Acclimation of potato plants to polyethylene glycol-induced water deficit I. Photosynthesis and metabolism

The acclimation of photosynthesis and metabolism in response to water deficit is characterized using hydroponically grown potato plants (Solanum tuberosum cv. Désirée). Plants were subjected to a reduced water potential of the nutrient solution by adding 10% (w/v) PEG 6000. PEG-treated plants were retarded in growth. Leaves which had been fully developed before the PEG treatment and leaves grown during the PEG treatment showed different phenotypes and biochemical and physiological properties. Photosynthesis of all leaves decreased during the whole treatment. However, the decrease of photosynthesis in the two types of leaves had different causes indicated by differences in their metabolism. Leaves which were fully developed at the beginning of the PEG treatment began to wilt starting from the leaf rim. The apoplastic ABA content increased, coinciding with a decreased stomatal conductance. Increased energy charge of the cells indicated impaired chloroplastic metabolism, accompanied by a decrease of amounts of chloroplastic enzymes. The apoplastic and the symplastic ABA content were increased during water deficit and because ABA was concentrated in the cytosolic compartment it is suggested that ABA is involved in decreasing photosynthetic enzyme contents in old leaves. Young leaves, grown after the imposition of water deficit, were smaller than control leaves and had a curly surface. In young leaves apoplastic and cytosolic ABA contents were identical with control values. Carboxylation efficiency of photosynthesis was decreased, but the water use efficiency remained unchanged. Metabolic data of the photosynthetic pathways indicate a down-regulation of chloroplastic metabolism. It is concluded that in young leaves photosynthesis was non-stomatally limited. This limitation was not caused by ABA.     

Adaptation versus shock response to polyethylene glycol-induced low water potential in cultured potato cells

We compared long-term adaptation versus short-term or shock response of potato ( Solanum tuberosum ) cells to polyethylene glycol (PEG)-induced low water potential. Potato cells, which were allowed to adapt gradually to a decreasing water potential, were able to grow actively in a medium containing 20% PEG. In contrast, no appreciable gain in dry weight was observed in potato cells shocked by abrupt transfer to the same medium. PEG-adapted cells were also salt-tolerant, as they were able to proliferate in a medium supplemented with 200 m M NaCl. No visible ultrastructural changes of mitochondria or proplastids were observed in adapted cells at values of low water potential (about −2.0 MPa), which caused membrane disruption and appearance of lipid droplets in unadapted cells. ABA cellular content increased 5-fold in PEG-shocked cells but no significant increase was found in PEG-adapted cells. The intracellular content of free proline increased 12.5 times over the basal level in PEG-adapted cells and 6.5 times in PEG-shocked cells. As shown by in vivo protein labeling, shock conditions strongly inhibited protein synthesis, which was completely recovered in PEG-adapted cells. Osmotin, a protein associated with salt adaptation in tobacco, was constitutively expressed at a high level in PEG-adapted cells and accumulated in PEG-shocked cells only three days after the transfer in a medium supplemented with 20% PEG. Proline and osmotin accumulation were coincident with the increase in cellular ABA content in PEG-shocked cells, but not in PEG-adapted cells. These data suggest that this hormone is mainly involved in shock response rather than long-term adaptation.     

Salt-tolerance in plants. II. In vitro translation of m-RNAs from salt-tolerant and salt-sensitive plants on wheat germ ribosomes. Responses to ions and compatible organic solutes

Abstract Messenger RNA from salt-sensitive and salt-tolerant plants Triticum aestivum. Beta vulgaris, Pisum sativum, Chenopodium album and Atriplex nummularia was translated in vitro in a wheatgerm translation system. The optimal monovalent and divalent ion concentrations for translation were independent of the salt tolerance of the plants from which the m-RNAs were derived. Translation was optimal in 100 120 mol m⁻³ potassium acetate and 1.5–2.0 mol m⁻³ Mg²⁺. Substitution of Na⁺ for K⁺, or of Cl⁻ for acetate, was inhibitory. The pattern of polypeptides synthesized from cytoplasmic m-RNAs of salt-sensitive and salt-tolerant plants remained constant in all the conditions examined. The effects of adding the ‘compatible' organic solutes glycine-betaine and mannitol were examined in the wheat-germ system primed with RNA from the leaves of Triticum aestivum or Beta vulgaris. The rate of translation, the optimum ionic concentrations and the distribution of polypeptide products were maintained in organic solute concentrations of up to 500 mol m⁻³. Proline above 300 mol m⁻³ and surcose above 100 mol m⁻³ did inhibit translation. The results indicate that translation in plants is unlikely in cytoplasmic K⁺ concentrations exceeding 180 mol m⁻³, but would proceed in the presence of up to 500 mol m⁻³ mannitol or glyinebetaine, or of up to 300 mol m⁻³ proline.     

Organ-specific distribution and subcellular localisation of ascorbate peroxidase isoenzymes in potato (Solanum tuberosum L.) plants

Summary. Ascorbate peroxidase (EC 1.11.1.11), a heme-containing homodimeric protein, is a hydrogen peroxide-scavenging enzyme, playing an important role in plants in order to protect them from oxidative stress, thus adverting cellular damage. Several ascorbate peroxidase isoenzymes have been reported but the understanding of their physiological role still depends on a better knowledge of their precise localisation within plant organs. Immunocytochemistry techniques were performed in order to elucidate the peroxisomal and cytosolic ascorbate peroxidase distribution within tissues of leaves and sprouts of potato plants. The peroxisomal isoenzyme was found to have a broad distribution in sprouts, but a differential one in leaves, being restricted to the spongy parenchyma. This differential expression may be associated to the mesophyll asymmetry and the diverse physiological processes that occur in it. The cytosolic isoenzyme was not detected in leaves under the used conditions, probably because it is present in low amounts in these tissues. The results obtained in sprouts were at least curious: cytosolic ascorbate was found to be adjacent to the amyloplasts. Given these results, it is possible to state that apart from their similarity, these two isoenzymes reside in different organelles and seem to take part in different physiological processes as suggested by their organ- and tissue-specific distribution. Correspondence and reprints: Plant Functional Biology Department, Institute for Cell and Molecular Biology, University of Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.     

Morphogenesis of potato plants in vitro. II. Endogenous levels,distribution, and metabolism of IAA and cytokinins

The levels of endogenous IAA and cytokinins (zeatin, zeatin riboside, isopentenyladenine, and isopentenyladenosine) were determined in potato plants cultured in vitro under red light (R) and blue light (B) on medium with or without hormones. On medium without hormones in B, plants contained much higher cytokinin levels, particularly in leaves and roots, and also slightly elevated IAA levels. Kinetin in the medium in B changed the distribution of cytokinins and significantly increased IAA level in roots. In R, the presence of kinetin led to an increased cytokinin level in the whole plant, while the IAA level was slightly lower. IAA in the medium in B decreased cytokinin level in all plant parts, while the IAA level did not change significantly. In R, the presence of IAA in the medium led to a moderate increase of CK level and to a significant increase in IAA level, especially in roots. Uptake of 1-¹⁴C-IAA and of ³H-zeatin was generally higher in B than in R. Higher percentage of IAA taken up in B was converted to conjugates in the roots. Metabolism of ³H-zeatin was similar in R and B with only slight differences in metabolite amounts.Thus, in all experimental situations in which tuber formation was stimulated, IAA level in roots and stolons rose significantly, stressing the importance of an IAA gradient for tuber formation.     

Cell membrane stability and biochemical response of cultured cells of groundnut under polyethylene glycol-induced water stress

Cell membrane stability (CMS) in suspension cultures of two groundnut cultivars was studied under polyethylene glycol(PEG)-induced water stress. There was a negative relationship between PEG concentration in the medium and membrane stability measured as electrolyte leakage. The CMS values in the cell cultures correlated well with the whole plant tissue and permitted the differentiation of cultivars based on their known response to drought stress. The cell membrane stability was lower (more electrolyte leakage) when cells were grown in culture as compared to the intact plant tissue. Kadiri-3, the drought tolerant cultivar maintained higher CMS than JL-24, the drought susceptible one. With increasing PEG levels the concentration of Potassium in cultured cells declined in both cultivars. However, Kadiri-3 maintained higher K values than JL-24 accompanied with greater cell membrane stability. Total soluble sugars also increased with increasing stress in both cultivars; the increase being higher in Kadiri-3. There was no significant change in the total free amino acids but proline accumulated markedly in both varieties. However, no relationship was found between proline levels and CMS. The results demonstrated that CMS test can also be used under in vitro conditions to differentiate the drought tolerant and susceptible cultivars and the cellular K level has a positive relationship with membrane stability.     

Salt-tolerance in plants. I. Ions, compatible organic solutes and the stability of plant ribosomes

Abstract Polysomes and ribosomes recovered from a number of plant species were tested for stability when incubated at 25°C in salt solutions in the absence of ATP and initiation factors. Stability was assessed by sucrose density gradient analysis. The stability was inversely proportional to salt concentrations above 125 mol m⁻³ KCl. Polysomes were less stable in the presence of Na⁺ than K⁺ salts, and were much less stable in Cl⁻ than in acetate salts. Polysomes from Triticum aestivum. Hordeum vulgare, Capsicum annuum, Helianthus annuus. Pisum sativum, Atriplex nummularia, Beta vulgaris, Cladophora sp., Enteromorpha sp. and Corallina cuvieri were similarly sensitive to KCl. Polysomes from Ulva lactuca were more sensitive than the other species. Cytoplasmic and plastid polysomes from T. aestivum were similarly unstable in 500 mol m⁻³ KCl. Unprogrammed ribosomal subunit couples from T. aestivum, B. vulgaris and U. lactuca showed Mg²⁺-dependent conformational instability and dissociation in KCl. Slight differences in ribosomal stability were observed between species, but these were unrelated to the salt tolerances of the plants. The ‘compatible’ organic solutes, glycinebetaine and proline, failed to reduce ion-induced instability. Ribosome yield and polysome profiles were similar in leaves of B. vulgaris containing significantly different levels of both Na⁺ and Cl⁻ after growth in media containing 50 or 200 mol m⁻³ NaCl. The results are consistent with the hypothesis that plants maintain a cytoplasmic solute environment that is compatible with ribosomal stability.     

Expression of an abscisic acid-binding single-chain antibody influences the subcellular distribution of abscisic acid and leads to developmental changes in transgenic potato plants

Potato (Solanum tuberosum L. cv. Désirée) plants were transformed to express a single-chain variable-fragment antibody against abscisic acid (ABA), and present in the endoplasmic reticulum at to up to 0.24% of the soluble leaf protein. The resulting transgenic plants were only able to grow normally at 95% humidity and moderate light. Four-week-old plants accumulated ABA to high extent, were retarded in growth and their leaves were smaller than those of control plants. Leaf stomatal conductivity was increased due to larger stomates. The subcellular concentrations of ABA in the chloroplast, cytoplasm and vacuole, and the apoplastic space of leaves were determined. In the 4-week-old transgenic plants the concentration of ABA not bound to the antibody was identical to that of control plants and the stomates were able to close in response to lower humidity of the atmosphere. A detailed analysis of age-dependent changes in plant metabolism showed that leaves of young transformed plants developed in ABA deficiency and leaves of older plants in ABA excess. Phenotypic changes developed in ABA deficiency partly disappeared in older plants.     

Brassinosteroids improve photosystem II efficiency,gas exchange,antioxidant enzymes and growth of cowpea plants exposed to water deficit

Water deficit is considered the main abiotic stress that limits agricultural production worldwide. Brassinosteroids (BRs) are natural substances that play roles in plant tolerance against abiotic stresses, including water deficit. This research aims to determine whether BRs can mitigate the negative effects caused by water deficiency, revealing how BRs act and their possible contribution to increased tolerance of cowpea plants to water deficit. The experiment was a factorial design with the factors completely randomised, with two water conditions (control and water deficit) and three levels of brassinosteroids (0, 50 and 100 nM 24-epibrassinolide; EBR is an active BRs). Plants sprayed with 100 nM EBR under the water deficit presented significant increases in Φ_PSII, q_P and ETR compared with plants subjected to the water deficit without EBR. With respect to gas exchange, P _N, E and g _s exhibited significant reductions after water deficit, but application of 100 nM EBR caused increases in these variables of 96, 24 and 33%, respectively, compared to the water deficit + 0 nM EBR treatment. To antioxidant enzymes, EBR resulted in increases in SOD, CAT, APX and POX, indicating that EBR acts on the antioxidant system, reducing cell damage. The water deficit caused significant reductions in Chl a, Chl b and total Chl, while plants sprayed with 100 nM EBR showed significant increases of 26, 58 and 33% in Chl a, Chl b and total Chl, respectively. This study revealed that EBR improves photosystem II efficiency, inducing increases in Φ_PSII, q_P and ETR. This substance also mitigated the negative effects on gas exchange and growth induced by the water deficit. Increases in SOD, CAT, APX and POX of plants treated with EBR indicate that this steroid clearly increased the tolerance to the water deficit, reducing reactive oxygen species, cell damage, and maintaining the photosynthetic pigments. Additionally, 100 nM EBR resulted in a better dose–response of cowpea plants exposed to the water deficit.     

Regulation of sucrose and starch metabolism in potato tubers in response to short-term water deficit

To investigate the effect of water stress on carbon metabolism in growing potato tubers (Solanum tuberosum L.), freshly cut and washed discs were incubated in a range of mannitol concentrations corresponding to external water potential between 0 and −1.2 MPa. (i) Incorporation of [¹⁴C]glucose into starch was inhibited in water-stressed discs, and labeling of sucrose was increased. High glucose overrode the changes at low water stress (up to −0.5 MPa) but not at high water stress. (ii) Although [¹⁴C]sucrose uptake increased in water-stressed discs, less of the absorbed [¹⁴C]sucrose was metabolised. (iii) Analysis of the sucrose content of the discs confirmed that increasing water deficit leads to a switch, from net sucrose degradation to net sucrose synthesis. (iv) In parallel incubations containing identical concentrations of sugars but differing in which sugar was labeled, degradation of [¹⁴C]sucrose and labeling of sucrose from [¹⁴C]glucose and fructose was found at each mannitol concentration. This shows that there is a cycle of sucrose degradation and resynthesis in these tuber discs. Increasing the extent of water stress changed the relation between sucrose breakdown and sucrose synthesis, in favour of synthesis. (v) Analysis of metabolites showed a biphasic response to increasing water deficit. Moderate water stress (0–200 mM mannitol) led to a decrease of the phosphorylated intermediates, especially 3-phosphoglycerate (3PGA). The decrease of metabolites at moderate water stress was not seen when high concentrations of glucose were supplied to the discs. More extreme water stress (300–500 mM mannitol) was accompanied by an accumulation of metabolites at low and high glucose. (vi) Moderate water stress led to an activation of sucrose phosphate synthase (SPS) in discs, and in intact tubers. The stimulation involved a change in the kinetic properties of SPS, and was blocked␣by protein phosphatase inhibitors. (vii) The amount of ADP-glucose (ADPGlc) decreased when discs were incubated on 100 or 200 mM mannitol. There was a strong correlation between the in vivo levels of ADPGlc and 3PGA when discs were subjected to moderate water stress, and when the sugar supply was varied. (viii) The level of ADPGlc increased and starch synthesis was further inhibited when discs were incubated in 300–500 mM mannitol. (ix) It is proposed that moderate water stress leads to an activation of SPS and stimulates sucrose synthesis. The resulting decline of 3PGA leads to a partial inhibition of ADP-glucose pyrophosphorylase and starch synthesis. More-extreme water stress leads to a further alteration of partitioning, because it inhibits the activities of one or more of the enzymes involved in the terminal reactions of starch synthesis. Received: 26 August 1996 / Accepted: 5 November 1996     

Maturation and subcellular compartmentation of potato starch phosphorylase.

The subcellular localization and maturation of starch phosphorylase (EC 2.4.1.1) was studied in developing potato tubers. The enzyme is localized inside the stroma of amyloplasts in young tubers, whereas in mature tubers it is found within the cytoplasm in the immediate vicinity of the plastids. A phosphorylase cDNA clone was isolated and used in RNA gel blot experiments to demonstrate that phosphorylase mRNAs are of the same size and abundance in both young and mature tubers. In vitro translation of mRNAs followed by immunoprecipitation with a phosphorylase antiserum indicates that the enzyme is synthesized as a higher molecular weight precursor in both young and mature tubers. The presence of a transit peptide at the N terminus of the protein was confirmed by the sequencing of the phosphorylase cDNA clone. The transit peptide has several structural features common to transit peptides of chloroplast proteins but contains a surprisingly large number of histidine residues. The mature form of the enzyme is present in both young and mature tubers, suggesting that a similar processing of the transit peptide may take place in two different subcellular locations.     

The use of Flory-Huggins theory in interpreting partitioning of solutes between organic liquids and water.

Solute partitioning data for dilute solutions have almost invariably been interpreted by equating experimental values of -RT in Kx (wherein Kx is the mole fraction partition coefficient) to delta mu infinity, the standard Gibbs energy change for solute transfer from one solvent to another. Recently, it has been alleged that this relation is insufficiently general. Instead, the statistical mechanical Flory-Huggins (FH) theory has been recommended for use, because it is designed to account for disparities in molecular size between solute and solvent. Our examination of the thermodynamics of partitioning shows that: (1) The customary interpretation is not only entirely correct (providing only that the solute is dilute), but is model-independent. (2) The dilute limit of the FH theory is seen to agree entirely with the usual interpretation of -RT in Kx, once certain misnomers are cleared away. (3) The use of FH theory being urged upon us in fact serves only to extract from delta mu infinity (the latter quite correctly determined as -RT in Kx) the contact part of delta mu infinity in order to obtain information on hydrophobic interactions. Some caveats are cited concerning such use of the FH statistical mechanical model.     

Polyethylene glycol-induced mammalian cell hybridization: effect of polyethylene glycol molecular weight and concentration.

The effects of polyethylene glycol (PEG) molecular weight and concentration on mammalian cell hybridization were studied. The peak hybridization-inducing activity with all grades of PEG from 400-6000 was found to occur in the concentration range of 50-55%. However, changes in concentration were seen to have different quantitative effects with different grades of PEG. For monolayer fusions, PEG 1000 at 50% seems to be the optimal combination of PEG molecular weight and concentration, in terms of both efficiency of hybridization and relative insensitivity to dilution effects.     

Acclimation and adaptive responses of woody plants to environmental stresses

The predominant emphasis on harmful effects of environmental stresses on growth of woody plants has obscured some very beneficial effects of such stresses. Slowly increasing stresses may induce physiological adjustment that protects plants from the growth inhibition and/or injury that follow when environmental stresses are abruptly imposed. In addition, short exposures of woody plants to extreme environmental conditions at critical times in their development often improve growth. Furthermore, maintaining harvested seedlings and plant products at very low temperatures extends their longevity. Drought tolerance: Seedlings previously exposed to water stress often undergo less inhibition of growth and other processes following transplanting than do seedlings not previously exposed to such stress. Controlled wetting and drying cycles often promote early budset, dormancy, and drought tolerance. In many species increased drought tolerance following such cycles is associated with osmotic adjustment that involves accumulation of osmotically active substances. Maintenance of leaf turgor often is linked to osmotic adjustment. A reduction in osmotic volume at full turgor also results in reduced osmotic potential, even in the absence of solute accumulation. Changes in tissue elasticity may be important for turgor maintenance and drought tolerance of plants that do not adjust osmotically. Water deficits and nutrient deficiencies promote greater relative allocation of photosynthate to root growth, ultimately resulting in plants that have higher root:shoot ratios and greater capacity to absorb water and minerals relative to the shoots that must be supported. At the molecular level, plants respond to water stress by synthesis of certain new proteins and increased levels of synthesis of some proteins produced under well-watered conditions. Evidence has been obtained for enhanced synthesis under water stress of water-channel proteins and other proteins that may protect membranes and other important macromolecules from damage and denaturation as cells dehydrate. Flood tolerance: Both artificial and natural flooding sometimes benefit woody plants. Flooding of orchard soils has been an essential management practice for centuries to increase fruit yields and improve fruit quality. Also, annual advances and recessions of floods are crucial for maintaining valuable riparian forests. Intermittent flooding protects bottomland forests by increasing groundwater supplies, transporting sediments necessary for creating favorable seedbeds, and regulating decomposition of organic matter. Major adaptations for flood tolerance of some woody plants include high capacity for producing adventitious roots that compensate physiologically for decay of original roots under soil anaerobiosis, facilitation of oxygen uptake through stomata and newly formed lenticels, and metabolic adjustments. Halophytes can adapt to saline water by salt tolerance, salt avoidance, or both. Cold hardiness: Environmental stresses that inhibit plant growth, including low temperature, drought, short days, and combinations of these, induce cold hardening and hardiness in many species. Cold hardiness develops in two stages: at temperatures between 10° and 20°C in the autumn, when carbohydrates and lipids accumulate; and at subsequent freezing temperatures. The sum of many biochemical processes determines the degree of cold tolerance. Some of these processes are hormone dependent and induced by short days; others that are linked to activity of enzyme systems are temperature dependent. Short days are important for development of cold hardiness in species that set buds or respond strongly to photoperiod. Nursery managers often expose tree seedlings to moderate water stress at or near the end of the growing season. This accelerates budset, induces early dormancy, and increases cold hardiness. Pollution tolerance: Absorption of gaseous air pollutants varies with resistance to flow along the pollutant’s diffusion path. Hence, the amount of pollutant absorbed by leaves depends on stomatal aperture, stomatal size, and stomatal frequency. Pollution tolerance is increased when drought, dry air, or flooding of soil close stomatal pores. Heat tolerance: Exposure to sublethal high temperature can increase the thermotolerance of plants. Potential mechanisms of response include synthesis of heat-shock proteins and isoprene and antioxidant production to protect the photosynthetic apparatus and cellular metabolism. Breaking of dormancy: Seed dormancy can be broken by cold or heat. Embryo dormancy is broken by prolonged exposure of most seeds to temperatures of 1° to 15°C. The efficiency of treatment depends on interactions between temperature and seed moisture content. Germination can be postponed by partially dehydrating seeds or altering the temperature during seed stratification. Seed-coat dormancy can be broken by fires that rupture seed coats or melt seedcoat waxes, hence promoting water uptake. Seeds with both embryo dormancy and seed-coat dormancy may require exposure to both high and low temperatures to break dormancy. Exposure to smoke itself can also serve as a germination cue in breaking seed dormancy in some species. Bud dormancy of temperate-zone trees is broken by winter cold. The specific chilling requirement varies widely with species and genotype, type of bud (e.g., vegetative or floral bud), depth of dormancy, temperature, duration of chilling, stage of plant development, and daylength. Interruption of a cold regime by high temperature may negate the effect of sustained chilling or breaking of bud dormancy. Near-lethal heat stress may release buds from both endodormancy and ecodormancy. Pollen shedding: Dehiscence of anthers and release of pollen result from dehydration of walls of anther sacs. Both seasonal and diurnal pollen shedding are commonly associated with shrinkage and rupture of anther walls by low relative humidity. Pollen shedding typically is maximal near midday (low relative humidity) and low at night (high relative humidity). Pollen shedding is low or negligible during rainy periods. Seed dispersal: Gymnosperm cones typically dehydrate before opening. The cones open and shed seeds because of differential shrinkage between the adaxial and abaxial tissues of cone scales. Once opened, cones may close and reopen with changes in relative humidity. Both dehydration and heat are necessary for seed dispersal from serotinous (late-to-open) cones. Seeds are stored in serotinous cones because resinous bonds of scales prevent cone opening. After fire melts the resinous material, the cone scales can open on drying. Fires also stimulate germination of seeds of some species. Some heath plants require fire to open their serotinous follicles and shed seeds. Fire destroys the resin at the valves of follicles, and the valves then reflex to release the seeds. Following fire the follicles of some species require alternate wetting and drying for efficient seed dispersal. Stimulation of reproductive growth: Vegetative and reproductive growth of woody plants are negatively correlated. A heavy crop of fruits, cones, and seeds is associated with reduced vegetative growth in the same or following year (or even years). Subjecting trees to drought during early stages of fruit development to inhibit vegetative growth, followed by normal irrigation, sometimes favors reproductive growth. Short periods of drought at critical times not only induce formation of flower buds but also break dormancy of flower buds in some species. Water deficits may induce flowering directly or by inhibiting shoot flushing, thereby limiting the capacity of young leaves to inhibit floral induction. Postharvest water stress often results in abundant return bloom over that in well-irrigated plants. Fruit yields of some species are not reduced or are increased by withholding irrigation during the period of shoot elongation. In several species, osmotic adjustment occurs during deficit irrigation. In other species, increased fruit growth by imposed drought is not associated largely with osmotic adjustment and maintenance of leaf turgor. Seedling storage: Tree seedlings typically are stored at temperatures just above or below freezing. Growth and survival of cold-stored seedlings depend on such factors as: date of lifting from the nursery; species and genotype; storage temperature, humidity, and illumination; duration of storage; and handling of planting stock after storage. Seedlings to be stored over winter should be lifted from the nursery as late as possible. Dehydration of seedlings before, during, and after storage adversely affects growth of outplanted seedlings. Long-term storage of seedlings may result in depletion of stored carbohydrates by respiration and decrease of root growth potential. Although many seedlings are stored in darkness, a daily photoperiod during cold storage may stimulate subsequent growth and increase survival of outplanted seedlings. For some species, rapid thawing may decrease respiratory consumption of carbohydrates (over slowly thawed seedlings) and decrease development of molds. Pollen storage: Preservation of pollen is necessary for insurance against poor flowering years, for gene conservation, and for physiological and biochemical studies. Storage temperature and pollen moisture content largely determine longevity of stored pollen. Pollen can be stored successfully for many years in deep freezers at temperatures near −15°C or in liquid nitrogen (−196°C). Cryopreservation of pollen with a high moisture content is difficult because ice crystals may destroy the cells. Pollens of many species do not survive at temperatures below −40°C if their moisture contents exceed 20–30%. Pollen generally is air dried, vacuum dried, or freeze dried before it is stored. To preserve the germination capacity of stored pollen, rehydration at high humidity often is necessary. Seed storage: Seeds are routinely stored to provide a seed supply during years of poor seed production, to maintain genetic diversity, and to breed plants. For a long time, seeds were classified as either orthodox (relatively long-lived, with capacity for dehydration to very low moisture contents without losing viability) or recalcitrant (short-lived and requiring a high moisture content for retention of viability). More recently, some seeds have been reclassified as suborthodox or intermediate because they retain viability when carefully dried. True orthodox seeds are preserved much more easily than are nonorthodox seeds. Orthodox seeds can be stored for a long time at temperatures between 2° and −20°C, with temperatures below −5°C preferable. Some orthodox seeds have been stored at superlow temperatures, although temperatures of −40°, −70°, or −196°C have not been appreciably better than −20°C for storage of seeds of a number of species. Only relatively short-term storage protocols have been developed for nonorthodox seeds. These treatments typically extend seed viability to as much as a year. The methods often require cryopreservation of excised embryos. Responses to cryopreservation of nonorthodox seeds or embryos vary with species and genotype, rate of drying, use of cryoprotectants, rates of freezing and thawing, and rate of rehydration. Fruit storage: Storing fruits at low temperatures above freezing, increasing the CO₂ concentration, and lowering the O₂ concentration of fruit storage delays senescence of fruits and prolongs their life. Fruits continue to senesce and decay while in storage and become increasingly susceptible to diseases. Both temperate-zone and tropical fruits may develop chilling injury characterized by lesions, internal discoloration, greater susceptibility to decay, and shortened storage life. Chilling injury can be controlled by chemicals, temperature conditioning, and intermittent warming during storage. Stored fruits may become increasingly susceptible to disease organisms. Fruit diseases can be controlled by cold, which inhibits growth of microorganisms and maintains host resistance. Exposure of fruits to high CO₂ and low O₂ during storage directly suppresses disease-causing fungi. Pathogens also can be controlled by exposing fruits to heat before, during, and after storage. Scald that often develops during low-temperature storage can be controlled by chemicals and by heat treatments.     

Activated oxygen production and detoxification in wheat plants subjected to a water deficit programme

Wheat plants (Triticum durum L. cv. Ofanto) were grown in acontrolled environment. In one set, control plants were regularlywatered; the other set of plants was subjected to two waterdeficit periods obtained by withholding water and rewateringto field capacity at the end of the first period. After bothperiods of stress, water potential (     

Acclimation to long-term water deficit in the leaves of two sunflower hybrids: photosynthesis, electron transport and carbon metabolism

The influence of long-term water deficit on photosynthesis, electron transport and carbon metabolism of sunflower leaves has been examined. Water deficit was imposed from flower bud formation up to the stage of full flowering in the field on two sunflower hybrids with different drought tolerance. CO2 assimilation and stomatal conductance of the intact leaves, determined at atmospheric CO2 and full sunlight (1500-2000 mol quanta m^-2 s^-1), decreased with water deficit. Maximum quantum efficiency of PSII (Fy/Fm) and relative quantum yield of PSII (II) determined under similar experimental conditions, did not change significantly in severely stressed leaves. The strong inhibition of the plateau region of the light response curve, determined at high CO2 (5%) in water-deficient sunflower leaves, indicates that photosynthesis is also limited by non-stomatal factors. The decreased slope and the plateau of the CO2 response curves show that the capacity of carboxylation and RuBP regeneration decreased in severely stressed intact leaves. Rubisco specific activity decreased in severely stressed leaves, but Rubisco content increased under prolonged drought. The increase of Rubisco content was significantly higher in leaves of the drought-tolerant sunflower hybrid indicating that a higher Rubisco content could be one factor in conferring better acclimation and higher drought tolerance.