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In recent years, there has been an increasing tendency towards the natural dyes due to the increasing awareness of toxicity and serious health hazards of synthetic dyes. Natural dyes find use in the colouring of textiles, drugs, cosmetics, etc.., owing to their non- toxic effects; they are also used for colouring various food products. The present paper is based on extensive survey. Collection of ethanobotanical information and Biodiversity of Natural dye yielding plants in Thanjavur district. It has also been known that the natural dyes are not harmful and eco – friendly. The study report 30 dye yielding plant species belong to 21 families. Along with their botanical name, common name, family, general description, season of flowering & fruiting, traditional uses, plant parts used, colour and application of dye used are mentioned this paper. Apart from, medicinal uses, it also describes preparation of dyeing stuff. Use mordant and specific dye utilization to serve various purposes, including several of the new dye resources. The use of Natural dye has diminished over generations due to dearth of documentation and in old civilization the ayurvedic medicine says natural dyes contain plenty of medicinal properties. Keeping these points in view the present study will be carried out to the biodiversity and selected dye yielding plant were applied in cotton cloth. These studies revealed that these economically important dye- yielding plants need to be conserved.