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Bud formation from tobacco nicotiana tabacum cultivar wisconsin 38 pith tissue grown in vitro cytochemical auto radiographic and cyto photometric analysis



Bud formation from tobacco nicotiana tabacum cultivar wisconsin 38 pith tissue grown in vitro cytochemical auto radiographic and cyto photometric analysis



Annales des Sciences Naturelles Botanique et Biologie Vegetale 16(1): 43-150



The origin and modalities of bud formation at the basal region of tobacco pith explants grown in vitro were investigated by cytochemistry, autoradiography and the 2 wave-length method of cytophotometry. Characteristic features of the pith were 1st examined at 4 different levels in the whole plant. In the same tissue, studies were done in vitro on the phenomenon of dedifferentiation. Pith differentiation in vivo is accompanied by an early endopolyploidization. In pith meristem, amounts of nuclear DNA vary from 2C to 4C; contents of 8C are already present at 300 .mu.m below this meristem whereas, in the pith nuclei at the basal end of stems of 1 m-height, they vary from 4C to 32C. Explants excised at 10 to 20 cm from the apex are able to produce vegetative buds, are mostly polyploid and heterogeneous from the nuclear point of view. Before the beginning of the dedifferentiation in vitro, DNA and starch synthesis increases in all the cells; nuclear fragmentations in 2 or 3 pieces occur, affecting nuclei of ploidy higher than 8C and localized at random in the explants. The cells at the basal end of the explants do not divide. Mitosis precedes cell divisions which are not synchronized. Cells capable of dividing are 2n, 8n or, more frequently, 4n. Comparative studies of divided and non-divided cells affirm that nuclear fragmentation and multinucleate cells are not involved in the redivisions of a differentiated tissue. In vitro, nuclear modifications, like an hyperploidization of non-totipotent cells, are induced. Buds are initiated either from the cambium or the redifferentiated parenchymatous cells of the callus. These buds are 2n or, more often, 4n. Only 1 aneuploid bud, without leaves, was observed. Not all the plant cells are totipotent and, contrary to the general hypothesis, there is no exclusive reestablishment of the diploid state during the dedifferentiation. The role of starch in dedifferentiation and organogenesis is discussed. Comparisons relative to DNA content, starch, polyphenols and oxidative activities were also made between regenerated buds in vitro and apical meristems of plants in vivo.

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Accession: 004862397

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