330 T. Venkatesan and S. K. Jalali
pooled. The standard procedure was followed to
determine carboxylesterase activity (CE).
Isoenzyme Analysis
CE activity was determined for the different
strains of T. chilonis in nondenaturing PAGE.
Homogenates from each strain stained for 1-Nap-
thyl acetate. Native PAGE was carried out using
mini vertical gel system and 7.5 % acrylamide
(wt: vol) gel at pH 8.5 and stacking gel of 3 %
acrylamide (wt: vol). Km values were calculated
for the lab susceptible and tolerant strains for the
substrate 1-Napthyl acetate, the substrate con-
centrations 0.025 to 1 mM were used.
Microsomal Cytochrome C Reductase and
p-Nitroanisole O-demethylase
Sigma reagent assay kit used for the preparation
of microsomes. Sixty milligram each strains of
T. chilonis adults were homogenized in a micro
pestle and mortar with 2 ml of PBS pH 7.5 con-
taining 0.1 mM EDTA.
Microsomal Cytochrome p-Nitroanisole
O-demethylase
This assay measures the reduction of cytochrome
c by NADPH—cytochrome c reductase in the
presence of NADPH. The absorption spectrum
of cytochrome c changes with oxidation/reduc-
tion state. Upon reduction a sharp absorption
peak is monitored by the increase of cytochrome
c absorbance at 550 nm. Cytochrome c reductase
assay was performed by following the manufac-
ture protocol.
Field Efficacy of the Multiple
Insecticides and High Temperature
Tolerant (MIHTTS) and Susceptible
Strain of T. chilonis on Cotton, Tomato,
and Cabbage Pests in Comparison to
Insecticides Alone
Cotton
First Year A field trial was conducted at Sirsa,
Haryana and crop was raised during Kharif
(monsoon) season of 2007. The variety sown
was CICR—2 in 10.0 acre with MIHTTS. The
comparison was made with susceptible strain and
farmer’s practice in one acre each. Each treat-
ment was separated by 200 m of barren land at
all the locations. In tolerant strain plots imida-
cloprid, neem oil, and lambda-cyhalothrin was
sprayed during the season, in susceptible strain
released plots confidor, monocrotophos + cyper-
methrin, fenvalerate, lambda-cyhalothrin (two
sprays), and neem oil was sprayed, in farmers’
practice plot, insecticides sprayed were imida-
cloprid, monocrotophos + cypermethrin, fenval-
erate, lambda-cyhalothrin (two sprays), endosul-
fan, and neem oil were sprayed once in a week
and tabulated the results.Second Year A field trial was conducted at
Sirsa, Haryana, in collaboration with National
Centre for Insect Pest Management (NCIPM),
New Delhi during 2008. The variety sown was
CICR—2 in 10.0 acre area. The comparison was
made with susceptible and farmer’s practice in
one acre each. Each treatment was separated by
200 m of barren land at all the locations. The
insecticides sprayed were imidacloprid, lambda-
cyhalothrin, monocrotophos + cypermethrin, fen-
valerate, endosulfan, and neem oil were sprayed
once in a week and tabulated the results.
Trichogramma chilonis (MIHTTS and suscep-
tible strain) was released in treatments T 1 and T 2
@ 150,000/ha/release in the form of parasitized
eggs to cover the egg laying period of H. armig-
era. The release of parasitoids commenced with
moth capture in pheromone traps. Eight days old
parasitized cards were released at weekly interval
from August to September 2007. The harvesting
continued till mid November 2007.
Larval population was recorded at each place
in 10 subplots in each treatment. Twenty plants
in each subplot at each location were observed
for the larval population. Thus, 200 plants were
observed each time to record larval population
per treatment. The percent boll damage, good
and bad open bolls, and yield were recorded at
harvest. Data on larval population, percent boll
damage and yield data were subjected to one-way
ANOVA and means were separated by CD values
at 5 %, wherever ANOVA was significant.