Polyamine depletion enhances oil body mobilization through possible regulation of oleosin degradation and aquaporin abundance on its membrane.

Oil body (OB) mobilization, a crucial event associated with early seedling growth, is delayed in response to salt stress. Previous reports suggest that careful regulation of polyamine (PA) metabolism is essential for salt stress tolerance in plants. Many aspects of PA-mediated regulation of metabolism have been uncovered. However, their role in the process of OB mobilization remains unexplored. Interestingly, the present investigations reveal a possible influence of PA homeostasis on OB mobilization, while implicating complex regulation of oleosin degradation and aquaporin abundance in OB membranes in the process. Application of PA inhibitors resulted in the accumulation of smaller OBs when compared to control (-NaCl) and the salt-stressed counterparts, suggesting a faster rate of mobilization. PA deficit also resulted in reduced retention of some larger oleosins under controlled conditions but enhanced retention of all oleosins under salt stress. Additionally, with respect to aquaporins, a higher abundance of PIP2 under PA deficit both under control and saline conditions, is correlated with a faster mobilization of OBs. Contrarily, TIP1s, and TIP2s remained almost undetectable in response to PA depletion and were differentially regulated by salt stress. The present work, thus, provides novel insights into PA homeostasis-mediated regulation of OB mobilization, oleosin degradation, and aquaporin abundance on OB membranes.

Polyamine homeostasis modulates plasma membrane- and tonoplast-associated aquaporin expression in etiolated salt-stressed sunflower (Helianthus annuus L.) seedlings.

Salt stress adversely affects plants by causing osmotic and ionic imbalance. Cellular osmotic adjustment occurs by modulation of water fluxes. Polyamines (PAs) are often advocated to be involved in osmoregulation during stressful conditions, and thus, they serve as potential "osmolytes." Aquaporins (AQPs), the water-transporting channels, are expected to play crucial roles in osmoregulation. Present investigations on etiolated sunflower seedlings demonstrate a possible correlation between PA homeostasis and maintenance of water balance, as a function of modulation of the abundance of two major AQP subfamilies: PIP2 (plasma membrane intrinsic protein 2) and TIP1 (tonoplast intrinsic protein 1). Salt stress (120 mM NaCl) restricts growth of sunflower seedlings and induces reduction in relative water content (RWC). This accompanies enhanced abundance of PIP2s and TIP1s in seedling roots and that of TIP1s in cotyledons, as revealed by Western blot analysis of AQP isoforms and also their imaging by confocal laser scanning microscopy (CLSM). Raising seedlings in the presence of 500 µM of DFMA (DL-α-difluoromethylarginine) or DFMO (DL-α-difluoromethylornithine), which are potent inhibitors of PA biosynthesis enzymes (arginine decarboxylase (ADC) and ornithine decarboxylase (ODC), respectively), significantly promotes root extension, irrespective of NaCl stress, and results in further lowering of salt-induced reduction in RWC in roots and cotyledons. This correlates with enhanced accumulation of both PIP2s and TIP1s in seedling roots, but not in cotyledons. Present work, therefore, implicates PA homeostasis in the maintenance of water status of sunflower seedlings, possibly via regulation of abundance and distribution of AQP isoforms associated with the plasma membrane and tonoplast.

Nitric oxide modulates polyamine homeostasis in sunflower seedling cotyledons under salt stress.

Free polyamine (PA) titers in plants may be regulated through reversible conjugate formation and/or through modulation of their synthesis, transport and degradation. PA signaling involves the well-acknowledged signaling molecule, nitric oxide (NO), which functions in diverse biological processes. Present investigations demonstrate the influence of salt stress (120 mM NaCl) and exogenous NO donor (250 µM Diethylenetriamine, DETA) on PA homeostasis of 2 d old, etiolated sunflower (Helianthus annuus L.) seedling cotyledons as a long-distance signaling response. Significantly enhanced intracellular spermine (Spm) accumulation was observed in seedling cotyledons under salt stress and in response to NO donor, the increase being more pronounced in seedlings treated with NO, evidently as a result of upregulation of the PA biosynthetic enzymes - arginine decarboxylase (ADC) and S-adenosylmethionine decarboxylase (SAMDC) - as revealed by Western blot and confocal imaging (CLSM). Moreover, salt stress induced the activity of polyamine oxidase (PAO), a PA catabolic enzyme, while NO lowered its activity in salt-stressed seedling cotyledons. NO, thus, appears to assist the seedlings in adapting to salt stress by positively regulating PA homeostasis through regulation of PA distribution between free, conjugated and bound forms, increased accumulation of PA biosynthetic enzymes and lowering the rate of PA catabolism.