Browsing by Author "Edupulapati Nandini"

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    Bio- elicitation of secondary metabolites in callus cultures of black pepper (Piper nigrum L.)
    (Department of Molecular Biology and Biotechnology, College of Agriculture , Vellayani, 2024-02-21) Edupulapati Nandini; Asha, S
    The present study on “Bio-elicitation of secondary metabolites in callus cultures of black pepper (Piper nigrum L)” was carried out at the Department of Molecular Biology and Biotechnology, College of Agriculture, Vellayani, during the period from 2021-2023. The objective of the study was development of callus culture of black pepper (Piper nigrum L.) for secondary metabolite production and its bio-enhancement using derivatives of Piriformospora indica. Callus cultures were successfully initiated from the leaves and mature ripe berries of the Panniyur 1 variety. The use of 0.1% mercuric chloride for 3 minutes proved effective in achieving healthy, contamination-free callus initiation from leaf explants, while a duration of 5 minutes was found to be effective for healthy callus initiation from berry explants. Callus induction was carried out using Murashige and Skoog's (MS) medium, incorporating various combinations and concentrations of plant growth regulators (PGR). The percentage of callus induction varied depending on the type of explant and media composition used. In this study, it was observed that leaf tissue exhibited a higher responsiveness to callus induction compared to berry. Out of the four treatments, the callus induction was observed on MS media supplemented with 5.0 mg/L Napthelene Acetic Acid (NAA) and MS media supplemented with 1.0 mg/L 6benzyladenine (BA). Among these, the best response and friable callus was noted from MS media with 5.0 mg/L NAA, with 75% callus induction percentage from leaf explant within 4weeks. Better Callus proliferation with mean diameter of 1.094cm was also found in this media. Callus cultures from leaves were subjected to 24 hrs, 48 hrs and 72hrs of elicitation with Piriformospora indica cell wall extract (PiCWE) (1:1) and culture filtrate (PiCF) (1:1) and the effect of elicitation on secondary metabolite production was studied by detecting the presence of piperine by High Performance Liquid Chromatography. The maximum improvement in the accumulation of piperine was observed in callus cultures at early stages (at 24hrs) of elicitation with P. indica cell wall extract and P. indica culture filtrate. Meanwhile, P. indica CWE and CF treatment at 48hrs and 72hrs treatment was found lower performance compared with 24hrs of bioelicitation. The expression profiling of piperine synthase and Pip CoA ligase genes showed upregulation of the piperine synthase (2.49fold), and pip CoA ligase genes (63.87fold) in the 24hrs treatment with PiCF in leaf derived callus, Berry derived callus showed upregulation of piperine synthase gene with PiCWE treatment at 24 hrs with a fold change of (21.06) while pip coa ligase showed fold change of 5.13. The expression of genes were found to be decreased in later stages. The results of the study indicated that P. indica culture filtrate and cell wall extract acts as effective elicitors in boosting the production of secondary metabolites in the leaf and berry derived callus cultures of black pepper (Piper nigrum L.) respectively. But the expression of key genes and subsequent production of piperine decreased in the leaf callus, indicating the possibility of site-specific regulation of metabolites. Future studies will be directed towards the elucidation of tissue specific regulation of metabolites in black pepper.