PLANT PHYSIOLOGY , Vol 111, Issue 4 1243-1250, Copyright © 1996 by American Society of Plant Biologists

DEVELOPMENT AND GROWTH REGULATION

Competence for Regeneration during Tobacco Internodal Development (Involvement of Plant Age, Cell Elongation Stage, and Degree of Polysomaty)

LJW. Gilissen, M. J. van Staveren, J. C. Hakkert and MJM. Smulders
DLO-Centre for Plant Breeding and Reproduction Research, Droevendaalsesteeg 1, P.O. Box 16, NL-6700 AA Wageningen, The Netherlands

This study deals with internodal development in vegetative plants of Nicotiana tabacum cv Samsun NN and its reflection in changes of the cellular competence for regeneration. During elongation of the internodes, the cells of the epidermis, subepidermis, and cortex exclusively expanded and increased their DNA content cell type specifically, generally from 2C to 4C. Cells with the 8C DNA content were found mainly among the cortex cells of mature internodes. The frequency of shoot regeneration (directly from subepidermal and epidermal cells together) on thin cell layer explants increased to an optimum along with elongation of the internodes and decreased in mature internodes along with aging. The frequencies of diploid shoots among the regenerants from elongating and mature internodes were high (88 and 75% on the average, respectively), indicating that most cells that had achieved the 4C DNA content generally retained the G2 phase of the diploid cell cycle. Shoots regenerated from explants of young plant material mainly had a vitrified appearance. The occurrence of this type of malformed growth was already determined by the physiological state of the cells in the internode and did not interfere with their acquisition of competence. Vitrification was unrelated to the degree of polysomaty of the internodal tissue. Using the occurrence of tetraploid root regenerants (from intermediate cortex-derived callus), up to a frequency of 50%, we show that the position in the plant where a majority of the 4C cortex cells switched to the G1 phase of the tetraploid cell cycle was at the transition from the elongation phase to the mature phase.