De novo shoot organogenesis (DNSO) is a procedure commonly used for the in vitro regeneration of shoots from a variety of plant tissues. Shoot regeneration occurs on nutrient media supplemented with the plant hormones cytokinin (CK) and auxin, which play essential roles in this process, and genes involved in their signaling cascades act as master regulators of the different phases of shoot regeneration. In the last 20 years, the genetic regulation of DNSO has been characterized in detail. However, as of today, the CK and auxin signaling events associated with shoot regeneration are often interpreted as a consequence of these hormones simply being present in the regeneration media, whereas the roles for their prior uptake and transport into the cultivated plant tissues are generally overlooked. Additionally, sucrose, commonly added to the regeneration media as a carbon source, plays a signaling role and has been recently shown to interact with CK and auxin and to affect the efficiency of shoot regeneration. In this review, we provide an integrative interpretation of the roles for CK and auxin in the process of DNSO, adding emphasis on their uptake from the regeneration media and their interaction with sucrose present in the media to their complex signaling outputs that mediate shoot regeneration.
15 September 2015 in Plant Cell, Tissue and Organ Culture (PCTOC)
An efficient in vitro shoot regeneration method from leaf explants of apple cultivars Golden Delicious and Melrose by optimization of regeneration medium, explant type and orientation, dark pre-treatment, and gelling agent is presented. Murashige and Skoog’s regeneration medium containing 22 μM thidiazuron (TDZ) and 1.5 μM indole-3-butyric acid (IBA) (M2 medium) was superior for regeneration as well as for subsequent shoot multiplication in both cultivars, providing regeneration frequency of 95% or higher in the best combination with other factors. Pre-incubation in the dark proved to be an essential factor for regeneration. The use of agar as a gelling agent provides satisfactory regeneration frequency compared with media gelled with Phytagel TM . Leaf explants of cv. Melrose with adaxial surface in contact with M2 medium and those of cv. Golden Delicious orientated contrary regenerated the highest mean number of shoots per explant. Under optimal conditions, a maximal index of shoot-forming capacity of 11.44 and 6.30 for ‘Melrose’ and ‘Golden Delicious’, respectively, was achieved. Regenerated shoots were successfully rooted and acclimated ex vitro.
Plant reactions to stress vary with development stage and fitness. This study assessed the relationship between light and chilling stress in Arabidopsis acclimation. By analysing the transcriptome and proteome responses of expanding leaves subjected to varying light intensity and cold, 2251 and 2064 early response genes and proteins were identified, respectively. Many of these represent as a yet unknown part of the early response to cold, illustrating a development‐dependent response to stress and duality in plant adaptations. While standard light promoted photosynthetic upregulation, plastid maintenance, and increased resilience, low light triggered a unique metabolic shift, prioritizing ribosome biogenesis and lipid metabolism and attenuating the expression of genes associated with plant immunity. The comparison of early response in young leaves with that in expanded ones showed striking differences, suggesting a sacrifice of expanded leaves to support young ones. Validations of selected DEGs in mutant background confirmed a role of HSP90‐1, transcription factor FLZ13, and Phospholipase A1 (PLIP) in response to cold, and the PLIP family emerged as crucial in promoting acclimation and freezing stress tolerance. The findings highlight the dynamic mechanisms that enable plants to adapt to challenging environments and pave the way for the development of genetically modified crops with enhanced freezing tolerance.
A plant’s main mechanism to diminish the effects caused by high free radical levels generated during high irradiance is the synthesis of various secondary metabolites. In addition to interspecies differences, their concentrations may be influenced by genetic, ontogenic, morphogenetic or environmental factors. We investigated the influence of genetic (genotypes from different natural habitats) and environmental (contrasting light regimes as well as successive parts of the vegetation period) variability on the accumulation of 10 selected phenolic compounds (phenolic acids, flavonoids, and xanthones) in Iris variegata genotypes. Genotypes originated from either sun-exposed or shaded natural habitats were transplanted to two experimental light treatments (high light intensity with a higher R/FR ratio and low light intensity with a lower R/FR ratio). Significant impacts of both genetic and environmental seasonal variability (spring, summer and fall during the vegetation period) on phenolic compound profiles were detected. Their highest amounts were detected in spring. The magnitude of difference between light treatments (high vs. low light intensity) and the direction of this change varied depending on the secondary compound class. Phenotypic correlations among the 10 analyzed secondary metabolites differed across the experimental light treatments and their number decreased from spring to fall.