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Dedifferentiation and cell-cycle re-entry. Totipotency is a characteristic of most plant species, reflecting their potential to build whole new individuals from a group of cells that become reprogrammed under favourable conditions. This attribute has been exploited successfully in tissue culture to establish regeneration protocols of various organs. Even highly specialized plant cells can retain full totipotency, as demonstrated, for example, by the stomatal guard cells of sugar beet ( Beta vulgaris ) from which plants can be regenerated [6]. Irrespective of the tissue type that is used as the source material, at least three main steps towards plant regeneration are required: cellular dedifferentiation, cell-cycle re-entry and induction of redifferentiation.Dedifferentiation and cell-cycle entry are intimately linked, making it difficult to disentangle both processes at the molecular level. Significant progress in our knowledge has come from studies with protoplast cultures. Plant protoplasts can be obtained from intact tissues by enzymatic removal of the cell wall and, under appropriate conditions, they can be regenerated into intact plants. However, before they can re-enter the cell cycle, protoplasts that originate from differentiated tissues need to undergo an abrupt switch from a fully differentiated to a dedifferentiated status. Evidence indicates that this transition is accompanied by a change in the chromatin structure, thereby altering the portion of the genome that is accessible for transcription (euchromatin) versus the portion that is repressed (heterochromatin) [7].At the onset of cell division, the proportion of euchromatin increases considerably. It is thought that the retinoblastoma (RB)-E2F pathway is involved in this process, as indicated by the change in chromatin structure of E2F target genes following the dedifferentiation of protoplasts [8]. The RB-E2F pathway is well known for its anticipated role in the activation of the G1-S-specific E2F-DP transcription factors ([Box 3]), but also for its role in remodelling the chromatin structure [9]. Dynamic changes in chromatin structure are primarily orchestrated by post-transcriptional modifications of the histones in nucleosomes [10]. The best-characterized type of histone modification in plants is acetylation by histone acetyltransferases. In maize (Zea mays ), the histone deacetylase RPD3I has been shown to be physically associated with the retinoblastoma-related protein RBR [11]. Together, both proteins probably cooperate in repressing genes that are required for cell-cycle entry [12] (Fig. 1).
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