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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 Evaluation of some introduced rootstocks for peach, plum and apricot(YSPU, 2013) Nagi, Manpreet; Krishan KumarThe present investigations entitled “Evaluation of some introduced rootstocks for peach, plum and apricot” were conducted in Department of Fruit Science, Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan. Eight exotic Prunusrootstocks (Cadaman, Citation, GF 31, GF 677, Ishtara, Manicot, Montclar and Rubira) along with two local seedling rootstocks viz., Wild Apricot and Wild Peach were assessed for their ability to propagate by conventional vegetative methods, their graft success with commercial scion cultivars and their tolerance to drought and cold stress conditions. Exotic rootstocks namely Montclar (39.9 %) and Rubira (25.8 %) were more amenable to propagation through IBA treated hardwood cuttings than Ishtara (13.6 %), GF 31(13.4 %), Citation (8.6 %) and Wild Peach (10.8 %). As many as three rootstocks Manicot, Cadaman and Wild Apricot did not respond to multiplication through hardwood cuttings. These rootstocks followed similar trend on propagation through softwood cuttings except the fact that the success rate was relatively lower than that achieved through hardwood cuttings. Overall, IBA treatment of 2000 ppm and 1000 ppm was found to result in maximum rooting in Montclar through hardwood (39.9 %) and softwood (26.6 %) cuttings, respectively. The success of multiplication throughstooling varied from very good (Rubira, Manicot, GF 31), good (Citation, GF 677) and low to very low in Wild Peach and Wild Apricot. Three rootstocks Montclar, Ishtara and Cadaman did not record any success in propagation through stooling. Maximum success was recorded in Rubira with nectarine cv. Snow Queen (50.0 %); in Manicot with apricot cv. Ema (43.3 %) and plum cv. Santa Rosa (50.0 %); in Montclar with plum cv. Red Beaut (43.3 %) and peach cv. July Elberta (43.3 %); in Ishtara with peach cv. July Elberta (60.0 %) and nectarine cv. Mayfire (50.0 %); in GF 31 with apricot cv. Ema (23.3 %), nectarine cv. Mayfire (40.0 %) and plum cv. Frontier (46.6 %); in Citation with peach cv. July Elberta (46.6 %); in GF 677 with plum cv. Santa Rosa (53.3 %); in Wild Peach with nectarine cv. Mayfire (53.3 %) and in Wild Apricot with apricot cv. NewCastle (43.3 %) and plum cv. Red Beaut (43.3 %). Exotic clonal rootstocks, on the whole, showed better graft success with scion cultivars than local seedlingrootstocks. Rootstocks namely Manicot, Montclar, Ishtara, Citation and Wild Apricot were observed as more drought tolerant than Rubira, GF 31, GF 677 and Wild Peach, whereas Manicot, GF 31, Ishtara and Cadaman were found to be more cold hardy than Rubira, Montclar,GF 677 and Citation.ThesisItem Open Access Studies on epidemiology and management of pink canker (Corticium salmonicolor Berk. & Br.) in apple(DYSPU, 2013) Durga Prashad; Ved RamPink canker (Corticium salmonicolor Berk. and Br.) in apple is one of the major limiting factor in its cultivation affecting both yield and fruit quality in Himachal Pradesh. Present investigations were undertaken with an objective to study the prevalence of the disease, role of abiotic environmental factors in disease development and to devise suitable disease management strategies. Pink canker was found to occur in moderate to severe form in different apple growing areas of Mandi, Shimla, Kullu and Sirmour districts of Himachal Pradesh. Based on morphological, cultural and molecular characters, the pathogen was identified as Corticium salmonicolor Berk. and Br. Moderate temperatures (19 + 2o C) coupled with high RH (80%) favoured disease development. Conidial germination and germ tube length was high in Mandi isolate. Out of nine cultivars only, Tydeman Early Worcester was found moderately susceptible under field conditions. Ace Spur, Spur Winter Banana and Granny Smith exhibited moderately resistant reaction under flask condition. Under congenial condition, the pathogen infected leaves and fruits of Royal Delicious. Among integration of fungicide and plant oils, integration of Contaf + Brassica juncea var. cunefolia and combinaion of Score + Brassica juncea var. cunefolia provided maximum wound recovery (92.94% and 91.86% respectively) and callus formation (>10 mm). The combination of Contaf + Cow urine + Melia azedarach + Vitex negundo + Artimisia roxburghiana + Juglans regia + Roylea elegans and Score + Cow urine + Melia azedarach + Vitex negundo + Artimisia roxburghiana + Juglans regia + Roylea elegans in white paint exhibited maximum wound recovery (87.17% and 85.81% respectively) and callus formation (>10 mm) during 2011-12. Among combination of fungicides + antagonists, significantly maximum wound recovery (80.47%) was recorded in Contaf + Pseudomonas sp. followed by a combination of Contaf + Trichoderma hamatum (79.95%).ThesisItem Open Access Vegetation dynamics of chir pine forests along altitudinal gradient in Giri catchment of Himachal Pradesh(YSPU, 2013) Mahato, Debasis; Gupta, B.The present investigation entitled “Vegetation dynamics of Chir pine forests along altitudinal gradient in Giri catchment of Himachal Pradesh” was carried out in Giri catchment, a component of Giri-Bata catchment in Himachal Pradesh located between 30o 33¢ 48² and 31o 16¢ 08² N latitude and 77o 02¢ 32² to 77o 38¢ 22² E longitude. This catchment is distributed in Shimla, Sirmaur and Solan districts of Himachal Pradesh and is comprised of 135 subwatersheds. The climatic condition of experimental sites ranges from tropical/sub-tropical through sub-humid to sub-temperate climate and clay to sandy soils. In each experimental site three sample plots of size 0.1 ha for trees, two plots of 10mx10m were marked to study shrubs and in each sample plot, 6 quadrates of 50cmx50cm were harvested to study herbs characteristics. Composition, density (plants/ha), basal area (m2/ha), biomass (q/ha), carbon stock (q/ha) of different vegetation layers and soil properties of grasslands and Chir pine forests for herb and shrub layers were analyzed in 42 experimental sites of 14 representative subwatersheds during 2010-2011. The data were analyzed by using factorial RBD, Duncan test, variability analysis through statistical techniques viz., measures of dispersion, Bertlett’s test of significance and TWINSPAN, whereas cluster analysis was carried out by CAP (Version 4.0) software. The floristic composition comprised of 4%, 5%, 7%, 21%, 25%, and 38% trees , sedges, legumes, grasses, shrubs and forbs respectively under Chir pine forest. However, grasslands consist of 7% sedges, 8% legumes, 25% grasses, 27% shrubs and 33% forbs. Herbage density (tillers/m2) in grasslands ranged form 926.65-1445.34, 851.32-1256.03 and 603.33-1128.01 for E1 (900-1300m), E2 (1300- 1700m) and E3 (1700-2100m) at different Silting Yield Index (SYI). The E1C1 (1295.58) reported significantly higher herbage density of grasslands than others. In Chir pine forests, the herbage density varied from 744.00 to 1279.34, 609.34 to 1086.65 and 420.68 to 827.99 for E1, E2 and E3, respectively at different SYI classes. The herbage density in Chir pine forests was significantly higher at E1C1 than other interactions. The shrub density varied from 1666.67 to 2866.67, 1300.00 to 2399.99 and 1466.67 to 2699.99 in grasslands, whereas, 2300.01 to 5066.67, 1466.67 to 4199.99 and 1400.01 to 3400.00 in Chir pine forests for E1, E2 and E3, respectively at different SYI classes. Herbage basal area (cm2/m2) varied from 40.10 to 56.01, 36.34 to 49.61, 25.21 to 45.93 in grasslands and 32.81 to 54.09, 28.27 to 49.39, 18.56 to 37.43 in Chir pine forest for E1, E2 and E3, respectively at different SYI classes. The shrub basal area (m2/ha) varied from 0.137 to 1.719, 0.136 to 0.685, 0.383 to 1.632 in grasslands and 0.240 to 2.338, 0.181 to 1.136, 0.299 to 2.223 in Chir pine forests for E1, E2, E3, respectively at different SYI classes. The C. montanus at E1 (IVI= 116.98) and E2 ( IVI=146.65), while, H. contortus, at E3 (IVI= 118.02) were dominant herbage species in grasslands. In Chir pine forests, dominant species was T. anathera with IVI 155.46, 186.77 and 180.68 at E1, E2 and E3, respectively. In grasslands the dominant shrub species at E1 was C. carandus, at E2 was M. africana while, at E3 was B. lycium. In Chir pine forests, M. africana at E1 and C. carandus at E2 while B. lycium at E3 were the dominant shrub species. The aboveground, belowground and total herbage biomass for were (28.44 to 46.13, 21.37 to 43.23, 14.75 to 41.80), (15.61 to 27.93, 11.11 to 27.87, 7.39 to 27.53) and (44.05 to 70.29, 32.48 to 68.17, 22.14 to 61.59) at E1, E2 and E3, respectively in grasslands. In Chir pine forests, the aboveground (23.25 to 44.07, 18.58 to 42.14, 18.02 to 37.75), belowground (11.96 to 22.01, 9.15 to 19.57, 7.95 to 17.70) and total herbage biomass (35.21 to 66.08, 27.73 to 61.33, 26.30 to 54.17) for E1, E2 and E3 were recorded. The shrub aboveground (4.621 to 9.884, 3.699 to 7.212, 3.741 to 9.302), belowground (0.969 to 3.184, 0.550 to 2.163, 0.757 to 2.197) and total (5.75 to 11.70, 4.59 to 8.83 and 4.56 to 11.50) biomass were recorded in grasslands for E1, E2 and E3. In Chir pine forests at E1, E2 and E3, shrub biomass for aboveground (9.295 to 19.810, 4.538 to 11.838, 5.469 to 18.018), belowground (1.151 to 3.972, 0.768 to 3.405, 0.940 to 3.125) and total (9.82 to 23.78, 6.11 to 14.16 and 6.71 to 21.14) were recorded. Average total biomass was highest at E2 (1399.04) as compared to E1 (1244.92) and E3 (1000.43). Total carbon stock (herb+shrub+soil) in grassland ranged from 308.69 to 520.16, 380.70 to 539.17, 429.75 to 550.27 for E1, E2 and E3, respectively. In Chir pine forests, the total carbon stock (herb+shrub+tree+soil) varied from 964.80 to 1263.58, 1138.69 to 1305.66, and 828.27 to 1170.01 for E1, E2 and E3, respectively at different SYI classes. The soil pH in grassland varied from 5.99 to 7.71, 6.03 to 7.56 and 6.17 to 7.68 whereas, in Chir pine forests 5.94 to 7.01, 5.91 to 6.93 and 6.23 to 6.84 at E1, E2 and E3, respectively. In grasslands, the bulk density was 1.06 to 1.22, 1.03 to 1.19 and 0.95 to 1.15, while, in Chir pine forests, 1.00 to 1.22, 0.96 to 1.12 and 0.93 to 1.05. In grassland SOC varied from 1.16 to 2.29, 1.52 to 2.47 and 1.81 to 2.75 whereas, at Chir pine forests it ranged from 1.68 to 2.48, 1.96 to 2.84 and 2.33 to 2.84 at E1, E2 and E3, respectively.ThesisItem Open Access Reproductive biology, molecular profiling and biochemical analysis of Terminalia chebula (Retz.)(DYSPU, 2013) Sankanur, Mahantappa; N. B. SinghThe present investigation entitled “Reproductive biology, molecular profiling and biochemical analysis of Terminalia chebula (Retz.)” was carried out at Regional Horticultural Research Station, Jachh, Kangra and in the Department of Tree Improvement and Genetic Resources, Dr. Y. S. Parmar University of Horticulture & Forestry Nauni, Solan (HP) during 2010-2013. Reproductive biology of six T. chebulagenotypes revealed that Kothi (G 1), Paragpur 2 (G 3 ), Kallar (G4), Bhella (G 5) and Tamber (G 6 ) initiated most of their phenological events earlierthan Paragpur 1 (G 2 ) genotype. The flower buds busted after the leaves started to emerge for all the genotypes. Among genotypes each spike is on an average 6.79 ± 0.45 cm and produces 40.67 ± 3.36 flowers acropetally over a period of 6–7 days. On thebasis of sizes and development stages, the flower buds were assorted into seven different stages. The stigma protrudes out of the calyx during the mature bud stage. It is receptive since then and continues until the evening of the 3 rd day. The pollen–ovule ratio is 10,890:1. Anin-vitropollen germination percentage of freshly collectedpollen was higher in 30% sucrose with Brewbaker and Kwack’s medium (BKM). Fresh pollen viability percentages were generally high (above 70 per cent) for most of the genotypes. -20 0 C temperature with controlled humidity was found effective in long term storage of pollens for breeding programmes. The manual pollinations performed for autogamy, geitonogamy and parthenocarpy did not set fruit. Whereas, those performed for the xenogamous mode set fruit (65.76 ± 10.4%) and in open-pollinations the fruit set was 7.83 ± 0.78%. The flowers were foraged during daytime by 42 species of insects representing bees, wasps, bugs, flies, butterflies and beetles. Crossability pattern studies revealed thatgenotypes under study were cross compatible shouldbe involved in intraspecific breeding programme. In all the fourteen successful crosses between different genotypes were obtained. Per cent successful cross was highest in cross Bhella (G5) × Paragpur 1 (G 2) that was 2.14 per cent. The data for various nursery traits viz., collar diameter, plant height and number of leaves for various crosses was recorded when seedlings were 8 months old. Collar diameter for various crosses ranged between 1.36 mm to 3.38 mm. maximum value for collar diameter was recorded for cross Bhella (G5) × Kallar (G 4 ) i.e.3.38 mm. The maximum plant height was 20.80 cm which was recorded for cross Paragpur 2 (G 3) × Paragpur 1 (G2 ). The maximum number of leaves was found in the cross Paragpur 2 (G 3 ) × Paragpur 1 (G 2 ). Among the six genotypes various qualitative morphological descriptors studied viz.,crown of tree, branching type, bark colour, leaf colour, arrangement, shape, tip shape, base, margin, flower type, flower colour, fruit base and tip and seed colour did not show any variation at all except forthat fruit shape. Fruit samples collected from sixdifferent genotypes showed variation in their shapes, bases, colours, physical dimensions and chemical characteristics. Fruit shapes were of obovoid to ellipsoidal obovoid. While, the fruit base were, varied from round - broad. Fruit colour were Yellow green group – 144 A and 144 B. Fruits from Paragpur 1 (G 2 ) showed maximum values of fresh fruit weight (44.41 g), fresh pulp weight/fruit (42.08 g), fresh fruit length (6.46 cm), fresh pulp/kernel ratio (18.06) and dry pulp/kernel ratio (5.39). The present study confirms the need for domestication of T. chebulato be based on two ideotypes, one for fruit flesh [Paragpur 1 (G 2 )] and the other for seed trait [Paragpur 2 (G 3)]. Twenty five RAPD and twelve ISSR primers were effective in revealing polymorphisms among different genotypes of T. chebula. RAPD exhibited 96.76 per cent polymorphism among six genotypes, out of the total, 124 scorable bands, 120 showed polymorphism and 4 bands exhibited monomorphism. Total numbers of amplified and polymorphic fragments generated per ISSR primer revealed 97.92 per cent polymorphism among genotypes. Genotype Paragpur 2 (G 3) came as outliner as revealed by both ISSR study and combined data (RAPD and ISSR). Crossability pattern studies revealed that genotypes under study were cross compatible should be involved in intraspecific breeding programme. The overall profile of various macro and micro minerals as well as other chemical constituents shows T. chebula fruits as highly nutritious. T. chebula fruits should be regularly, used either in the raw form or in the form of 'Jams' and 'Murebba’ so that these fruits become an important part of our diet to supplement human dietary requirements. Signature of MajorThesisItem Open Access Effect of planting dates on growth, flowering and seed production of selected winter annuals(DYSPU, 2013) Sharma, Priyanka; Gupta, Y.C.The present investigation entitled, “Effect of planting dates on growth, flowering and seed production of selected winter annuals.” was carried out at experimental farm of the Department of Floriculture and Landscaping, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan (H.P) during 2010 and 2011. The studies were conducted on six winter annuals namely; annual chrysanthemum (Chrysanthemum coronarium L.), candytuft (Iberis amara L.), godetia or satin flower (Godetia grandiflora Lindl.), helichrysum or paper flower (Helichrysum bracteatum Andr.), snapdragon (Antirrhinum majus L.), sweet william (Dianthus barbatus L.). The experiment was laid out in RBD keeping planting dates as treatments with four replications. Planting was done at an interval of 15 days starting from September 17 in both the years. Plantings dates were; September 17, October 2, October 17, November 1, November 16 and December1. Findings revealed that in case of annual chrysanthemum (Chrysanthemum coronariumL.), maximum plant height (120.29 cm), plant spread (55.50 cm), number of side stems per plant (16.23), duration of flowering (41.43 days), number of flowers per stem (21.98), number of heads per plant (308), number of seeds per head (243.95), seed yield per plant (77.40 g), 1000 seed weight (1.67 g) and benefit cost ratio (14.24) were recorded when planting was done on September 17. Earliest visible flower bud formation (60.08 days), flowering (95.08 days) was also recorded September 17 planting; however head formation was earliest (158.53 days) in December 1 planting. In candytuft (Iberis amara L.) also, September 17 planting resulted in maximum plant height (34.81 cm), plant spread (33.23 cm), number of side stems per plant (6.25), duration of flowering (41.43 days), number of flower clusters per stem (15.46), number of siliquae per plant (3467.72), seed yield per plant (10.25 g), 1000 seed weight (2.18 g) and benefit cost ratio (1.62). Earliest visible flower budformation (55.43days), flowering (77.80 days) was also recorded September 17 planting; however siliqua formation was earliest(157.58 days) in December 1 planting. In case of godetia (Godetia grandifloraLindl.), maximum plant height (68.35 cm), plant spread (50.66 cm), stem length (58.38 cm), duration of flowering (33.91 days), number of flowers per stem (central and side; 110.08 and 15.74), number of capsules per plant (327.50), number of seeds per capsule (90.69), seed yield per plant (9.18 g) and benefit cost ratio (1.21) were recorded when planting was done on September 17. Earliest visible flower bud formation (121.36 days), flowering (151.53 days) and capsule formation (205.42 days) were observed in December 1 planting. In helichrysum (Helichrysum bracteatumAndr.), maximum plant height (92.88 cm), plant spread (46.00 cm), number of side stems per plant (12.35), duration of flowering (45.00 days), numberof flowers per stem (6.02), flower size (5.63 cm),number of heads per plant (65.38), number of seeds per head (534.93), seed yield per plant (24.84 g), 1000 seed weight (0.77 g) and benefit cost ratio (3.07) were recorded when planting was done on September 17. Earliest visible flower bud formation (62.28 days), flowering (100.44 days) was also recorded September17 planting; however head formation was earliest (162.01 days) in December 1 planting. In case of snapdragon (Antirrhinum majusL.), maximum plant height (91.83 cm), plant spread(36.78 cm), stem length (82.00 cm), duration of flowering (39.97 days), number of florets per stem (31.70), number of pods per stem (30.87), number of seeds per pod (390.76), seed yield per plant (8.44 g), 1000 seed weight (0.126g) and benefit cost ratio (1.59) were recorded when planting was done on September 17. Earliest visible flower bud formation (70.18 days) and flowering (107.35 days) was recorded in September 17 planting, however, earliest capsule formation (182.48 days) was observed in December 1 planting. In case of sweet william (Dianthus barbatusL.), maximum plant height (63.54 cm), plant spread (33.32 cm), stem length (56.30 cm), durationof flowering (41.98 days), number of flowers per stem (central and side; 109.42 and 27.58), number of capsules per plant (373.75), number of seeds per capsule (40.85), seed yield per plant (10.75 g) and benefit cost ratio (1.75) were recorded when planting was done on September 17. Earliest visible flower bud formation (71.15 days), flowering (99.50 days) and capsule formation (153.28 days) were observed in September 17 planting.ThesisItem Open Access Characterization of plant growth regulators produced by fluorescent Pseudomonas species and their role in control of replant problem of apple and pear(DYSPU, 2013) Sharma, Shweta; Mohinder KaurIsolation, identification and characterization of native fluorescent Pseudomonas isolates for multifarious plant growth activities from normal and replant site of apple and pear orchard was done for selection of best plant growth regulators producing strains. These were screened out for plant growth promoting activities like production of plant growth regulators, phosphate solubilization, antifungal, siderophores, HCN, ammonia and lytic enzymes. Plant growth regulators are important secondary metabolites produced by fluorescent Pseudomonas sp. such as auxins, gibberellic acid and cytokinins which play significant role in increasing the root surface and length that may help in early establishment of plant in replant site. On the basis of PGPR activities, ten isolates were genotypically characterized. Two best isolates (An-1-kul and An-13-Kul) were selected for 16S rRNA sequencing. The sequence of the 16S rRNA gene is used as a molecular clock to estimate relationships among bacteria and to identify an unknown bacterium to the genus or species level). RAPD studies were also carried out with ten selected isolates which showed the best relatedness among isolates which were isolated from same location. Three regulators auxins, gibberellins and cytokinins were extracted and purified from twobest selected strains of Pseudomonas aeruginosaAn-1-kul and An-13-kul and were purified and characterized by TLC, Sephadex G-25, Dowex-50 column chromatography, high performance liquid chromatography (HPLC) and their specific bioassays. Partial purified cytokinins and auxins extracted from these two strains An-1-kul and An-13-kul were also evaluated by tissue culture bioassay on callus formation and shoot regeneration in broccoli explant and root regeneration in in vitrodeveloped shootlets of cabbage respectively. Three strains of Pseudomonas aeruginosaAn-1-kul and An-13-kul, An-4-shr were used individually and their consortia for treatment of apple and pear seedlings before plantation in replant field at Bajaura and Sharontha respectively. The performance of apple and pear seedlings was much better in terms of plant establishment, growth promotion in terms of plant height, number of nodesand number of branches, chlorophyll content of leaves and NPK of rhizospheric soil over their respective control after nine and twelve months of plantation. These strains can be further exploited and recommended for the management of replant problem of apple and pear after conducting more field trials in replant sites . These characterized strains of Pseudomonas aeruginosa can have great importance in the field of horticulture, especially in the treatment of replant problem of apple and pear especially in Himachal Pradesh.ThesisItem Open Access Agrobacterium mediated cry 1Aa gene transfer in punica granatum L. cv. Kandhari Kabuli(DYSPU, 2013) Verma, Vipasha; Kanwar, KamleshThe present investigation aims at “Agrobacterium mediated cry 1Aa gene transfer in Punica granatum L. cv. Kandhari Kabuli”. A valuable plant regeneration and genetic transformation protocol was developed for Punica granatum L. cv. Kandhari Kabuli using both mature and juvenile explants. Regeneration from mature explants (leaf and nodal segment) procured from 8 years old selected tree and juvenile (cotyledon and hypocotyl) explants excised from 14 to 15 days old in vitro germinated seedlings ofPunica granatum L. cv. Kandhari Kabuli was carried out through indirect and direct method. Calli were initiated from mature leaf, cotyledon and hypocotyl explants. The best media for callus induction from mature (leaf) and juvenile explants were MS medium supplemented with 20.0 µM NAA + 10.0 µM BA and MS medium supplemented with 12.5 µM NAA+15.0 µM BA, respectively. The highest percentage of callus was obtained from cotyledon (84.56%) explants followed by hypocotyl (77.71%) and leaf (76.72%) explants. The calli thus obtained from mature (leaf) and juvenile (cotyledon and hypocotyl) explants showed highest differentiation on MS medium supplemented with 8.0 µM BA + 2.5 µM kinetin + 2.5 µM NAA and MS medium supplemented with 11.0 µM BA + 2.5 µM NAA. The highest percentage of direct adventitious shoot bud induction from mature explants such as leaf (42.95%) and nodal segment (57.80%) was observed on MS medium supplemented with 10.0 µM BA + 2.5 µM NAA and MS medium supplemented with 9.0 µM BA. Solid MS medium supplemented with 9.0 µM BA + 8.0 µM kinetin + 2.5 µM NAA resulted in highest per cent shoot regeneration from both cotyledon (69.60%) and hypocotyl (58.80%) explants. Cotyledon explants was found to be most responsive explants for regeneration through direct as wells as indirect method. The adventitiousshoots obtained were rooted on ½ strength MS medium containing 500 mg l -1 activated charcoal. Sixty five percent of plantlets were successfully established in earthen pots containing soil and sand (1:1). Different explants such as leaf, nodal segment, cotyledon and hypocotyl were transformed using Agrobacterium tumefaciens strain EHA105 harbouring pBinAR-1Aa plasmid carrying cry1Aa (insect resistance gene) and neomycin phoshotransferase-II (nptII) marker through both indirect and direct standardized regeneration protocols. Factors affecting transformation frequency such as age of seedlings, pre-conditioning duration, wounding of explants, immersion duration, co-cultivation duration, presence of acetosyringone and osmoprotectants (betaine HCl and proline) and pH of infection and co-cultivation medium, explants type were studied. Both age of seedlings and pre-conditioning of explants affected putative transformation frequency. Co-cultivation of all the explants (wounded mildly with needle) for 2 days after immersion in Agrobacteriumsuspension for 5 minutes was found to be optimal for transformation. Inclusion of acetosyringone (100 µM) and osmoprotectants (12 µM betaineHCl and 10 µM proline) in the infection and co-cultivation medium (adjusted at 5.8 pH) led to an increase in putative transformation frequency. By applying optimized transformation conditions, the highest putative transformation frequency of 15.12% was obtained with cotyledon explants among all the explants through indirect organogenesis. On the other hand nodal segment showed highest putative transformation frequency of 9.08% as compared to all the other three explants through direct organogenesis. Efficient selection was obtained and escapes were prevented when kanamycin was used at 50 mg l -1 concentration. The putative transgenic shoots were rooted on selective rooting medium and hardened in plastic cups containing sterilized sand. PCR analysis showed integration of cry1Aa and npt-II gene in 55% of the putative transgenic plantlets. Subsequent RT-PCR analysis showed expression of cry1Aa gene in 100% of the PCR confirmed transformed plantlets. Signature of MajorThesisItem Open Access Production, purification and characterization of cellulase free xylanase from Cellulosimicrobium cellulans in solid state fermentation of apple pomace and its application in pulp biobleaching(YSPU, 2013) Walia, Abhishek; Shirkot, C.K.Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is first report on actinomycete from Cellulomonas genera to produce cellulase-free xylanase, which is an important industrial enzyme used in the pulp and paper industry. Strain CKMX1 was characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16Sr DNA and found to be Cellulosimicrobium cellulans CKMX1. Cultural conditions and process parameters i.e. type of medium, particle size of carbon source, incubation period, temperature, initial pH, inoculum size, yeast extract, NH4NO3, urea, peptone, CMC, MgSO4 and CaCO3 were optimized using one factor at a time approach and xylanase activity was increased to 570.0 U/g DBP. CMCase, avicelase, FPase and -glucosidase activities were not detected, highlighting the novelty of the xylanase enzyme produced by CKMX1. Further optimization of enzyme production was carried out using central composite design following response surface methodology with eight independent variables which resulted in very high levels of xylanase (1027.65 U/g DBP). The optimization resulted in 3.1-fold increase of xylanase production, compared with the lowest xylanase production of 331.50 U/g DBP. The enzyme was purified by gel permeation and anion exchange chromatography and had a molecular mass of 29 kDa. Xylanase activity was maximum at pH 8.0 and 60ºC. The enzyme was somewhat thermostable, retaining 50% of the original activity after incubation at 50ºC for 30 min. The xylanase had Km and Vmax values of 26.4 mM and 2,000 μmol/min/mg protein, respectively. All metal ions except HgCl2, CoCl2 as well as CdCl2 were well tolerated and did not adversely affect xylanase activity. The deduced internal amino acid sequence of C. cellulans CKMX1 xylanase by MALDI-TOF MS resembled the sequence of acetyl xylan esterase of the Thermotoga sp. EMP (Accession. no. WP_008192031). Molecular cloning was done by using pGEM-T easy vector and sequence of xylanase gene of C. cellulans CKMX1 showed maximum homology (98%) with xylanase gene of Cellulosimicrobium sp. (Accession no. FJ859907.1). The hydrolytic products of the insoluble oat spelt xylan incubated with xylanase were identified by xylose standards using HPLC. Xylanase effectively hydrolyzed xylan and the hydrolysis by xylanase produced mainly xylose as the main product after 5 min incubation time. Cellulase-free xylanase from C. cellulans CKMX1 under C-EP-D sequence has been shown to bring about a 12.5% reduction of chlorine, decrease of 0.8 kappa points (40%) and gain in brightness was 1.42 % ISO points in 0.5% enzyme treated pulp as compared to control where no enzyme pre-treatment was given, when enzymatically prebleached pulp was charged with 7.4% of total chlorine. From the present studies it is clear that C. cellulans CKMX1 xylanase is having the characteristic suited for an industrial enzyme (xylanases that are active and stable at elevated temperatures and alkaline pH).ThesisItem Open Access Studies on construction of frame work genetic linkage map of Stevia rebaudiana Bertoni using molecular markers(YSPU, 2013) Sharma, Neha; Kaur, RajinderThe present investigation on Stevia rebaudiana was carried out with the objective to construct frame work genetic linkage map of stevia by employing multiple marker systems. Before starting the work on linkage map construction, genetic diversity was analyzed amongst the available genotypes/ clones/ accessions/ morphotypes of Stevia rebaudiana to confirm two parents with differences in rebaudioside-A and stevioside content. This study represents the first genetic linkage study of Stevia rebaudiana comprising of multiple marker systems together. Among the collected 16 accessions polymorphism was studied using 27 RAPD, 26 ISSR and 50 EST-SSR. Results were analyzed in the form of dendrograms, similarity, disimmilarity matrices and polymorphism information content. For linkage map construction, F2 population was used as a mapping population. To survey the polymorphism among contrasting parents 170 RAPD, 26 ISSR and 89 EST SSR were employed and it was observed that 36 RAPD, 10 ISSR and 33 EST – SSR primers were found to be polymorphic. These primers were then used for the genotyping of the mapping population. Phenotyping was carried out by using High Performance Liquid Chromatography (HPLC) of parents as well as segregating population. Both genotypic and phenotypic data were used to construct a linkage map using MAPMAKER/EXP ver 3.0b. A total of four linkage groups were constructed spanning a distance of 927.3 cM with an average distance between loci as 16.29 cM. First linkage group (LG1) comprised of 33 markers, second linkage group (LG2) contained 6 markers and third (LG3) and fourth group (LG4) had 16 and two markers, respectively. On QTL identification, a total of 53 QTL locations were found for both trait 1 (rebaudioside-A) and trait 2 (stevioside). Among 53 QTL locations of trait 1, two major QTLs were found for rebaudioside-A viz., in the marker interval of L67- L71 (ISSR HB-11) - (IISRS-3-L) in LG1 and in the marker interval L38 - L40 (Sigma-5383-027)- (Sigma-5383-029) in LG3 at LOD of 2.5 and 2.7, respectively.