Isolation of a FLORICAULA/LEAFY putative orthologue from Chenopodium rubrum and its expression during photoperiodic flower induction.

The short day plant (SDP) Chenopodium rubrum L. (ecotype 374) has been a model plant for physiological studies on photoperiodic flower initiation for many years. Using reverse transcription-polymerase chain reaction (RT-PCR) we identified a C. rubrum putative orthologue of the FLORICAULA/LEAFY genes from Antirrhinum majus and Arabidopsis thaliana, referred to as CrFL. Kinetics of the expression of CrFL in the apical part of C. rubrum during flower induction was followed using semi-quantitative RT-PCR. Expression of CrFL in vegetative apices was relatively high and started to decrease after 6 h of darkness (critical photoperiod). It reached its minimum between the 9th and the 12th hour of the 12-h inductive dark span, stayed at low levels for the next 6 h and increased again after the flower induction was completed. Our results indicate that expression of CrFL is regulated by photoperiod and that it is important both in the vegetative state and during flower development.

Changes in the geometry of the apical meristem and concomitant changes in cell wall properties during photoperiodic induction of flowering in Chenopodium rubrum.

• A putative role for local forces at the surface of the apical meristem for plant organogenesis has been postulated in various studies. To correlate changes in morphogenesis to altered local forces, we followed the geometry of the apical meristem during photoperiodic flower induction in Chenopodium rubrum. • The shape of the apical meristem was determined using cryo-scanning electron microscopy. Cell wall properties on the surface of the dome were visualized using uplight polarization microscopy. • A subtle depression at the summit of the apical dome, typical for the vegetative state, became rounded during the early phase of flower induction. The observed changes were quantified as an increase in the ratio of height to diameter of the dome. In parallel, the properties of cell walls on the surface of the dome changed. • The changes in geometry of the apical meristem were attributed to water movement in the meristem paralleled by a change in cell wall properties at the top of the dome. The results support the hypothesis that local changes in surface tension precede initiation of organ primordia.

Plasma membrane H+-ATPase is involved in auxin-mediated cell elongation during wheat embryo development.

Previous investigations suggested that specific auxin spatial distribution due to auxin movements to particular embryonic regions was important for normal embryonic pattern formation. To gain information on the molecular mechanism(s) by which auxin acts to direct pattern formation in specific embryonic regions, the role of a plasma membrane (PM) ATPase was evaluated as downstream target of auxin in the present study. Western-blot analysis revealed that the PM H(+)-ATPase expression level was significantly increased by auxin in wheat (Triticum aestivum) embryos (two-three times increase). In bilaterally symmetrical embryos, the spatial expression pattern of the PM H(+)-ATPase correlates with the distribution pattern of the auxin analog, tritiated 5-azidoindole-3-acetic acid. A strong immunosignal was observed in the abaxial epidermis of the scutellum and in the epidermal cells at the distal tip of this organ. Pseudoratiometric analysis using a fluorescent pH indicator showed that the pH in the apoplast of the cells expressing the PM H(+)-ATPase was in average more acidic than the apoplastic pH of nonexpressing cells. Cellulose staining of living embryos revealed that cells of the scutellum abaxial epidermis expressing the ATPase were longer than the scutellum adaxial epidermal cells, where the protein was not expressed. Our data indicate that auxin activates the proton pump resulting in apoplastic acidification, a process contributing to cell wall loosening and elongation of the scutellum. Therefore, we suggest that the PM H(+)-ATPase is a component of the auxin-signaling cascade that may direct pattern formation in embryos.