Effects of co-expressing the plant CDK inhibitor ICK1 and D-type cyclin genes on plant growth, cell size and ploidy in Arabidopsis thaliana.

The cyclin-dependent kinase (CDK) plays a crucial role in regulating the cell cycle of eukaryotic organisms including plants. From previous studies, it is known that ICK1, the first plant CDK inhibitor identified in Arabidopsis plants, interacts with Arath;CycD3;1 (CycD3) and Arath;CDKA;1 (Cdc2a). Overexpression of ICK1 has major effects on cell division, plant growth, and morphology. In this study, approaches were taken to determine the effects on transgenic 35S::ICK1 Arabidopsis plants of introducing another gene that could potentially modulate the activity of ICK1. F1 plants were obtained by crossing 35S::ICK1 plants with wild type (Wt) and transgenic plants expressing 35::GUS, 35S::CycD3, 35S::CycD2, or 35S::antiICK1 ( antiICK1 refers to antisense- ICK1). The major effects on plant growth and morphology observed in the 35S::ICK1 plants were partially reversed in the F1 plants from the crosses [35S::ICK1 x 35S::CycD2] and [35S::ICK1 x 35S::CycD3], and completely restored in the F1 plants from the cross [35S::ICK1 x 35S::antiICK1]. This observation was further supported by the results of ploidy analysis and structural characterization. Overexpression of CycD2 and CycD3 had the opposite effect on leaf cell size to the overexpression of ICK1. In addition, in ICK1-overexpressing plants, the CycD2 and CycD3 transcript levels increased, indicating a possible feedback regulation. The present results demonstrate that the interactions between ICK1 and D-type cyclins previously observed by the yeast two-hybrid and in vitro techniques are biologically relevant. These results illustrate the possibility of modifying plant growth and architecture dynamically by adjusting the levels of positive and negative cell-cycle regulators.

Control of petal and pollen development by the plant cyclin-dependent kinase inhibitor ICK1 in transgenic Brassica plants.

The cyclin-dependent protein kinases (CDKs) have a central role in cell cycle regulation and can be inhibited by the binding of small protein CDK inhibitors. The first plant CDK inhibitor gene ICK1 was previously identified in Arabidopsis thaliana. In comparison to known animal CDK inhibitors, ICK1 protein exhibits unique structural and functional properties. The expression of ICK1 directed by the constitutive CaMV 35S promoter was shown to inhibit cell division and plant growth. The aim of this study was to determine the effects of ICK1 overexpression on particular organs and cells. ICK1 was expressed in specific tissues or cells of Brassica napus L. plants using two tissue-specific promoters, Arabidopsis AP3 and Brassica Bgp1. Transgenic AP3-ICK1 plants were morphologically normal except for some modified flowers either without petals or with petals of reduced size. Surprisingly, petals of novel shapes such as tubular petals were also observed, indicating a profound effect of cell division inhibition on morphogenesis. The cell size in the smaller modified petals was similar to that in control petals, suggesting that the reduction of petal size is mainly due to the reduction of cell numbers and that the inhibition of cell division does not necessarily lead to an increase in cell size. Transgenic Bgp1-ICK1 plants were normal morphologically; however, dramatic decreases in seed production were observed in some plants. In those plants, the ability of pollen to germinate and pollen nuclear number were affected. These results are discussed in relation to the cell cycle and plant development.