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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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Subject: Biotechnology × Author: GAMBHIR, GEETIKA × End date: 2015 × Start date: 2013 ×
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    ThesisItemOpen Access
    STUDIES ON Agrobacterium-MEDIATED INSECT RESISTANCE GENE TRANSFER IN CABBAGE (Brassica oleracea L. var. capitata) AND MOLECULAR ANALYSIS OF REGENERATED PLANTLETS
    (2013) GAMBHIR, GEETIKA; SRIVASTAVA, D.K.
    ABSTRACT Genetic transformation studies were carried out to standardize a protocol for insect resistance gene (cryIAa) transfer in cabbage (Brassica oleracea L. var. capitata cv. Pride of India). 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. cotyledon, hypocotyl, leaf and petiole Cotyledon and hypocotyl explants were used from seven to nine days old aseptically grown seedlings whereas, leaf and petiole explants were procured from glass house 20-25 days old grown seedlings of cabbage. Hypocotyl explants showed better shoot regeneration as compared to other explants. High efficiency shoot regeneration was obtained in cotyledon (91.11 %), hypocotyl (94.44 %), leaf (91.11 %) and petiole (88.88%) explants on MS medium supplemented with 0.33mg/l TDZ + 79.7 mg/l Adenine, 0.22mg/l TDZ + 0.088mg/l IAA, 0.22mg/l TDZ + 0.02mg/l NAA and 0.33mg/l TDZ + 0.02mg/l NAA, respectively.MS medium supplemented with 0.10mg/l NAA was found best for root regeneration from in vitro developed shoots. The regenerated plantlets were acclimatized successfully on cocopeat. Kanamycin sensitivity experiment was carried out to study the effect of antibiotic on relative growth of leaf and petiole tissues and to select transgenic shoots during transformation experiment. Kanamycin sensitivity (10-60mg/l) was checked by fresh weight of the 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 cotyledon and hypocotyl explants of cabbage and found no much effect of cefotaxime on regeneration potential. Effect of different concentrations of cefotaxime and kanamycin (50mg/l) were studied on the growth of agrobacterial cells and regeneration potential of cotyledon and hypocotyl tissues after cocultivation. In both the explants the growth of agrobacterial cells were controlled at concentration of 400mg/l cefotaxime and maximum per cent shoot regeneration in cotyledon (35.55 %) and hypocotyl (48.15 %) was obtained on MS medium supplemented with 400mg/l cefotaxime, respectively. Effect of preculturing and cocultivation was studied on the transformation frequency. Preculturing of cotyledon and hypocotyl explants for 72 hours and co-cultivation with agrobacterial cells for 48 hours worked out to be the best treatment as it gave the highest transformation frequency (4.66 %) and (14.50 %) in respective explants. Effect of different concentrations of acetosyringone was studied in cotyledon and hypocotyl explants to enhance the transformation frequency. The maximum percent shoot regeneration (18.66 %) and (32.00 %) was obtained from cotyledon and hypocotyl explants cultured on shoot regeneration medium containing 100μM acetosyringone at standardized preculturing and cocultivation time interval i.e. 72 hours and 48 hours. The presence/integration of transgene (cryIAa) into the genome of cabbage was confirmed by PCR using gene specific primers and Southern blot analysis using radioactive labelled DNA probe. The Southern blot analysis has also been used to confirm copy number of transgene into the genome of cabbage. For PCR analysis, 40 putative transgenic shoots were randomly selected and out of 40 putative transgenic shoots/plantlets, 20 shoots were found to be +ve for the presence/integration of transgene i.e. cryIAa into the genome of cabbage. For Southern blot analysis 10 RT-PCR +ve shoots were selected. Out of the 10 PCR +ve shoots, 5 shoots were confirmed +ve for integration of transgene cryIAa into the genome of cabbage with 1 to 3 copies of gene insertion. The confirmation of expression of the transgene cryIAa into the genome of cabbage 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 cabbage (Brassica oleracea L. var. capitata cv. Pride of India) has been standardized.