Browsing by Author "Sharma, Sneh"
Now showing 1 - 9 of 9
Results Per Page
Sort Options
ThesisItem Open Access ASSESSMENT OF GENETIC FIDELITY OF IN VITRO RAISED PLANTS OF GLORIOSA SUPERBA L. THROUGH MOLECULAR MARKERS(COLLEGE OF HORTICULTURE AND FORESTRY, DR Y S P UHF, NERI, HAMIRPUR, 2020-06-22) Devi, Bandna; Sharma, SnehThe present investigation on the “Assessment of genetic fidelity of in vitro raised plants of Gloriosa superba L. through molecular markers” was carried out in the Department of Biotechnology. Gloriosa superba L. is an important endangered medicinal plant belongs to family Colchicaceae. It is a perennial tuberous herb extensively scattered in tropical and sub-tropical parts of India. Due to the medicinal value, these plants are collected from wild as raw material for large scale medicinal industry, leading to overexploitation and it is the main reason to include this plant species in the Red Data Book. Hence, to overcome this problem micropropagation is an alternate and effective method. An efficient in vitro regeneration protocol has been established using shoot tips and tuber explants. Explants were sterilized using different concentrations (0.05-1.0 w/v) of mercuric chloride for varying time exposure. Explants were cultured on MS medium containing different concentrations and combinations of cytokines (BAP, NAA and Kn). The best shoot induction was observed on MS medium supplemented with 2.0 mg/l BAP in case of tuber explants and in case of shoot tips explants best shoot induction was observed on MS medium supplemented with 1.5mg/l BAP and 0.5 mg/l NAA. Best rooting was obtained from shoots cultured on half-strength of MS medium fortified with 2.0 mg/l IBA. In vitro raised shoots were transferred to hardening media containing different combinations of substrates. Among all the combinations, sand: soil: vermicompost (1: 2: 1) produced the highest survival rate of 73.33%. A large number of plants can be produced in vitro under aseptic conditions, but there is always a danger of producing somaclonal variants by tissue culture technology. Thus, the genetic fidelity of micropropagated clones was evaluated by using random amplified polymorphic DNA (RAPD) and inter specific sequence repeat (ISSR) analysis. During the study a total of 17 primers were screened, out of which 2 RAPD and 2 ISSR primers produced clear, distinct and reproducible amplicons. Fragments were scored for the presence of band as 1 (band present) and absence of band as 0 (band absent) and thus a matrix was obtained and further analyzed with NTSYS-pc software by using Jaccard’s coefficient to calculate the similarity matrix for mother and in vitro raised plants. Dendrogram was plotted using UPGMA between mother plant and in vitro plants of both, RAPD and ISSR, markers. A preliminary phytochemical analysis among the mother and in vitro raised plants showed a higher yield of secondary metabolites. The in vitro regeneration protocol provides a basis for germplasm conservation and also harnesses the various secondary metabolites of medicinal importanceThesisItem Open Access Biosynthesis and characterization of silver nanoparticles using S. mukorossi extract possessing antimicrobial potential(College of Horticulture and Forestry Dr YSP UHF, Neri, Hamirpur(H.P.), 2023-11-28) Thakur, Sapna; Sharma, SnehThe present study on the “Biosynthesis and characterization of silver nanoparticles using S.mukorossi extract possessing antimicrobial potential” were carried out in Department of Biotechnology, College of Horticulture and Forestry (Dr. Y.S. Parmar University of Horticulture and Forestry), Neri, Hamirpur (H.P) during 2022-2023. Sapindus mukorossi is an extremely valuable cultivated medicinal plant which is mostly found in hilly regions of India. In the lists of herbs, it is a popular herb in Ayurveda and is used as an important ingredient in cleansers and shampoos. It is well known for its folk medicinal values and known by several names like washnut, soapnut, soapberry, dodan and ritha. The use of this plant promoted considerable attention because of their various biological and pharmacological activities. The major compound isolated from genus Sapindus are saponins, triterpeniods, fatty acids and flavonoids are well known for their antimicrobial, antidiabetic, cytotoxic, fungicidal, and anti-inflammatory activities. The study was undertaken for biosynthesis and characterization of silver nanoparticles derived from S. mukorossi pericarp extract. The sample was collected from village Sheglagalu, Chail Chowk, Distt. Mandi (H.P). Synthesis was carried out using different solvents viz., methanol, chloroform and aqueous. It was found that distilled water acts as a better solvent for the synthesis of silver nanoparticles. The characterization of prepared silver nanoparticles was assessed using distinct techniques namely UV-vis spectroscopy, X-ray diffraction (XRD), Fourier transmission infrared (FTIR) and High resolution- transform electron microscopy (HR-TEM). The antioxidant activity of aqueous extract and biosynthesized silver nanoparticles was evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The different concentrations of sample ranging from (100-1000 µg/ml) was prepared and mixed with 3 ml of DPPH solution. The maximum antioxidant activity was confirmed at 1000 µg/ml of concentration that is 91 % and 88 %, respectively. The antimicrobial property of silver nanoparticles derived from S. mukorossi extract was evaluated against four strains viz., P. aeruginosa, E. coli, B. subtilis and S. aureus. The maximum antimicrobial property was seen against gram-negative bacterium P. aeruginosa, E. coli then gram-positive bacterium B. subtilis and S. aureus. These results suggest that silver nanoparticles made from S. mukorossi extract may beused as antimicrobial and antioxidant agents in several fields, including medicine, agriculture, and personal care products.ThesisItem Open Access Green Synthesis, Characterization and Application of Silver Nanoparticles using Cauliflower (Brassica oleracea var. botrytis L.) extract(College of Horticulture and Forestry Dr YSP UHF, Neri, Hamirpur(H.P.), 2023-11-28) Prashar, Ankita; Sharma, SnehThe present investigation on the “Green Synthesis, Characterization and Application of Silver Nanoparticles using Cauliflower (Brassica oleracea var. botrytis L.) extract.” were carried out in the Department of Biotechnology, College of Horticulture and Forestry (Dr. Y.S. Parmar University of Horticulture and Forestry), Neri Hamirpur (HP) during 2022-2023. Silver nanoparticles have gained significant attention in various fields due to their unique physical, chemical, and biological properties. Nanotechnology offers the possibility of modifying the materials used in conventional packaging for food and enables the introduction of fresh ideas for quality control, food safety, identification of pathogens by mechanical means, and barrier properties. Nanoparticles are also carried out in relation to food, its manufacturing, processing, and specially packaging, which is intimately tied to the shelf-life of food production. The cauliflower is a rich source of predominantly S-methylcystein sulfoxide, sinigrin, and glucobrassisin, which have predominant anti-carcinogenic properties. The planting material were collected from experimental farm in Department of Vegetable science at College of Horticulture and Forestry Neri, Distt. Hamirpur, Himachal Pradesh. The aim of this study was to synthesize silver nanoparticles from cauliflower extract and investigate the antibacterial activity and evaluation of biosynthesized nanoparticles in post-harvest. The UV-vis spectrum shows the absorption peak at 412 nm which confirms the synthesis pf AgNPs. FTIR spectroscopy confirms the presence of bio-compound functional groups on the surface of AgNPs. The particle size of AgNPs were found to be 10-50 nm in HR-TEM. In XRD, AgNPs determined to be cubic crystal structure with face centered. The synthesized silver nanoparticles from cauliflower showed antibacterial activity against B. subtilis, E. coli, P. aeruginosa and S. aureus. These properties utilized to reduce the bacterial load on vegetables, thereby inhibiting the spoilage and extending the shelf life of vegetables. The biosynthesized silver nanoparticles can act as barrier against moisture loss, gas exchange, and microbial contamination, thus preserving the freshness of vegetables and increasing their shelf life. The antioxidant activity of biosynthesized silver nanoparticles and aqueous extract of B. oleracea var. botrytis was carried out by using 2,2-diphenyl-1-picryhydrazyl (DPPH) assay. The different concentrations of AgNPs and aqueous extract (50, 100, 150, 200, 250, 300 µg/mL) mixed with 3 mL of DPPH. The result revealed that the antioxidant activity of biosynthesized silver nanoparticles and aqueous extract of B. oleracea var. botrytis was increased at 200 µg/mL and maximum antioxidant of solutions observed 80.04 % and 69.07 % at 300 µg/mL due to the presence of polyphenols, vitamin A, beta carotene and sulphrophane.ThesisItem Open Access “In vitro propagation and phytochemical assessment of Andrographis paniculata.”(College of Horticulture and Forestry Neri, Hamirpur (H.P.), 2021-11-10) Thakur, Neha; Sharma, SnehThe present investigation on the “In vitro propagation and phytochemical assessment of Andrographis paniculata” was carried out in the Department of Biotechnology. Andrographis paniculata is an important endangered medicinal plant belongs to family Acanthanaceae. It is a annual herbaceous plant found commonly in India, China and Sri Lanka. Conventional vegetative propagation of this important medicinal plant is very difficult and too slow to meet the commercial quantities required and it is not possible to produce large number of clones. Hence, to overcome this problem micropropagation is an alternate and effective method. An efficient in vitro regeneration protocol has been established using shoot tips and nodal explants. Explants were surface sterilized using 0.1% concentration of mercuric chloride for varying time exposure (1-7 min.). Explants were cultured on MS medium containing different concentrations of cytokinins such as BAP and Kn singly. The best shoot induction was observed on MS medium supplemented with 0.5 mg/l BAP both in case of shoot tip and nodal explants which results in 80.19% and 81.36% of shoot regeneration. Best rooting was obtained from shoots cultured on MS medium fortified with 0.2 mg/l IBA. In vitro raised shoots with roots were transferred to hardening media containing different combinations of substrates. Among all the combinations, sand: soil: vermicompost (1: 2: 1) produced the highest survival rate of 62%. A preliminary qualitative phytochemical analysis helps us to identify the type of secondary metabolites present in plant leaves of three extracts used, aqueous extract shows the presence of all phytoconstituents in it and thus proves to be best solvent for phytochemical estimation and quantitative phytochemical analysis. Among the mother and in vitro plants was carried out to compare the concentration of different phytoconstituents of mother and in vitro raised plants of Andrographis paniculata using aqueous extract. The genetic assessment of micropropagated clones was evaluated by using Random Amplified Polymorphic DNA (RAPD). Out of 7 primers used, 3 showed 75.6% similarity with the mother plant.ThesisItem Open Access In vitro propagation and screening of anti-oxidant potential of valerian jatamasi- a high valued medicinal herb(COLLEGE OF HORTICULTURE AND FORESTRY, DR Y S P UHF, NERI, HAMIRPUR, 2021-01-16) Sharma, Sakshi; Sharma, SnehThe present investigation “In vitro propagation and screening of antioxidant potential of Valeriana jatamansi- a high value medicinal herb” were carried out in the Department of Biotechnology. Valeriana jatamansi is an important medicinal plant belonging to the family Caprifoliaceae, used for insomnia, leprosy, ulcers, convulsions, jaundice, cardiac debility, dry cough, asthma, seminal weakness, chronic fever etc. due to presence of various metabolites and having sedative and anxiolytic property. Being highly medicinal, the species is constantly uprooted from nature for trade. As a result, availability of the species in its natural habitat is decreasing. Besides, propagation through seeds is also very poor. The present studywas henceforth aimed to develop an efficient, rapid and reproducible protocol for in vitro establishment along with analyzing antioxidant activity. In first experiment, surface sterilization protocol was standardized using Sodium hypochlorite and mercuric chloride as sterilant. Among all surface sterilization treatment, combination of Sodium hypochlorite and Mercuric chloride gave maximum survival rate in leaf (80.33±0.33) and nodal segment (77.00 ±0.57) explants. Maximum callus induction frequency (88.33± 0.33) was achieved on MS medium with 10.0 mg/l BAP and 2.0 mg/l NAA from leaf explants. MS medium fortified with BAP 4.0 (mg/l) + TDZ (1.0 mg/l) + NAA (0.5 mg/l) showed highest frequency (74.33 ± 0.66) of shoot induction from callus in 17 days with 15 ± 0.57 number of shoots per explant. Maximum direct shoot induction frequency from nodal explant was achieved on MS medium enriched with BAP (5.0 mg/l) + TDZ (3.0 mg/l) + NAA (1.0 mg/l) in 18 days. Microshoots inoculated on MS media supplemented with 4.0 mg/l BAP, 2.0 mg/l TDZ, 1.0 mg/l NAA and 0.5 mg/l GA3) showed maximum frequency of shoot multiplication with average shoot length (4.36 ± 0.08 cm) and average shoot number (14.00 ± 0.57). A 75.00 ± 1.15 % rooting with significantly high mean root number (8.00 ± 0.57) and root length was achieved in full strength MS medium, supplemented with same concentration i.e. 4.0 mg/l BAP, 2.0 mg/l TDZ, 1.0 mg/l NAA and 0.5 mg/l GA3) combination. A separate medium for root initiation was not required. Maximum plantlets survive after 1 year of acclimatization. RAPD and ISSR markers used to confirmed genetic stability of in vitro raised plants by showing 100 % monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant. There is lack of information on antioxidant activity of in vitro raised plants of V. jatamansi leaf part extract. In second experiment, aim was to measure antioxidant activity of mother plant and in vitro raised plants of V. jatamansi in methanol and diethyl ether extract. The methanol (63.07 ± 0.92) and diethyl ether (41.35 ± 1.88) extract showed highest antioxidant activity in mother plant in comparison to in vitro raised plants of V. jatamansi. The study has practical implications as it will be helpful to meet out industrial as well as domestic demand. In addition, it will ensure conservation of the species by providing the uniform quality planting material.ThesisItem Open Access Isolation and purification of proteins from Tinospora cordifolia against Pseudomonas aeruginosa(College of Horticulture and Forestry Dr YSP UHF, Neri, Hamirpur(H.P.), 2023-05-14) Parul; Sharma, SnehThe present investigation on the “Isolation and purification of proteins from Tinospora cordifoliaagainst Pseudomonas aeruginosa.” were carried out in the Department of Biotechnology, College of Horticulture and Forestry (Dr. Y.S. Parmar University of Horticulture and Forestry), Neri Hamirpur (HP) during 2021-2022. Tinospora cordifolia is a deciduous shrub. It is a well-recognized for its medicinal properties in Indian Ayurveda system. It is one of the most versatile shrub commonly known as “giloy”. The plant gained attention due to its various biological activities such as anti-diabetic, anti-microbial, anti-allergic, anti-oxidant and anti-cancer activities. The plant parts contains various chemical constituents such as alkaloids, steroids, glycosides and polysaccharides. The study was undertaken for isolation and purification of antibacterial protein of Tinospora cordifolia collected from Hamirpur district (Neri), Himachal Pradesh. Analysis was done by stem ethanolic, methanolic, extract of samples taken from Neri village. It was found that methanol act as better solvent for extraction of proteins. Ammonium sulphate precipitation of methaolic crude extract showed that 60-70% precipitation was found best for precipitation of proteins. Further, 60-70% precipitates was subject for dialysis followed by ion exchange chromatography. The fractions obtained after ion exchange chromatography was assayed by antibacterial activity against test organism. The active fractions were pooled together and carried out for SDS-PAGE. The total protein content present in methanolic crude extract was 2.056 mg/ml. The ammonium sulphate precipitation showed 10.833±0.928 mm zone of inhibition and after purification the antibacterial activity increased to 15±0.289 mm. The partial characterization of proteins revealed that the antibacterial proteins was stable at optimal pH 8, heat stable at 40 ℃ temperature and stable at low salt concentration. For future aspects, the stem extract and protein can be further exploited for its other properties such as its sequencing.ThesisItem Open Access Metabolites fingerprinting of Ocimum basilicum cell suspension culture in response to methyl jasmonate elicitation(College of Horticulture and Forestry Neri, Hamirpur (H.P.), 2022-06-08) Bhandari, Suhasini; Sharma, SnehThe present investigation on the “Metabolites fingerprinting of Ocimum basilicum cell suspension culture in response to methyl jasmonate elicitation was carried out in the Department of Biotechnology. Ocimum basilicum L. of the family Lamiacea is a herbaceous species rich in aromatic essential oils and valuable for its pharmaceutical, aromatic and culinary properties. The medicinal, culinary, and aromatic value depends on their bioactive phytochemical constituents. Extraction of these secondary metabolite from naturally grown whole plants on a commercial basis involves large scale crop cultivation. Hence, to overcome this problem plant cell culture provides an alternative approach to whole plant for the production of high value secondary metabolites. An efficient in vitro establishment of cell suspension culture has been established using leaves as explants. Explants were sterilized using different concentration of sodium hypochlorite and mercuric chloride. The best uncontaminated cultures were observed on sodium hypochlorite (2%) for 15 min and mercuric chloride (1%) for 1min. Explants were cultured on MS medium containing different concentrations and combinations of cytokinin (2,4-D, IAA, NAA, BAP, TDZ and Kn). The best callus induction was observed on MS medium supplemented with 2,4-D (1.0 mg/l)+ Kn (0.50 mg/l).i.e 80%. The maximum biomass accumulation (11.4±0.06 g/l DW) was observed on 18th day of exponential phase of Ocimum basilicum cell suspension culture. The maximum production of extracllular phenolic content in cell suspension were at 4 day (6.65±0.15 mg GAE/g) of cell suspension culture whereas the intracellular phenolic content were maximum on day (3.75±0.28 mg GAE/g). After elicitation, biomass accumulation was observed highest at (25 µM) methyl jasmonate (14.11±0.35 g/l DW) and hence, increased the biomass accumulation by 31% respectively. Highest phenolic content (6.37±0.45 mg GAE/g) were obtained on 150 µm (MeJA) when elicited on the 12 th day and harvested at 24 hours and 96 hours post elicitation. It was obtained that exposure to 150 µm MeJA at 24 hours and 96 hours post elicitation increased the phenolic content by 74% and 88% compared to control cultures. Fingerprinting analysis of Ocimum basilicum phytoconstituents were done using LC-MS technique. A total of thirty two compounds were identified on basis of MS m/z ratio and retention time (RT). Those compounds are rosmarinic acid, eugenol, oleanolic acid, palmitic acid, linoleic acid, luteolin, germacrene, sinapic acid, stearic acid, methyl chavicol, crisilineol, vicenin, methyl eugenol, carvacrol, orientin, ascorbic acid, rutin, lupanone, nerolidol, xanthomicrol, basilmoside, hymenoxin, chlorogenic acid, nepetoidin A, linalool, luteolin, methyl cinnamate, cirsimartin, terpineol, methyl acetate, pinene and apigenin 7,4,-dimethyl which was responsible for the medicinal properties of Ocimum basilicum. For future aspects, the isolation and purification of bioactive molecules and evaluation of their pharmacological activity from the crude sample are recommended as future research.ThesisItem Open Access Quantification of bioactive molecules and evaluation of anti-microbial potential of Tinospora cordifolia Miers(College of Horticulture and Forestry Neri, Hamirpur (H.P.), 2021-12-04) Rajvanshi, Ashish Kumar; Sharma, SnehThe present investigation on the “Quantification of bioactive molecules and evaluation of antimicrobial potential of Tinospora cordifolia Miers.” were carried out in the Department of Biotechnology, College of Horticulture and Forestry (Dr. Y.S. Parmar University of Horticulture and Forestry), Neri Hamirpur (HP) during 2020-2021. Tinospora cordifolia is an angiosperm belonging to the Menispermaceae family and is a division of Magnoliophyta, class Magnoliopsida, and order of Ranunculaceae. It is a wide deciduous, glabrous, rapidly ascending shrub with several coiling branches extending approximately 3-4 feet in height and roughly 1 foot long. Tinospora cordifolia is a well-recognized and widely distributed traditional plant that is used successfully in Indian Ayurveda medicine. It has shown many promising biological activities such as antioxidative, antimicrobial, antihyperglycemic, anti-inflammatory, osteoprotective, hepatoprotective, antidiarrheal and antistress effects. It also contains many secondary plant metabolites, such as terpenes, alkaloids, flavonoids, steroids, and glycosides. The study was undertaken for evaluating the bioactive molecules and antimicrobial potential of Tinospora cordifolia collected from different districts of Himachal Pradesh. Analysis was done by using HPTLC of each root, stem and leave ethanolic, methanolic, chloroform and aqueous extract of samples taken from two villages of 3 districts. On the basis of RF value the districts Kangra sample was found to posses highest RF value i.e., 0.97 and the sample with highest RF value was selected for LC -MS. In result found that 7 number of compounds were present. Those seven compounds are berbrine, magnoflorine, menisperine, 20b-hydroxyecdysone, palmatine, 2deoxy-20b-hydroxyecdysone and columbaine which was responsible for the medicinal properties of Tinospora cordifolia. Considering the vast potentiality of Tinospora cordifolia as a source for antimicrobial drugs with reference to antibacterial, antifungal and anticandida agents, a systematic investigation was undertaken to screen roots, stem and leaves samples of Tinospora cordifolia collected from different Districts of Himachal Pradesh for its activity against various pathogens. Ethanolic, methanolic, aqueous and chloroformic extracts were tested but methanolic leaf extract of Tinospora cordifolia showed the maximum inhibitory activity against tested pathogens i.e S. aureus, E. coli, P. aeruginosa and Aspergillus niger. MIC of leaf methanolic extract against Staphylococcus aureus and Pseudomonas aeruginosa was observed and found to be 4.7mg/ml and 3.2mg/ml. For future aspects, the isolation and purification of bioactive molecules and evaluation of their pharmalogical activity from the crude sample are recommended as future researchThesisItem Open Access Somatic Embryogenesis And Cryopreservation Of Walnut (Juglans Regia L.) And Pecan (Carya Illinoensis Kock.)(Dr Yashwant Singh Parmar University of Horticulture and Forestry;Solan, 2001) Sharma, Sneh; Sharma, S.K.