1a 8±0.1a 0.384 20 11±0.09a 9±0.09a 0.666
15ppm 20 9±0.09 a
11±0.09 a 0.666 20 10±0.11 a
10±0.11 a 1.00
Cis-hexanol 1ppm 20 14±0.1 a
6±0.1a 0.07 20 13±0.11 a
7±0.11 a 0.186
5ppm 20 16±0.09 a
4±0.09 b 0.004 20 14±0.1 a
6±0.1a 0.07
10ppm 20 17±0.8 a
3±0.8b 0.0004 20 16±0.09 a
4±0.09 b 0.004
15ppm 20 12±0.1a 8±0.1a 0.384 20 9±0.09a 11±0.09a 0.666
b-caryophyllene 1ppm 20 10±0.11a 10±0.11a 1.00 20 9±0.09a 11±0.09a 0.666
5ppm 20 12±0.1 a
8±0.1a 0.384 20 13±0.11 a
7±0.11 a 0.186
10ppm 20 11±0.09 a
9±0.09 a 0.666 20 11±0.09 a
9±0.09 a 0.666
15ppm 20 9±0.09 a
11±0.09 a 0.666 20 8±0.11 a
12±0.11 a 0.384
* For each odour source, different letters indicate significant differences from control (Paired t test).

Figure-3 : Behavioral responses (mean per cent response) of Coccinella transversalis Fab (a) male and (b) female to synthetic
HIPVs in Y-tube olfactometer. Asterisks indicate significant differences over the control ( Paired t test: N = 20, *P =
0.05, **P= 0.01, ***P= 0.001, ****P =0.0001).
 Rakesh Kumar et al., 3017

Table-5 : Behavioral responses of Coccinella transversalis Fab. in Y-tube olfactometer to different combinations of synthetic
HIPVs.
Synthetic HIPVs Concen- Male (mean %±SE response) Female (mean %±SE response)
tration N Source Control P value N Source Control P value
b-caryophyllene + 1ppm 20 8±0.11a 12±0.11a 0.384 20 9±0.11a 11±0.11a 0.666
Cis-hexanol (1:1) 5ppm 20 1.00 0.186
10±0.11a 10±0.11a 20 13±0.11a 7±0.11a
10ppm 20 13±0.11 a
7±0.11 a 0.186 20 13±0.11 a
7±0.11 a 0.186
15ppm 20 10±0.11 a
10±0.11 a 1.00 20 14±0.1 a
6±0.1a 0.07
Methyl salicylate+ 1ppm 20 12±0.1 a
8±0.1 a 0.186 20 11±0.11 a
09±0.11 a 0.666
Cis-hexanol (1:1) 5ppm 20 a a 0.07 a a 0.07
14±0.1 6±0.1 20 14±0.1 6±0.1
10ppm 20 15±0.10 a
5±0.10 b 0.02 20 16±0.09 a
4±0.09 b 0.004
15ppm 20 16±0.09a 4±0.09b 0.004 20 15±0.10a 5±0.10b 0.02
Methyl salicylate+b- 1ppm 20 9±0.11 a
11±0.11 a 0.666 20 9±0.11 a
11±0.11a 0.666
caryophyllene (1:1) 5ppm 20 0.384 0.07
12±0.1a 8±0.1a 20 14±0.1a 6±0.1a
10ppm 20 15±0.10 a
5±0.10 b 0.02 20 16±0.09 a
4±0.09 b 0.004
15ppm 20 15±0.10a 5±0.10b 0.02 20 10±0.11a 10±0.11a 1.00
b-caryophyllene + 1ppm 20 14±0.1a 6±0.1a 0.07 20 15±0.10a 5±0.10b 0.02
Cis-hexanol + Methyl
5ppm 20 15±0.10 a
5±0.10 b 0.02 20 15±0.10 a
5±0.10 b 0.02
salicylate (1:1:1)
10ppm 20 17±0.8 a
3±0.8 b 0.0004 20 16±0.09 a
4±0.09 b 0.004
15ppm 20 18±0.07a 2±0.07b <0.001 20 17±0.8a 3±0.8b 0.0004
* For each odour source, different letters indicate significant differences from control (Paired t test).

Figure-4 : Behavioral responses (mean per cent response) of Coccinella transversalis Fab. (a) male and (b) female to different
combinations of synthetic HIPVs in Y-tube olfactometer. Code for treatments: CA+HE, b-caryophyllene +
Cis-hexanol (1:1); MeSA+HE, Methyl salicylate+ Cis-hexanol (1:1); MeSA+CA, Methyl salicylate+ b-caryophyllene
(1:1); CA+HE + MeSA, b- caryophyllene + Cis-hexanol + Methyl salicylate (1:1:1). Asterisks indicate significant
3018 Frontiers in Crop Improvement, Volume 9 (Special Issue-VII) December 2021
salicylate + Cis-hexanol (1:1) at 1 ppm, 10 ppm, methyl
salicylate (MeSA) + b-caryophyllene (1:1) at 1 ppm, 5
ppm, 15 ppm and methyl salicylate (MeSA)+
â-caryophyllene + Cis-hexanol (1:1:1) at 15 ppm failed to
elicit significant responses (Fig.-2). Similarly, among the
different combinations of HIPVs tested, methyl salicylate
+ Cis-hexanol (1:1) at 5 ppm, 10 ppm and methyl
salicylate (MeSA)+ b-caryophyllene + Cis-hexanol (1:1:1)
at 1 ppm, 5 ppm, 10 ppm and 15 ppm elicited significantly
higher responses over the control in C. septempunctata
Linn., female. On the contrary, â-caryophyllene +
Cis-hexanol (1:1) at 1 ppm, 5 ppm, 10 ppm, 15 ppm,
methyl salicylate + Cis-hexanol (1:1) at 1 ppm, 15 ppm,
methyl salicylate (MeSA) + â-caryophyllene (1:1) at 1
ppm, 5 ppm, 10 ppm and 15 ppm failed to elicit significant
responses (Fig.-2). The present findings conform to those
made by Zhu and Park (2005) and James (2005), who
reported methyl salicylate as highly attractive to adults of
C. septempunctata Linn.
The responses of C. transversalis Fab., male and
female to synthetic HIPVs presented in Table-4. C.
transversalis Fab., male was attracted by methyl
salicylate 10 ppm (N = 20, P = 0.004), Cis-3-hexenyl
acetate 5 ppm (N =20, P= 0.02) and Cis-hexanol 5 ppm
(N = 20, P = 0.004), 10 ppm (N = 20, P= 0.0004). Methyl
salicylate at 1, 5, 15 ppm, Cis-3-hexenyl acetate at 1, 10,
15 ppm, Cis-hexanol at 1, 15 ppm, while limonene and
â-caryophyllene at all concentrations tested, failed to
attract significantly higher C. transversalis Fab., males
over the control. Similarly, C. transversalis Fab., female
responded to methyl salicylate at 5 ppm (N= 20, P = 0.02)
and Cis-hexanol at 10 ppm (N = 20 P = 0.004), while
methyl salicylate at 1, 10, 15 ppm and Cis-hexanol at 1, 5,
15 ppm failed to elicit significant responses. Similarly, all
concentrations of limonene and â-caryophyllene also
failed to elicit significant responses over the control
(Table-4. Fig.-3). The responses of C. transversalis Fab.
male and female to different combinations of synthetic
HIPVs are presented in Table 5. Interestingly, significantly
higher responses of C. transversalis Fab. male over the
control were recorded from methyl salicylate (MeSA)+
â-caryophyllene + Cis-hexanol (1:1:1) at 5 ppm (N = 20, P
= 0.02), 10 ppm (N = 20, P = 0.0004), 15 ppm (N = 20, P =
=0.001), methyl salicylate + Cis-hexanol (1:1) at 10 ppm
(N = 20, P = 0.02), 15 ppm (N = 20, P = 0.004) and methyl
salicylate (MeSA) + â-caryophyllene (1:1) at 10 ppm (N =
20, P = 0.02), 15 ppm (N = 20, P = 0.02). In contrast,
â-caryophyllene + Cis-hexanol (1:1) at 1, 5, 10, 15 ppm,
methyl salicylate + Cis-hexanol (1:1) at 1, 5 ppm, methyl
salicylate (MeSA) + â-caryophyllene (1:1) at 1, 5 ppm and
Methyl salicylate (MeSA) + b-caryophyllene + Cis-hexanol
(1:1:1) at 1 ppm failed to elicit significant responses
(Table-5, Fig.-4). Similarly, significantly higher responses
of C. transversalis Fab. female over the control were
recorded from methyl salicylate (MeSA)+ b-caryophyllene
+ Cis-hexanol (1:1:1) at 1 ppm (N = 20, P = 0.02), 5 ppm
(N = 20, P = 0.02), 10 ppm (N = 20, P = 0.004), 15 ppm (N
= 20, P = 0.0004), methyl salicylate + Cis-hexanol (1:1) at
10 ppm (N = 20, P = 0.004), 15 ppm (N = 20, P = 0.02)
and methyl salicylate (MeSA) + b-caryophyllene (1:1) at
10 ppm (N = 20, P = 0.004). Conversely, b-caryophyllene
+ Cis-hexanol (1:1) at 1, 5, 10, 15 ppm, methyl salicylate +
Cis-hexanol (1:1) at 1, 5 ppm and methyl salicylate
(MeSA) + b-caryophyllene (1:1) at 1, 5, 15 ppm were
failed to elicit significant responses (Table-5, Fig.-4). Our
findings revealed that, C. transversalis Fab. male was
attracted by methyl salicylate 10 ppm, Cis-3-hexenyl
acetate 5 ppm and Cis-hexanol 5, 10 ppm. Methyl
salicylate at 1, 5, 15 ppm, Cis-3- hexenyl acetate at 1, 10,
15 ppm, Cis-hexanol at 1, 15 ppm, while limonene and
â-caryophyllene at all concentrations tested, failed to
attract significantly higher C. transversalis Fab. males
over the control. Similarly, C. transversalis Fab. female
responded to methyl salicylate at 5 ppm and Cis-hexanol
at 10 ppm, while methyl salicylate at 1, 10, 15 ppm and
Cis-hexanol at 1, 5, 15 ppm failed to elicit significant
responses. Similarly, all concentrations of limonene and
â-caryophyllene also failed to elicit significant responses
over the control (Fig. 3). Interestingly, significantly higher
responses of C. transversalis Fab. male over the control
were recorded from methyl salicylate + â-caryophyllene +
Cis-hexanol (1:1:1) at 5 ppm, 10 ppm, 15 ppm, methyl
salicylate + Cis-hexanol (1:1) at 10 ppm, 15 ppm, and
methyl salicylate (MeSA) + b-caryophyllene (1:1) at 10
ppm, 15 ppm. In contrast, b-caryophyllene + Cis-hexanol
(1:1) at 1, 5, 10, 15 ppm, methyl salicylate + Cis-hexanol
(1:1) at 1, 5 ppm, methyl salicylate (MeSA) +
b-caryophyllene (1:1) at 1, 5 ppm and methyl salicylate
(MeSA) + â caryophyllene + Cis-hexanol (1:1:1) at 1 ppm
failed to elicit significant responses (Fig. 4). Similarly,
significantly higher responses of C. transversalis Fab.
female over the control were recorded from methyl
salicylate + â-caryophyllene + Cis-hexanol (1:1:1) at 1
ppm, 5 ppm, 10 ppm, 15 ppm, methyl salicylate +
Cis-hexanol (1:1) at 10 ppm, 15 ppm and methyl
salicylate (MeSA) + â-caryophyllene (1:1) at 10 ppm.
Conversely, â-caryophyllene + Cis-hexanol (1:1) at 1, 5,
10, 15 ppm, methyl salicylate + Cis-hexanol (1:1) at 1, 5
ppm and methyl salicylate (MeSA) + b-caryophyllene (1:1)
at 1, 5, 15 ppm were failed to elicit significant responses
(Fig.-4). Maeda et al., 2015 also reported that the
mixtures of synthetic herbivore induced plant volatiles are
attractive to Stethorus punctum picipes (Casey) than the
signal volatile compound tested. As per our knowledge a
detailed survey of the literature shows that no previous
study on the response of herbivore induced plant
 Rakesh Kumar et al.,
volatile’s components on C. transversalis Fab. has been
reported. Methyl salicylate has been demonstrated to be
attractive predatory bug Anthocoris nemoralis as well as
minute pirate bugs, Orius tristicolor; bigeyed bugs,
Geocoris pallens and lady beetle stethorus punctum
(Drukker et al., 2000; James, 2003). In an another study
Salamanca et al. 2017 reported attraction of predatory
coccinellid ladybeetle Hippodamia convergens to Methyl
salicylate under laboratory and greenhouse conditions.
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