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EVALUATION OF INSECTICIDE RESISTANCE PROFILE IN Spodoptera litura (FABRICIUS) POPULATIONS THROUGH BIOLOGICAL, BIOCHEMICAL AND MOLECULAR DIAGNOSIS
(PAU Ludhiana, 2013) Harpreet Kaur Cheema; B. K., KANG
Evaluation of insecticide resistance profile in Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) populations was done through biological, biochemical and molecular diagnosis. Population from Sangrur was found to be the least susceptible for most of the insecticides, except for chlorpyriphos, chlorantraniliprole and novaluron which were least toxic to population from Bathinda, and pyridalyl which was least toxic to population from Ludhiana. Population from Hoshiarpur was found to be the most susceptible for all insecticides except acephate and pyridalyl to which Bathinda population was most susceptible. Thiodicarb among the conventional group of insecticides and chlorantraniliprole among the newer insecticides were found to be the most toxic against all the test populations. The synergistic effect of TPP (6.16- fold), PBO (3.72- fold) and DEM (1.73- fold) in the least susceptible population suggested the involvement of esterases and mixed function oxidases in providing resistance to various insecticides. Synergistic effect of PBO (6.21- fold) on the fenvalerate- selected population followed by TPP (5.74- fold) and DEM (1.89- fold) indicated the possible role of mixed function oxidases, esterases and to some extent glutathione S- transferases in imparting resistance to pyrethroids in S. litura. Multiple detoxification enzymes and AChE insensitivity were found to be involved in imparting resistance in S. litura towards various insecticides. Selection with fenvalerate increased the activities of MFOs, esterases and GST by 1.84, 1.73 and 1.3 times in S. litura proving their role in pyrethroid-mediated resistance. Activities of MFO, esterases and insensitivity of AChE can be used as biochemical tools for monitoring insecticide resistance in S. litura. RAPD markers established the presence of inter and intra population variations that might be related to insecticide resistance. Alignment between COI sequences of fenvalerate- selected (SGR-Sel) and susceptible types (SGR) identified existence of a single base/ nucleotide polymorphism at position 421 represented by substitution of T in SGR-Sel with C in SGR type. Based on this polymorphism, COI based specific primers were developed, tested and validated for differentiating amongst these S. litura types.
Open Access
UPTAKE AND METABOLISM OF IMIDACLOPRID IN SUGARCANE AND ITS PERSISTENCE IN SOIL
(PAU Ludhiana, 2012) Smriti Sharma; Balwinder Singh
The uptake and metabolism of imidacloprid in sugarcane leaves and soil were studied following application of imidacloprid @ 20 and 80 g a.i. ha-1. Samples of sugarcane leaves and soil were collected at various time intervals and analyzed by using High Performance Liquid Chromatograph (HPLC) and confirmed by Liquid Chromatograph –Mass Spectrometry (LCMS). Total residues of imidacloprid and its metabolites were found to be 4.97 and 12.99 mg kg-1 in leaves collected 7 days after the application of imidacloprid @ 20 and 80 g a.i. ha-1 respectively. These residues declined to below the detectable limit at 90 days after the application at both the dosages. The samples of sugarcane juice and trash did not reveal the presence of imidacloprid residues at harvest. In soil samples total residues of imidacloprid were found to be 4.29 and 7.81 mg kg-1 collected 7 days after the application @ 20 and 80 g a.i. ha-1 respectively. These residues could not be detected at 90 days after the application at both the dosages. In laboratory experiment, residues of imidacloprid in sandy loam, loamy sand and clay loam after 7 day of its application @ 100 mg kg-1 were found to be 69.74, 72.66 and 77.20 mg kg-1, respectively. The persistence of imidacloprid was almost similar in all type of soils. In sterilized soil, Bacillus alkaninitricus was found to be more efficient than B. aerophilus and their consortium while, in unsterilized soil, bacterial consortium proved to be more efficient in imidacloprid degradation.
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DEVELOPMENT OF TOBACCO CATERPILLAR, Spodoptera litura (FABRICIUS) (LEPIDOPTERA: NOCTUIDAE) ON DIFFERENT CULTIVARS OF CABBAGE
(PAU Ludhiana, 2012) Jasjinder Kaur; Inderpal Singh)
The development of tobacco caterpillar, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) was studied during 2012-13 on cabbage cultivars viz. Pride of India, Pusa Drum Head, KGMR-1, Golden Acre and Pusa Mukta at the Entomological Laboratory, PAU, Ludhiana. There were significant differences in incubation period and hatchability of eggs with a minimum incubation period on Pusa Mukta (4.27 ± 0.09 days) and maximum on Pride of India (4.87 ± 0.10 days). Egg hatchability was maximum on Pusa Mukta (86.66 ± 0.82%) and minimum on Pride of India (81.76 ± 1.06%). There were significant differences in duration of larval instars on different cultivars. Total larval duration was minimum on Pusa Mukta (15.55 ± 0.12 days) and maximum on Pride of India (20.09 ± 0.14 days). The larval survival was minimum on Pride of India (71.00 ± 1.29%) and maximum on Pusa Mukta (89.00 ±1.29%). Pre-pupal and pupal periods were observed as minimum on Pusa Mukta (2.26 ± 0.02 and 6.38 ± 0.03 days, respectively) and maximum (2.89 ± 0.10 and 9.08 ± 0.05 days, respectively) on Pride of India. The male and female pupal weight was maximum on Pusa Mukta (0.362 ± 0.003g and 0.364 ± 0.004g, respectively) and minimum on Pride of India (0.349 ± 0.003g and 0.351 ± 0.003g, respectively). Pupal survival was maximum on Pusa Mukta (92.00 ± 1.83%) and minimum on Pride of India (80.00 ± 2.42%) and Pusa Drum Head (80.00 ± 2.08%). No significant differences were observed in pre-oviposition, oviposition, post-oviposition periods; adult longevity and fecundity on all the test cultivars. The female and male sex ratio ranged from 1.22:1 to 1.86:1 on different test cultivars. The total life duration of S. litura varied significantly among different cultivars with maximum on Pride of India (36.93 ± 0.25 days) and minimum on Pusa Mukta (28.46 ± 0.54 days). The extent of damage in terms of leaf area consumed/ larva was maximum on Pusa Mukta (158.28 ± 0.89 cm²) and minimum on Pride of India (139.82 ± 0.69 cm²). The Consumption Index (CI), Approximate Digestibility (AD), Relative Growth Rate (RGR) and Efficiency of Conversion of Ingested food (ECI) were found to be maximum on Pusa Mukta (2.20 ± 0.04, 88.10 ± 1.04%, 0.20 ± 0.01 and 9.18 ± 0.37%, respectively). However, Pride of India recorded minimum CI (1.06 ± 0.02) and AD (76.10 ± 1.74%) with comparatively lower RGR (0.09 ± 0.01) and ECI (8.28 ± 0.33%), and thus proved least suitable for the development of S. litura.
Open Access
CROP LOSSES BY BLISTER BEETLE (Mylabris pustulata Thunberg) (Coleoptera: Meloidae) ON PIGEONPEA AND MUNGBEAN AND ITS MANAGEMENT
(PAU Ludhiana, 2012) Gurjeet Singh; Ravinder Singh
Studies on seasonal incidence, crop losses, host preference and management of blister beetle (Mylabris pustulata Thunberg) (Coleoptera: Meloidae) on pigeonpea and mungbean were carried out at Punjab Agricultural University, Ludhiana during Kharif 2010 -11. The crop losses studies were carried out under net house conditions as well as under field conditions while the management studies were made under field conditions. The experiments on seasonal abundance revealed that blister beetle started appearing in the 4th week of August with peak activity period during the 4th week of September in pigeonpea and mungbean under Punjab conditions. The blister beetles were more active during morning and evening hours on both the crops. The activity during noon hours was relatively low. Studies on crop losses under field conditions showed that blister beetle caused up to 36.20 per cent damage to flowers and 53.22 per cent losses in yield at 1.96 beetles per plant in pigeonpea and 15.80 per cent damage to flowers and 35.90 per cent losses in yield at 1.3 beetles per plant in mungbean crop. The investigations on crop losses under net house conditions revealed that M. pustulata caused maximum reduction up to 54.18 per cent in pod setting, 20.15 per cent in seed setting and 65.00 per cent in grain yield @ 2 beetles per plant in pigeonpea. However, in mungbean crop, M. pustulata caused maximum reduction up to 67.14 per cent in pod setting, 26.65 per cent seed setting and 75.29 per cent in grain yield @ 4 beetles per plant. Host preference studies carried out under laboratory conditions revealed that M. pustulata showed highest preference towards pigeonpea (87.22 per cent flowers eaten), followed by urdbean (43.52 per cent flowers eaten) and mungbean (30.38 per cent flowers eaten). Amongst varieties, pigeonpea variety AL 201, mungbean variety ML 818 and urdbean variety Mash 338 were more preferred than other varieties of these pulse crops. The studies on management of M. pustulata showed that deltamethrin 2.8 EC @ 500 ml ha-1 > cypermethrin 25 EC @ 250 ml ha-1 > chlorpyriphos 20 EC @ 3750 ml ha-1 were the most effective insecticides among all the treatments in both pigeonpea and mungbean.
Open Access
TOXICITY OF DIFFERENT INSECTICIDES AGAINST COTTON MEALY BUG, Phenacoccus solenopsis Tinsley
(PAU Ludhiana, 2010) Amit Punj; A. K., Dhawan
The studies on the evaluation of LC50 of insecticides against mealy bug, Phenacoccus solenopsis population collected from cotton belt during vegetative, flowering and maturity phase were conducted at Entomological Farm, PAU, Ludhiana. The new molecule, spirotetramat was found to be most toxic to first instar, third instar nymphs and adult females. Among the conventional insecticides, profenophos and thiodicarb was proved to be the most toxic. The order of toxicity was spirotetramat > clothianidin > profenophos > thiodicarb > buprofezin > chlorpyriphos > acephate. In another experiment, the persistence of different insecticides at recommended concentrations against first instar, third instar nymphs and adult females was estimated. Based on the index of persistence toxicity, the order of effectiveness for the first instar nymphs of mealy bug was profenophos (645.40) > thiodicarb (619.60) > buprofezin (572.52) > spirotetramat (430.00) > acephate (400.17) > clothianidin (376.83) > chlorpyriphos (330.17). Similarly, for third instar nymphs the order of effectiveness was profenophos (589.18) > thiodicarb (562.93) > buprofezin (548.76) > spirotetramat (395.01) > acephate (384.18) > clothianidin (359.24) > chlorpyriphos (314.53). Similarly, for the adult females the order of effectiveness was profenophos (523.23) > buprofezin (478.77) > thiodicarb (435.13) > spirotetramat (330.10) > acephate (305.00) > clothianidin (290.34) > chlorpyriphos (260.19). The third experiment was conducted to study the effect of host of mealy bug like bitterweed, cocklebur and carpet weed on the efficacy of promising insecticides. Based on LC50 values, profenophos was found to be extremely toxic to first instar, third instar nymphs and adult females of mealy bug on the above mentioned weed hosts in comparison to thiodicarb.
Open Access
EVALUATION OF NAPIER GRASS (Pennisetum sp.) AS A TRAP CROP IN THE MANAGEMENT OF Chilo partellus (Swinhoe) ON MAIZE
(PAU Ludhiana, 2010) Bhupinder Singh Sidhu; Uma, Kanta
The first experiment on the differential development and survival of Chilo partellus on different genotypes of Napier grass viz; Copricon, K-56351, K-56379 and M-285303, conditions at 27.08 ± 2.35 °C and 75.91 ± 5.83 per cent relative humidity. After 4 days of larval release on different genotypes of Napier grass, Napier millet and PMH 1, the larval survival of C. partellus among different treatments was maximum(73.33%) on PBN-342. The larvae could not survive on M-285303, K-56351 and K-56379 after 19 days of their release. After 21 days, the larvae could survive only on Copricon and maize (PMH 1). The developmental period from hatching to adult formation on Copricon and maize was 32.48 and 36.01 days, respectively and survival was 4.0 and 34.0 per cent, respectively. From these studies, it was inferred that different genotypes of Napier grass and Napier millet were not suitable for the development and survival of C.partellus. The second experiment was conducted on the ovipositional preference of female moth of C. partellus for different genotypes of Napier grass, Napier millet and PMH 1. The female moths laid significantly more number of egg clusters per plant in all the treatments in comparison to maize. The genotypes of Napier grass viz; Copricon, M-285303, K-56379, K-56351 and Napier millet, i.e. PBN-233 and PBN-342 received more number of eggs than maize showing that these genotypes were preferred than maize for oviposition. The third experiment was conducted to evaluate different genotypes of Napier grass and Napier millet as trap crop against C. partellus. The maize plots with two rows of all the different genotypes of Napier grass and Napier millet as trap crop on either side of plot suffered significantly less leaf injury and mean damage grade (range- 14.63 to 17.56% and 3.0 to 4.0) than the plots (37.85 to 45.33% and 5.0 to 7.0 m.d.g) having treated maize and maize without any treatment and resulted in obtaining more maize yield in plots having two border rows of genotypes of Napier grass and Napier millet over the treatments with treated maize and maize without any border row or treatment.
Open Access
Incidence of insect pests and management of shoot fly, Atherigona spp. in spring sown maize.
(PAU Ludhiana, 2013) Jawala Jindal; Dulcha S., Brar
The studies on the incidence of insect pests and management of shoot fly, Atherigona spp. in spring sown maize were conducted during 2011 and 2012 in the Punjab. At Ludhiana and Hoshiarpur, shoot fly was recorded as the key pest of this crop, while the incidence of other insect pests of maize was of minor importance. At Ludhiana, the small brown planthopper, Laodelphax striatellus (Fallén) was observed for the first time on the crop and its highest counts were observed in 3rd (96.06 to 186.08/ sweep) in 2011 and 4th week of April (0.16 to 1.49/ plant) in 2012. The predatory coccinellids were also observed in correspondence to the higher population of L. striatellus during 2011. The maximum egg counts of Atherigona spp. were observed at 10 DAG (0.19 to 0.52 eggs/ plant ) in 2011 and at 17, 7 and 12 DAG (0.15 to 0.79 eggs/ plant ) in end January, mid and end February sown crops, respectively during 2012. The deadhearts due to shoot fly damage were more in end February (11.58 & 17.72 %) followed by mid February (9.73 & 16.85 %) and least in end January sown crop (5.79 & 8.98 %) with grain yield loss of 27.93 & 37.97, 38.21 & 38.66; and 38.99 & 45.04 per cent, respectively during 2011 and 2012. Based on deadhearts incidence, the genotypes JH 3459 (10.49 %) & JH 3956 (10.95 %); CM 143 (15.16 %), PMH 1 (15.35 %) & JH 31244 (16.70 %); and LM 16 (21.48 %), PMH 2 (21.52 %) & Parkash (23.57 %) were found to be least, intermediate and highly susceptible to shoot fly, Atherigona naqvii Steyskal. The proportion of deadhearts out of total shoot fly incidence was minimum in JH 3459 & JH 3956.Whereas LM 16, PMH 2 & Parkash had lower larval & pupal periods; and higher larval survival, male & female pupal weight & fecundity. Thus, revealed the presence of tolerance and antibiosis mechanism of plant resistance in test genotypes against A. naqvii. The more leaf length, leaf width, stem girth and reducing sugars content were found related to susceptibility in the test genotypes and thus, could be useful in screening of maize germplasms against A. naqvii. The seed treatment with imidacloprid 600 FS @ 6 ml/ kg seed was found to be highly effective for the management of shoot fly in the spring sown maize.
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VIRULENCE OF SYMBIOTIC BACTERIA ASSOCIATED WITH ENTOMOPATHOGENIC NEMATODES FOR INSECT PEST MANAGEMENT
(PAU Ludhiana, 2013) Rajwinder Kaur; Vikas Jindal
The studies on the virulence of symbiotic bacteria associated with EPNs were carried out on five EPN strains (N-12, N-13, N-14, N-15 & N-17) isolated form Punjab soils. Bacterial isolates (1, 2, 3, 4 & 5) from local EPN strains produced brick red coloured colonies and were non-bioluminescent. The infective juveniles (IJs) of each EPN strain developed into adults within 72 hours, when fed on respective bacterial isolates. This confirmed the association between bacterial isolates and respective EPN strains. The dendrogram developed based on RAPD-PCR analysis established that isolates 2, 4 and 5 are very closely related (94% similarity). However, isolates 1 and 3 (65% similar) are different from other three isolates. Based on sequence homology of 16S rDNA region of all bacterial isolates, the bacterial isolate 1 was identified as Delftia acidovorans, isolate 2 as Brucella spp., isolate 3 as Pectobacterium cypripedii, isolates 4 and 5 as Chryseobacterium indologenes. As all the bacterial isolates were different from most commonly occurring symbiotic bacteria (Xenorhabdus and Photorhabdus) with EPNs, thus four local EPN strains were identified based on ITS (internal transcribed spacer) region. All local EPN strains showed a maximum identity with new EPN genera viz. Heterorhabditidoides sp. RG081015 (88% similarity) followed by Rhabditis sp. Tumian-2007 and Heterorhabditidoides chongmingensis. This is the first record of a new species/genera of EPN isolated from India, which is very closely related to Heterorhabditidoides based on ITS region. In the bioassay experiment, none of the bacterial isolates was found effective for management of tobacco caterpillar, Spodoptera litura (Fabricius).
Open Access
Persistence and Metabolism of Imidacloprid in rice (Oryza sativa Linn.)
(PAU, 2014) Akoijam, Romila; Singh, Balwinder
The persistence and metabolism of imidacloprid in rice was studied by following application of imidacloprid @ 20 and 80 g a.i. ha-1. Samples of paddy leaves and soil were collected from each treatment at varying time intervals and analyzed by High Performance Liquid Chromatograph (HPLC). The initial deposits of imidacloprid were found to be 4.57 and 13.94 mg kg-1 in leaves collected 0 day after its application @ 20 and 80 g a.i. ha-1 respectively. The residues declined to below the determination limit of 0.01 mg kg-1 in samples collected 60 and 90 days after its application at recommended and four times the recommended dosages, respectively. The samples of rice grains, bran, straw and husk did not reveal the presence of imidacloprid residues at harvest. In soil samples, total residues of imidacloprid and its etabolites were found to be 0.44 and 1.61 mg kg-1 collected at 0 day following application @ 20 and 80 g a.i. ha-1 respectively. These residues could not be detected at 60 and 90 days after its application at corresponding dosages. In laboratory conditions, the levels of total imidacloprid in sandy loam, loamy sand and clay loam at 0 day of its application @ 100 mg kg-1 were found to be 80.01, 83.64 and 89.00 mg kg-1, respectively. The persistence of imidacloprid was found to be more in clay loam followed by loamy sand and sandy loam soil. In sterilized soil, Bacillus aerophilus was found to be more efficient as compared to Bacillus alkalinitrilicus in degradation of imidacloprid.

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