It has been suggested that plant phytochromes are autophosphorylating serine/threonine kinases. However, the biochemical properties and functional roles of putative phytochrome kinase activity in plant light signalling are largely unknown. Here, we describe the biochemical and functional characterization of Avena sativa phytochrome A (AsphyA) as a potential protein kinase. We provide evidence that phytochrome-interacting factors (PIFs) are phosphorylated by phytochromes in vitro. Domain mapping of AsphyA shows that the photosensory core region consisting of PAS-GAF-PHY domains in the N-terminal is required for the observed kinase activity. Moreover, we demonstrate that transgenic plants expressing mutant versions of AsphyA, which display reduced activity in in vitro kinase assays, show hyposensitive responses to far-red light. Further analysis reveals that far-red light-induced phosphorylation and degradation of PIF3 are significantly reduced in these transgenic plants. Collectively, these results suggest a positive relationship between phytochrome kinase activity and photoresponses in plants.
During wound healing, cells migrate with electrotactic bias as a collective entity. Unlike the case of the electric field (EF)-induced single-cell migration, the sensitivity of electrotactic response of the monolayer depends primarily on the integrity of the cell–cell junctions. Although there exist biochemical clues on how cells sense the EF, a well-defined physical portrait to illustrate how collective cells respond to directional EF remains elusive. Here, we developed an EF stimulating system integrated with a hydrogel-based traction measurement platform to quantify the EF-induced changes in cellular tractions, from which the complete in-plane intercellular stress tensor can be calculated. We chose immortalized human keratinocytes, HaCaT, as our model cells to investigate the role of EF in epithelial migration during wound healing. Immediately after the onset of EF (0.5 V/cm), the HaCaT monolayer migrated toward anode with ordered directedness and enhanced speed as early as 15 min. Cellular traction and intercellular stresses were gradually aligned perpendicular to the direction of the EF until 50 min. The EF-induced reorientation of physical stresses was then followed by the delayed cell-body reorientation in the direction perpendicular to the EF. Once the intercellular stresses were aligned, the reversal of the EF direction redirected the reversed migration of the cells without any apparent disruption of the intercellular stresses. The results suggest that the dislodging of the physical stress alignment along the adjacent cells should not be necessary for changing the direction of the monolayer migration.
Hyphessobrycon eques is a famous fish for ornamental fish market and aquarium. They are inhabit in regions of Amazon and Paraguay River basin. Serpae fishs were investigated 2-3 males are chased to female, and then males attempted to simulate the females abdomen. After fertilization, eggs were kept in incubators at 28 o C. The fertilized eggs had adhesive and demesal characteristics and had a mean diameter of 0.92 ± 0.01 mm. Larvae hatched at 16 hrs post fertilization. The hatched larvae averaged 2.90 ± 0.16 mm in total length (L T). Complete yolk sac resorption and mouth opening occurred on the third day post hatching. At 45 days post hatching, the larvae were 12.5 ± 1.60 mm L T and had reached the juvenile stage.
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