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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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20136
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Subject: Molecular Biology and Biotechnology × End date: 2013 × Start date: 2013 × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    STUDIES ON CHARACTERIZATION AND PURIFICATION OF α-AMYLASE INHIBITOR FROM BEAN (Phaseolus vulgaris L .) CULTIVAR
    (2013) ASHU, RANI; NATH, AMARJEET K.
    ABSTRACT Seven bean (Phaseolus vulgaris L.) cultivars of Himalayan region were analyzed for α-amylase inhibitor activity. α-Amylase inhibitor from seed flour of Phaseolus vulgaris L. showed highest activity in 0.1 M phosphate buffer (pH 7.6) contaning 0.15 NaCl in 1 hours of extraction time. Among the seven cultivars of Phaseolus vulgaris L. Triloki cultivar was found to have maximum total and specific inhibitor activity. The α–amylase inhibitor from seeds of Triloki cultivar was partially purified to 15.25 fold. The specific inhibitor activity increased from 0.19 in crude extract to 3.01 in Gel filtration (Sephadex G-100) chromatography. A single band of the purified inhibitor was obtained by Native–PAGE. SDS-PAGE revealed the purified inhibitor to be a monomer with molecular weight of 25,000 daltons. The nature of inhibition was found to be of non-competitive type as determined Dixon’s plot. The inhibitor was found to be heat labile was stable up to 30°C and retained 55.51 percent activity at 70°C temperature. Inhibitor was found to have two pH optima of 5 and 7.6. The purified inhibitor was found to have inhibitory activity against α–amylase extracted from larvae of Corcyra cephalonica. 100 percent larval mortality of Corcyra cephalonica was observed after 11 days when they were fed on wheat flour mixed with 252 µg of partially purified α- amylase inhibitor. Partially purified α–amylase inhibitor was also found to inhibit the activity of α-amylases of Corcyra cephalonica larvae, Aspergillus oryzae, Human salivary and Bacillus spp. Molecular characterization of bean cultivar Triloki was done by amplification of its genomic DNA using primer OPS-02. The number of bands amplified was 2.
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    ThesisItemOpen Access
    STUDIES ON IN VITRO PROPAGATION OF BLUEBERRY(VACCINIUM CORYMBOSUM L.) - A MEDICINALLY IMPORTANT FRUIT SPECIES
    (2013) SAPNA, KUMARI; KAUR, RAJINDER
    ABSTRACT A protocol for in vitro propagation was developed for Vaccinium corymbosum, a medicinally important fruit species. The sterilized explants (buds) cultured on MS and WPM supplemented with 5.0 mg/l 2-iP + 1.0 mg/l GA3 and 2.5 mg/l zeatin + 1.0 mg/l GA3, respectively gave best results for in vitro shoot bud establishment. Out of the two media found best for bud establishment, WPM supplemented with 2.5 mg/l zeatin + 1.0 mg/l GA3 proved to be the best for in vitro shoot multiplication. Also comparison of different growth regulators viz., zeatin, 2-iP and BAP in both MS as well as WPM was studied and out of these three different growth regulators, zeatin was found to be best for in vitro shoot multiplication. Leaves were found to be the best explants for callus induction from both in vivo and in vitro explants on both MS as well as WPM supplemented with 0.25 mg/l TDZ + 0.05 mg/l IBA in each case. The callus obtained with both in vivo and in vitro leaf explants was compact nodular soft in texture and green in color. For shoot regeneration from callus, it was found that in vivo shoot segments derived callus when cultured on WPM medium supplemented with 2.0 mg/l zeatin showed best results. Root primordium formation was observed on basal WPM supplemented with different concentrations of IBA viz., 1.0 mg/l and 2.0 mg/l and also with 0.2 % activated charcoal. Ex vitro rooting was carried out and 80.00% of survival rate was obtained on rooting mixture containing cocopeat : perlite (1:1).
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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.
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    ThesisItemOpen Access
    ASSESSMENT OF GENETIC DIVERSITY IN Aloe vera L. AMONG DIFFERENT PROVINCES OF H.P.
    (2013) RANA, SABINA; KANWAR, KAMLESH
    ABSTRACT The present investigation “Assessment of genetic diversity in Aloe vera L. among different provinces of H.P.” was undertaken using morphological, biochemical, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers. On the basis of morphological and biochemical characters very low variation was found among selected genotypes. A total of 30 RAPD primers and 6 ISSR primers were screened and out of 30 only 24 RAPD and all the 6 ISSR primers gave amplification. In RAPD analyses, a total of 91 bands were amplified, out of which 82 were polymorphic in nature however in case of ISSR markers, 21 polymorphic bands were observed from a total of 24 bands. The amplified products from both the analysis ranged from 100 to 3000 bp and 250 to 1500 bp respectively. Maximum number of bands 107 was scored with primer OPE-10 during RAPD analysis and 114 bands with primer hb-13 during ISSR studies. The similarity coefficient value ranged from 0.62 to 0.91 on the basis of dendrogram. Cluster analysis, using UPGMA, SAHN clustering (NTSYS-pc ver. 2.0) grouped the genotypes into 4 main clusters. C1 genotype showed its distinct nature by sidelining at 0.62 similarity index value leaving behind 65-91% genetic similarity among the remaining genotypes. On the basis of dendrogram formed by ISSR markers 57-100% of genetic similarity was obtained forming two clusters representing 3 and 21 genotypes in each, thereby depicting 80% similarity in minor and 65% similarity in major cluster. Hence the study showed existence of genetic relatedness within the species.
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    ThesisItemOpen Access
    BIOPROSPECTING OF THERMOTOLERANT BACTERIA FROM HOT WATER SPRINGS OF H.P. FOR PRODUCTION OF THERMOSTABLE PROTEASE ENZYME
    (2013) ZEBA; SHIRKOT, POONAM
    ABSTRACT Thermophiles are the organisms which are adapted to live at high temperatures. The survival of these organisms at high temperature is possible due to the thermostability of their enzymes. Many thermostable enzymes such as Taq DNA polymerase, aldolase, amylase, lipase, protease, cellulase, RNA polymerase etc. find a number of commercial applications because of their thermostability. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are very important in terms of discovering new industrial enzymes. Keeping in view, hot water springs of Himachal Pradesh could serve as a good source for new thermophilic microorganisms with novel industrially important properties. Therefore aim of the present study was the isolation and characterization of thermotolerant bacteria from hot water springs of Himachal Pradesh for production of thermostable protease enzyme. Four hot water springs viz., Tattapani , Manikaran, Khirganga and Vashisht were purposely selected for the present studies. Samples consisted soil, water, rock matting and pebbles were collected from each hot water spring. The pH and temperature of the four thermal springs were ranged from 4.1-6.0 and 51-105oC respectively. The chloride, sulphate, total hardness, calcium hardness, CaCO3 hardness and magnesium content ranged from 490-2673, 12-50, 147-483, 30.0-144.2, 65-264 and 7.53-26.73 mg/l respectively. Forty seven bacterial isolates were isolated from the samples using skim milk medium. All the bacterial isolates were studied for various morphological characters and on the basis of ability of formation of zone of clearance on skim milk medium, forty bacterial isolates were selected, which were further investigated for biochemical characters. Quantitative screening was done to select one bacterial isolate showing maximum thermostable protease activity. MCW220 bacterial isolate was found to show maximum thermostable protease activity of 33.4 U/l and was selected further for molecular characterization. Genomic DNA was isolated from the selected bacterial isolate MCW220. PCR amplification of the isolated DNA was carried out using universal primers for 16S rDNA gene and an amplicon of 1200 bp was obtained. Sequencing of the PCR product was done using same primers. Sequence of the MCW220 bacterial isolate so obtained was found to be 1469 bp. In silico analysis using BLASTn showed 97% homology of the query sequence with Aneurinibacillus thermoaerophilus strain L420-91. A total of 20 sequences were mined and these sequences were used to compare the 16S rDNA sequence of the test isolate MCW220. Alignment score was highest for Aneurinibacillus thermoaerophilus strain L420-91 16S ribosomal RNA, partial sequence (NR_029303.1). Phylogenetic analysis based on nucleotide sequences using NJ method was achieved via phylip 3.68 and EXOMETM HORIZON. Aneurinibacillus thermoaerophilus strain MCW220 was studied further for enzymatic studies. Optimization of culture conditions for growth and thermostable protease activity of isolate Aneurinibacillus thermoaerophilus strain MCW220 was carried out. The crude extracellular extract was produced using optimized conditions and further partially purified using ammonium sulphate precipitation technique. The procedure yielded 0.435 g protein with 0.70 fold purification with a percent yield 67.5 %. Molecular weight of the partially purified enzyme was found to be 40 kDa using SDS-PAGE gel.
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    ThesisItemOpen Access
    PLANT REGENERATION, GENETIC TRANSFORMATION AND EXPRESSION OF FOREIGN GENE IN LETTUCE (Lactuca sativa L.)
    (2013) SHARMA, POOJA; SRIVASTAVA, D.K.
    ABSTRACT The investigation was carried out to standardize a protocol for plant regeneration, genetic transformation and expression of foreign gene in lettuce (Lactuca sativa L.). Plant regeneration studies were carried out using two types of explants viz leaf and petiole. The leaf explants shows high percentage of shoot regeneration (82.44%) on MS medium supplemented with 0.25mg/l BAP and 0.10mg/l NAA as compared to petiole explants (64.18%) on MS medium supplemented with 0.50 mg/l kinetin + 0.25mg/l NAA. MS medium supplemented with 0.50mg/l IBA was found to be best for root regeneration (81.43%). 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 (npt-II) for selection in both bacteria and plant was used for co-cultivation experiment to transfer uid A (gus) and npt-II genes in lettuce. After co-cultivation only the transformed cells/ explants were able to grow on selective shoot regeneration medium (50mg/l Kanamycin and 500mg/l cefotaxime) whereas non-transformed cells/ explants turned brown and died on the selective medium. Transformation experiment could be scored as early as 3-4 weeks after selection. The effect of pre-culturing and co-cultivation was studied. Explants pre-cultured for 72 hours and co-cultivated for 48 hours resulted in improved transformation frequency. Putative transformed calli and shoot were obtained from leaf explants, which were able to grow on the selective medium containing 50mg/l Kanamycin. The presence of npt-II and uid A (gus) was confirmed by PCR using designed primers. Out of 8 putative transformed calli 5 have shown the amplification of npt-II and 4 shown the amplification of uid A (gus) genes. Expression of uid A (gus) gene was studied in these transformed calli by GUS assay using biochemical and histochemical methods. Five transformed calli which showed presence of npt-II and four transformed calli which showed presence uid A (gus) genes were also GUS positive. A protocol for plant regeneration and genetic transformation in lettuce tissues has been standardized.