Stress responses of tobacco cells to high temperature and salinity. Proline accumulation and phosphorylation of polypeptides

Stress responses to high temperatures (35–48°C) and salinity (170–340 mM NaCl) and thermotolerance were investigated for the salt-sensitive wild type and the salt-tolerant N_rE_s-1 strain of Nicotiana sylvestris L. using suspension-cultured cells. Under saline conditions, N_rE_s-1 strain cells accumulated proline, polyamine (putrescine and spermidine) and betaine in contrast to wild-type cells. The simultaneous treatment of salt-tolerant cells with high temperature (40°C) and NaCl (170 or 340 mM ) resulted in a transient overproduction of proline accompanied by an increase in their thermotolerance. At the high temperature, the synthesis of polypeptides and the accumulation of heat shock protein HSP70 mRNA were not affected by salinity. The higher thermotolerance of the NaCl-tolerant cells could not be related to osmoprotecting sugar-starch interconversions but could rather be associated with selective phosphorylation of several polypeptides (23–24, 27, 31–32, 47 kDa) prior to the accumulation of proline. The possible role of polyamines and polypeptide phosphorylation in this respect is discussed.     

Protaphorura tricampata, a euedaphic and highly permeable springtail that can sustain activity by osmoregulation during extreme drought

We have investigated drought physiology of soil dwelling springtails since water availability is a key environmental factor governing their performance, and predictions of climate change suggest increased frequency and intensity of summer droughts. Here we show in field and laboratory experiments that the typical euedaphic springtail, Protaphorura tricampata, can survive extreme drought and remain active in soils where the water potential is much lower than equivalent to normal osmotic pressure of springtails. Euedaphic springtails (i.e. species living in deeper soil layers) have an extraordinary ability to up-regulate osmotic pressure of body fluids and prevent water loss in soils where the water potential has dropped to well below the permanent wilting percentage of plants. The ability to regulate osmotic pressure of body fluids is based on accumulation of compatible osmolytes such as sugars and free amino acids. Alanine was the most important osmolyte in P. tricampata and accumulated to concentrations of about 300 μmol g⁻¹ dry weight. It is suggested that alanine also serves as a non-toxic storage of ammonia during drought periods where the normal urine production is hampered. The results presented here show, contrary to convention, that high cuticular permeability is not necessarily accompanied by poor drought tolerance, and is not a good predictor of drought susceptibility.     

Proton-Decoupled 31P Magnetic Resonance Spectroscopy Reveals Osmotic and Metabolic Disturbances in Human Hepatic Encephalopathy

Abstract: Quantitative proton and quantitative proton-decoupled ³¹P magnetic resonance spectroscopy (MRS) of the brain was performed in 16 patients with liver disease (10 with and six without chronic hepatic encephalopathy) and four patients with hyponatremia, as well as 20 age-matched normal subjects. Patients with hepatic encephalopathy were distinguished from controls by significant reduction in levels of cerebral nucleoside triphosphate (2.45 ± 0.20 vs. 2.91 ± 0.21 mmol/kg of brain; p < 0.0003), inorganic phosphate ( p < 0.03), and phosphocreatine ( p < 0.04). In addition of increased levels of cerebral glutamate plus glutamine and decreased concentrations of myo -inositol, patients with hepatic encephalopathy showed a reduction of total visible choline and of glycerophosphoryl-choline (0.67 ± 0.13 vs. 0.92 ± 0.20 mmol/kg of brain in controls; p < 0.005) in ¹H MRS, and of glycerophosphoryl-ethanolamine (0.40 ± 0.12 vs. 0.68 ± 0.12 mmol/kg of brain in controls; p < 0.0003) in proton-decoupled ³¹P MRS. Of the reduction of "total choline," 61% was accounted for by glycerophosphorylcholine, a cerebral osmolyte. Similar metabolic abnormalities were seen in hyponatremic patients. The results are consistent with disturbances of cerebral osmoregulation and energy metabolism in patients with chronic hepatic encephalopathy.     

Dehydrins: Emergence of a biochemical role of a family of plant dehydration proteins

A number of proteins have been identified that typically accumulate in plants in response to any environmental stimulus that has a dehydrative component or is temporally associated with dehydration. This includes drought, low temperature, salinity and seed maturation. Among the induced proteins, dehydrins (late embryogenesis abundant [LEA] D-II family) have been the most commonly observed, yet we still have an incomplete knowledge of their fundamental biochemical role in the cell. Current research trends are changing this situation: immunolocalization and in vitro biochemical analyses are, through analogies to other more fully characterized proteins and molecules, shaping our understanding. In brief, dehydrins may be structure stabilizers with detergent and chaperone-like properties and an array of nuclear and cytoplasmic targets. Recent progress on the mapping of dhn genes and the inheritance of freezing tolerance in barley and other Triticeae species tentatively points to dehydrins as key components of dehydration tolerance.     

Brain Amino Acids During Hyponatremia In Vivo: Clinical Observations and Experimental Studies

Hyponatremia is a highly morbid condition, present in a wide range of human pathologies, that exposes patients to encephalopathic complication and the risk of permanent brain damage and death. Treating hyponatremia has proved to be difficult and still awaits safe management, avoiding the morbid sequelae of demyelinizing and necrotic lesions associated with the use of hypertonic solutions. During acute and chronic hyponatremia in vivo, the brain extrudes the excessive water by decreasing its content of electrolytes and organic osmolytes. At the cellular level, a similar response occurs upon cell swelling. Among the organic osmolytes involved in both responses, free amino acids play a prominent role because of the large intracellular pools often found in nerve cells. An overview of the changes in brain amino acid content during hyponatremia in vivo is presented and the contribution of these changes to the adaptive cell responses involved in volume regulation discussed. Additionally, new data are provided concerning changes in amino acid levels in different regions of the central nervous system after chronic hyponatremia. Results favor the role of taurine, glutamine, glutamate, and aspartate as the main amino acid osmolytes involved in the brain adaptive response to hyponatremia in vivo. Deeper knowledge of the adaptive overall and cellular brain mechanisms activated during hyponatremia would lead to the design of safer therapies for the hyponatremic patient.     

Desiccation increases sucrose levels in Ramonda and Haberlea, two genera of resurrection plants in the Gesneriaceae

A few genera of angiosperms are known as 'resurrection plants' since their leaves withstand complete desiccation. In many organisms, including some resurrection plants, desiccation tolerance is associated with the accumulation of special carbohydrates. We examined whether this is also true for the two European angiosperm genera of resurrection plants, Ramonda and Haberlea in the Gesneriaceae. Using gas chromatography, non-structural carbohydrates were determined as a percentage of the dry weight in leaves of Ramonda nathaliae subjected to various desiccation regimes. Sucrose was the predominant soluble carbohydrate in all samples, and its level steadily increased from 2 to 10% during desiccation. Starch amounted to ca 2% in control leaves and disappeared completely within 8 days of desiccation. Considerable amounts (1–2.5%) of raffinose and smaller amounts of its precursor galactinol (1-a-galactosyl- myo -inositol) were present in control leaves; these carbohydrates showed only minor changes upon desiccation. Similar results were obtained when excised leaves of Ramonda nathaliae, Ramonda myconi and Haberlea rhodopensis were subjected to desiccation. These data indicate that sucrose accumulation is connected to desiccation tolerance in Gesneriaceae; the presence of raffinose may be a pre-adaptation since this sugar prevents crystallization of sucrose during drying.     

Molecular cloning and characterization of a pea chitinase gene expressed in response to wounding,fungal infection and the elicitor chitosan

The fungicidal class I endochitinases (E.C.3.3.1.14, chitinase) are associated with the biochemical defense of plants against potential pathogens. We isolated and sequenced a genomic clone, DAH53, corresponding to a class I basic endochitinase gene in pea, Chil. The predicted amino acid sequence of this chitinase contains a hydrophobic C-terminal domain similar to the vacuole targeting sequences of class I chitinases isolated from other plants. The pea genome contains one gene corresponding to the chitinase DAH53 probe. Chitinase RNA accumulation was observed in pea pods within 2 to 4 h after inoculation with the incompatible fungal strain Fusarium solani f. sp. phaseoli, the compatible strain F. solani f.sp. pisi, or the elicitor chitosan. The RNA accumulation was high in the basal region (lower stem and root) of both fungus challenged and wounded pea seedlings. The sustained high levels of chitinase mRNA expression may contribute to later stages of pea's non-host resistance.     

Exogenously supplied glycine betaine in spinach and rapeseed leaf discs: compatibility or non-compatibility?

CMS, cell membrane stability
GB, glycine betaine
PEG, polyethylene glycol
TTC, 2,3,5-triphenyltetrazolium chloride

When leaf discs of spinach ( Spinacia oleracea cv. Junius) and rapeseed ( Brassica napus var. oleifera cv. Samourai) were incubated in the light in the presence of glycine betaine (GB), they accumulated GB at a very high level. In comparison with the spinach leaf explants, the uptake of GB by rapeseed tissues was restricted, probably by the destabilizing effects exerted by GB in this plant material. In contrast, the viability of spinach leaf discs, as assessed by their capacity to reduce 2,3,5-triphenyltetrazolium chloride (TTC), was not affected, suggesting that the GB taken up was compatible in the leaf tissues of the GB accumulator. In rapeseed leaf discs treated with GB, chlorophyll loss as well as significant changes in polyamine content were induced, leading to a dramatic increase of the putrescine/(spermidine + spermine) ratio. In contrast, this ratio remained constant in the GB treated spinach explants, suggesting that spinach has the capacity to stabilize polyamine metabolism in the presence of high amounts of GB. The treatment of spinach leaf discs with GB prior to application of osmotic or salt shocks provided protection from stress. A weak capacity to accumulate proline under stress conditions was partially suppressed. The protein content decreased while the free amino acid level increased independently of the presence of GB. It is concluded that GB behaves as a true compatible solute in spinach, which is a typical GB accumulator, and that GB is damaging when loaded into the leaf tissues of rapeseed, which do not normally accumulate GB.