Summary• Here, using light microscopy, we studied the cellular basis of growth of Arabidopsis cotyledons during postembryonic development.• Analysis of serial paradermal sections of developing cotyledons revealed that early growth involved cell enlargement and some cell division. The daughter cells remained tightly packed in growing clusters, which enabled us to monitor both divisions and cell enlargement.• By contrast to other epigeal cotyledons, cells of all mesophyll layers could enter division. The cells in the cotyledon margins displayed a higher proliferative activity; however, the orientation of the division plane did not permit cells at the margins to give rise to tissue internally, nor to generate new cell layers. Exogenous cytokinin stimulated both cell expansion and cell division in attached dark-grown cotyledons. Light stimulated cell growth. Higher-levels of endogenous cytokinins in the amp1 mutant were confirmed to enhance the light-stimulated process of cell enlargement in cotyledons of dark-grown seedlings.• This work demonstrates an approach to use Arabidopsis cotyledons as an experimental system and provides a framework for molecular-genetic analysis of cellular dynamics during the growth of this organ.

The spatial and temporal patterns of post-embryonal cell growth and cell division were characterised in excised cotyledons of vegetable marrow ( Cucurbita pepo L. var. giromontia Alef.) incubated in water. The concurrent roles of these two processes in cotyledon growth were determined using paradermal sections of the first palisade layer of developing cotyledons. Tissue specificity was observed in the pattern of cell division. The daughter cells derived from an initial cell, which had already differentiated before imbibition of the seeds, were tightly packed in a cluster, which enabled us to monitor cell division during early cotyledon development. Heterogeneity of cell size was recognised during the process of cell proliferation in the cluster, suggesting that cell division is uncoupled from control of cell size. There was significantly more cell division in the marginal part of the cotyledons than in other parts, suggesting high activity of the marginal meristem. Light enhanced cell and cotyledon enlargement, but had no effect on the number of divisions. This study elucidated the cellular basis of post-germinative Cucurbita cotyledon morphogenesis and development.

The temporal and spatial effects of exogenous cytokinins on both cell expansion and division activity in the plate meristem of cultured zucchini cotyledons were studied. N6-benzylaminopurine (1-100 microM) and N-(2-chloro-4pyridyl)-N'-phenylurea (4PU-30) (0.1-100 microM) greatly stimulated the cell growth and division. They provoked multiple cell cycles, formation of larger clusters of daughter cells and an increase of the final number of cells. Both cytokinins led to earlier achievement of final cotyledon size and shortened the cell doubling time. By contrast to the purine cytokinin, phenylurea cytokinin 4PU-30 enlarged the cotyledon predominantly in length. Zeatin and kinetin were less effective, particularly in stimulating cell expansion. In low concentrations, all cytokinins were more effective in stimulating division activity rather than expansion. The cells in the cotyledon margins displayed a higher division activity, especially when treated with exogenous cytokinins. The final cotyledon and cluster areas were not of the strict proportional dependence upon the number of their cells. These results provide a novel example where stimulated cell division fails to evoke a respective increase in the final organ size.

The jasmonates are well studied in the context of plant defence but increasingly are also recognised as playing roles in development. In many systems, jasmonates antagonise the effects of cytokinins. The aim of the present work was to elucidate interactions between methyl jasmonate and cytokinin (benzyladenine) in regulating growth of zucchini (Cucurbita pepo L., cv. Cocozelle, var. Tripolis) cotyledons, taking advantage of the ability to simultaneously quantify cell enlargement and division from paradermal sections of the first palisade layer. Growth regulators were applied to cotyledons, excised from dry seeds and grown in darkness. Cytokinin stimulated expansion and division whereas, surprisingly, jasmonate stimulated expansion but inhibited division. Jasmonate antagonised the stimulating effect of cytokinin on division but worked cooperatively with cytokinin in increasing expansion. However, expansion with jasmonate was more isotropic than with cytokinin. Jasmonate also stimulated the loss of cellular inclusions and soluble protein. Soluble proteins revealed a partial antagonism between jasmonate and cytokinin. These results illustrate the complex interplay between jasmonates and cytokinin in the regulatory network of cotyledon development following germination.