Nathanaël Prunet

When asked to provide a picture for the cover of the Flowering Newsletter, I picked this image of an Arabidopsis thaliana inflorescence expressing fluorescent reporters for two key regulators of flower development: APETALA3 (AP3), which promotes petal and stamen identity, and SUPERMAN (SUP), which encodes a transcriptional repressor that defines the boundary between stamens and pistil (Fig. 1). The choice was easy: it was an important breakthrough in my research on the role of SUP in the separation of stamens in whorl 3 and carpels in whorl 4; and among the images of flowers I have taken with a confocal microscope, it is also one my favourites aesthetically. The image won awards at the 2015 Nikon Small World and FASEB BioArt competitions and is published in Prunet et al. (2017).

Significance: The Arabidopsis thaliana flower is a complex structure that consists of discrete floral organs (sepals, petals, stamens, and carpels) that are separated by regions lacking organ growth called boundaries. The SUPERMAN ( SUP ) gene functions to define the boundary between the male organs (stamens) and female organs (carpels) in the flower. Previous work on boundary formation in plants has focused on growth repression, rather than on identity separation. Using live confocal imaging, we demonstrate that SUP functions by keeping the male and female developmental programs spatially and temporally separate, which is critical for the fertility of the flower. In addition, we show a second role of SUP in the timely termination of floral stem cells.

The study of plant growth and development has long relied on experimental techniques using dead, fixed tissues and lacking proper cellular resolution. Recent advances in confocal microscopy, combined with the development of numerous fluorophores, have overcome these issues and opened the possibility to study the expression of several genes simultaneously, with a good cellular resolution, in live samples. Live confocal imaging provides plant biologists with a powerful tool to study development, and has been extensively used to study root growth and the formation of lateral organs on the flanks of the shoot apical meristem. However, it has not been widely applied to the study of flower development, in part due to challenges that are specific to imaging flowers, such as the sepals that grow over the flower meristem, and filter out the fluorescence from underlying tissues. Here, we present a detailed protocol to perform live confocal imaging on live, developing Arabidopsis flower buds, using either an upright or an inverted microscope.

There are only three grand theories in biology: the theory of the cell, the theory of the gene, and the theory of evolution. Two of these, the cell and gene theories, originated in the study of plants, with the third resulting in part from botanical considerations as well. Mendel's elucidation of the rules of inheritance was a result of his experiments on peas. The rediscovery of Mendel's work in 1900 was by the botanists de Vries, Correns, and Tschermak. It was only in subsequent years that animals were also shown to have segregation of genetic elements in the exact same manner as had been shown in plants. The story of developmental biology is different – while the development of plants has long been studied, the experimental and genetic approaches to developmental mechanism were developed via experiments on animals, and the importance of genes in development (e.g., Waddington, 1940) and their use for understanding developmental mechanisms came to botanical science much later – as late as the 1980s.