Effect of Plant Resistance in Castor on Endolarval Parasitoid Snellenius Maculipennis (Szepligate) of Castor Semilooper Achaea Janata L

Available online at www.ijpab.com Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 DOI: http://dx.doi.org/10.18782/2320-7051.6950 ISSN: 2320 – 7051 Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) Research Article

Effect of Plant Resistance in Castor on Endolarval Parasitoid Snellenius maculipennis (Szepligate) of Castor Semilooper Achaea janata L.

K. Chakkani Priya*, K. V. Hari Prasad, N. C. Venkateswarlu and V. Umamahesh Agricultural entomology, Acharya N.G Ranga Agricultural University, Lam, Guntur, Andhra Pradesh *Corresponding Author E-mail: [email protected] Received: 16.06.2018 | Revised: 20.07.2018 | Accepted: 27.07.2018

ABSTRACT A study on correlation between biological parameters of S. maculipennis with biophysical (epicuticular wax content) and biochemical (total free amino acids, proteins, carbohydrates and phenols) constituents of castor genotypes revealed that wax content was significantly negatively correlated with per cent parasitisation (-0.445*); total free amino acids were significantly negatively correlated with per cent parasitisation (-0.702**) and egg and larval period (- 0.546**); the proteins were significantly positively correlated with per cent parasitisation (0.938**); carbohydrates were significantly positively correlated with per cent parasitisation (0.957**) and significantly negatively correlated with adult longevity (-0.622**) and phenols were significantly negatively correlated with per cent parasitisation (-0.759**) and egg and larval period (-0.486*).

Key words: Snellenius maculipennis, Achaea janata, Genotypes, Correlation, Parasitisation.

INTRODUCTION which India, China, Brazil, USSR, Thailand, Castor, Ricinus communis L. is one of the most Ethiopia and Philippines are major castor important commercial, non-edible oilseed crop growing accounts for majority of the world's in Euphorbiaceae family. The genus, "Ricinus" production (http://www.commoditiescontrol. is derived from Latin word meaning "dog tick" com). because of it's seed resemblance to the India is the world's single largest common pest of dog. It is reported to have producer of castor seed accounting for more originated in the tropical belt of both India and than 85% followed by China with about 7% Africa. Castor is cultivated around the world and Brazil with about 5% of world castor seed because of the commercial importance of its output. There has been a consistent increase in oil. Historically, Brazil, China and India have output primarily on account of rise in levels in been the key producing countries meeting India. In recent years, however, Brazil and global requirements. It is cultivated in 30 China have experienced stagnation in Castor different countries on commercial scale, of crop (http://www.commoditiescontrol.com).

Cite this article: Chakkani Priya, K., Hari Prasad, K.V., Venkateswarlu, N.C. and Umamahesh, V., Effect of Plant Resistance in Castor on Endolarval Parasitoid Snellenius maculipennis (Szepligate) of Castor Semilooper Achaea janata L., Int. J. Pure App. Biosci. 6(5): 243-256 (2018). doi:

http://dx.doi.org/10.18782/2320-7051.6950

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 India is the world's single largest producer of Achaea janata (Linnaeus); castor shoot and castor seed accounting for more than 85% capsule borer, Dichocrocis punctiferalis followed by China with about 7% and Brazil (Guenee); hairy caterpillar, Euproctisfraterna with about 5% of world castor seed (Moore); tobacco caterpillar, Spodoptera litura output. There has been a consistent increase in (Fabricius) and sucking pests, such as output primarily on account of rise in levels in leafhopper, Amrasca biguttula biguttula India. In recent years, however, Brazil and (Ishida); white fly, Trialeurodes ricini (Misra); China have experienced stagnation in Castor thrips, Retithrips syriacus (Mayet) and mites, crop (http://www.commoditiescontrol.com). Tetranychus telarius (Linnaeus)19. Castor oil and its derivatives, besides Castor semilooper A. janata is a major being used in medicine, are also used in a wide defoliator and under severe infestations, range of sectors including agriculture, textile completely devour the green foliage, leaving industry, paper industry, plastic engineering, only the veins and enforce the farmers to re- rubber and pharmaceuticals4. sow the crop. It causes yield reduction to the Castor oil is also used in many veterinary extent of 20 to 23%9. medicines, as an emollient, soothing medium Castor semilooper, A. janata is when dropped into the eyes of animals after regulated by hymenopteran endolarval removal of foreign bodies. In eri silk- parasitoid Snellenius (=Microplitis) producing areas, castor leaves are fed to eri maculipennis (Szepligate) (Hymenoptera: worms. After extraction of oil, castor cake is Braconidae) which is cosmopolitan in valued as manure. It contains 6.4% N, 2.5% distribution8. Under field conditions, it is phosphoric acid and 1% K and some capable of parasitizing up to 77.31% of micronutrients. Castor is also valued for its semilooper population29. The early instars of anti-termite properties. Consequently, there A. janata were attacked by the parasitoid and has been a steady increase in the demand for parasitized larvae did not feed and died later castor oil and its products in the world market on40. owing to their renewable nature, Indiscriminate use of pesticides leads biodegradability and eco-friendliness4. to development of resistance in insects to Castor (Ricinuscommunis L.) is insecticides, toxic effect on natural enemies, cultivated around the world because of the pest resurgence and pesticide residues on food commercial importance of its oil. India is and adverse effect on human beings and the world’s largest producer of castor seed and causes environmental pollution23. These meets most of the global demand for castor oil. negative externalities, though cannot be It contributes about eight lakh tonnes of castor eliminated altogether, their intensity can be seed and three lakh tonnes of castor oil and minimized through development, sharing 65% and 51% respectively in the dissemination and promotion of alternative world total production. Gujarat, Rajasthan and technologies such as integrated pest Andhra Pradesh are the major castor growing management as well as good agronomic states in India. In Andhra Pradesh, castor practices rather relaying solely on chemical occupied about 0.33 lakh ha area in 2016-17 pesticides. Integrated Pest Management (IPM) and production was estimated about 0.16 lakh means a careful consideration of all available tonnes against 0.51 lakh ha area and 0.29 lakh pest control techniques and subsequent tonnes production in 2015- integration of appropriate measures that 16(https://www.indiastat.com). discourage the development of pest Both biotic and abiotic stresses are the populations by keeping pesticides and other major constraints for decline in castor interventions to levels that are economically production. Insect pests are the major biotic justified and reduce or minimize risks to factors in lowering of castor yield. The most human health and the environment. important insect pests are castor semilooper, Copyright © Sept.-Oct., 2018; IJPAB 244

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 One of the important components of integrated RH during 2017-18. Adults of A. janata were pest management is the use of natural enemies collected from college farm and Regional as biocontrol agents. This has many Agricultural Research Station, Tirupati and advantages over the traditional method of released into oviposition cages (32 cm × 30 cm chemical control34. With regard to parasitoids × 30 cm) for mating and were provided with as biocontrol agents, they assume immense 20% honey solution mixed with vitamin - E importance as they account for 87% tablets in cotton swabs. The petioles of castor suppression of insect pest in nature in leaves were kept in conical flasks by placing comparison to 12% by predators and 1% by the cut end of petioles dipped in water pathogens20. containing 10% sugar solution, these leaves Host plant resistance along with acted as substrate for oviposition and old natural enemies and cultural practices is a leaves were replaced with fresh leaves as and central component of any pest management when required. After one or two days of strategy. Resistance in the form of plant exposure, leaves having eggs of A. janata were defence is a physiochemical characteristic of shifted to another separate plastic troughs of plant, which influence the behaviour, biology, 25 cm diameter and 10 cm height where the survival and reproduction of insects38. eggs were allowed to hatch. Fresh leaves were The characteristics of plant structure provided adlib to the newly hatched larvae and and texture can affect herbivores, natural the rearing troughs were cleaned regularly to enemies of herbivores, and their interaction. keep a healthy stock culture. These effects may be genetically variable Pupae formed from the stock culture among plants and induced in individual plants were collected and kept in separate oviposition by herbivore attack, and are mediated by cages for adult emergence for further rearing primary plant attributes (i.e., nutritional quality as described above. The first instars were used and physical structure) and defense-related for all the experiments in the present products (i.e., secondary chemicals and plant investigations. volatiles). The study of tri-trophic interactions Maintenance of nucleus culture of is importantin order to understand natural parasitoid, Snelleniusmaculipennis enemies interactions and to manipulate these The stock culture of S. maculipennis was interactions in pest control1. Herbivore- maintained at Insectary, Department of induced plant volatiles have been suggested to Entomology, S.V. Agricultural College, function as indirect defence signals that attract Tirupati at 25 ± 2°C, 75 ± 5% RH. Initially, natural enemies of herbivores. Several insect parasitized larvae of A. janata with cocoons parasitoids are known to exploit such plant- were collected from college farm and Regional provided cues to locate their hosts13. Agricultural Research Station, Tirupati and Generally, parasitoids are host specific were kept in Petri plates of 15 cm diameter. and parasitize the host in a density dependent Adults emerged from these cocoons were manner2, and when exploited successfully this released into oviposition cages for mating and can be an effective way to reduce the were provided with 5% honey solution mixed frequency of pesticide applications and also with proteinex powder (Nutricia International reduced environmental pollution41. Pvt Ltd. India) has ingredients namely Vitamin A, D, E, K, B complex group, Folic MATERIAL AND METHODS Acid, Biotin etc. and several minerals Maintenance of stock-culture of semilooper like iron, calcium, phosphorus and choline as A. Janata well, dipped in cotton swab as food material. The stock culture of A. janata was maintained Second instar larvae of A. janata were in the Insectary, Department of Entomology, released on castor leaves and were offered to S.V. Agricultural College, ANGRAU, adults of S. maculipennis for oviposition in Tirupati, Andhra Pradesh at 25 ± 2°C, 75 ± 5% oviposition cages. The cut end of petioles of Copyright © Sept.-Oct., 2018; IJPAB 245

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 castor leaves were dipped in water in a conical degrees of resistance to A. janata33 were flask containing 10% sugar solution. After grown in plastic pots of 23 cm diameter and 20 exposing the larvae for 24 to 48 hrs to the cm height during kharif 2017 under net house adult parasitoids, the parasitised larvae were conditions at 25-39oC and 48-80% RH . The collected and reared separately in plastic seeds of these germplasm were procured from troughs of 20 cm diameter and provided with Regional Agricultural Research Station, castor leaves till cocoon formation. After Palem, Telangana, India and The Indian formation of the cocoon from parasitized Institute of Oilseeds Research (IIOR), larvae, parasitized larvae with cocoons still Hyderabad, Telangana, India. The substrate attached to its body were kept in Petri plates of used for growing of plants was red soil : 15 cm diameter for further rearing as described compost in 3 : 1 ratio. above. The parasitoids were reared and The seeds of test plants were treated with multiplied for two generations to obtain fungicides (Mancozeb 2g Kg1) to prevent sufficient number of parasitoids before using seed-borne fungal diseases. All the normal them for the experiments. agronomic practices were followed for pot Pot culturing of castor genotypes with culturing of plants. Leaves from these castor different blooms. genotypes at peak vegetative stage were used Three genotypes of castor viz., DPC-9 (Green, in the experiments. Staggered planting of zero bloom, susceptible), 48-1 (Red, double castor genotypes was done for continuous bloom, moderately resistant) and GCH-4 (Red, supply of leaves during the period of triple bloom, resistant) showing different experimentation.

Table 1: Bloom characters of castor genotypes used in the present investigation Genotype Bloom nature Character

DPC-9 Green zero No bloom or waxy material on any part of the plant.

Bloom on stems, petioles and lower surface of 48-1 Red double leaves but not on the upper surface of the leaves. Bloom on every part of plant such as stems, GCH-4 Red triple petioles, upper and lower surface of leaves.

Studies on biology of S. maculipennis on A. replications. After two days, five pairs (five janata larvae, reared on castor genotypes males and five females) of parasitoids were with different blooms. released in each cage and were allowed to The biology of S. maculipennis was studied oviposit on A. janata larvae feeding on excised under laboratory conditions of 25 ± 2˚C leaves of castor with different wax blooms. temperature and 75 ± 5% RH on excised After exposing the host larvae for 24 to 48 hrs leaves of three castor genotypes with different to the adult parasitoids, the parasitized larvae blooms viz., DPC-9, 48-1 and GCH-4. The were collected and reared in plastic troughs petioles of each castor genotype leaves were separately, till cocoon formation and dipped in water in separate conical flasks parasitized larvae with cocoons still attached containing 10% sugar solution. These conical to its body were kept in separate plastic boxes flasks containing excised leaves of castor of size 8.5 cm × 4.5 cm × 2.5 cm for adult genotypes were kept in three separate emergence. After adult emergence they were oviposition cages. provided with 5% honey mixed with proteinex Ten first instar larvae of semilooper A. powder dipped in cotton swab as food Janata were released on the excised leaves of material. Data was recorded on egg+larval each genotype and were left for feeding on the duration, number of cocoons formed, cocoon leaves for two days. There were total of eight duration and adult longevity of S. Copyright © Sept.-Oct., 2018; IJPAB 246

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 maculipennis on A. janata reared on castor After two days of larval feeding, five pairs of genotypes having different wax blooms. adult parasitoids were released inside the cage Studies on role of castor wax blooms on and allowed to oviposit (free choice condition) ovipositional behavior of S. maculipennis on the larvae of A. janata feeding on different A fully developed leaves from each green zero castor genotypes with different blooms. There bloom, red double bloom and red triple bloom were total of eight replications. After exposing were excised upto petiole and cut ends of the larvae for 24 to 48 hrs to the adult petiole was placed in conical flask with 10% parasitoids, the parasitized larvae were sugar solution and this setup was used for collected and reared separately in plastic oviposition preference of S. maculipennis. troughs and provided with leaves of respective Ten first instar A. janata larvae were castor genotypes till cocoon formation. released on leaves each of zero, double and Emerged adults from cocoons, were provided triple bloom. The larvae were allowed to feed with 5% honey solution mixed with proteinex powder dipped in cotton swab as food on the leaves of respective genotypes for two material. days. Care was taken to prevent movement of Oviposition preference of S. maculipennis larvae from leaves of one genotype to another towards A. janata larvae fed on castor by keeping conical flasks (with leaves) on a 20 genotypes with different blooms viz.,DPC-9, cm diameter trough underneath and were 48-1 and GCH-4 under free choice condition arranged in such a way that the leaves do not were recorded and expressed as per cent touch each other. parasitisation.

No. of larvae parasitized Per cent parasitisation =  100 Total no. of larvae released

Studies on biochemical constituents of water. The test tubes were then heated in castor genotypes with different blooms. boiling water bath for 20 min. Then five ml of The biochemical constituents such as protein, diluents was added and the contents were phenol, total free amino acids, and total mixed. After 15 minutes intensity of the purple carbohydrates were estimated at Institute of colour was read against a reagent blank in a Frontier Technology (IFT), Regional colorimeter at 570 nm. A standard curve was Agricultural Research Station, Tirupati. The prepared using leucine and it was linear within procedures and protocols are enlisted as the range (R2 = 0.984). The total free amino below. A total of eight replications were used acids of the leaf sample was then calculated by for estimating the biochemical constituents of substituting the sample Optical Density values castor genotypes with different blooms in the formula derived from the graph (Y = viz.,DPC-9, 48-1 and GCH-4. 0.012X – 0.202), where Y = Optical Density Total free amino acids (µg/g) value of the sample and X = concentration (μg Total free amino acids was estimated by ml-1) of free amino acids. Then the adopting the method suggested by Sadasivam concentration of free amino free amino acids and Manickam32, Moore and Stein24. 500 mg in (µg ml-1) was converted to concentration of of leaf sample was taken and ground in five ml free amino acids in µg/g for different entries. of 80% ethanol. The homogenate was Estimation of proteins (mg/g) centrifuged at 6000 rpm for 30 min at 20oC. The protein content in fresh leaves of all the The supernatant obtained was used for three castor genotypes such as DPC-9, 48-1 estimation of free amino acids. 0.1 ml of the and GCH-4 were estimated as per the method extract was taken and to it one ml of by Wildman and Bonner43. 250 mg of leaf Ninhydrin solution was added, followed by sample was macerated with ten ml of making up the volume to two ml with distilled phosphate solution. The contents were Copyright © Sept.-Oct., 2018; IJPAB 247

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 centrifuged at 3000 rpm for about ten min and carbohydrates in (µg ml-1) was converted to then the supernatant was collected followed by concentration of carbohydrates in g/g for volume make up to 25 ml with distilled water. different entries. One ml of supernatant was pipette out and five Phenol content (%) ml of alkali copper tartarate was added to it. Phenol content in the castor leaves was The solution was kept as such for 30 min for estimated by adopting the method suggested colour development. 0.5 ml of phenol reagent by Sadasivam and Manickam32, Malik and was then added and the Optical Density value Singh22, Muthukumaran25. One g of the leaf of the sample was measured at 660 nm in sample was ground in ten time volume of 80% Spectrophotometer. A standard curve was ethanol with a pestle and mortar. Then the prepared by using bovine serum albumin and it homogenate was centrifuged at 10000 rpm for was linear within the range (R2 = 0.969). The 20 min, supernatant was saved. Then the protein content of the sample was calculated residue was extracted with five times volume by substituting the sample Optical Density of 80% ethanol and centrifuged. Then the values in the formula derived from the graph supernatants were pooled and evaporated to (Y = 0.0065X – 0.014), where Y = Optical dryness. The residue was dissolved in ten ml Density value of the sample and X = of distilled water. 0.4 ml of the sample extract concentration (µg ml-1) of the protein. Then was pipetted into test tubes and the volume the concentration of proteins in (µg ml-1) of was made upto three ml with distilled water leaf sample was converted to concentration of followed by addition of 0.5 ml of folin- proteins in mg/g for different entries. ciacalteau reagent. After three min, two ml of Total carbohydrates (g/g 20% sodium carbonate was added and the test Total carbohydrates was estimated by adopting tubes were kept in boiling water bath for one the method suggested by Sadasivam and min, cooled and the absorbance was measured Manickam (1961), Hedge and Hofreiter12. 50 against a reagent blank at 650 nm. A standard mg of leaf sample was taken in a 50 ml test curve was prepared by using catechol and it tube and 2.5 ml of 2.5 N HCl was added to it. was linear within the range (R2 = 0.963). The The test tubes were kept in hot water bath phenol content of the sample was calculated (100oC) for three hours and cooled to room by substituting the sample Optical Density temperature. Solid sodium carbonate was then values in the formula derived from the graph added to the test tubes followed by volume (Y = 0.397X + 0.013), where Y = Optical make upto 25 ml using distilled water. The test Density value of the sample and X = tubes were then kept at room temperature for concentration (μg ml-1) of the phenol. Then the 20 min without any disturbance. For analysis concentration of phenols in μg ml-1 was one ml of the aliquot was taken, cooled on ice, converted to concentration of phenols in % for and four ml of ice cold anthrone reagent was different entries. added to it. The test tubes were then kept in Studies on biophysical constituent (wax) of hot water bath for eight min and cooled castor genotypes with different blooms. rapidly. The absorbance of the resultant green The biophysical constituent i.e., wax was was measured against a reagent blank at 630 estimated at Department of Crop Physiology, nm. A standard curve was prepared using S.V. Agricultural College, Tirupati. glucose and it was linear within the range (R2 The wax content in fresh leaves of all the three = 0.987). The total carbohydrates of the leaf castor genotypes viz., DPC-9, 48-1 and GCH-4 sample was then calculated by substituting the with different blooms were estimated as per sample Optical Density values in the formula the method suggested by Eberconet al.6, with derived from the graph (Y = 0.006X – 0.023), suitable modifications. where Y = Optical Density value of the sample and X = concentration (µg ml-1) of carbohydrate. Then the concentration of Copyright © Sept.-Oct., 2018; IJPAB 248

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 Principle RESULTS AND DISCUSSION Colour change produced due to the reaction of Biology of endolarval parasitoid (Snellenius wax with acidic K2Cr2O7 reagent was maculipennis) of castor semilooper Achaea measured at 590 nm by calorimetry. janata. Reagent The present study on the biology of S.

Acidic K2Cr2O7 reagent: 20 g K2Cr2O7 was maculipennis Szep. revealed that the newly dissolved in 40 ml of distilled water and then emerged adult parasitoid mated on the day of mixed with one litre H2SO4 and heated to clear emergence and the oviposition commenced solution. from the succeeding day. Twenty four hours Procedure after parasitisation, the larvae became The leaf discs (10 no’s each of 9 cm diameter) sluggish, however it continued feeding at a were immersed in 15 ml chloroform for 15 slower rate till 5-6 days after parasitisation sec. The extract was filtered and evaporated when the parasitoid grub emerged from lateral, over boiling water bath till there was no smell posterior end of the host insect body. The grub immediately started forming the cocoon and of chloroform. Five ml of the reagent K2Cr2O7 attached itself to the posterior ventral portion was added to this extract and placed on boiling of the host insect. After the emergence of the water bath for 30 min. After cooling, 12 ml of parasitoid grub the host survived for 4 to 5 distilled water was added to the resultant days but stopped feeding and died later. extract. The Optical Density was measured at Duration of egg+larva of S maculipennis 590 nm after 30 min. Sorghum wax collected While studying the biology of S. maculipennis from leaf sheaths was dissolved in chloroform on A. janata larvae, it was observed that the and used as standard wax. female parasitoid thursts eggs into the host Standard curve body (in hemolymph), from the dorsal side of Standard solution: 100 mg of sorghum wax the abdominal segments. The egg after was collected from leaf sheaths of 60 days old hatching remains in the hemolymph of host sorghum plants. The same was dissolved in body and feeds there in. When the grub 100 ml of chloroform. completes its larval period, final instar cuts Working standard: One ml of standard solution through the body wall of the host larvae on the was dissolved in 100 ml of chloroform to get lateral side, comes out and spins a cocoon minimum concentration of 10 µg in 1 ml. around its body within five hours. The grub Different concentration of wax, i.e., was apodous, spindle shaped and transparent. 20, 40, 60, 80 and 100 µg was prepared by Longest durations of egg+larval stages taking different volume of 2, 4, 6, 8 and 10 ml of S. maculipennis (8.02 days) was recorded in of working standard solution in different test A. janata larvae reared on GCH-4. Shorter tubes and evaporated over boiling water bath duration of 5.09 days was recorded on 48-1 till there was no smell of chloroform. Five ml followed by DPC-9 (6.72 days) which were on of the reagent was added to this working par with each other (Table 4.1). standard and placed on boiling water bath for Number of cocoons formed 30 min. After cooling, 12 ml water was added More number of cocoons of S. maculipennis to the resultant working standard. After 30 (9.63) was formed on A. janata larvae reared min, the Optical Density was measured at 590 on DPC-9. Less number of cocoons of S. nm. To get the standard curve Optical Density maculipennis (7.75) was formed on A. janata values were plotted against concentration. larvae reared on 48-1 followed by GCH-4 By comparing the Optical Density (8.75) which were significantly different from value of the sample in the standard curve, the each other (Table 4.1). leaf epicuticular wax content in the sample Duration of cocoon of S. maculipennis (seven replications) was computed and The cocoon was initially grey in colour and expressed in µg per unit of cm2 area. later turned brown. The cocoons on one side Copyright © Sept.-Oct., 2018; IJPAB 249

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 was firmly attached to rear end of the host recorded on DPC-9 followed by 48-1 (4.36 larvae and other side to the leaf surface days) which were on par with each other rendering the host immobile. (Table 4.1). Longest cocoon duration of S. Duration of total life cycle of the parasitoid, maculipennis (5.33 days) was recorded on A. S. maculipennis janata larvae when reared on DPC-9. Shortest Longest duration of total life cycle of duration of 4.11 days was recorded on GCH-4 parasitoid S. maculipennis (18.50 days) was followed by 48-1 (4.81 days) which were on recorded when reared from larvae of A. janata par with each other (Table 4.1). fed on GCH-4 and shortest duration of total Longevity of adult parasitoid, S. life cycle of 14.26 days was recorded when maculipennis reared on 48-1 followed by DPC-9 (15.69 Longevity of adult S. maculipenniswas days) which were on par with each other calculated as the day when they were emerged (Table 4.1). from cocoons till death. The adult parasitoid Our results were in conformity with was short, black in colour. The first half of the the Prabhakar and Prasad28 who reported that abdomen was glossy yellowish in colour. egg and larval, cocoon, adult longevity, and Longest adult longevity of S. total life cycle of S. maculipennis on A. janata maculipennis (6.38 days) was recorded when larvae as 6.25 ± 0.96 days,5.08± 0.90 days, 3.5 reared on larvae of A. janata fed on GCH-4 ± 1.02 days and 14.83 ± 2.83 days and shortest adult longevity of 3.64 days was respectively.

Table 2: Biological parameters of endolarval parasitoid S. maculipennis on castor semilooper A. janata larvae reared on castor genotypes with different wax blooms Egg and larval Number of Pupal period Adult longevity Total life cycle Genotype period (days) cocoons formed (days) (days) (days) DPC-9 6.72ab 9.63a 5.33a 3.64b 15.69ab (Zero bloom, Susceptible) 48-1 5.09b 7.75ab 4.81ab 4.36b 14.26b (Double bloom, Moderately resistant) GCH-4 8.02a 8.75a 4.11b 6.38a 18.50a (Triple bloom, Resistant) LSD at 0.01 1.90 1.11 1.18 1.83 3.14 CV % 20.31 8.99 17.60 26.92 13.75 Values followed by same letter are not significantly different at 0.01 level.

Role of castor wax bloom on ovipositional 65% was recorded on A. janata larvae when behavior ofS. maculipennis. reared on DPC-9 and was offered for Adults of S. maculipennis were allowed to parasitisation. Lowest parasitisation of 13.75% oviposit on A. janata larvae reared on DPC-9 was recorded on A. janata larvae when reared (Green, zero bloom, susceptible), 48-1 (Red, on 48-1 and was offered for parasitisation double bloom, moderately resistant) and GCH- followed by GCH-4 (18.75 %) which were on 4 (Red, triple bloom, resistant) under free par with each other (Table 4.2.1). choice condition. Highest parasitisation of

Table 3: Parasitisation of A. janata larvae by S. maculipennis reared on castor genotypes with different wax blooms under free choice condition Genotype Per cent parasitisation DPC-9 65a (Zero bloom, Susceptible) (53.82) 48-1 13.75c (Double bloom, Moderately resistant) (21.48) GCH-4 18.75bc (Triple bloom, Resistant) (25.54) LSD at 0.01 9.14 CV % 19.86 Values followed by same letter are not significantly different at 0.01 level and the values in parenthesis are transformed values.

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 Biochemical and biophysical constituents of susceptible genotypes of rice was found higher castor genotypes with different blooms compared to resistant genotypes ( 2.97 to 4.2 Biochemical constituents viz., total free amino mg/g). acids, protein content, carbohydrates, phenol Carbohydrate content content and biophysical constituent i.e., wax Highest amount of carbohydrate (0.24 g/g) from leaves of castor genotypes with different was recorded from the leaves of DPC-9. blooms namely DPC-9 (Green, zero bloom, Lowest amount of carbohydrate (0.15 g/g) was susceptible), 48-1 (Red, double bloom, found in GCH-4 followed by 48-1 (0.17 g/g). moderately resistant) and GCH-4 (Red, triple All were significantly different from each bloom, resistant) were estimated as per other (Table 4.3). protocols in section of material and methods, Phenol content and are presented in (Table 4.3). Highest amount of phenol (11.39%) was Biochemical constituents recorded from the leaves of 48-1 followed by Total free amino acids (TFA) GCH-4 (7.17%). The lowest amount of phenol Highest amount of total free amino acids (5.34%) was found in DPC-9. All were (307.45 µg/g) was recorded from the leaves of significantly different from each other (Table 48-1 followed by GCH-4 (203.43 µg/g). 4.3). Our results were in conformity with Lowest amount of total free amino acids Sharma and Bharadwaj39, who found that (175.76 µg/g) was found in DPC-9. All were amounts of polyphenols was highest in significantly different from each other (Table genotypes which are resistant to Helicoverpa 4.3). armigera. Protein content Biophysical constituent Highest amount of protein (0.57 mg/g) was Wax content recorded from the leaves of DPC-9. Lowest Highest amount of wax (1.43 µg/cm2) was amount of protein (0.30 mg/g) was found in recorded from the leaves of GCH-4 followed 48-1 followed by GCH-4 (0.37 mg/g). All by48-1 (1.20 µg/cm2) which were significantly were significantly different from each other different from each other. The lowest amount (Table 4.3). The results were in accordance of wax (0.98 µg/cm2) was found in DPC-9 with Kumar et al.18, who reported that amount (Table 4.3). of proteins (5.19 to 6.78 mg/g) in all

Table 4: Biochemical and biophysical constituents of castor genotypes with different wax blooms

Total free Proteins Carbohydrates Phenols Wax(µg/ Genotype Amino acids (mg/g) (g/g) (%) cm2) (µg/g)

DPC-9 c c a a 5.34 b (Zero bloom, Susceptible) 175.76 0.57 0.24 0.98 (13.36) 48-1 (Double bloom, Moderately 11.39a 307.45a 0.30c 0.17b 1.20ab resistant) (19.72)

GCH-4 b b b c 7.17 a (Triple bloom, Resistant) 203.43 0.37 0.15 1.43 (15.53)

LSD at 0.01 0.58 0.04 0.01 0.39 0.44 CV % 0.18 7.68 5.33 3.45 23.56 Values followed by same letter are not significantly different at 0.01 level and the values in parenthesis are transformed values.

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 Correlation study of different significant negative correlation with larval developmental periods of S. maculipennis on feeding11. A. janata larvae reared on castor genotypes In the present investigations, it was with different wax blooms and per cent observed that total free amino acids and parasitisation in free choice condition with phenols had a significant negative correlation biochemical and biophysical constituents of with and per cent parasitisation. At the same castor genotypes with different wax blooms time proteins and carbohydrates had a Biochemical constituents significant positive correlation with per cent Total free amino acids were significantly parasitisation. negatively correlated with per cent Phenols which is a plant secondary parasitisation (-0.702**) and egg and larval metabolite is known to exert a negative effect period (-0.546**) and for the rest of other on growth and metabolism of insect biological parameters the correlations were herbivores. Several reports have shown non significant. negative effect of phenols on insect herbivore, The protein content were significantly at the same time proteins and carbohydrates positively correlated with per cent tend to act as phagostimulants to most of parasitisation (0.938**) and for the rest of insect herbivores which increases the other biological parameters the correlations susceptibility of host plants to insect were non significant. herbivores. The results were in close 42 Carbohydrates were significantly association with Tilakchand . who reported positively correlated with per cent that protein (0.8292 mg/g) content manifested parasitisation (0.957**) and significantly positive and significant correlations while negatively correlated with adult longevity (- phenol (-0.9459 mg/g) content recorded 0.622**) and for the rest of other biological significantly negative correlation with larval population of H. armigera. According to parameters the correlations were non Sharma and Singh37 plant biochemical significant. characters in okra i.e., total sugar and crude Phenols were significantly negatively protein were positively correlated with fruit correlated with per cent parasitisation (- borer infestation, whereas total phenols had a 0.759**) and egg and larval period (-0.486*) negative correlation. Rath and Nayak31, and for the rest of other biological parameters revealed that the lower level of protein in fruits the correlations were non significant. The of tomato varieties were responsible for the results were on par with the findings of low susceptibility of tomato varieties to fruit Ghumare and Mukherjee10, who reported that borer and high amount of phenol were lower the amount of total phenolics, better the responsible for resistance. Rashmi and performance of Spodoptera lituraand have Vemadevaiah30 reported significantly higher inverse relationship with preference and larval amount of total phenol in bollworm tolerant 25 growth. Muthukumaran reported that phenol genotype Sahana (6.25 mg/g) in all the stages content was higher in resistant varieties and it than Laxmi (5.60 mg/g) indicated its positive exerted a significant negative correlation with role in pest tolerance. Presence of lesser larval feeding and negatively correlated with number of survived larvae in Sahana than fruit worm H. armigera infestation and leaf Laxmi indicated its biochemical basis of caterpillar S. litura Fab. incidence in tomato. tolerance, the genotype Sahana has higher total Biochemical contents of the tomato such as phenols content than Laxmi. Muthukumaran25 non-reducing sugar and phenols present in found that phenol content was high in resistant higher quantities were found negatively varieties of tomato against fruit worm, H. correlated with fruit worm H. armigera armigera (Hubner) and exerted a significant infestation35,5,36. Further, phenol content was negative correlation with larval feeding and higher in resistant accession and it exerted a negatively correlated with fruit worm H.

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 armigera infestation and leaf caterpillar S. secondary metabolites or might have resulted litura F. incidence in tomato. in storage excretion where the phenols might Biophysical constituent. have been stored in fat bodies in the insect Wax content were significantly negatively hemolymph. Nishio27 reported that larvae of correlated with per cent parasitisation (- the monarch butterfly Danaus plexippusL. 0.445*) and for the rest of other biological utilize a large volume of gut fluid as a critical parameters the correlations were non factor in the processing of cardenolides significant. ingested from the milkweed Asclepias In the present investigation, wax humistrata host plants. Larvae can regurgitate content had significant negative correlation gut fluid readily, giving them cardenolide-rich with per cent parasitisation. It was also shield that can be thrust at predators. These observed from Table 4.2.1 that the per cent bitter-tasting steroids reappear in the teneral parasitisation of A. janata larvae by S. adult and are concentrated in the wing scales maculipennis was highest when larvae were and hemolymph, providing the imago with a allowed to feed on DPC-9 (Green, zero bloom, readily available defence against avian susceptible) with no wax bloom as compared predators such as blue jay Cyano cittacristata. to that of 48-1 (Red, double bloom, The parasitoid used in the present moderately resistant) and GCH-4 (Red, triple investigation i.e., S. maculipennis deposits bloom, resistant). eggs within hemolymph and it has been In the present investigation, proteins postulated by many workers that adult and carbohydrates had a significant positive parasitoids generally tries to choose the best correlation with per cent parasitisation. host for its offspring during oviposition. Low phenol (5.34%) and wax (0.98 Karowe and Schoonhoven17, reported that µg/cm2) contents in susceptible genotype vigorous hosts feed more and therefore DPC-9 had positive effect on the host insect A. produce more tissue and/or hemolymph on janata, which might have resulted in highest which the parasitoids chooses to oviposit. per cent parasitisation when larvae reared on In our present investigation, larvae of DPC-9 were offered for parasitisation to adult A. janata feeding on castor genotypes with parasitoid S. maculipennis in free choice, dual more phenols probably were least preferred for choice and no choice condition. parasitisation by the adults of S. maculipennis When A. janata larvae were reared on as because of phenols in the bodies a moderately resistant genotype 48-1 having (hemolymph) of host larvae rendered them more phenol content (11.39%), the host larvae unpalatable or unsuitable for egg laying. either might have detoxified the plant

Table 5: Correlation of different developmental periods of S. maculipennis on A. janata larvae reared on castor genotypes with different wax blooms and per cent parasitisation in free choice condition with biochemical and biophysical constituents of castor genotypes with different wax blooms Biochemical constituents of castor genotypes with different wax blooms Correlation of developmental Total free Proteins Carbohydrates Phenols (%) Wax content periods of S. maculipennis amino acids (mg/g) (g/g) (µg/cm2) (µg/g) Per cent Pearson -0.702** 0.938** 0.957** -0.759** -0.445* parasitisation correlation Egg and larval Pearson -0.546** 0.167 -0.124 -0.486* 0.105 period correlation Pearson Cocoon period -0.062 0.274 0.39 -0.124 -0.294 correlation Pearson Adult longevity -0.045 -0.388 -0.622** 0.025 0.313 correlation Pearson Total life cycle -0.397 -0.038 -0.328 -0.336 0.15 correlation ** Correlation is significant at the 0.01 level (2- tailed). * Correlation is significant at the 0.05 level (2- tailed).

Chakkani Priya et al Int. J. Pure App. Biosci. 6 (5): 243-256 (2018) ISSN: 2320 – 7051 Summary and Conclusion positively correlated with per cent Effect of wax bloom of castor genotypes on parasitisation (0.957**) and significantly the biology of endolarval parasitoid negatively correlated with adult longevity (- Snellenius maculipennis 0.622**), phenols were significantly More number of cocoons (9.63) of S. negatively correlated with per cent maculipennis were formed on A. janata larvae parasitisation(-0.759**) and egg and larval reared on DPC-9 and less number of cocoons period (-0.486*) and for the rest of other (7.75) were formed on A. janata larvae reared biological parameters the correlations were on 48-1 followed by GCH-4 (8.75). Durations non significant. of egg and larval stages of S. maculipennis in A. janata larvae was longest on GCH-4 (8.02 REFERENCES days) and shortest on 48-1 (5.09 days). 1. Agrawal, A.A., Mechanisms, ecological Cocoon duration of S. maculipennis on A. consequences and agricultural implications janata larvae reared on DPC-9 was longest of tri-trophic interactions.Current Opinion (5.33 days) and shortest on GCH-4 (4.11 in Plant Biology. 3: 329–335 (2000). days).Adult longevity of S. maculipennis on A. 2. Annecke, D.P. and Moran, V.C., Insects janata reared on GCH-4 was longest (6.38 and Mites of Cultivated Plants in South days) and shortest on DPC-9 (3.64 days). Africa. Butterworths, Durban, South Duration of total life cycle of S. maculipennis Africa. 338 (1982). on A. janata reared on GCH-4was longest 3. Annual Report - DOR. Directorate of Oil (18.50 days) and shortest on 48-1 (14.26 days). Seeds Research, Hyderabad.1-2 (1997). Role of castor wax bloom on ovipositional 4. Annual Report – DOR. Directorate of Oil behavior of S. maculipennis Seeds Research, Hyderabad.1-2 (2003). Under free choice condition highest 5. Dhakshinamoorthy, G., Studies on parasitisation of 65% was recorded on A. resistance of tomato against fruit borer, janata larvae, reared on susceptible genotype Helicoverpaarmigera (Hubner). M.Sc. DPC-9. Lowest parasitisation of 13.75% was (Ag.) Thesis, Annamalai University. India recorded on onA. janata larvae, reared on (2002). moderately resistant genotype 48-1, followed 6. Ebercon, A., Blum, A and Jordan, W.R., A by resistant genotype GCH-4 (18.75%). rapid colorimetric method for epicuticular Correlation between biological parameters, wax content of sorghum leaves. Crop per cent parasitisation of S. maculipennison Science. 17: 179-80 (1977). A. janata with biophysical and biochemical 7. Food and Agriculture Organisation of the constituents of castor genotypes United Nations.Integrated Pest Correlation study of different developmental Management.http://www.fao.org/agriculture periods of S. maculipennis on A. janata larvae /crops/thematicsitemap/ theme/pests/ipm/en/ reared on castor genotypes with different wax 8. Gaikwad, B.B. and Bilapate, G.G., Field blooms and per cent parasitisation in free life-tables and key mortality factors of choice condition with biochemical and Achaea janata Linn. on castor. Animal biophysical constituents of castor genotypes Sciences. 98(5): 331-339 (1989). with different wax blooms revealed that wax 9. Gaikwad, B.B. and Bilapate, G.G., content was significantly negatively correlated Parasitization of Achaea janata and with per cent parasitisation (-0.445*), total free estimation of losses on castor.Journal of amino acids were significantly negatively Maharashtra Agricultural Universities. correlated with per cent parasitisation (- 17: 195-196 (1992). 0.702**) and egg and larval period (-0.546**), 10. Ghumare, S.S. and Mukharjee, S.N., the proteins were significantly positively Performance of SpodopteralituraFabricius correlated with per cent parasitisation on different host plants: influence of (0.938**), carbohydrates were significantly nitrogen and total phenolics of plants and Copyright © Sept.-Oct., 2018; IJPAB 254

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