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«D. ANITHA KUMARI THESIS SUBMITTED TO THE ACHARYA N.G RANGA AGRICULTURAL UNIVERSITY COLLEGE OF AGRICULTURE, RAJENDRANAGAR IN PARTIAI FULFILLMENT OF ...»

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inflorescences (30 cm long) with few leaves were brought from the field and placed in a conical flask filled with water. Five pairs of 2-day old moths were released inside the cage. Moths are provided with sucrose solution in a conon swab through out the experiment. After releasing the moths in the cages, the moths were allowed to oviposit for three nights on the test plants. To avoid predation by the ants, tangle foot glue was smeared on all the four legs of the cages. Observations were recorded on the number of eggs laid on each inflorescence placed in a cage. The moths were allowed to oviposit on the test entries for three consecutive nights. Each experiment was replicated five times. Data were subjected to analysis of variance using completely randomized design (Plate 1).

Oviposition preference under dual-choice conditions 3.2.2.2

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studied by keeping a test variety with a susceptible check, ICPL 87 inside the wooden cage as described above. The inflorescence (30 cm long) were obtained from the field. Five inflorescences each of the test variety and the susceptible check were kept in two conical flasks separately at the comer inside the cage. Five pairs of two day old moths were released inside the cage. The moths were provided with sucrose solution in a cotton swab. To avoid predation by the ants, tangle foot " glue was applied to all the four legs of the wooden cage. The experiment was replicated five times. Significance of difference between the two test genotypes was compared

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Oviposition non-preference under multi-choice conditions...

Non-preference for oviposition under multi-choice conditions was studied by keeping all the 12 genotypes inside a wooden cage (80 x 70 x 60 cm) placed inside a growth chamber The growth chamber was maintained at 26" C during the day and 20°C during night Relative humidity was 709'0, and photopenod

- 12 hours Inflorescences of the test genotypes were brought from the fic!d and kept in a conlcal flask filled with water Comcal flasks (containing the inflorescences) of all the test genotypes were arranged inside the wooden cage in completely random~zedblock design Th~rtypars of two day old adults were released inside the cage Moths were provided with sucrose solution in a cotton swab The moths were allowed to oviposit on the test entries for three consecutive nights To avoid predation by the ants, tangle foot glue was applied to all the four legs of the wooden table (Plate 3) Observations were recorded on the number of eggs laid on

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250 m plast~c l cups Ten neonate larvae were released on the leaves with the help of a fine camel hair brush. A moistened filter paper was attached to the inner side of the lid and the plastic cups were covered immediately. The plastic cups were kept in the lab at 27+Z°C and 45 to 65%RH. The leaves were changed every alternate days.

From fifth day onwards, the larvae were reared individually in cups to avoid cannibalism. Larval weights and mortality were recorded on 5, 10 and 15 days after release. Data on larval and pupal period, adult emergence, and pupal weights were also recorded. The experiment was conducted in completely randomized design.

There were 5 replications. Data was subjected to ANOVA.

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pods. Therefore, neonate H,armigera larvae were fed on the flowers for 5 days, and then transferred to pods of respective pigeonpea genotypes. The flowers and pods were kept in petri plates with a moistened filter paper attached to the lid. The petri plates were kept in the laboratory at 27+2OC and 45 -65% RH.Larval weights were recorded on 5, 10, and 15 days after release. Data was also recorded on larval and pupal period, pupal weights, and adult emergence. The experiment was conducted in a completely randomized design with five replications, and 10 larvae were released on each replication. Data was subjected to ANOVA. Data on larval weight, larval mortality on 10' day after initiation, larval and pupal period, pupal weight and adult emergence of the above two experiments were subjected to principle component analysis.

Standardisation of artificial diet impregnated with lyophilized...

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impregnating lyophilized plant material into the artificial diet. Leaves of 12 genotypes were collected from two month old plants raised under field conditions.

Leaves were removed from the plant at the growing point, and freeze dried in a lyophilizer for 36 h to avoid changes in chemical composition of the leaves, and then powdered in a Willey Mill to 80 mesh size.

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artificial diet on survival and development of H,armigera: For this purpose 0, 5, 10, 15, and 20 g of pigeonpea leaf powder ICPL 87 (susceptible check) and ICPL 332 (resistant check) was added in 250 ml- artificial diet. Pigeonpea leaf powder was soaked in 100 ml of warm water (70°C) and blended with fraction-A (Table 3 and Plate 4) for two minutes. Agar was boiled in 80 ml of water (Fraction-B), cooled to 40°C, and then poured into the blender containing Fraction -A.

Formaldehyde was added, finally and all the constituents blended for three minutes.

Each treatment was replicated three times (a small cup of 50 ml capacity containing 20 ml diet). One first instar larva was released in each cup.





Observations on larval mortality and larval weights were recorded on 10' day. Observations were also recorded on pupal weights, per cent pupation, per cent adult emergence, and larval and pupal periods. Data was subjected to ANOVA.

Table 3: Composition of artificial diet impregnated with lyophilized leaf powder

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10, 15, and 20 g of pigeonpea pod powder ICPL 87 (susceptible check) and ICPL 332 (resistant check) was added in 250 ml- artificial diet. Pigeonpea pod powder

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for two minutes. Agar was boiled in 80 ml of water (Fraction-B), cooled to 40' C, and then poured into the blender containing Fraction -A. Formaldehyde was added, finally and all the constituents blended for three minutes. Each treatment was replicated three times (a small cup of 50 ml capacity containing 20 ml diet). One first instar larva was released in each cup.

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10' day. Observations were also recorded on pupal weights, per cent pupation, per cent adult emergence, and larval and pupal periods. Data was subjected to ANOVA.

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For each genotype two treatments with different proportions of chickpea flour and pigeonpea leaf powder (65: 10 :: chickpea flour : pigeonpea leaf powder) were tested.

The preparation of diet was same as above. Each treatment was replicated 3 times.

One first instar larva was released in each cup and the cups were placed in the rearing room In the rearing room, temperature was maintained at 28 f. 1°C,

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For each genotype two treatments with different proportions of chickpea flour and pigeonpea pod powder (65:lO :: chickpea flour : pigeon pea pod powder) were tested. The preparation of diet was same as above. Each treatment was replicated 3 times. One first instar larva was released in each cup and the cups were placed in the rearing room. In the rearing room, temperature was maintained at 28 f 1°C, 60 - 70% RH and photoperiod of 12 h.

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per cent pupation, per cent adult emergence, and larval period and pupal periods were recorded. The data were subjected to analysis of variance. Data on larval weight, larval mortality on lo* day after initiation, larval and pupal period, pupal weight and adult emergence of the above two experiments were subjected to principle component analysis.

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scissor and immediately placed in a 250 rnl cup with 3% agar agar. Ten neonate larvae were released with the help of a fine camel hair brush. Observations on larval weights and larval mortality were recorded after five days. The experiment was conducted in a completely randomized design with 12 treatments and 5 replications.

Data was subjected to ANOVA (Plate 5).

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Trichomes are the most common morphological structures which play an important role in insect host plant interaction in pigeonpea and the variation in their forms and Functions quite often are associated with plant resistance to insect attack (Southwood, 1986). Hence the study was carried out to identify different types of trichomes and their density in 12 different pigeonpea genotypes.

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collecting a minimum of 15 pods and flowers from each accession and there were three replications. The material was preserved in a fixation (Acetic acid, absolute alcohol 1 3 ) and examined under a Zeiss stereomicroscope (Carl Zeiss Inc., Thom Nood, NY) at a magnification of 32X with an ocular measuring grid density of trichomes was recorded based on mean of 15 pods.

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Estimation of nitrogen, phosphorus and potassium contents was done by collecting flowers in the field during flowering stage of the crop. The flowers were subjected to lyophilisition and powdered. The flower powder was then used for analysis: Similar procedure was followed for pods also. Nitrogen was estimated by microkjeldahls method (Jackson, 1967), phosphorus by Ammonium meta venadata yellow colour method (Jackson, 1967) and potassium by flame photometer method (Jackson, 1967). Estimation of proteins was calculated by multiplying the nitrogen content with factor 6.24.

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pigeonpea, the material was extracted with hot aqueous-ethyl alcohol. On treatment with phenol sulphuric acid, the sugars produced a stable and sensitive golden yellow col6r. The absorbance of the golden yellow color was measured at 490 nm, which was used to estimate percentage of total soluble sugars present in leaves and pods.

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raised in the field, and were oven dried for 12 h. The oven-dried material was then powdered in a Willey mill, and defatted by using hexane. Ethyl alcohol (go%), 5% phenol (5 g phenol dissolved in water and volume made up to 100 ml), 96% sulphuric acid (specific gravity 1.84), glucose (w/v) standard (stock solution: 1000

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a 100 m volumetric flask, and volume made up to 100 ml, to have the final l concentration of 125 pglml) were used for estimating the total soluble sugars.

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boiling test tube, to which 25 ml of hot 80% ethanol was added. The mixture was vigorously shaken on a vortex mixer. The material was allowed to settle for 30 minutes and the supernatant was filtered by passing through a Whatman No. 41 filter paper. This step was repeated thrice for complete extraction of sugars. By placing the extract on hot sand bath, ethanol was evaporated completely. After complete removal of ethanol, 3 ml water was added to dissolve the contents. One rnl aliquot from the above solution was pipetted into a test tube, and 1 rnl of 5% phenol and 5 ml of 96% sulphuric acid were added. The mixture was shaken vigorously on a vortex mixer. The tubes were allowed to cool in cold water. A blank was prepared by taking 1 ml water. Absorbance of the golden yellow color was red at 490 nnl using Spectronic 2 1.

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125pg of glucose) were prepared from the working standard, and their absorbance was read by taking 1 ml aliquats.

Percent total soluble sugars were calculated by using the formula:

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prepared by placing pods of known surface area into 500 ml of hexane or methanol, and stirred for 120 seconds with a glass rod. Each extract was then gravity filtered before being evaporated under vacuum to dryness. Extracts of the pod surfaces of ICPL 87 were re-dissolved in either hexane or methanol so that 100 p1 of solution contained a quantity of extract equivalent to 3.46 cm2 of pod surface (the area of glass fibre disc). The larvae were presented with a naturally occumng concentration

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discs were air dried for 24 h. Subsequently, each disc was weighed and placed into separate plastic petri dish (9 cm diameter) along with a pre-weighed, untreated disc.

Both the discs were moistened with 100 @ of distilled water as the larvae does not feed on dry discs. One third instar larva was placed in each Petri dish. The experiment was replicated 20 times. After 24 h the larvae were removed from petri dishes, and the glass fibre discs were dried and re-weighed. All discs were kept in growth chambers maintained at 12 W27'C (light: 12h.20°C (dark) (Plate 6).

Feeding (FI) and antifeedant activity were calculated using the formula given below:

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flowers, and pods of similar age group and free from insect eggs and larvae were collected from the field and brought to the laboratory. The preference of the larvae of H. armigera to the leaves, flowers, and pods of 12 genotypes were studied under no-choice, dual-choice and multi-choice conditions.

Feeding preference under no-choice conditions 3.2.7.1

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and one 3" instar larva was released in each petri dish. The extent of leaf feeding was rated visually on a 1-9 damage rating scale after 24 and 48 h. This procedure was repeated for all the 12 genotypes. There were 5 replications for each genotype.

Similar procedure was followed for leaves and flowers as well.

Feeding preference under dual-choice conditions 3.2.7.2

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leaf of a susceptible genotype (ICPL 87) and a leaf of the test genotype in a petri dish arena (9 cm). A single 3" instar larva was released in each petri dish. The extent of leaf feeding was recorded visually on a 1-9 damage rating scale, after 24 and 48h. The procedure was repeated with all genotypes. Similar procedure was followed for leaves and flowers as well.

Feeding preference under multi-choice conditions 3.2.7.3

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