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Open Access
Study of heavy metals toxicity on biomolecules composition, antioxidant system and genotoxicity in Eudrilus eugeniae (Kinberg, 1867)
(CCSHAU, 2019) Shefali; Gupta, R.K.
The present study was carried out to analyze the toxicity of arsenic and chromium on E. eugeniae. Paper contact toxicity test was used to determine LC50. LC50 of arsenic and chromium against adult E. eugeniae was 2.09% and 0.13%, respectively that confirms the higher toxicity of chromium as compared to arsenic. The biomolecule composition and antioxidant response in the earthworms, E. eugeniae, exposed to heavy metals were investigated. The carbohydrate, lipid and protein content were decreased with exposure to heavy metals. The changes in the levels of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were used as functional biomarkers to determine the effect of metals on earthworms. DNA damages were evaluated on the earthworm’s coelomocytes using the comet assay. After an exposure into soils, arsenic and chromium revealed to be genotoxic at higher doses after 28 d of exposure.
Open Access
Molecular characterization and management of Tyrophagus putrescentiae (Schrank) in button mushroom, Agaricus bisporus (Lange)
(CCSHAU, 2018) Arvind; Gulati, Rachna
Molecular characterization of acaricide susceptible, resistant and untreated Tyrophagus putrescentiae on mushroom clusters having a similarity coefficient of 0.45. Cluster one was represented by untreated (control) and resistant population sharing a similarity coefficient of 0.68 where as the cluster 2 was represented by susceptible population alone. Preliminary primers screening produced 22 polymorphic bands (70.96% polymorphism) and 9 monomorphic bands (29.03%). Methanolic seed and leaf extracts of P. pinnata showed acaricidal activity against T. putrescentiae. In Direct spray and Treated arena bioassay method, maximum mortality of 82, 76 % and 75.33, 63.33% was observed at 1.0 % concentration of seed and leaf extract, respectively. Both the extracts were equally effective against T. putrescentiae when direct spray on mites was done. The mortality was 59.63 and 58.15 percent in seed and leaf extract spray, respectively. Under Treated arena bioassay, seed extract was statistically more effective against T. putrescentiae (52.22 % mortality) as compared to leaf extract (43.56 % mortality). The LC50 value in Direct spray Treated arena bioassay method was 0.77, 0.63 and 0.85 and 0.64% in leaf and seed extract of P. pinnata. Among the solvents, hexane was most potent as 8, 5.66 and 0.33 mites were recovered from 0.5, 1 and 1.5 percent concentration after 36 h as compared to 38.66 mites in control. Bioefficacy studies conducted in compost bags under direct spray showed that among the treatments, Propargite (0.05%) significantly was more effective in reducing T. putrescentiae number (0.78 mites) than P. pinnata leaf extract at 0.752% (30 mites), 0.852% (18.89 mites) and 0.952% (12.22 mites) concentrations. P. pinnata seed extract at 0. 738% reduced the population to 5.88 mites which was at par with Propargite treatment.The role of T. putrescentiae as vector of dry bubble and green mould disease caused by fungus Verticillium fungicola and Trichoderma harzianum, respectively was also ascertained during the present study. V. fungicola reared T. putrescentiae population on Agaricus bisporus spread the disease within 8 days culture plates and fruiting bodies. Similarly, T. harzianum reared T. putrescentiae population on Agaricus bisporus spread the green mould disease to culture plates and fruiting bodies.A peak in population (47.40, and 60.80 mites) on V. fungicola and T. harzianum was recorded after 12 and 14 days, respectively which was statistically higher than the mites recorded at other observation periods.
Open Access
Comparative impact of insecticides on bacterial and acetylcholinesterase activity of Eisenia fetida (Savigny) and Pheretima posthuma (Kinberg)
(CCSHAU, 2017) Yadav, Jyoti; Dharambir Singh
The present study was carried out to analyze the toxicity of chlorpyrifos and carbofuran on E. fetida and P. posthuma. Paper contact toxicity test was used to determine LC50. LC50 of chlorpyrifos against adult E. fetida and P. posthuma was 0.25% and 0.11%, respectively. While LC50 of carbofuran against adult E. fetida and P. posthuma was 5.14% and 4.11%, respectively that confirms the greater sensitivity of P. posthuma towards insecticide induced toxicity in comparison to E. fetida. Lower LC50 value of chlorpyrifos confirms its higher toxicity as compared with carbofuran. Total 52 bacterial isolates were isolated from the gut of worms, out of which six resistant gut bacterial species having agricultural importance were identified as Sphingobacterium sp., Citrobacter freundii, Citrobacter murliniae, Bacillus australimaris and Burkholderia paludis. Inhibition in AChE activity due to chlorpyrifos and carbofuran exposure was observed at 24h. While E. fetida exposed to carbofuran tried to maintain AChE level, which may be seen as strategy to cope up with toxicity after prolonged exposure.
Open Access
Evaluation of management practices against Polyphagotarsonemus latus (Banks) and its role as vector of leaf curl virus in chilli
(CCSHAU, 2018) Monika; Gulati, Rachna
Under in vitro 35.00 to 67.27 and 33.89 to 63.89 percent mortality of P. latus was recorded in Direct Spray Bioassay, with 2.5 to 10.0 % and 1.25 to 10.0 % seed and leaf extract concentrations of P. pinnata, respectively. LC50 values of methanolic seed and leaf extract of P. pinnata were 5.36 and 5.31, respectively for mobile stages of P. latus. Under field conditions, the percent reduction in P. latus was from 40.60 to 72.55 percent with all the treatments concentration of P. pinnata. Under in vivo P. pinnata seed extract 10.0% (72.55%), was the most potent in reducing the mite population followed by P. pinnata leaf extract 10.0% (65.10 %), P. pinnata seed extract 7.5 % (62.66 %), P. pinnata seed extract 5.4 % (55.49%), P. pinnata leaf extract 7.5 % (54.98 %), P. pinnata seed extract 5.0 % (48.33%), P. pinnata leaf extract 5.3 % (47.58%), P. pinnata leaf extract 5.0 % (40.16%) and Pyriproxyfen 5% + Fenpropathrin 15% EC (1ml/l H2O) (37. 83%), respectively. Studies on seasonal incidence of P. latus revealed a peak in its population on chilli during second fortnight of October during the respective crop seasons in both the years under field conditions. Strata wise, top strata harboured significantly more number of mites followed by middle and bottom strata. A comparison of mulch treatments showed that Organic mulch 12t/ha was most effective in reduction of P. latus population (79.30 %), followed by Black/Silver polythene sheet (78.50 %), Black/White polythene sheet (78.30 %), Black polythene sheet (77.50 %), Organic mulch 9t/ha (74.70 %) and Organic mulch 6t/ha (65.30 %) treatment as compared to control. In terms of percent increase in yield, Organic mulch 12t/ha caused highest increase (57.90 % over control) followed by Black polythene sheet mulch, Black/Silver polythene sheet mulch, Black/White polythene sheet mulch, Organic mulch 9t/ha and Organic mulch 6t/ha showing 47.49, 46.72, 45.17, 42.47 and 34.36 percent increase over control. P. latus showed non-transmissible nature of virus. Thus it indicated that mite is not a carrier vector for transmission of leaf curl virus into chilli plant.
Open Access
Molecular characterization of Beauveria bassiana (Balsamo) vuillemin and its bioefficacy against Tetranychus urticae Koch (Acari: Tetranychidae)
(CCSHAU, 2011) Geroh, Monika; Gulati, Rachna
Investigations were carried out to assess the pathogenicity of Beauveria bassiana against Tetranychus urticae on okra. Molecular characterization of six strains of B. bassiana was done by RAPD- PCR to know the genetic similarity among them. The yield of genomic DNA of B. bassiana was higher in Lysis Buffer method (2.10 to 10.50μg/ ml) as compared to CTAB method (2.34 to 7.10μg/ ml). Absorbance ratio of all strains varied from 1.60 to 2.20. Eleven random primers generated 19 (82.16%) polymorphic and 4 (17.39%) monomorphic bands. The size of PCR amplified products ranged from 500 to 4000 bp. The genetic similarity of the strains varied from 50 to 70 percent. Under in vitro 42 to 94 and 34.1 to 66.25 percent mortality of T. urticae was recorded in Direct Spray and Treated Food Bioassay, respectively with 1 × 105 to 1 × 1012 conidia ml-1 conecntrations of B. bassiana. LC50 values of B. bassiana were in the range of 3 × 105 to 2.6 × 1011 conidia ml-1 and LT50 values varied from 52.12 to 141.53 h for eggs, larvae, nymphs and adults in both the bioassay methods. Direct Spray Bioassay was more effective as compared to Treated Food Bioassay. Under field conditions, the percent reduction in T. urticae was from 51.03 to 65.18 percent with all the strains. Hb- Hyderabad (65.18%), was the most potent in reducing the mite population followed by ITCC- 4668 (62.36%), ITCC- 5408 (58.55%), ITCC- 6063 (54.61%), ITCC- 5549 (52.48%) and ITCC- 4513 (51.03%). There was no effect of B. bassiana spray S. punctillum population as their numbers in B. bassiana (strain Hb- Hyderabad) treatments (1.43 beetles/ leaf) and control (1.17 beetles/ leaf) were statistically comparable. Higher yield of okra fruits was recorded in B. bassiana treated plots (581.43 g/ plot) as compared to untreated plots (577.66 g/ plot). The Chlorophyll content was also higher in B. bassiana treated leaves (0.289 to 0.492 μg/ ml) as compared to untreated plots (0.166 μg/ ml).
Open Access
Studies on the pollination mechanisms in different varieties of plum (Prunus domestica L.)
(CCSHAU, 2011) Nidhi; Sihag, R.C.
Insects are associated with flowering plants since their evolution on the earth. While some of them act as their pests, others are either pollinators or parasites and predators of the pests. Although a variety of insects frequent the entomophilous (insect pollinated) crop blossoms for their food rewards (nectar and pollen), only bees belonging to the genus Apis outnumber others and are efficient pollinators. Plums are richest sourse of antioxidants, they provide protection against diseases. The study was made on floral biology and phenology, diversity, relative abundance and pollinating efficiencies of insects visiting the blossoms of three different varieties of plum and the role of different modes of pollination in fruit set of the three varities of plum. Flowering habit of all the three varieties of this study viz. Alu Bokhara, Titron and Kala-Amritsari was similar. Floral structure was studied under dissecting microscope revealed the presence of perigynous condition in the flowers of plum. Abundance of insect visitors studied at hourly and weekly intervals which was maximum during peak flowering period of this plant. Twelve insect species visited the crop of plum; out of these five species i.e. Apis dorsata, A.mellifera, A.florea, A.cerana and Dipterans were the main pollinators.Among the hymenopterous insects, Apis.mellifera visited maximum number of flower per minute, followed by A.cerana, A.dorsata, A.florea and Dipterans Among all the insect visitors, Apis dorsata remained active for the longest duration on plum flower followed by, A.mellifera, A.cerana A.florea and Dipterans respectively. Out of the three varieties Titron was the most prefered variety by the insect visitors followed by Kala-Amritsari and Alu-Bokhara.Pollinating efficiencies of the pollinators were derived on the basis of their abundance, foraging rate, mean activity duration and loose pollen grains on the body of insect. Among the five insect pollinators Apis dorsata came out to be on the first place and seemed to be the best pollinator of this crop .Other pollinators with relatively lower ranking in descending order of pollinating efficiency were A.mellifera, A.cerana A.florea.Studies on fruit set parameters revealed that plum flowers were highly benefited from insect visits, and insect pollination greatly enhanced its fruit set. Crop without insects produced very low fruit set. It was further revealed that plum is a cross-pollinated and entomophilous plant.The structure shape and fertility status of the flower clearly revealed that it has honeybee pollinated flowers and mellitophily pollination syndrome existed in plum.
Open Access
Studies on the pollination mechanisms in different varieties of amla (Emblica officinalis Gaertn.)
(CCSHAU, 2011) Saini, Reena; Sihag, R.C
Amla (Emblica officinalis Gaertn.) is an important minor fruit crop of India from the family Euphorbiaceae. It is known for its highly nutritious and tremendous medicinal value. The present study was made on the floral biology and pollination ecology of this plant. Three varieties of amla viz. Chakaiya, NA-7 and NA-10 were selected for this study. Floral structure of the three varieties revealed that amla consisted of unisexual, minute flowers with monoadelphous stamen and a hypogynous ovary. The sex ratio varied highly in three varieties and was 58.95:1 (male:female flowers) for NA-7, 60.33:1 for Chakaiya and 88.21:1 for NA-10. All the three varieties of amla blossomed during March- April in 2009 and 2010 and the flowering period persisted for a month. Twelve insect species from different orders visited the flowers of amla on all the three varieties in 2009 and 2010 and the major insect visitors included the hymenopterans namely Apis dorsata, Apis mellifera and Apis florea and a dipteran Sarcophaga. The rest of the insect visitors which included a wasp (Polistes hebraeus), a bee (Halictus sp.), flies (Eristalis sp., Episyrphus sp., Syrphus sp., Syritta sp.) and a butterfly (Psichotoe duvauceli) were clubbed under “other insect pollinators.” Abundance of insect visitors was studied at one and a half hour and weekly intervals and it was maximum during peak flowering period of this plant. The insect visitors predominately made more visits on male flowers than on female flowers as the male flowers were the richest source of pollen and the nectar was scarce in female flowers. Apis dorsata had the highest foraging rate followed by Apis mellifera, Apis florea, Sarcophaga sp. and other insect pollinators. Among all the insect visitors, Apis dorsata remained active for the longest duration on amla flower in all the three varieties. The abundance and the activity duration of insect visitors was maximum on NA-7 followed by Chakaiya and NA-10. Pollinating efficiencies of the pollinators were derived on the basis of their abundance, foraging rate, mean activity duration and loose pollen grains on the bodies of different insect pollinators. Among the five insect pollinators Apis dorsata was found to be the most efficient pollinator of this crop followed by Apis mellifera, Apis florea, Sarcophaga sp. and other insect pollinators on all the three varieties in 2009 and 2010. Studies on yield parameters revealed that amla flowers were highly benefited from insect visits and insect pollination greatly enhanced its fruit yield. It was further revealed that amla is a cross-pollinated and entomophilous plant.
Open Access
Isolation and identification of associated proteins/allergens in mite, Tyrophagus putrescentiae (Schrank) and its management
(CCSHAU, 2016) Anita; Gulati, Rachna
Investigations were carried out to assess the incidence of Tyrophagus putrescentiae in granaries, flour mills and domestic storage. In granaries, first peak in the month of October (12.33 mites/50g residual dust) and in flour mills and domestic storage, it was recorded in the month of November (30.06, 18.06 mites/50g wheat flour). At all the sites, second peak was recorded in the month of May (23.40, 33.26 and25.13 mites/50 g sample, respectively). Significantly more number of mites (19.00, 17.00 mites/50g grains) were recorded in open type storage (wheat grains stored on the floor of a room) than in metallic bins. Mite incidence showed significant positive correlation with maximum temperature, sunshine hours and negative correlation with evening relative humidity in granaries, flour mills and domestic storage structures. In the latter two survey sites, significant negative correlation between mite incidence and morning relative humidity was also observed. Significantly higher number of mites were recorded in flour mills (24.83 mites/ 50g sample) followed by domestic storage structures (15.84 mites/50g sample) and granaries (9.56 mites/50g sample). Maximum number of mites (64.72 mites/50g sample) were recorded during summer season (April- June), followed by winter (60.26 mites/50g sample; October- January), rainy (44.93 mites/50g sample; July- September) and spring (30.98 mites/50g sample; February-March) season. Seven unknown protein of 90 kDa, 87 kDa, 68 kDa, 50 kDa, 36 kDa, 26 kDa and 16 kDa molecular weight were isolated from T. putrescentiae body. wheat flour infested with T. putrescentiae showed a significant higher count of protein, fat acidity, free fatty acids, peroxide value whereas total antioxidant activity decreased in infested flour. The pathogenicity of Beauveria bassiana against T. putrescentiae was evaluated. Under in vitro 20 to 80.5 and 17 to 74.5 percent mortality of T. putrescentiae was recorded in Direct Spray and Treated Observation arena Bioassay, respectively with 1 × 105 to 1 × 1012 conidia ml-1 concentrations of B. bassiana. LC50 values of B. bassiana were in the range of 1.83×109 to 4.45× 1011 conidia ml-1 and LT50 values varied from 1.05 to 48.17 h for eggs, larvae, nymphs and adults in both the bioassay methods. Direct Spray Bioassay was more effective as compared to Treated Food Bioassay.
Open Access
Management of Tyrophagus putrescentiae Schrank (Acari: Acaridae) in Oyster mushroom, Pluerotus sajor-caju and its molecular characterization
(CCSHAU, 2017) Komal; Gulati, Rachna
The studies on population buildup of Tyrophagus putrescentiae on compost and fruiting body of Pleurotus sajor-caju revealed that irrespective of initial inoculums level mite number decreased significantly at each observation period. No mites were recorded after 24, 24, 30 days and 24, 34, 38days on wheat straw and wheat straw plus calcium sulphate based compost at initial inoculums level of 10, 20 and 30 T. putrescentiae pairs, respectively. The weight loss of the compost was 0 to 10.3 percent in these levels. Percent loss in total sugar, reducing sugar, non reducing sugar, starch, protein content of compost was in the range of 13.3 to 52.1%, 11.3 to 27%, 1.2 to 4.07%, 14.2 to 35.7%, 45 to 71%, respectively. In fruiting bodies, a peak in population (41.66, 59.00, 83.00 mites/10g fruiting body) was recorded at initial inoculums level of 10, 20 and 30 T. putrescentiae pairs. Weight loss of the fruiting body was 0.00 to 20.5 percent in these levels. Significant reduction in total sugar, reducing sugar, non reducing sugar, starch, protein content of fruiting bodies was recorded. It was 0.134, 0.055, 0.070, 0.010, 0.279 mg/10g fruiting body at 30 mite pairs as compared to 0.162, 0.075, 0.099, 0.029, 0.741mg/10g fruiting body in control. Percent reduction in T. putrescentiae population with Withania somnifera, Azadirachta indica, neem oil and neem cake was 82.20 to 100, 43.64 to 100, 23.00 to 100 and 25.60 to 100 percent after 45 days post-treatment. Higher concentrations (2 and 1%) were significantly more effective as these showed no T. putrescentiae counts as compared to initial pre treatment count (50 mite pairs). Molecular characterization of four T. putrescentiae populations was done by RAPD- PCR to know the genetic similarity among them. The yield of DNA from T. putrescentiae population collected from soil samples of horticulture, forestry plantations and vegetable crops and wheat flour ranged from 876to 2000 μg/ml. A total of 208 bands were generated for T. putrescentiae where 204 bands (98.07%) were polymorphic and 4 1.92%) bands were monomorphic. Among the four populations range of polymorphism was 87.5 to 100 percent. The T. putrescentiae populations collected from soils of vegetable and forestry plantations were found to be the most diverse with a minimum similarity value of 0.475.

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