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Extraction, Characterization and Tissue Culture of Plant Derived Indigo From Indigofera tinctoria-Preliminary Studies

Chandra Prabha M N, Sanchita Sharma

Abstract


Indigo is the most preferable of all natural dyes for its high demand worldwide because of the blue denim. Natural Indigo dye is eco friendly, non carcinogenic and do not pose hazard to the environment as compared to synthetic dyes. Although a growing demand for naturally derived indigo dye exists, with increased use of synthetic dyes the plant species is now on the verge of being endangered. Plant tissue culture technique is an effective alternative for micropropagation and invitro production of dye. No research on the mass cultivation and dye extraction of Indigo dye from Indigofera tinctoria plant has been reported. The present study reports the extraction and characterization of plant derived indigo dye from Indigofera tinctoria. In vitro propagation has been attempted for rapid propagation of Indigofera tinctoria. Crude dye was obtained after overnight fermentation of Indigofera tinctoria plant leaves either in water or ammonia. Extracted dye was characterized by UV-Vis spectrophotometry and High Pressure Liquid Chromatography (HPLC). Water based extraction method gave almost similar peak value (608nm) as the indigo standard (607 nm) with least noise as compared to ammonia based extraction. HPLC analysis of the crude indigo extract showed 2.9% indigo content per 5 mg crude dye, which is higher than indigo content reported in other species such as Polygonum tinctorium, Isatis tinctoria etc. Initiation of callus was done from the leaf disc explants. Callus was initiated successfully on medium containing: 0.5-3 mg/l 1-Naphthaleneacetic acid (NAA) alone and in combination with 0.5-2mg/l 6-Benzylaminopurine (BAP) and 0.5-2mg/l kinetin (Kin). The leaf explants proliferated into multiple callus on induction media after four weeks of culture. After the callus was fully grown, it was subcultured onto a suitable medium for morphogenesis. Keywords: Callus Induction, Dye extraction, Indigofera tinctoria, Tissue culture, Spectophotometry, HPLC.

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References


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