Plant Tissue Cult. 4(2) : 93-103, 1994

High-frequency Plant Regeneration from Fused Protoplasts between Brassica oleracea L. and B. campestris L.

Mohammad Mofazzal Hossain¹ and Susumu Yazawa

Laboratory of Vegetable and Ornamental Horticulture, Faculty of Agriculture, Kyoto University, Sakyoku, Kyoto-606, Japan

Key words: Regeneration frequency, Auxin, Cytokinin, Somatic hybrid

Abastract

A protocol has been developed to obtain high-frequency plant regeneration from PEG-mediated fused protoplasts of Brassica oleracea L. cvs. Yoshin and Ruby ball, and B.campestris L. cvs. Kenshin, Kaga and Eishin, using simple MS-based medium. The optional concentration of 2,4-D in liquid medium for the growth of fused protoplasts was between 4.52 and 11.30 ?M. Osmolarity decreasing from 0.5 to 0.2 M was appropriate for colony proliferation. 2,4-D (4.52 to 9.05 ?M)with BA (2.22 to 4.40 ?M) was the best combination for callus induction. The addition of zeatin riboside (2.85 ?M) and BA (4.40 ?M) into regeneration medium enhanced plant regeneration (37.14%). Regeneration frequency could be increased up to 60.4% using 2,4-D (4.52 ?M) and BA (2.22 ?M) in callus induction medium, and zeatin riboside (2.85 ?M) and BA (4.40 ?M) in regeneration medium.

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