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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: Molecular Biology and Biotechnology × Author: SHARMA, SHIKHA × End date: 2013 × Start date: 2013 × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    PLANT REGENERATION AND GENETIC TRANSFORMATION STUDIES IN PEA (Pisum sativum L.) TISSUES
    (2013) SHARMA, SHIKHA; SRIVASTAVA, D.K.
    ABSTRACT An efficient protocol for in vitro plant regeneration and genetic transformation has been developed in pea (Pisum sativum L. var. Lincon) tissues. Hypocotyl, root, leaf and cotyledon were used as explants for in vitro plant regeneration studies in pea. The explants were excised from 10-12 days old in vitro grown seedlings and placed on shoot induction medium. High frequency shoot regeneration from hypocotyl (81.45%), root (83.53%) and cotyledonary node (72.76%) explants was obtained on MS + 4.5 mg/l BAP + 1.6 mg/l NAA, MS + 2.0 mg/l TDZ and MS + 4.5mg/l BAP + 1.8 mg/l IBA, respectively. MS medium supplemented with 0.20 mg/l IBA was found to be best for root regeneration (63.33%) from in vitro developed shoots. The pea plantlets were able to regenerate 6-7 weeks. Regenerated plantlets were successfully acclimatized. The high frequency regeneration system served as an excellent tool for the establishment of an efficient transformation method for pea. For genetic transformation, disarmed Agrobacterium tumefaciens LBA 4404 strain containing a reporter β- glucuronidase [uid A (gus)] gene in binary vector (pBI 121) system along with kanamycin resistance gene (nptII) for selection in both bacteria and plant was used for co- cultivation experiment to transfer uid A (gus) and npt-II genes in pea. After co-cultivation only the transformed cells were able to grow on selective shoot regeneration medium containing 50mg/l Kanamycin and 500mg/l cefotaxime, whereas non-transformed cells/explants turned brown to black and died on the selective medium. Pre incubation of 48 hrs and cocultivation of 48 hrs was found optimum as it gave maximum transgenic shoot regeneration on selective medium. The transformation frequency was low in hypocotyl explants (2.66%) as compared to root explants (3.55%) on selective shoot regeneration medium (50mg/l Kanamycin and 500mg/l cefotaxime). The putative transformants were randomly selected for the amplification of npt-II and gus genes with specific designed primer by polymerase chain reaction. Out of 10 putative transformed calli and 3 shoot, 6 calli and a shoot have shown the amplification of uid A (gus) genes. These positive samples were then further analysed by PCR using designed primers for npt-II genes. Out of 6, only 3 of the transformed calli showed the presence of npt-II genes in pea genome. The expression of gus gene was analyzed by using biochemical and histochemical techniques of GUS assay. The gus gene was expressed in the PCR positive transformed calli of pea. A protocol for genetic transformation in pea with npt-II and gus genes has been standardized.