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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 Effect of tree spacing and organic manures on growth and yield of vegetable crops under Melia composita Willd. based agroforestry system(YSPU, 2015) Bhat, Suhail Ahmad; Pant, K.S.The present investigations entitled “Effect of tree spacing and organic manures on growth and yield of vegetable crops under Melia composita Willd. based agrisilviculture system” was carried out at Agroforestry Research Farm, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh for two consecutive years, 2012 and 2013. The experiment was carried out on the existing 6 years old Melia composita Willd. based agroforestry system. The experiment was laid out by using Randomized Block Design (factorial) with seven different doses of organic manures, T1 FYM (5 t ha-1), T2 FYM (7.5 t ha-1), T3 FYM (10.5 t ha-1), T4 vermicompost (2.5 t ha-1), T5 vermicompost (3.5 t ha-1), T6 vermicompost (5 t ha-1), T7 control (no manure) with three replications and two distinct tree spacings viz. 8 m x 5 m and 8 m x 4 m under Melia and one open condition (without trees). The pooled result revealed that yield and growth performance of winter season vegetable crops was almost same for both spacings and open spacing. In summer season crop yield reduction upto 45% and 50% was observed in capsicum under tree spacings 8m x 5m and 8m x 4m respectively when compared to sole cropping. Similarly 70% and 74% fruit yield reduction was also observed in tomato under tree spacings 8m x 5m and 8m x 4m respectively as compared to sole cropping. The difference in crop yield during winter season was found to be negligible. Among pea crops, maximum pod yield was recorded in spacing 8m x 4m which was significantly at par with 8m x 5m whereas, minimum pod yield was recorded in sole crop. However in cauliflower, maximum curd yield was recorded in sole crop and minimum curd yield was recorded in tree spacing 8m x 4m which was, however, statistically at par with spacing 8m x 5m. On the other hand optimum use of organic manure benefited the intercrop by improving the growth of plants even in the presence of trees as compared to sole plots during both Kharif and Rabi crop season. In most of the cases vermicompost @ 5 t ha-1 showed highest crop yield followed by Vermicompost @ 3.5 t/ha and FYM @ 10 t ha-1. On contrary with that, control plots (no manures) recorded minimum crop yield, during both the cropping seasons. Use of organic manures increased the growth of vegetable crops, higher doses of organic manures were found more effective in improving the growth of vegetable crops in presence and absence of Melia composita trees. The findings of present study suggested that cultivation of vegetable crops in combination with trees like Melia composita, especially during winter season may be recommended for better economic return from the agroforestry systems. However, during summer season decrease in growth and yield parameters of tomato and capsicum within the agroforestry system probably indicates intense competition for critical resources like water, nutrients, photo synthetically active radiation. Soil chemical properties like soil organic carbon, pH, EC and nutrients availability were improved under agroforestry system than sole crop system. The availability of nutrients like N, P, K were high where, higher doses of organic manures were applied in agroforestry system as well as in sole crop system. It also suggested that the agroforestry system have higher potential of producing biomass (woody and nonwoody) than monocropping system.ThesisItem Open Access Biological and molecular characterization of apple stem pitting virus(YSPU, 2015) Brakta, Ajay; Thakur, P.D.Surveys conducted during 2011 and 2012 in different apple growing areas of Himachal Pradesh, Uttarakhand and Jammu and Kashmir revealed viral disease incidence ranging from 3 to 60 per cent. Chlorotic spots coupled with necrotic lesions on apple leaves were the predominant symptoms. Orchards located at Regional Horticulture Research Station, Mashobra and Dhangvi village of Kotkhai area were selected for conducting biological and serological detection of apple stem pitting virus (ASPV). Serological detection through DAC and DAS-ELISA resulted in the detection of apple stem pitting virus (ASPV) either alone or in mixed infection with apple chlorotic leaf spot virus (ACLSV), apple mosaic virus (ApMV) and apple stem grooving virus (ASGV). Biological detection of one of the ASPV seropositive isolate on herbaceous hosts resulted in the production of symptoms on Chenopodium quinoa, C. amaranticolor, Nicotiana tabacum var. White Burley and Phaseolus vulgaris. Detection on woody indicators (Malus pumila Spy 227 and Jay Darling) under field conditions through double grafting of inoculators and indicator budwood resulted in the production of typical viral symptoms on leaves in Jay Darling and Spy 227indicators. Graft incompatibility and necrotic symptoms were produced at the graft union of Spy 227 indicator followed by decline and dieback. Leaf samples drawn during March to May months were found suitable for the ELISA of ASPV whereas petals were the best source in addition to seropositive detection of ASPV in anthers and sepals. Association of ASPV with viral symptoms in apple was also confirmed by RT-PCR assays. Internal control primers along with coat protein gene specific primers were used to overcome the problem of false negative results. Molecular characterization of full coat protein gene of 3 ASPV isolates, 5 ASGV isolates and partial characterization of coat protein gene of 10 ACLSV isolates was carried out and the resultant sequences were then submitted to NCBI. Phylogenetic analysis of sequences showed the presence of variability among isolates and confirmed that there is no correlation between the geographic origin and genetic diversity of these isolates, which does not allow drawing conclusion on their origin and dispersion. Serological indexing resulted in the selection of 13 trees of 7 cultivars free from infection of ASGV, ACLSV, ASPV and ApMV in ELISA test of 36 symptomless trees of 12 cultivars.ThesisItem Open Access Studies on Marssonina blotch pathogenesis and its management through novel methods(YSPU, 2015) Phurailatpam, Sumitra; Sharma, J.N.Marssonina blotch caused by Marssonina coronaria is one of the most important disease of apple particularly in Himachal Pradesh. Present investigations were undertaken with the objectives to carry out detailed study on Marssonina blotch pathogenesis in apple, investigate the effect of Marssonina blotch on the physiological parameters of the host plants and develop novel methods of disease management through induced resistance. Based on morphological and molecular characters, the isolates collected from different locations of Himachal Pradesh and Uttarakhand associated with Marssonina blotch of apple were confirmed to be belonging to Marssonina coronaria. Colonies of all the isolates were dark brown to black without aerial mycelium with colony diameter ranging from 9 to 14 mm. Conidia were hyaline, ampule shaped, one septate, constricted at the septum and guttulate, and the conidial size ranged from 3-8×12-28 μm. Molecular characterization of the seven isolates of Marssonina spp. showed 96-100% homology to with Marssonina coronaria. Phylogenetic tree depicted that all the isolates were close clustered to Chinese isolates deposited earlier in the NCBI. Pathogenesis studies revealed highest percentage of conidia germination of Marssonina coronaria on the leaves of Starking Delicious cultivar and minimum in Granny Smith. Whereas, shortest incubation and latent periods were recorded in Starking Delicious leaves showing that the cultivar was susceptible to all the isolates tested, while Tydeman’s Early Worcester showing moderately susceptible and Granny Smith moderately resistant reaction. Effect of Marssonina coronaria infection on host physiology studies of different apple cultivars revealed that reducing and non-reducing sugar contents were more in uninoculated leaves of all the test cultivars which reached maximum with time lapse. Total phenolic content, polyphenol oxidase, peroxidase and PAL activity, auxins and gibberellins were more in inoculated plants during all the sampling intervals in all the three test cultivars. Induced resistance studies against Marssonina blotch pathogen on potted plants by using systemic acquired resistance (SAR) chemicals and bioagents revealed that salicylic acid and acibenzolar-S-methyl @ 100 ppm were found to be at par and most effective in all the cultivars tested with minimum disease severity (1.08 to16.50 %) and maximum disease control (66.77 to 88% ) in the polyhouse.Whereas, among bioagents tested, Pseudomonas flourescens @ 10-5 dilution was the best with minimum disease severity (1.33 to 18.50 %) and maximum disease control (61.45 to 83.33 %). To know the mechanism of induced resistance in all the cultivars tested after treatment with SAR inducers and bioagents, post-inoculation with the pathogen showed that the biochemical parameters (reducing and non- reducing sugar contents; total phenolic content, peroxidase, polyphenol oxidase , PAL activity; -l, 3-glucanase and chitinase activity and ROS production) were highest in salicylic acid treated leaves and minimum in dipotassium phosphate; whereas in bioagent treated leaves they were recorded in highest levels in case of Pseudomonas flourescens and minimum in Bacillus subtilis. Further, on comparison among the cultivars, it was found that their contents were higher in resistant Granny Smith leaves than that in the susceptible cultivar Starking Delicious.ThesisItem Open Access Studies on effect of plant growth promoting rhizobacteria and GA3 on plant growth, fruiting and soil health of strawberry (Fragaria x ananassa Duch.) cultivar Chandler(YSPU, 2015) Seema Kumari; Mehta, KuldeepThe present investigations entitled “Studies on the effect of plant growth promoting rhizobacteria and GA3 on plant growth, fruiting and soil health of strawberry cultivar Chandler” were conducted in the experimental area of Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, during the year 2013 and 2014. The studies were conducted under two different experiments, with the objectives to study the effect of plant growth promoting rhizobacteria on plant growth, yield, quality, leaf nutrient status and soil health, and to study the effect of GA3 and its combinations with plant growth promoting rhizobacteria on plant growth, yield, quality and leaf nutrient status. The experiments were laid out in randomized block design consisting 35 treatments along with control and replicated thrice. In the present investigation, Experiment-1 included the application of five different isolates of plant growth promoting rhizobacteria (S1: Bacillus licheniformis CKA 1, S2: Bacillus subtilis CB 8 A, S3: Bacillus sp. RG1, S4: Bacillus sp. S1 and S5: Bacillus sp. S2) to the strawberry plants through root dip (at the time of planting), foliar application (20 days before expected flowering) and root dip+ foliar application methods, while in Experiment- II, different concentrations of GA3 (25, 50 and 75 ppm) alone or in combinations with these isolates applied as foliar application. Among the different treatment combinations, S1, S3 and S4, alone applied as root dip + foliar application methods or in combination with GA3 @ 75 ppm as foliar application method, were recorded to be most effective isolates with respect to plant height, spread and number of crowns per plant, whereas S4 and S5 was effective in increasing leaf area and number of runners per plant. All the isolates alone or in combination with GA3 resulted in early flowering, harvesting, longest flowering and harvesting durations and yield as compared to the control. The number of fruits and yield per plant were recorded highest with the individual application of S1, S2 and S4 through root dip + foliar application method or in combination with 75 ppm GA3. In case of fruit quality, the isolates S1, S3 and S5 resulted in increased fruit quality viz., fruit weight, size, TSS, acidity, sugars and ascorbic acid. The TSS and sugars were maximum when these isolates were applied alone through root dip or in combination with 25 ppm GA3. The increase in GA3 concentrations resulted in decreased TSS and sugar content, and increase in acidity and ascorbic acid content. The anthocyanin content was maximum with the application of S1 and S2 through root dip method or in combination with 50 ppm GA3 but showed a decline in content at higher GA3 concentration. Results further showed that the leaf nutrient status (N, P, K, Ca, Mg, Zn, Fe, Cu and Mn) were also increased with the application of S1, S2 and S5 when applied through root dip + foliar application method or in combination with GA3 @ 75 ppm. However, improved soil health and soil biological properties (available N, P, K, Zn, Fe, Cu, Mn, total rhizobacteria and endo-rhizobacteria, phosphate solubilizing, siderophore producing and nitrogen fixing bacteria) were also observed with the application of these isolates when applied through root dip + foliar application methods.ThesisItem Open Access Phytophagous mites and their natural enemies in different horticultural ecosystems of Himachal Pradesh(YSPU, 2015) Singh, Vijay; Chauhan, UshaOf the seventy nine species recorded in the present study, twenty one species were of phytophagous mites and fifty eight were natural enemies. Phytophagous mites were Tetranychus urticae Koch, T. ludeni Zacher, T. sp. nr. ludeni, T. hypogeae Gupta, T. neocaledonicus Andre, T. udaipurensis Gupta & Gupta, T. sayedi Baker & Pritchard, T. macfarlanei Baker & Pritchard, T. sp. nr. macfarlanei, Panonychus ulmi (Koch), P. citri (McGregor), Eotetranychus frosti McGregor, Eutetranychus orientalis (Klein), Bryobia praetiosa Pritchard & Baker, Porcupinychus abutiloni Anwarullah, Brevipalpus essigi Baker, B. cucurbitae Mohansundaram, B. euphorbiae Mohansundaram, Aculus malus Zaher & Abou Awad, Aceria litchi (Keifer) and Tarsonemus sp. Of fifty eight natural enemies, forty six were predatory mites and twelve species were of predatory arthropods. Predarory mites were Amblyseius aerialis (Muma), A. largoensis (Muma), A. cucurbitae Rather, A. herbicolus (Chant), A. multidentatus (Swirski & Shechter), A. guajavae Gupta, Euseius prasadi (Chant & McMurty), E. eucalypti Ghai & Menon, E. sp. nr. ahaioensis, E. alstoniae Gupta, E. finlandicus (Oudemans), E. neococcinae Gupta, E. sp. nr. coccineae, E. delhiensis (Narayanan & Kaur), E. chitradurgae Gupta, N. paspalivorus (DeLeon), N. longispinosus (Evans), N. sp. nr. neoghanii, Typhlodromus (Anthoseius) hadii Chaudhri, T. (A.) mori Gupta, T. (A.) pruni Gupta, T. (A.) sp. nr. channabasavannai, T. (A.) himalayensis Gupta, T. (A.) darjeelingensis Gupta, T. (A.) sp. nr. darjeelingensis, Phytoseius roseus Gupta, P. maldahensis Gupta, P. sp. nr. maldahensis, P. intermedius Evans & Mcfarlanne, P. crinitus Swirski & Shechter, P. corniger Wainstein, P. kapuri Gupta, Agistemus fleschneri Summers, A. terminalis (Quayle), A. obscura Gupta, A. sp. nr. obscura, A. sp. nr. macrommatus, A. sp. nr. heterophylla, Tydeius cumini Gupta, Pronematus fleschneri Baker, Acarus gracilis Margaret, Tyrophagus putrescentiae (Schrank), Anystus baccarum (Linnaeus), Walzia darjeelingensis Gupta, Lasioseius sp., Leptus sp. Eleven species were new record. T. urticae, T. (A.) mori and T. (A.) himalayensis were relatively abundant on fruits, T. urticae, Euseius prasadi and E. delhiensis on ornamental and T. urticae, A. largoensis and E. finlandicus on vegetable crops. Maximum number of prey and predatory mites were recorded on fruits (48) followed by vegetables (25) and ornamentals (17). Maximum numbers of natural enemies were also recorded on fruits (36) as compare to vegetables (21) and ornamentals (17). The most dominant species on apple were T. urticae, P. ulmi, T. (A.) himalayensis and T. (A.) mori, whereas, on rose were T. urticae, Feltiella sp. and E. prasadi, on carnation were, T. urticae, T. ludeni and S. sexmaculatus, on cucumber were T. macfarlanei and A. largoensis and on tomato were T. ludeni and N. sp. nr. neoghanii. T. (A.) hadii and Oligota sp. were the new record on apple and rose in the state.ThesisItem Open Access Variability and management studies of Botrytis cinerea causing grey mould in gladiolus(YSPU, 2015) Kaur, Gurvinder; Chandel, SunitaBotrytis Grey Mould (Botrytis cinerea Pers. ex Fr.) in gladiolus is one of the major limiting factors 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, variability among the Botrytis isolates on the basis of cultural, morphological, pathological and molecular parameters. The role of environmental factors in disease development, germplasm screening and devising of suitable disease management strategies were elucidated. Botrytis grey mould was found to occur in moderate to severe form in different gladiolus growing areas of Solan, Shimla, Mandi, Bilaspur, Kullu and Sirmour districts of Himachal Pradesh. The average disease incidence and severity in two years of surveys (2012- 2013) ranged between 14.83 to 64.92 and 2.98 to 33.33 per cent, respectively. Based on the cultural, morphological and molecular characterization, out of fifteen isolates, fourteen were identified as Botrytis cinerea and one was identified as Botrytis fabae. Mycelial growth and sporulation was recorded maximum on PDA media at temperature of 20oC however sporulation was obtained excellent at temperatures 10 and 15oC. Sclerotia formation was more at low temperature compared to high temperatures (30oC). Fructose and asparagines were adjudged best as carbon and nitrogen sources for multiplication of the pathogens. There was variation in among the isolates in respect to physiological studies. Molecular variability was carried by using 45 RAPD primers, out of which only 25 primers were amplified and showed 100 per cent polymorphism. DNA amplification was done by using 16 ISSR primers, only 14 primers were amplified and gave 143 polymorphic bands and showed 100 per cent polymorphism. Moderate temperature (22±2oC) coupled with high RH (80%) favoured the disease development under natural epiphytotic conditions. Esta Bonita, Grand Prix and Amesterdam cultivars were found to be resistant against grey mould under natural conditions. In vitro effect of different fungicides on mycelial growth of B.cinerea revealed that two fungicides i.e., Quintal and Contaf gave no mycelial growth with cent per cent inhibition at all the concentrations tested. Amongst different bioresources, minimum growth was recorded in Allium sativum and Azadirachta indica. Amongst 15 plant oil peppermint, geranium, palmarosa, citronella and oregano completely inhibited the growth of B. cinerea. Integrated disease management studies revealed that the treatments, Quintal+ Allium sativum + Neem oil and Contaf + Allium sativum + Neem oil gave minimum disease severity and maximum disease reduction of 94.40 and 93.19 per cent, respectively. Plant growth parameters viz., plant height, spike length,no. of flowers/ plot and yield also increased with application of these combinations in field conditions during the years 2012-2013.ThesisItem Open Access Studies on reproductive success of Aconitum heterophyllum under domestication(YSPU, 2015) Vinod Kumar; Raina, RavinderThe present investigation entitled “Studies on reproductive success of Aconitum heterophyllum under domestication” was carried out both in field as well as in laboratory during 2010-15. The field studies were conducted at twenty locations (10 wild and 10 domesticated) spread across different regions of Himachal Pradesh and at Medicinal and Aromatic Plants Research Farm, Shilly located at an altitude of 1550 m; Latitude N 30054’30”; Longitude E 77007’30”. The laboratory studies were conducted in the laboratory of the Department of Forest Products, College of Forestry, Dr. Y S Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.). These studies were divided into six experiments namely (i) Inter-population studies on vegetative parameters under wild and domesticated conditions, (ii) Inter-population studies on floral biology under wild and domesticated conditions, (iii) Chromosomal studies ,(iv) Studies on sexual behaviour under wild and domesticated conditions, (v) Inter population studies on seed quality and germination and (vi) Molecular studies. Vegetative characters as well as tuber yield were positively impacted by domesticated conditions which makes this species amenable for cultivation. Non significant variation in seed output was observed between domesticated and wild conditions indicating no effect of domestication on seed output. Maximum seed germination of 67.78% was recorded in seed sourced from population VIID (Nichar, Kinnaur), no significant difference was noticed in seed germination amongst the domesticated and wild populations. However of the twenty wild and domesticated populations, two populations under each condition, (IW) Sisoo and (VIW) Puikar, Lahaul & Spiti under wild condition; Solang Nalla, Kullu (XD) & Sisoo, Lahaul & Spiti (IID) under domesticated conditions were found to be best on the basis of morphological as well as yield characters. Based on floral characters like flower colour, no. of sepals, colour of anthers and no. of follicles per flower, population IIW (Chansal) appears to be unique amongst all wild populations. The flowers of this polulation were dark blue coloured in sharp contrast to bluish purple in all other populations. RAPD analysis also indicated this population to be genetically unique as it formed a singlet in dendrogram. Flowers were observed to be dichogamous (protogamy) and no difference in ploidy level was observed in all studied populations (2n=2x=16). Genetic diversity among the populations was confirmed by both RAPD and ISSR markers, with some populations found to be genetically similar. RAPD analysis revealed that populations VW (Moral Danda, Shimla) and IXW (Kalatop, Chamba) are genetically similar whereas population IIW (Chansal, Shimla) formed a singlet indicating its genetic uniqueness. ISSR markers analysis revealed that populations VIIID (Topi Nalla, Chamba) and IID (Sisoo, Lahaul & Spiti) are also genetically similar. Both the marker systems have shown that populations (i.) populations VIID (Nichar) & VIIIW (Chot Kanda), (ii) populations VIIID (Topi Nalla) & IID (Sisoo), (iii.) populations VW (Moral Danda) & VID (Katgaon) are genetically closely placed, though their sites of occurrence happen to be quite far away from each other. The observations indicate suitability of Aconitum heterophyllum for domestication to be grown as a sole crop which improves its yield capacity with regard to its tuber (economic part) and seed (for propagation). The study has indicated that: i) Under domestication, the yield characters of Aconitum heterophyllum improve making it suitable for cultivation. ii) Domesticated populations like XD (Solang Nalla), ID (Shillaru, Shimla), IIID (Jagatsukh) and VIID (Nichar) were found to be the best as compared to others in terms of tuber yield. iii) No significant difference in seed yield was observed under domestication conditions indicating that domestication does not adversely impact seed production potential. iv) The population growing at Chansal (IIW), needs to be conserved as it was found to be unique on the basis of morphological as well as molecular characters.ThesisItem Open Access Effect of tree spacing and organic manures on growth, flower and seed production of floriculture crops under Grewia optiva Drummond. based agroforestry system(YSPU, 2015) Prem Prakash; Pant, K.S.The present investigation entitled “Effect of tree spacing and organic manures on growth, flower and seed production of floriculture crops under Grewia optivaDrummond. based agroforestry system” was carried out in the experimental farm of Department of Silviculture and Agroforestry, Dr.Y.S.Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.) for consecutive two years during 2012-2014. The experiment was laid out in split plot design. The study was aimed at exploring the possibilities of successful cultivation of floriculture crops as intercrop under Grewia optivaalong with the different doses of organic manures used. The experiment comprised of four tree spacing (1m, 2m, 3m and open) and eight doses of organic manures (5.00 kg FYM,3.00 kg vermicompost,5.00 kg municipal solid waste, PGPR, 5.00 kg FYM + PGPR , 3.00 kg vermicompost + PGPR , 5.00 kg municipal solid waste + PGPR and control i.e. without manures ) in combination with and without Grewia. The results revealed that winter and summer annuals can be grown successfully grown under agroforestry system. The use of vermicompost and FYM in combination with PGPR at appropriate doses was found beneficial for minimizing competition for critical resources and enhancing the productivity of floriculture crops. Vermicompost @2.00 kg/plot + PGPR found to be the best dose followed by FYM @ 5.00 kg/ plot +PGPR. Alone PGPR was not as much effective, while in combination with FYM, Vermicompost, FYM and Municipal solid waste it significantly increases the growth, flower and seed parameters of the winter and summer annuals. It was observed that winter annuals i.e. Dianthus barbatus L. and Godetia grandiflora L. performed well under Grewia optiva in compare to summer annuals i.e. Callistephus chinensis L. Nees and Gomphrena globosa L. Godetia grandiflora got the maximum seed yield under agroforestry system as well as in open condition. Flower parameters of Dianthus barbatus found to be the best under agroforestry system. Growth, flower andseed production parameters were also found to be higher in open condition as compared to beneath tree canopy. Among 3 spacing of tree 3m spacing found to be best for overall growth, flower and seed production of floriculture crops. The present study reveals that the maximum leaves and branch wood biomass was recorded in 3m spacing. Soil physical and chemical properties found to improve under agroforestry system as compare to open (tree less area). The availability of nutrients N, P and K was high where, applied with organic manures in agroforestry system as well as in sole crop. The benefit cost ratio of growing floriculture crop with Grewia optivaranged from 1.66 to 3.78 where it was 1.55 to 3.53 for sole condition. Integrating flower crops with Grewia optiva offers an excellent opportunity for diversificationand high income generating than the monoculture. It will also open new vistas for the research to evaluate the performance of flower crop under tree canopy.ThesisItem Open Access Bioprospecting of thermophilic bacteria from hot water springs of Himachal Pradesh for laccase enzyme production(YSPU, 2015) Sharma, Ruchika; Shirkot, PoonamLaccase enzyme has acquired the status of ‘green catalyst’ as it possesses remarkable bioremediation potential along with numerous applications in effluent detoxification, degradation of textile dyes, herbicide and insecticide degradation, wine clarification, enzymatic conversion of chemical intermediates, biosensors and organic synthesis. In the present study, significantly high diversity of laccase producing bacteria from hot water springs of Himachal Pradesh was assessed. A total of 118 laccase producing thermophilic bacterial isolates were obtained from 200 hot water spring samples using TYM containing 5 mM guaiacol which were morphologically characterized. These were rescreened on the basis of their ability to oxidise tannic acid, dimethoxyphenol and syringaldazine leading to selection of 50 laccase producing thermophilic bacterial isolates, which were characterized biochemically. Eighteen thermophilic bacterial isolates exhibiting maximum laccase activity of 0.0007-0.0038 U/l were selected for further, molecular characterization using RAPD-PCR and 16S rrna gene technology. In silico analysis of 16S rrna gene sequences led to identification of these bacterial isolates and they were found to belong to genus Bacillus, Aneurinibacillus and Pseudomonas, as Bacillus licheniformis strain RSV20, Bacillus licheniformis strain RSM8, Bacillus licheniformis strain RSV10, Bacillus sonerensis strain RSM17, Bacillus sonerensis strain RSV8, Bacillus licheniformis strain RSP1, Bacillus licheniformis strain RSP2, Bacillus licheniformis strain RSP3, Bacillus licheniformis strain RSP7, Bacillus sonerensis strain RSP5, Bacillus sonerensis strain RSP11, Aneurinibacillus thermophilus strain RSP13, Bacillus aerius strain RSP4, Bacillus aerius strain RSP9, Bacillus subtilis strain RSP8, Bacillus amyloliquifacience strain RSP10, Bacillus pumilis strain RSP12 and Pseudomonas taiwanensis strain RSP6. On the basis of maximum laccase enzyme activity Bacillus licheniformis strain RSM8 was selected for production and purification of the laccase enzyme. Maximum extracellular enzyme production was achieved at 60°C, pH 9.0 and 24 hrs incubation with 5 mM guaiacol, 5 % tryptone and 3 % yeast extract in combination with nitrogen source. Crude extracellular thermolaccase enzyme preparation was purified by ammonium salt precipitation (50-90%) followed by gel filtration and ion exchange chromatography which showed 15.21 yield and 10.5 fold purification. The purified enzyme had optimal activity at pH 9.0 and 60 °C, and 16.22 μM Km value. The molecular weight of thermolaccase in the present study was found to be 72.5 kDa. However activity was inhibited by sodium azide and DTT. Bacillus licheniformis strain RSM8 as well as its enzyme preparations were investigated for their ability to decolourize dyes which are the potential contributors of water pollution. Six different synthetic dyes were decolourized RBBR (68 %), congo red (86 %), indigo carmine (73 %), brilliant blue (40 %), bromophenol blue (51 %) and aniline blue (54 %) when treated with the crude enzyme preparation of Bacillus licheniformis strain RSM8. And partially purified enzyme preparation of Bacillus licheniformis strain RSM8 showed greater decolourization of dyes comparatively RBBR (74 %), congo red (91 %), indigo carmine (80 %), brilliant blue (60 %), bromophenol blue (64 %) and aniline blue (67 %). The purified enzyme was successfully immobolized using adsorption method in calcium alginate beads with 76% immobolization percentage and immobolized laccase enzyme beads were studied for their ability to degrade dyes. The stability and reusability of the immobilized enzyme system has the potential to make the entire treatment process inexpensive. Bacillus licheniformis strain RSM8 enzyme preparations was investigated for phytotoxicity evaluation of three dyes viz., Congo red, RBBR and Indigo carmine and each of enzyme treated dyes for Phaseolus mungo and Calendula officinalis and Tagetes patula plant species respectively, under in vitro conditions and Phaseolus mungo with Congo red dye under in vivo conditions. Significant germination inhibition, a slower rate of plumule and radicle seedlings growth was observed for Congo red, RBBR and Indigo carmine dyes as compared to enzyme treated dyes. An extracellular laccase producing gene has been isolated using degenerate primer based on the copper I and II conserved site of laccase enzyme, from the hot water spring bacteria, Bacillus licheniformis strain RSM8 followed by determination of the amino acid which were translated from nucleotide sequence and encodes a polypeptide comprised of 50 amino acids showeng 97 % identity with the amino acid sequences of bacterial laccases i.e. copper oxidase [Bacillus licheniformis]. Further multiple sequence alignment using MULTALIN and structure prediction using Phyre1 & 2 revealed conserved histidine residues.