Plant Tissue Cult. 8(1) : 91-99, 1998 (December)

Protoplast Isolation and Regeneration from Cell Cultures in Some Cultivated and Wild Species of Nicotiana

P. K. Das and N. Mandal

Bidhan Chandra Krishi Viswavidyalaya, Mohanpur ? 741 252, West Bengal, India

Key words: Nicotiana, Cultivated sp., Wild sp., Protoplast, Regeneration

Abastract

Calluses initiated in 2, 4-D (0.4 mg/1) supplemented MS medium (MX) from leaf tips of two cultivated (tabacum, rustica) and two wild species of Nicotiana (Glutinosa, gossei) were to initiate cell suspension cultures. A large number of protoplasts were harvested following the treatment with three different enzyme mixtures. Among them cellulase, macerozyme and drieslase combination proved to be the best as this treatment yielded maximum viable protoplasts in a shorter period of time. Protoplasts of two uncommon but genetically important species N. glutinosa and N .gossei showed division in MX + 5% glucose at a frequency comparable to that observed with two cultivated species, N. tabacum and N. rustica. Plants of all the four species were reproducibly regenerated from protoplast-derived cultures. MS basal medium supplemented with IAA and Kn in 1.6 : 1 proportion registered their superiority in inducing shoots in all the four species. However, MS medium with BA recorded dicisively high response for shoot regeneration only for gossei and tabacum.

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