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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 IN VITRO SELECTION OF CELL LINES IN Punica granatumL. (DARU) AGAINST BACTERIAL BLIGHT(UHF,NAUNI, 2013) GARIMA, KUMARI; KANWAR, KAMLESHABSTRACT The present investigation aims at “In vitro selection of cell lines in Punica granatum L. (Daru) against bacterial blight. Cotyledon and hypocotyl segments of 3 weeks old in vitrogerminated seedlings and mature leaf were used as explants. For leaf explants the sterilization protocol was standardized with 0.2 % bavistin treatment for 10 minutes and 0.5% sodium hypochloride treatment for 5 minutes. Callus induction and plantlet regeneration varied with explant type and growth regulators. The best callus induction medium for all the explants was MS medium supplemented with 2.0 mg/l BA and 4.0 mg/l NAA. Hypocotyl and cotyledon were more responsive explant and showed 96.67 and 85.00 per cent callus induction. The best medium for callus differentiation and shoot bud induction fromcotyledon derived callus was MS medium supplemented with BA (2.0 mg/l), Kinetin (1.5 mg/l) and GA 3 (3.0 mg/l), while best medium for shoot bud induction from leafderived callus was MS medium containing BA (2.0 mg/l) and Kinetin (0.5 mg/l). Cotyledon showed better regeneration (81.67 percent) as compared to leaf explant (48.33per cent). No regeneration was observed in hypocotyl derived callus explants. Rooting of in vitro raised shoots was done on half strength MS medium supplemented with 0.04 % charcoal. The well rooted plantlets were acclimatized in autoclaved sand. Cell line selection was done by using bacterial culture filtrate of Xanthomonas axonopodis pv. punicae as a selective agent. Resistant lines were selected at 40 per cent level of culture filtrate after two cycles of selection. Multiplication and shoot regeneration from selected calli was obtained on previously standardized medium. Fresh weight of callus increased progressively upto third subculture passage while shoot bud induction from selected calli was observed only after third subculturing. The selected microshoots were rooted on the rooting medium. After in vitro testing of shoots regenerated from selected calli 4 resistant plantlets were obtained.ThesisItem Open Access In vitro PROPAGATION AND CONSERVATION OF Swertia chirayita(UHF,NAUNI, 2013) SHAILJA; KANWAR, KAMLESHAbstract A protocol for in vitro propagation and conservation was developed for Swertia chirayita, an endangered medicinal plant. The sterilized explants (leaves) cultured on MS medium supplemented with 0.1 mg/l NAA and 3.0 mg/l BA gave best results for in vitro callus induction. Shoot regeneration was obtained from the callus on the same medium. The in vitroshoots cultured on MS medium supplemented with 2.5mg/l BA and 0.1 mg/l Kinetin gave best results for in vitro shoot multiplication. The MS medium supplemented with 0.1 mg/l NAA and 3.0 mg/l BA medium was found to be thebest for direct shoot regeneration from in vitroleaves. 80.30% root induction from in vitro grown shoots was obtained on half strength MS medium supplemented with 400 mg/l activated charcoal and 0.1 mg/l NAA. In vitro conservation was carried out by using two different approaches namely slow growth by changing media composition (sucrose and abscisic acid), at low temperature and cryopreservation following vitrification.With increase in concentration of sucrose and ABA decrease in growth of in vitroshoots was observed. No shoot multiplication with average leaf size of 0.35 cm and shoot length 0.67 cm was observed on half MS containing 90 g/l sucrose. Similarily, in case of media containing half strength MS salts and 3.0 mg/l ABA showed no shoot multiplication,0.83 cm average leaf size and 0.83 cm shoot length. At low temperature the in vitro shoots incubated at 4 o C, showed 100% retrieval, with 1.00 cm average number of shoots, 0.86 cm shoot length and 0.34 cm leaf size. In vitro shoots incubated at 10 o C, showed 100% retrieval, with 1.00 cm average number of shoots, 0.76 cm shoot length and 0.23 cm leaf size. During studies the vitrified shoot gave retrieval of 42.33% when precooled at 4 o C while only 22.37% vitrified shoots were retrieved from those precooled at 10 o C.ThesisItem Open Access STUDIES ON DEVELOPMENT OF GENIC-SSRs IN RASPBERRY (Rubus ellipticusSmith.) AND THEIR TRANSFERABILITY ACROSS RELATED SPECIES(UHF,NAUNI, 2013) THAKUR, RASHMI; KAUR, RAJINDERABSTRACT Rubus ellipticus Smith. commonly known as ‘Yellow Himalayan raspberry’ is an important member of Rosaceae family with high medicinal importance having high . In the present study EST-SSR markers were datamined for R. ellipticusand were used for crosstransferability studies. EST sequences of R. ellipticus /Rubus were downloaded from NCBI website (www.ncbi.nlm.nih.gov/nucest). Seven EST sequences for R.ellipticusand 3184 for other Rubusspecies, R. ulmifoliusand R.idaeuswere obtained. ESTs containing SSR motifs were extracted out using an online tool, SSRIT (www.gramene.org/db/searches/SSRtool). None of the R.ellipticusESTs contained any SSR motif, so EST sequences obtained for R.ulmifoliusand R.idaeuswere used for SSR extraction. SSR primers were designed from the EST-SSR containing sequences using PRIMER 3 software (www.frodo.wimit.edu/primer3/) and 20 primers were custom synthesized. SSR studies was carried out using ten Rubus species (four R. ellipticus collections of different geographical origin, R. ulmifolius, R. hypargyrus, R. panniculata, R.nutans, R.macilentusand R.strigosus). To study polymorphism and transferability among the ten Rubus species, DNA was isolated from young leaves of all the ten species using CTAB method (Doyle and Doyle, 1987). The polymorphism study among ten Rubus accessions was carried out with the 20 custom synthesized Rubus primers. All 20 primers showed amplification, with polymorphism of 98.36%. The transferability studies were also carried out using already used 20 polymorphic peach primers, which had shown transferability of 95% to four genotypes of apple and rose each, all belonging to Rosaceae family. Jaccard’s similarity matrix was developed and dendrogram was generated using NTSYSpc ver.2.02h to establish the percent similarity among the ten Rubus accessions. Two clusters ‘A’ and ‘B’ were obtained. R. strigosusin cluster ‘A’ was found to diverge from rest of the nine accessions, all grouped under cluster ‘B,’ revealing high percentage of variability of R. strigosus from rest of the nine species. Maximum similarity was found between R.ellipticusIII and R.macilentus. Thus EST-SSRs used in the present study revealed a high level of polymorphism in the ten Rubusaccessions. Also interspecific and intergeneric cross transferability was established among these accessionsThesisItem Open Access MOLECULAR CHARACTERIZATION OF LETTUCE (Lactuca sativa L.) GENOTYPES USING RAPD-PCR(UHF,NAUNI, 2013) SHARMA, SHUBHANGI; SRIVASTAVA, D.K.Abstract The present studies on ‘‘Molecular characterizationof lettuce (Lactuca sativa L.) genotypes using RAPD” was carried out to study the genetic relationship among various genotypes. The aim of this study was to investigate genetic relationship among different genotypes of lettuce. 25 lettuce genotypes namely 1-(CGN 10944-2), 2-(CGN-14651), 3-(CGN-05169), 4-(UHF-Sel.-07), 5-(CGN 10944-3), 6-(CGN-05198), 7-(UHF-Sel.-03), 8-(CGN-05167), 9-(CGN-20721), 10-(CGN-04987), 11-(CGN 10944-1), 12-(CGN-04990), 13-(CGN-04933), 14-(CGN-04934), 15-(CGN-14629), 16-(UHF-Sel.-04), 17-(UHF-Sel.-04), 18-(CGN-19009), 19-(CGN-09373), 20-(CGN-11358), 21-(Great lakes), 22-(CGN-19088), 23-(CGN-04543), 24-(UHFSel.-01) and 25-(UHF-Sel.-02) were selected for the study. A totalof 45 primers were tried to generate RAPD profile, out of these reproducible patterns were obtained with 22 primers. A total of 87 bands were obtained out of which all were polymorphic and seven primers have shown unique bands with the specific genotypes. The percentage of total polymorphism was 100% in RAPD. Similarity index was computed based on Jacquard coefficient and used for cluster analysis based on UPGMA. RAPD technology could be useful for identification of different accessions as well as accessing the genetic similarity among different genotypes of lettuce.ThesisItem Open Access TECHNOLOGY REFINEMENT FOR MICROPROPAGATED Aloe vera L. AND ASSESSMENT OF GENETIC FIDELITY(2013) SHARMA, SWATI; KANWAR, KAMLESHABSTRACT The present investigation aims at “Technology refinement for micropropagated Aloe vera L. and assessment of genetic fidelity”. Surface sterilized shoot buds cultured on control (MS + 3% sucrose + 0.8% agar-agar ) and low cost medium (MS + 3% table sugar + 10% tapioca pearls) supplemented with 2.0 mg/l, 0.5 mg/l and 0.2 mg/l NAA which showed 81.66% and 79.33% establishment respectively. Both control as well as low cost medium were found at par with each other having 81.99% and 81.80% shoot regeneration respectively. 77.67% rooting and 4.33 average roots per shoot were found on 1/4 MS + 3% sucrose + 0.8% agar-agar and 78.00% rooting and 3.67 average roots per shoot on 1/4 MS + 10% tapioca pearls + 3% table sugar. The study has resulted in the reduction cost of the medium by substituting agar-agar and sucrose with tapioca pearls and table sugar. Further, morphological parameters showed that there is similarity between in vitro raised plants. During the RAPDs studies, 17 random decamer primers were used. Total 55 scorable bands were obtained out of which 52 bands found monomorphic and 3 were polymorphic in nature. The similarity coefficient value ranged from 0.94 to 1.00. Similarity of 98% from dendrogram has been observed which was constructed on similarity matrix.ThesisItem Open Access MERISTEM CULTURE OF APPLE CULTIVAR ‘OREGON SPUR-II’(2013) VIVEK, MANU; MODGIL, MANJUABSTRACT In the present study, an attempt was made to eliminate ACLSV, ApMV, ASGV and ASPV from apple cultivar ‘Oregon Spur-II’. Thermotherapy was carried out at 37-40°C for 4 weeks followed by culturing of meristems of different sizes. During establishment of explants, highest survival percentage (62.35%) and proliferation (30.68%) was recorded during summer season, due to less browning and contamination. However, size of meristems and position of buds from where meristems were excised also influenced their survival. The meristems of size 0.6 – 0.7 mm were found to be the most appropriate for maximum establishment i.e., 36.36 %. Meristems excised from buds positioned on distil portions of actively growing shoots showed better results. MS medium supplemented with BA (1.0 mg/l), IBA (0.05 mg/l) and GA3 (0.1 mg/l) resulted in 56.62% establishment of explants, while maximum number of meristems proliferated with low BA (0.5 mg/l), IBA(0.08mg/l) and same GA3 concentration. Two to four fold multiplication was observed. Virus indexing of shoots raised from different sizes of meristems was carried out and found that 0.3- 0.6mm size was able to eliminate ACLSV, ApMV, ASGV and ASPV. However, some of 0.5-0.6 mm sized shoots were found infected with ACLSV. Larger meristems could not completely eliminate the viruses under study.ThesisItem Open Access Biotechnological approaches for management of plant virus(es) in tomato (Solanum lycopersicum L.)(2013) SIRCAIK, SHABNAM; BHARDWAJ, S.V.ABSTRACT Present investigations entitled “Biotechnological approaches for management plant virus(es) in tomato (Solanum lycopersicum L.)” were undertaken to retrieve virus tested tomato through meristem tip culture and through use of inhibitors of plant origin. Tomato was selected as a test crop due to presence of prominent symptoms on plants (mosaic and necrosis on leaves) and their wide susceptibility to virus infection. Infected plants were collected and maintained under glass house conditions. These plants were indexed both biologically and serologically and found to be infected with PVMV. The virus cultures were also maintained under in vitro conditions by culturing apical and nodal segments from infected plants as explants on MS medium. The most suitable medium for establishment and shoot multiplication was found to be MS medium supplemented with 0.5 mg l-1 BAP. For root regeneration, half strength MS medium supplemented with 0.1 % activated charcoal and 1.0 mg l-1 NAA was found to be the best. Different sizes of meristem ranging between 0.1-1.0 mm were cultured on standardized MS medium with 0.1 mg l-1 BAP and 0.1 mg l-1 GA3. Survival of meristem and virus elimination was seen to be directly and indirectly proportional to the size of meristems. The virus was not detected in plants raised from meristem sizes ranging between 0.1- 0.4 mm. Three plants extracts (i.e Vitex negundo Linn., Prinsepia utilis Royle. and Tinospora cordifolia Willd.) at various concentrations were tried in vitro and in vivo conditions to retrieve virus free plants of tomato. Under in vitro conditions explants were found to be unable to grow on media charged with Vitex extract and browning of medium was observed whereas, the other two extracts have eliminated the virus at concentration 20 mg l-1. The extracts different concentrations (i.e. 5, 10, 15, 20 mg l-1 for spraying and 50, 100, 150, 200 mg l-1) were also tested under in vivo conditions for inhibition of the test virus and it was observed that on spraying Vitex and Tinospora extract inhibited the test virus at every concentration whereas, Prinsepia extract was not able to inhibit/eliminate the test virus. Only Vitex extract has retained its virus inhibiting properties on drenching.ThesisItem Open Access STUDIES ON TRYPSIN INHIBITOR IN LOCAL BEAN (Phaseolus vulgaris L.) CULTIVARS(2013) NEGI, PRATIBHA; NATH, AMARJIT K.ABSTRACT Trypsin inhibitor from seed flour of Phaseolus vulgaris L. showed highest activity in 0.1 M phosphate buffer (pH 7.5) in 3 hours of extraction time. Among the five local cultivars of Phaseolus vulgaris L. Baspa cultivar was found to have maximum total and specific inhibitor activity. The seed flour extracts of all the cultivars inhibited gut protease of Spodoptera littoralis larvae and highest inhibitory activity was found in Baspa cultivar. The inhibitor activity in Baspa cultivar was found to decrease during germination period (1-6 days). In developing seeds (8 to 64 DAF) it increased with seed maturation. The extacts of developing seeds inhibited gut protease of Spodoptera littoralis larvae and maximum inhibition was observed with seed flour extract of mature seeds (64 DAF). Trypsin inhibitor was partially purified from seed flour extract of Baspa cultivar to 3.94 fold with 50.37 per cent recovery by ammonium sulfate precipitation and gel filtration chromatography on Sephadex G-50 column. Linear increase in percent inhibition with increase in inhibitor concentration was observed till 71.The partially purified inhibitor was found to be heat stable and retained 82.35% activity of control at 77oC. It was found to have pH optimum of 7.5 and it exhibited non-competitive inhibition pattern. The inhibitor lost its activity on incubation with increasing concentrations (20-100Mm) of DTT thereby indicating the role of disulfide linkages in maintaining its stability and three dimensional structures. Feeding bioassay of Pieris brassicae larvae on leaf discs of cabbage coated with partially purified inhibitor showed decline in per cent survival of larvae with increase in inhibitor concentration. The larvae survived after bioassay experiments were shifted to normal leaf discs for further growth and development. In treatment, delay in pupation was observed, deformed pupa was formed and no adult emerged. In control, all the larvae formed normal pupa and adults. Pieris brassicae eggs treated with partially purified inhibitor showed 89.6 percent reduction in hatching as compared to control. Blood clotting time was found to increase by 11 minutes in treated human blood with partially (220μg inhibitor/2ml blood) as compared to control.ThesisItem Open Access STUDIES ON ISOLATION AND CHARACTERIZATION OF TRYPSIN INHIBITOR (TI) GENE FROM Dolichos biflorus L. (Kulth)(2013) REENA KUMARI; NATH, A.K.ABSTRACT Protease inhibitors are one of the most promising agents that confer resistance in plants against insect pests by inhibiting larval midgut proteases. Maximum extraction of trypsin inhibitor protein from seed flour of Dolichos biflorus L. was in 0.1 M phosphate buffer (pH 7.6) after four hours of extraction. Screening of Dolichos biflorus L. cultivars for trypsin inhibitor activity revealed maximum activity in HPK4 cultivar and further studies were conducted in this cultivar. Crude trypsin inhibitor of all cultivars inhibited midgut protease of P. brassicae larvae. Inhibitor activity was detected at early stages of seed development (3 days after flowering (DAF)) and it increased progressively with seed development (21 DAF to 60 DAF). Trypsin inhibitor activity decreased during seed germination as compared to dry seeds. Crude trypsin inhibitor extracted from developing and germinating seeds also inhibited larval midgut protease of S. littoralis. Neonate larvae of P. brassicae fed on cabbage leaf discs coated with 0.025-2.50 mg crude trypsin inhibitor caused 10–80 % larval mortality. The calculated LC50 value was 1.05 mg crude trypsin inhibitor and for 2.5 mg crude trypsin inhibitor the calculated LT50 value was 3.2 days. Leaf area eaten and faecal matter produced by treated larvae were significantly lower as compared to untreated controls. Larvae fed on leaf discs coated with 2.5 mg crude trypsin inhibitor for 5 days had significantly less total soluble protein in faecal matter and midgut trypsin activity as compared to untreated control. Significant reduction in egg hatching (75%) was observed in egg mass treated with 5.3mg of crude trypsin inhibitor of mature seeds. Trypsin inhibitor gene (309 bp) was amplified from cDNA synthesized from mature seeds of Dolichos biflorus L. HPK4 cultivar using designed primers. The amplified PCR product was cloned and sequenced. Sequence of Dolichos biflorus L. HPK4 cultivar trypsin inhibitor (DbTI) gene has been submitted to NCBI with Accession No. JQ259858. DbTI gene and its deduced amino acid sequence showed homology with Bowman-Birk inhibitors of Dolichos spp., Phaeolus spp., Vigna spp. and Glycine spp. The predicted molecular weight of deduced amino acid sequence was ~11.5 KDa and it had N terminal signal peptide of 19 amino acid residues. The secondary structure of deduced amino acid sequence of DbTI showed dominance of coils and sheets over alpha helix. Homology modelling was employed to predict the three dimensional structure of DbTI. Docking of trypsin enzyme and DbTI showed the inhibitor to be of non- competitive type.