Dehydrin metabolism is altered during seed osmopriming and subsequent germination under chilling and desiccation in Spinacia oleracea L. cv. Bloomsdale: possible role in stress tolerance.

Osmopriming improves seed germination performance as well as stress tolerance. To understand the biochemistry of osmopriming-induced seed stress tolerance, we investigated dehydrin (DHN) accumulation patterns at protein and transcript level (determined by immunoblotting and qPCR) during priming, and subsequent germination under optimal and stress conditions (i.e. chilling and desiccation) in spinach (Spinacia oleracea L. cv. Bloomsdale) seeds. Our data indicate enhanced germination performance of primed seeds is accompanied by increased accumulation of three dehydrin-like proteins (DLPs): 30, 26, and 19-kD. Moreover, 30, 26 and 19-kD DLPs that first only transiently accumulated during priming re-accumulated in response to stresses, suggesting an evidence for 'cross-tolerance', which is initially induced by priming and later recruited during post-priming germination under stresses. Study with CAP85, a spinach DHN, corroborates above observations at the gene-expression and protein accumulation level. Additionally, our results suggest that during seed germination and seedling establishment, CAP85 expression may be regulated by the interplay of two factors: seedling development and stress responses. In conclusion, our data suggest that 30, 26, and 19-kD dehydrin-like proteins and CAP85 may be used as potential biochemical/molecular markers for priming-induced stress tolerance in 'Bloomsdale' spinach.

Chen K ，Fessehaie A ，Arora R 《-》

Dynamics of the antioxidant system during seed osmopriming, post-priming germination, and seedling establishment in Spinach (Spinacia oleracea).

Osmopriming is a pre-sowing treatment that improves seed germination performance and stress tolerance. To understand osmopriming physiology, and its association with post-priming stress tolerance, we investigated the antioxidant system dynamics during three stages: during osmopriming, post-priming germination, and seedling establishment. Spinach seeds (Spinacia oleracea L. cv. Bloomsdale) were primed with -0.6 MPa PEG at 15°C for 8 d, and dried at room temperature for 2 d. Unprimed and primed germinating seeds/seedlings were subjected to a chilling and desiccation stresses. Seed/seedling samples were collected for antioxidant assays and germination performance and stress tolerance were evaluated. Our data indicate that: (1) during osmopriming the transition of seeds from dry to germinating state represses the antioxidant pathways (residing in dry seeds) that involve CAT and SOD enzymes but stimulates another pathway (only detectable in imbibed seeds) involving APX; (2) a renewal of antioxidant system, possibly required by seedling establishment, occurs after roughly 5 d of germination; (3) osmopriming strengthens the antioxidant system and increases seed germination potential, resulting in an increased stress tolerance in germinating seeds. Osmopriming-mediated promotive effect on stress tolerance, however, may diminish in relatively older (e.g. ~5-week) seedlings.

Chen K ，Arora R 《-》

Seed priming improves chilling tolerance in chickpea by modulating germination metabolism, trehalose accumulation and carbon assimilation.

Chilling stress is one of the major abiotic stresses affecting chickpea productivity worldwide. This study evaluated the potential role of seed priming in improving resistance to chilling stress in chickpea (cv. Punjab, 2008). The priming treatments involved soaking seeds of chickpea cultivar Punjab 2008 in either water for 8 h (on-farm priming), aerated water (hydropriming) for 18 h, or CaCl2 solution (ψs -1.25 MPa; osmopriming) for 18 h. Primed and untreated seeds were grown either at 18/15 °C (control) or 13/10 °C (chilling stress). Chilling stress suppressed the growth of chickpea while seed priming mitigated the adverse effects of chilling stress by improving stand establishment, growth, water relations, photosynthesis, α-amylase activity, sugar metabolism, antioxidant enzyme activity, membrane stability, and leaf accumulation of proline, nitrogen, potassium and soluble phenolics. Seed priming also improved the performance of chickpea under optimal (control) conditions. The overall order of improvement in resistance to chilling by using seed priming was osmopriming > hydropriming > on-farm priming. Osmopriming improved seedling dry weight, specific leaf area, leaf CO2 net assimilation rate, maximal photochemical efficiency of PSII, α-amylase activity, trehalose content and leaf relative water content by 10, 22, 17, 20, 73, 48 and 7%, respectively, relative to the non-primed control under chilling stress. Under optimal temperature conditions, the corresponding values were 30, 32, 16, 10, 83, 75 and 5%, respectively. Sugar metabolism, especially trehalose content, was strongly linked with stand establishment, photosynthesis, antioxidant potential (under chilling stress) and plant biomass. Overall, seed priming improved chickpea performance under both optimal temperature conditions and chilling stress through better germination metabolism and the accumulation of trehalose, which protected from oxidative damage and helped to maintain carbon assimilation and seedling growth.

Farooq M ，Hussain M ，Nawaz A ，Lee DJ ，Alghamdi SS ，Siddique KHM ... -  《-》

Enhanced expression of the proline synthesis gene P5CSA in relation to seed osmopriming improvement of Brassica napus germination under salinity stress.

Osmopriming is a pre-sowing treatment that enhances germination performance and stress tolerance of germinating seeds. Brassica napus seeds showed osmopriming-improved germination and seedling growth under salinity stress. To understand the molecular and biochemical mechanisms of osmopriming-induced salinity tolerance, the accumulation of proline, gene expression and activity of enzymes involved in proline metabolism and the level of endogenous hydrogen peroxide were investigated in rape seeds during osmopriming and post-priming germination under control (H2O) and stress conditions (100 mM NaCl). The relationship between gene expression and enzymatic activity of pyrroline-5-carboxylate synthetase (P5CS), ornithine-δ-aminotransferase (OAT) and proline dehydrogenase (PDH) was determined. The improved germination performance of osmoprimed seeds was accompanied by a significant increase in proline content. The accumulation of proline during priming and post-priming germination was associated with strong up-regulation of the P5CSA gene, down-regulation of the PDH gene and accumulation of hydrogen peroxide. The up-regulated transcript level of P5CSA was consistent with the increase in P5CS activity. This study shows, for the first time, the role of priming-induced modulation of activities of particular genes and enzymes of proline turnover, and its relationship with higher content of hydrogen peroxide, in improving seed germination under salinity stress. Following initial stress-exposure, the primed seeds acquired stronger salinity stress tolerance during post-priming germination, a feature likely linked to a 'priming memory'.

Kubala S ，Wojtyla Ł ，Quinet M ，Lechowska K ，Lutts S ，Garnczarska M ... -  《-》

Understanding the cellular mechanism of recovery from freeze-thaw injury in spinach: possible role of aquaporins, heat shock proteins, dehydrin and antioxidant system.

Recovery from reversible freeze-thaw injury in plants is a critical component of ultimate frost survival. However, little is known about this aspect at the cellular level. To explore possible cellular mechanism(s) for post-thaw recovery (REC), we used Spinacia oleracea L. cv. Bloomsdale leaves to first determine the reversible freeze-thaw injury point. Freeze (-4.5°C)-thaw-injured tissues (32% injury vs <3% in unfrozen control) fully recovered during post-thaw, as assessed by an ion leakage-based method. Our data indicate that photosystem II efficiency (Fv/Fm) was compromised in injured tissues but recovered during post-thaw. Similarly, the reactive oxygen species (O2 (•-) and H2 O2 ) accumulated in injured tissues but dissipated during recovery, paralleled by the repression and restoration, respectively, of activities of antioxidant enzymes, superoxide dismutase (SOD) (EC. 1.14.1.1), and catalase (CAT) (EC.1.11.1.6) and ascorbate peroxidase (APX) (EC.1.11.1.11). Restoration of CAT and APX activities during recovery was slower than SOD, concomitant with a slower depletion of H2 O2 compared to O2 (•-) . A hypothesis was also tested that the REC is accompanied by changes in the expression of water channels [aquaporines (AQPs)] likely needed for re-absorption of thawed extracellular water. Indeed, the expression of two spinach AQPs, SoPIP2;1 and SoδTIP, was downregulated in injured tissues and restored during recovery. Additionally, a notion that molecular chaperones [heat shock protein of 70 kDa (HSP70s)] and putative membrane stabilizers [dehydrins (DHNs)] are recruited during recovery to restore cellular homeostasis was also tested. We noted that, after an initial repression in injured tissues, the expression of three HSP70s (cytosolic, endoplasmic reticulum and mitochondrial) and a spinach DHN (CAP85) was significantly restored during the REC.

Chen K ，Arora R 《-》