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Reconstitution of Diploids from Leaf Tissue of Anther-derived Haploids in Tobacco¹

Normal diploid plants were regenerated from leaf midveins of cytologically verified, anther-derived haploid tobacco (Nicotiana tabacum L.) plants. Large numbers of haploids were obtained by culturing immature anthers on a chemically defined tissue culture medium. The haploids were verified cytologically soon after emergence frem the cultured anthers. They were screened under field, greenhouse, or growth chamber environments for disease resistance, chemical composition, and/or physical characteristics. Leaf midveins from selected haploid plants were trimmed of lamina and cultured on a chemicaliy defined medium. Leafy shoots emerged in about 3 weeks. Plants regenerated from midveins of young (less than fully expanded) leaves were all haploids. Those regenerated from aged leaves (those that remained on the plant 3 to 4 weeks after attaining full expansion) included some diploids and some haploids. The diploids were normal in appearance, self-fertile and produced diploid progeny.

Key Words: Tissue culture • Nicotina tabacum L. • Plant breeding • Cytology

² Plant Physiologist, Tobacco Physiology Investigations. Plant Science Research Division, Agricultural Research Service, US Department of Agriculture, and Associate Professor, respectively, Department of Agronomy, University of Kentucky, Lexington, Ky. 40506.

Received for publication August 26, 1971.

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