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Dr. Y. S. Parmar University of Horticulture & Forestry, Solan
Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, was established on 1st December, 1985 with the objective to promote education, research and extension education in the fields of Horticulture, Forestry and allied disciplines. Late Dr. Yashwant Singh Parmar, the first Chief Minister and the architect of Himachal Pradesh perceived the importance of Horticulture and Forestry to develop and improve the State economy which led to the establishment of this University. Its history lies in erstwhile Himachal Agricultural College, Solan, established in 1962 and affiliated to the Panjab University. It became one of the campuses of Agriculture Complex of Himachal Pradesh University on its formation in 1970. Consequent upon the establishment of Himachal Pradesh Krishi Vishvavidyalaya in 1978, this campus became its Horticulture Complex and finally in 1985, assumed the status of a State University, being the only University in the country engaged exclusively in teaching, research and extension in Horticulture and Forestry.
The University is located at Nauni in Solan District of Himachal Pradesh, 13 km from Solan on Solan-Rajgarh Road, at an elevation of 1300 metres above mean sea level. Solan town is situated on national highway (NH-22) and is well connected by train and bus services.
The University has four constituent colleges, out of which, two are located at the main campus Nauni, one for horticulture and the other for forestry, having 9 and 7 departments, respectively. The third College i.e., College of Horticulture & Forestry is located at Neri in Hamirpur District on Nadaun-Hamirpur state highway, about 6 Km from Hamirpur town and is well connected with bus service. The college offers three Undergraduate Degree Programmes i.e. BSc (Hons.) Horticulture, BSc (Hons.) Forestry and B. Tech. Biotechnology and MSc degree programme in a few subjects. The fourth college i.e. College of Horticulture and Forestry, Thunag (Mandi) is located at Thunag District Mandi. This college offer BSc (Hons.) Horticulture and BSc (Hons.) Forestry degree programme. In addition, there are five Regional Research Stations, 12 Satellite Stations and five Krishi Vigyan Kendras (KVKs) situated in different zones of the State.
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ThesisItem Open Access Bioprospecting of thermophilic bacteria from hot water springs of Himachal Pradesh for laccase enzyme production(YSPU, 2015) Sharma, Ruchika; Shirkot, PoonamLaccase enzyme has acquired the status of ‘green catalyst’ as it possesses remarkable bioremediation potential along with numerous applications in effluent detoxification, degradation of textile dyes, herbicide and insecticide degradation, wine clarification, enzymatic conversion of chemical intermediates, biosensors and organic synthesis. In the present study, significantly high diversity of laccase producing bacteria from hot water springs of Himachal Pradesh was assessed. A total of 118 laccase producing thermophilic bacterial isolates were obtained from 200 hot water spring samples using TYM containing 5 mM guaiacol which were morphologically characterized. These were rescreened on the basis of their ability to oxidise tannic acid, dimethoxyphenol and syringaldazine leading to selection of 50 laccase producing thermophilic bacterial isolates, which were characterized biochemically. Eighteen thermophilic bacterial isolates exhibiting maximum laccase activity of 0.0007-0.0038 U/l were selected for further, molecular characterization using RAPD-PCR and 16S rrna gene technology. In silico analysis of 16S rrna gene sequences led to identification of these bacterial isolates and they were found to belong to genus Bacillus, Aneurinibacillus and Pseudomonas, as Bacillus licheniformis strain RSV20, Bacillus licheniformis strain RSM8, Bacillus licheniformis strain RSV10, Bacillus sonerensis strain RSM17, Bacillus sonerensis strain RSV8, Bacillus licheniformis strain RSP1, Bacillus licheniformis strain RSP2, Bacillus licheniformis strain RSP3, Bacillus licheniformis strain RSP7, Bacillus sonerensis strain RSP5, Bacillus sonerensis strain RSP11, Aneurinibacillus thermophilus strain RSP13, Bacillus aerius strain RSP4, Bacillus aerius strain RSP9, Bacillus subtilis strain RSP8, Bacillus amyloliquifacience strain RSP10, Bacillus pumilis strain RSP12 and Pseudomonas taiwanensis strain RSP6. On the basis of maximum laccase enzyme activity Bacillus licheniformis strain RSM8 was selected for production and purification of the laccase enzyme. Maximum extracellular enzyme production was achieved at 60°C, pH 9.0 and 24 hrs incubation with 5 mM guaiacol, 5 % tryptone and 3 % yeast extract in combination with nitrogen source. Crude extracellular thermolaccase enzyme preparation was purified by ammonium salt precipitation (50-90%) followed by gel filtration and ion exchange chromatography which showed 15.21 yield and 10.5 fold purification. The purified enzyme had optimal activity at pH 9.0 and 60 °C, and 16.22 μM Km value. The molecular weight of thermolaccase in the present study was found to be 72.5 kDa. However activity was inhibited by sodium azide and DTT. Bacillus licheniformis strain RSM8 as well as its enzyme preparations were investigated for their ability to decolourize dyes which are the potential contributors of water pollution. Six different synthetic dyes were decolourized RBBR (68 %), congo red (86 %), indigo carmine (73 %), brilliant blue (40 %), bromophenol blue (51 %) and aniline blue (54 %) when treated with the crude enzyme preparation of Bacillus licheniformis strain RSM8. And partially purified enzyme preparation of Bacillus licheniformis strain RSM8 showed greater decolourization of dyes comparatively RBBR (74 %), congo red (91 %), indigo carmine (80 %), brilliant blue (60 %), bromophenol blue (64 %) and aniline blue (67 %). The purified enzyme was successfully immobolized using adsorption method in calcium alginate beads with 76% immobolization percentage and immobolized laccase enzyme beads were studied for their ability to degrade dyes. The stability and reusability of the immobilized enzyme system has the potential to make the entire treatment process inexpensive. Bacillus licheniformis strain RSM8 enzyme preparations was investigated for phytotoxicity evaluation of three dyes viz., Congo red, RBBR and Indigo carmine and each of enzyme treated dyes for Phaseolus mungo and Calendula officinalis and Tagetes patula plant species respectively, under in vitro conditions and Phaseolus mungo with Congo red dye under in vivo conditions. Significant germination inhibition, a slower rate of plumule and radicle seedlings growth was observed for Congo red, RBBR and Indigo carmine dyes as compared to enzyme treated dyes. An extracellular laccase producing gene has been isolated using degenerate primer based on the copper I and II conserved site of laccase enzyme, from the hot water spring bacteria, Bacillus licheniformis strain RSM8 followed by determination of the amino acid which were translated from nucleotide sequence and encodes a polypeptide comprised of 50 amino acids showeng 97 % identity with the amino acid sequences of bacterial laccases i.e. copper oxidase [Bacillus licheniformis]. Further multiple sequence alignment using MULTALIN and structure prediction using Phyre1 & 2 revealed conserved histidine residues.ThesisItem Open Access Bioprospecting of chlorpyrifos degrading bacteria for bioremediation of apple orchard soils of Himachal Pradesh(YSPU, 2015) Sharma, Anish Kumar; Shirkot, PoonamOrganophosphorus compounds are the most widely used insecticides, accounting for an estimated 34% of worldwide insecticide sales. Chlorpyrifos is a non-systemic organophosphorus insecticide, with broad spectrum insecticidal activity. Contamination of soil from pesticides as a result of their bulk handling at the farmyard or following application in the field or accidental release may lead occasionally to contamination of surface and ground water. The present study was carried out to explore the status of indigenous chlorpyrifos degrading bacteria in the apple orchard soils in the state of Himachal Pradesh, which harbours plenty of diverse forms of microbial life, which has remained largely unexplored till date. The pH and electrical conductivity (EC) of the thirty soil samples from apple orchards were recorded and ranged from 6.55- 7.58 and 0.24-0.39 dSm-1 respectively. The percent organic carbon (% OC), nitrogen (N), phosphorus (P) and potassium (K) content ranged from 0.66-0.116%, 302-486 Kg/ha, 12.8-23.6 Kg/ha and 255-371 Kg/ha respectively. A total of 72 bacterial isolates were isolated from 30 soil samples using enrichment technique and mineral salt medium. These 72 isolates were rescreened on selective medium i.e. eosin-methylene blue agar (EMBA) containing 50 mg/l chlorpyrifos and only 15 isolates were able to change the colour of colonies from creamish/yellowish/whitish/transparent to red, which was observed as an indication of chlorpyrifos degradative ability of these bacterial isolates. These 15 bacterial isolates were characterized biochemically alongwith organophosphorus hydrolase activity and 6 bacterial isolates exhibiting highest extracellular OPH activity of 0.012-0.165 U/ml were selected. Molecular characterization of selected isolates was carried out using RAPD-PCR and 16S rrna gene technology. Only 5 bacterial isolates were amplified by universal primers and in silico analysis of 16S rrna gene sequence led to identification of these bacterial isolates as Pseudomonas indoloxydans ASK3.2, Pseudomonas aeruginosa ASR1.1, Pseudomonas resinovorans AST2.2, Pseudomonas otitidis AST3.2 and Pseudomonas stutzeri ASM1.1. Two bacterial strains Pseudomonas indoloxydans strain ASK3.2 and Pseudomonas resinovorans strain AST2.2 exhibited mottled growth in form of colonies on agar plates fortified with 1600 mg/l concentration of chlorpyrifos and these two bacterial strains were selected for optimization and biodegradation experiments. In both strains highest OPH activity was observed at pH 8.0, temperature 37°C, after 48hrs in presence of glucose as carbon source. Activity of extracellular OPH enzyme in both the strains was found to be enhanced in presence of Co+2 metal ions, while Cu+2, Mg+2 and Fe+2 ions had showed lesser effect on OPH activity. GC-FID results confirmed that Pseudomonas indoloxydans strain ASK3.2 is a more efficient degrader as 82.72% chlorpyrifos was degraded successfully as compared to 43.90% by Pseudomonas resinovorans strain AST2.2. GC-MS analysis results confirm that peak area/height in both test samples was found to be lower than in control sample, indicating decreasing amount of chlorpyrifos due to its degradation into smaller intermediate compounds. Only one metabolite i.e. phosphoric acid, diethyl 3,5,6-trichloro-2-pyridyl ester (C9H11Cl3NO4P) with CAS no. 5598-15-2 was identified by NIST library search in both test samples at RT 11.10 and RT 11.11 respectively.ThesisItem Open Access Studies on chill units requirement in apple (Malus X Domestica Borkh.)(YSPU, 2015) Babita; Chauhan, P.S.The present investigations entitled “Studies on chill units requirement in apple (Malus X Domestica Borkh.)” were conducted in the experimental orchard of the Regional Horticulture Research and Training Station Mashobra, Shimla and in a private apple orchard in Aisha, Tehsil Sunni, Shimla District, during the year 2013 and 2014. The field experiment was laid out in randomized block design and replicated thrice. In experiment-1 the effective chill units (CU) and GDH°C requirements of apple at Mashobra, Shimla located 31.1°N with an elevation of 2286 m were estimated by adopting North Carolina model with base temperature (4.5°C) and (6.1°C). The estimated CU and GDH°C were 1220 and 8244 with base temperature (4.5°C), while using base temperature (6.1°C) estimated CU and GDH°C were 1222 and 5385 respectively. In experiment-2 the estimated CU and GDH°C were tested and compared with models. Using CU and GDH°C constants of all models to calculate full bloom dates of six years i.e. 2008-2014 at three locations of different elevations and compared with actual full bloom dates for different years. The difference of days of proposed models was compared with difference of days of other chill unit models. At Mashobra (2286m amsl) Shimla and Kotkhai (1815m amsl) location North Carolina (Proposed Model with base temperature 4.5°C) recorded minimum difference of two and five days, respectively. However, at Seobag (1330m amsl) Kullu location North Carolina model (base temperature 6.1°C) was found more accurate. In the experiment-3 the chilling requirement of five apple cultivars were estimated under control condition. Among all Red Fuji recorded the maximum (66.4%) and Golden Delicious recorded minimum (41%) bud burst. Red Fuji completed chilling requirement after completing 1100 CU. Cultivar, Scarlet Gala and Red Chief met the chilling requirement in between 1100 CU to 1200 CU, whereas, Starking Delicious and Golden Delicious met their chilling requirement in between 1200 CU and 1300 CU. In the experiment-4 the Starking Delicious apple trees were treated with different treatment combination of Dormex and HMO. The T5 (Dormex @ 1.0% + 2.0% HMO) was found to be the most effective in improving per cent flowering, reducing duration of flowering, increasing fruit set, improving fruit retention and enhancing maturity. However, T6 (Dormex @ 1.5% + 2.0% HMO) was significantly at par in all the parameters. The studies indicate that Dormex @ 1.0% + 2.0% HMO may be helpful improving per cent flowering, reducing duration of flowering, increased fruit set, improved fruit retention and enhanced maturity.ThesisItem Open Access Impact of Intensive Agriculture on Natural Resources and Vulnerability of Mountainous Rural Communities to Environmental Changes in Himachal Pradesh(YSPU, 2015) Ndungu, Charles Kimani; Bhardwaj, S.K.The present investigation entitled “Impact of intensive agriculture on natural resources and vulnerability of mountainous rural communities to environmental changes in Himachal Pradesh” was conducted during the years 2014 and 2015 in the Department of Environmental Science, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh. The study aimed at determining the impact of cropping systems on natural resources, assessing biophysical vulnerability of mountainous communities to environmental changes and identifying their adaptations to climate change. To determine the impact of cropping systems on natural resources like soil and groundwater quality, four cropping systems namely; vegetable, fruit, cereal and agroforestry were selected in mid-hills of Himachal and uncultivated land was considered as the control. In total there were five treatments which were replicated six times under randomized block design. To assess the vulnerability of the people to environmental changes and determine their adaptations to changing climate, 275 households were selected in the mid-hill region of the state and data on various indicators of vulnerability and adaptations were collected using a pretested questionnaire. The study used Principal Component Analysis (PCA) to assess household level vulnerability, while, logit regression was used to predict farmers’ adaptation to climate change. In the mid-hills of Himachal Pradesh, the cropping systems did not influence the soil properties like SOC, bulk density, pH and EC adversely. In surface soil, available NPK ranged from 303.76 to 555.45, 20.62 to 43.96 and 198.87 to 451.12 kg ha-1, respectively. Surface soil accumulation of As, Ni, and Zn was within WHO permissible limits for soil. In mid-hills, Pb and Cd exceeded by 0.34, 0.33, 0.17, 0.16, 0.06 and 0.11, 0.10, 0.08, 0.06, 0.01 mg kg-1 in surface soils under vegetable, fruit, cereal and agroforestry cropping systems and control, respectively over the prescribed permissible limit of 0.1 (Pb) and 0.01 (Cd) mg kg-1. Groundwater pH and EC ranged from 6.76 to 7.48 and 0.46 to 0.99 dSm-1, respectively. The COD and BOD values of groundwater ranged from 1.33 to 2.33 and 7.67 to 17.33 mg l-1. The groundwater chlorides, nitrates and sulphates ranged from 7.33 to 15.50, 1.39 to 9.98 and 19.75 to 58 mg l-1, respectively. Concentration of zinc and arsenic in groundwater was within permissible limits. In the region Pb and Cd exceeded by 0.17, 0.04 and 0.01, 0.01 mg l-1 in the groundwater under vegetable and fruit cropping systems, respectively over their corresponding prescribed limit of 0.1 and 0.01 mg l-1. Surface soil carbon density ranged from 10.71 to 20.60 Mg C ha-1 .The total soil carbon stock was in the range of 1505.82 to 10427.79 Gg. In mid-hills the frequent droughts and climate change enhanced the vulnerability of mountain people. In the region dependence on natural resource based income has increased the sensitivity of the people. The physical assets have enhanced the adaptive capacity of the people. The overall vulnerability index ranged from -4.08 to 4.59 and followed the trend; Kandaghat> Naggar> Solan >Kullu. The region is presently experiencing rise in temperature and decrease in rainfall. In mid-hills the factors such as farming experience, education, off farm income and access to credit and extension services significantly determined famer’s adaptation to changing climate. The study indicated that the cropping systems and development activities have started affecting soil and groundwater quality by increasing heavy metal concentration and biophysical vulnerability of the households in remote areas due to low adaptive capacity. Under such situations, fruit based cropping system was found to have the best soil carbon sequestration potential and further it has improved the physico-chemical properties of the soil and maintained its fertility status. In order to reduce biophysical vulnerability of the mountain people to changing situations, good practices for managing drought and reducing sensitivity of the people of remote areas by providing required infrastructure facilities and diverse fruit based cropping systems need to be encouraged.ThesisItem Open Access GENETIC STUDIES ON CARNATION (Dianthus caryophyllus L.)(2015) CHAUHAN, PRATIBHA; DHIMAN, S.R.ABSTRACT The present investigation entitled “Genetic studies on carnation (Dianthus caryophyllus L.)” was carried out at experimental farm of Department of Floriculture and Landscape Architecture, Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan (HP) during 2012 to 2014. The experiment was laid out in a RBD with three replications and data was recorded for the three flowering flushes. Fifty diverse genotypes including check (genotype ‘Master’) were evaluated for various growth and flowering parameters to assess the extent of variability, heritability, genetic divergence and the morphological characterization of these genotypes. Wide and significant variations for all the parameters were observed among the different carnation genotypes. Genotypes ‘Snow Storm’, ‘Don Pedro’, ‘Hermes’, ‘Cinderella’, ‘Kleos’, ‘UHFSCar Col.-1’ and ‘UHFSCar Col.-4’, were found to be superior with respect plant height, stem length and flower size. Early flowering was recorded in genotypes ‘UHFSCar Col.-9’, ‘UHFSCar Col.-11’, ‘Tempo’, ‘Madras’ and Liberty’, whereas, maximum number of flowers per plant was recorded in genotypes ‘UHFSCar Col.-9’, ‘Madame Colette’, ‘Liberty’, ‘Tamarind’ and ‘Baltico’. For the production of quality stems in terms of stem sturdiness, genotypes ‘Liberty’, ‘Golem’, ‘Dark Rendez-Vous’ and ‘Arka Flame’ were found best, whereas, maximum production of ‘A’ grade flowers based on stem length was recorded in genotypes ‘Snow Storm’, ‘Don Pedro’, ‘Happy Golem’ and ‘UHFSCar Col.-3’. Maximum to moderate variability estimates in terms of GCV, PCV, heritability, genetic advance and genetic gain was observed for characters such as calyx splitting, disease incidence, plant height, stem length, number of flower stems per plant, flower size and vase life indicating high genetic variability for these traits. Significant positive correlation of the desirable traits such as number of flower stems per plant, flower size, stem length etc., was found to be favourable for breeding as it helps in simultaneous improvement of correlated characters. Positive direct effect of plant height, vase life and calyx splitting on the number of flowers per plant revealed the true relationship between them and direct selection for these traits will be rewarding for the flower yield. Further, divergence studies indicated that hybridization between cluster-I and IV can be utilized for getting the superior recombinants in segregating generations they were having maximum inter cluster distance. Morphological characterization of 50 genotypes was done for 55 characters and the genotypes were then categorized accordingly and the results were summarized in the form of draft guidelines.ThesisItem Open Access STUDIES ON CHILL UNITS REQUIREMENT IN APPLE (Malus × domestica BORKH.)(2015) BABITA; CHAUHAN, P.S.ABSTRACT The present investigations entitled “Studies on chill units requirement in apple (Malus X Domestica Borkh.)” were conducted in the experimental orchard of the Regional Horticulture Research and Training Station Mashobra, Shimla and in a private apple orchard in Aisha, Tehsil Sunni, Shimla District, during the year 2013 and 2014. The field experiment was laid out in randomized block design and replicated thrice. In experiment-1 the effective chill units (CU) and GDH°C requirements of apple at Mashobra, Shimla located 31.1°N with an elevation of 2286 m were estimated by adopting North Carolina model with base temperature (4.5°C) and (6.1°C). The estimated CU and GDH°C were 1220 and 8244 with base temperature (4.5°C), while using base temperature (6.1°C) estimated CU and GDH°C were 1222 and 5385 respectively. In experiment-2 the estimated CU and GDH°C were tested and compared with models. Using CU and GDH°C constants of all models to calculate full bloom dates of six years i.e. 2008-2014 at three locations of different elevations and compared with actual full bloom dates for different years. The difference of days of proposed models was compared with difference of days of other chill unit models. At Mashobra (2286m amsl) Shimla and Kotkhai (1815m amsl) location North Carolina (Proposed Model with base temperature 4.5°C) recorded minimum difference of two and five days, respectively. However, at Seobag (1330m amsl) Kullu location North Carolina model (base temperature 6.1°C) was found more accurate. In the experiment-3 the chilling requirement of five apple cultivars were estimated under control condition. Among all Red Fuji recorded the maximum (66.4%) and Golden Delicious recorded minimum (41%) bud burst. Red Fuji completed chilling requirement after completing 1100 CU. Cultivar, Scarlet Gala and Red Chief met the chilling requirement in between 1100 CU to 1200 CU, whereas, Starking Delicious and Golden Delicious met their chilling requirement in between 1200 CU and 1300 CU. In the experiment-4 the Starking Delicious apple trees were treated with different treatment combination of Dormex and HMO. The T5 (Dormex @ 1.0% + 2.0% HMO) was found to be the most effective in improving per cent flowering, reducing duration of flowering, increasing fruit set, improving fruit retention and enhancing maturity. However, T6 (Dormex @ 1.5% + 2.0% HMO) was significantly at par in all the parameters. The studies indicate that Dormex @ 1.0% + 2.0% HMO may be helpful improving per cent flowering, reducing duration of flowering, increased fruit set, improved fruit retention and enhanced maturity.ThesisItem Open Access STORAGE QUALITY OF FRESH MUSHROOMS (Agaricus bisporus Lange.) AS AFFECTED BY VARIOUS PREAND POST-HARVEST TREATMENTS(2015) BIRLA, ZUBERI MOHAMED; THAKUR, K.S.ABSTRACT Studies on ‘Storage quality of fresh mushrooms (Agaricus bisporus Lange.) as affected by various preand post-harvest treatments’ were carried out in the Department of Food Science and Technology, Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Nauni, Solan during 2012-13 and 2013-14. The whole programme of work was divided into three separate experiments. In Experiment I(a) and Experiment I(b) various concentrations of different plant extracts and CaCl2 were applied to standardize the best post-casing treatments for production of quality mushrooms, which were then combined with various packaging materials to standardize the complete protocol of post-casing and packaging treatments under Experiment I(c). In Experiment II plant extracts and CaCl2 were tried as post-harvest treatments, whereas in Experiment III various coating materials were tested for their effect on storage quality of fresh mushrooms. All the post-casing treatments improved yield and physico-chemical quality characteristics of mushroom at harvest as well as during storage. Carbendazim @ 0.1 per cent in combination with 0.3 per cent CaCl2 applied at post-casing stage resulted in a yield of 2.26 kg/10kg bag of substrate and it was followed by 3.0 per cent garlic extract + 0.3 per cent CaCl2 treatment which produced 2.23 kg/10 kg bag as compared to 1.56 kg/10 kg bag under the control treatment. Garlic extract and CaCl2 treatments appeared to be more effective in retaining storage quality, as evaluated on the basis of reduction in physiological loss in weight (PLW), spoilage, total plate count (TPC), enzymatic browning and polyphenol oxidase activity. It was also more effective in retention of colour, moisture, pH, total soluble solids (TSS), proteins and total phenol contents during storage, as a result of which they were rated to be of higher overall acceptability, especially when mushrooms were packed in plastic moulded cups (punnets) and stored under refrigerated conditions. Plant extracts applied as post-harvest treatments were also effective in reducing deterioration in mushroom quality during storage with 3.0 per cent garlic extract + 0.3 per cent CaCl2 outperforming other treatments. The most effective postharvest coating treatment in retaining storage quality of mushrooms was the combination of 5.0 per cent calcium caseinate + 2.5 per cent glycerol + 0.25 per cent CMC + 0.125 per cent CaCl2, whereas chitosan acetate @ 1.5 per cent, Starlight @ 45 per cent and honey @ 10 per cent concentrations were effective to a lesser extentThesisItem Open Access STUDIES ON Agrobacterium- MEDIATED INSECT RESISTANCE GENE [cry1A(a)] TRANSFER IN CAULIFLOWER (Brassica oleracea L. var.botrytis)(2015) GAUR, AYESH; SRIVASTAVA, D.K.ABSTRACT Genetic transformation studies were carried out to standardize a protocol for insect resistance gene (cryIAa) transfer in cauliflower (Brassica oleracea L. var. botrytis cv. Pusa Snowball K1). Agrobacterium tumefaciens strain containing npt-II and cryIAa genes in binary vector pBin-1Aa was used for genetic transformation studies. Plant regeneration studies were carried out using four different types of explants viz. hypocotyl, cotyledon, petiole and leaf. High efficiency shoot regeneration was obtained in hypocotyl (96.00%), cotyledon (76.00%), petiole (76.00%) and leaf (56.00%) explants on MS medium supplemented with 0.44mg/l TDZ + 0.08mg/l IAA, 0.30mg/l BAP + 0.10mg/l NAA, 0.22mg/l TDZ and 0.77mg/l TDZ + 79.7mg/l Adenine, respectively. Hypocotyl explants showed better shoot regeneration as compared to other explants. MS medium supplemented with 0.10mg/l IBA was found best for root regeneration from in vitro developed shoots. The regenerated plantlets were acclimatized successfully on cocopeat and were morphologically similar and normal. Kanamycin sensitivity (10-60mg/l) was checked by observing fresh weight of the leaf and petiole explants which was measured at the interval of 7 days. From the relative growth of the explants it was found that the concentration as low as 10mg/l is toxic to the explants. Effect of different concentrations of cefotaxime was studied on the regeneration potential in leaf and petiole explants of cauliflower and found not much effect of cefotaxime on regeneration potential. Effect of different concentrations of cefotaxime and kanamycin (20mg/l) were studied on the growth of agrobacterial cells and regeneration potential of hypocotyl and petiole tissues after cocultivation. In both the explants the growth of agrobacterial cells were controlled at concentration of 300mg/l cefotaxime and maximum per cent shoot regeneration in hypocotyl (22.60%) and petiole (13.30 %) was obtained on MS medium supplemented with 300mg/l cefotaxime, respectively. Effect of preculturing and co-cultivation was studied on the transformation frequency. Preculturing of hypocotyl and petiole explants for 48 hours and co-cultivation with agrobacterial cells for 96 hours worked out to be the best treatment as it gave the highest transformation frequency (4.28 %) and (3.28 %) in respective explants. Effect of different concentrations of acetosyringone was studied in hypocotyl and petiole explants to enhance the transformation frequency. The maximum percent shoot regeneration was obtained from hypocotyl (9.23%) and petiole (10.13%) explants cultured on shoot regeneration medium containing 100μM acetosyringone and 125μM acetosyringone respectively at standardized preculturing and cocultivation time interval. The presence/integration of transgene (cryIAa) into the genome of cauliflower was confirmed by PCR using gene specific primers and Southern blot analysis using radioactive labelled DNA probe. The expression of the transgene (cryIAa) in cauliflower at transcriptional level was confirmed by reverse transcriptase-PCR and Real Time-PCR and at translational level by Bioassay. A protocol for high frequency plant regeneration and insect resistance gene transfer in cauliflower (Brassica oleracea L. var. botrytis cv. Pusa Snowball K1) has been standardized.ThesisItem Open Access STUDIES ON SITE CHARACTERISTICS, NATURAL REGENERATION STATUS AND NURSERY TECHNIQUES OF HAZELNUT (CORYLUS COLURNA L.) IN HIMACHAL PRADESH(2015) GUPTA, DINESH; SHARMA, D.P.ABSTRACT The present investigation “Studies on site characteristics, natural regeneration status and nursery techniques of hazelnut (Corylus colurna L.) in Himachal Pradesh.” was carried out in the Department of Silviculture and Agroforestry, Nauni, Solan, Himachal Pradesh, India, during 2011-2013. The study involved work on four different aspects viz; phytosociological, site and stand characteristics, natural regeneration status and standardization of seed and nursery techniques in hazelnut. Hazelnut was the dominant tree species in Mindal and Pattidhank forest, while Pinus wallichiana and Picea smithiana were dominant in Gajta and Sali forest. Total tree density varied from 445 to 535 per hectare and total basal area varied from 8783.35 cm2 to 5978.08 cm2 per hectare. Better regeneration success in Gajta and Sali forest might be attributed to better site quality w.r.t more organic carbon, soil moisture, available nitrogen, available phosphorus and available potassium. The study on seed and nursery techniques involved two types of experiments on effect of i) five stratification periods viz., 0, 20, 40, 60 and 80 days, three stratification temperatures viz., room temperature, out-door pit, 4 ±1 0C and 0 ±1 0C and treated with two gibberellic acid concentration viz., 0 (water only), 100 ppm GA3 and 200 ppm GA3 (CRD factorial) prior to actual sowing in laboratory, ii) three stratification medium viz. naked, sand and cow dung at six different temperatures and assessed seedling growth parameters under field condition (RBD factorial), to standardize techniques for large scale production of quality stock in the species. The out-door pit treatment, outclassed for all temperatures by registering maximum germinability viz., GP (46.28 %), GC (74.79 %), GE (33.17 %), GS (0.64), PV (0.44), MDG (1.65), GV (0.94) and GI (0.71). The combined effect of stratification period, temperature and gibberellic acid exhibited significantly maximum value of GP (96.67 %), GC (99.17 %), GE (76.67 %), GS (1.70), PV (1.27), MDG (3.45) GV (4.39) and GI (1.49) when seeds were stratified for 60 days in out-door pit and the subsequently treated with 200ppm GA3. While, under the field condition seeds stratified in sand for three week warm (25-280 C) followed by three week cold (30 C) were treated with 150ppm GA3 before sowing resulted in maximum germination (74.17%), plant height (14.94 cm), root length (27.50 cm), total dry weight (1.82 g) and stock quality index (0.53) when used for sowing. Experiment on cuttage propagation comprised of seven IBA formulation, two pre-conditioning treatment and two cutting portion i.e. upper and basal portion, involving a RBD factorial experiment with three replications in nursery under a shade net house. Girdling of cuttings of lower/basal portion when treated with 0.4 % IBA in combination with 3% captan + 3% sucrose-talc registered significantly maximum sprouting (76.67 %), rooting (41.67 %) and mean dry root weight (430.17 mg) in the spring season