Sharma, SnehDevi, Bandna2020-09-212020-09-212020-06-22https://krishikosh.egranth.ac.in/handle/1/5810151690The 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 importanceEnglishThesis