Protein synthesis in halophytes: The influence of potassium,sodium and magnesium in vitro

The amino acid (³⁵S-methionine) incorporating activity of an in vitro wheat germ translation system was found to be maximal in 80 to 125 mol m^–3 K with 2 to 4 mol m^–3 Mg both as the acetate. Substitution of Na for K, or chloride for acetate at concentrations above 80 mol m^–3 inhibited incorporation. When the K acetate concentration was raised to 200 mol m^–3, no incorporation of radioactive methionine occurred.Translation by polysomes extracted from leaf tissue of S. maritima, supplemented with postribosomal supernatant from wheat germ, showed activity which was optimal in the presence of 225 mol m^–3 K acetate and 8 mol m^–3 Mg acetate. However, the translation system was not directly comparable with the wheat germ system, as studies with an initiation inhibitor, aurintricarboxylic acid, suggested that the S. maritima system was essentially elongation-dependent, while initiation occurred in the wheat germ system.Elongation-dependent polysomal preparations were extracted from leaves of the glycophytes Pisum sativum, Triticum aestivum, Oryza sativa and Hordeum vulgare, and from the halophytes Atriplex isatidea and Inula crithmoides. Translation by polysomes from the salt-tolerant plants was optimal at higher K and Mg concentrations, than by polysomes from the glycophytes. Furthermore, NaCl was better able partially to substitute for the role of K in polysomal preparations from halophytes than glycophytes.     

Corn uptake and microbial immobilization of 15N-labeled urea-N in soil as affected by composted pig manure

The tolerance of 24 genotypes of barley was assessed by estimating their survival in saline conditions either in a glasshouse or in a controlled environment cabinet. Two cultivars, sensitive Triumph and resistant Gerbel, were picked for further study, which involved sequential harvesting of plants grown in a range of salinities. After about one month in 200 mol m^–3 sodium chloride, the sodium concentration in the roots and shoots of the sensitive Triumph was about 1.5 times that in the roots of resistant Gerbel. The addition of Na to the root medium reduced the potassium transport to the shoot in Triumph to a much greater extent than in Gerbel, so the K:Na ratio of Gerbel was twice that for Triumph, when averaged over all treatments and harvests. The sodium, potassium and chloride concentrations within the major subcellular compartments of the cortical cells of roots of Triumph and Gerbel were determined by X-ray microanalysis following freeze-substitution and dry-sectioning. The mean cytoplasmic sodium concentration (245 mol m^–3 analysed volume) in Triumph grown in 200 mol m^–3 NaCl for 15 d was almost 1.4 times greater than that in the resistant Gerbel: the potassium concentration in Gerbel showed a lower reduction than did that of Triumph. Another major difference between the two cultivars was the higher concentrations of sodium and chloride in the cell walls of Triumph than Gerbel: the sodium concentration in the cortical cell walls of the salt-sensitive cultivar was about 1.75 times that in the more salt-resistant cultivar. The exchange capacity of the cell walls of Gerbel was greater than that of Triumph. We hypothesise that ion transport to the shoot reflects cytosolic ion concentrations, with a more sensitive cultivar having a higher sodium concentration in its cytoplasm than a more resistant variety. It is noteworthy that the difference in the K:Na ratio between the shoots of Gerbel and Triumph after 15 days of exposure to 200 mol m^–3 NaCl was similar to the difference in their symplastic K:Na ratios.     

Combined effect of salinity and hypoxia in wheat (Triticum aestivum L.) and wheat-Thinopyrum amphiploids

The effects of sodium chloride salinity and hypoxia were studied in eight wheat lines and three wheat-Thinopyrum amphiploids in vermiculite-gravel culture. The lines were treated with either 100 or 150 mol m^–3 NaCl with and without hypoxia. Saline hypoxic conditions significantly reduced the vegetative growth, water use, grain and straw yields for all wheat varieties except the amphiploids, whereas NaCl or hypoxia alone had less pronounced effects. In addition, saline hypoxic stress reduced K⁺ concentration and increased significantly the Na⁺ and Cl^– concentrations in cell sap expressed from leaves. There was more Na⁺ and Cl^– accumulation in wheats than the amphiploids in hypoxic conditions at 150 mol m^–3 NaCl. Of the wheats, Pato was the most sensitive at all stress levels while aTriticum aestivum cv. Chinese Spring ×Thinopyrum elongatum amphiploid was the most tolerant of the three amphiploids.     

Effects of short-term salinity on leaf gas exchange of the fig (Ficus carica L.)

The influence of short-term salinity (day 1–day 2: 50 mol m^–3 NaCl, day 3–day 7: 100 mol m^–3 NaCl in the nutrient solution) on leaf gas exchange characteristics were studied in two fig clones (Ficus carica L.), whose root mass had been varied in relation to the leaf area. The stomatal conductance was diminished by NaCl in the first week of treatment. NaCl slightly reduced the calculated intercellular partial pressure of CO₂. The net photosynthetic rate of plants with many roots was stimulated by NaCl on some days of the first week of treatment, whereas the net assimilation rate of the plants with few roots remained unaltered or decreased by NaCl. Only the assimilation of the salt-treated plants of one clone for some days during the first week of treatment seemed to be influenced by stomatal conductance. Nonstomatal factors were primarily responsible for the changes in CO₂ uptake in response to salt and/or root treatment. The water use efficiency increased during several days of the first week of NaCl treatment. Decreased stomatal conductance, increased water use efficiency and stimualtion of the net CO₂ assimilation rate appear to enhance salt tolerance during the first few days of salinity. ei]H Lambers     

Salinity studies with drought-resistant species of Sporobolus

Summary Dry matter productivity under saline conditions was compared in 5 desiccation-tolerant resurrection grasses and one desiccation sensitive species, all in the genus Sporobolus. S. stapfianus was the most salt tolerant, requiring 215 mole NaCl m^-3 to reduce shoot dry matter increments to 50% of increments in plants not treated with salt. (This was comparable to published values for the salt tolerant grass Diplachne fusca.) S. lampranthus was salt sensitive, requiring 35 mol m^-3 for 50% control yields. S. festivus, S. aff. Fimbriatus, and the deisccation sensitive S.pyramidalis was moderately tolerant (150–170 mol m^-3). The moderate salt resistance of S. aff. fimbriatus was attributed mainly to exclusion of NaCl by roots. Salt export through leaf surfaces was a minor factor. Half of the leaf mesophyll cells survived 50 min immersion in 200 mol NaCl m^-3. Plants of S. aff. fimbriatus and S. pyramidalis tolerated a broad range of soil pH. Plants of 4 desiccation tolerant Sporobolus species survived air-dryness following 3 weeks pretreatment with salinities up to 200 mol m^-3     

Relationship between morphological and physiological responses to waterlogging and salinity in Sporobolus virginicus (L.) Kunth

The effects of waterlogging and salinity on morphological and physiological responses in the marsh grass Sporobolus virginicus (L.) Kunth were investigated in a 4×2 factorial experiment. Plants were subjected to four salinity levels (0, 100, 200 and 400 mol m^–3 NaCl) and two soil inundation conditions (drained and flooded) for 42 days. Flooding at 0 mol m^–3 NaCl caused initiation of adventitious surface roots, increased internal acration and plant height, induced alcohol dehydrogenase activity (ADH), and decreased belowground biomass and the number of culms per plant. Salinity increase from 0 to 400 mol m^–3 NaCl under drained conditions increased leaf and root proline concentrations and decreased photosynthesis, aboveground biomass, number of culms per plant and number of internodes per culm. Concurrent waterlogging and salinity induced ADH activity and adventitious surface roots but decreased plant height and aboveground biomass. Internal air space increased with waterlogging from 0 to 100 mol m^–3 NaCl but further increases in salinity to 400 mol m^–3 reduced air space. Combined waterlogging and salinity stresses, however, had no effect on photosynthesis or on the concentrations of proline in leaves or roots. These results are discussed in relation to the widespread colonization by S. virginicus of a wide range of coastal environments varying in soil salinity and in the frequency and intensity of waterlogging.     

Consequences of depletion of stratospheric ozone for terrestrial Antarctic ecosystems: the response of Deschampsia antarctica to enhanced UV-B radiation in a controlled environment

Mini UV lamps were installed over antarctic plants at Léonie Island, Antarctic peninsula, and shoot length measurements of Deschampsia antarctica were performed during the austral summer January–February 1999.We studied the response of the antarctic hairgrass, Deschampsia antarctica to enhanced UV-B. In a climate room experiment we exposed tillers of Deschampsia antarctica, collected at Léonie Island, Antarctic peninsula, to ambient and enhanced levels of UV-B radiation. In this climate room experiment with 0, 2.5 and 5 kJ m^–2 day^–1 UV-B_BE treatments we observed that length growth of shoots at 2.5 and 5 kJ m^–2 day^–1 UV-B_BE was markedly reduced compared to 0 kJ m^–2 day^–1 UV-B_BE. In addition, there was an increased number of shoots and increased leaf thickness with enhanced UV-B. The Relative Growth Rate (RGR) was not affected by UV-B, possibly because reduced shoot length growth by enhanced UV-B was compensated by increased tillering. Light response curves of net leaf photosynthesis of plants exposed to 5 kJ m^–2 day^–1 UV-B_BE did not differ from those exposed to 0 kJ m^–2 day^–1 UV-B_BE. The content of UV-B absorbing compounds of plants exposed to increasing UV-B did not significantly change.Mini UV-B lamp systems were installed in the field, to expose the terrestrial antarctic vegetation at Léonie Island to enhanced solar UV-B. In that study, the increment of shoot length of tagged plants of Deschampsia antarctica during the January-February 1999 at Léonie Island, was recorded and compared to shoot length growth under controlled conditions.The consequences of enhanced UV-B radiation as a result of ozone depletion for the terrestrial antarctic ecosytems are discussed.     

Salinity effect on plant growth and leaf demography of the mangrove, Avicennia germinans L.

We assessed the effect of salinity on plant growth and leaf expansion rates, as well as the leaf life span and the dynamics of leaf production and mortality in seedlings of Avicennia germinans L. grown at 0, 170, 430, 680, and 940 mol m⁻³ NaCl. The relative growth rates (RGR) after 27 weeks reached a maximum (10.4 mg g⁻¹ d⁻¹) in 170 mol m⁻³ NaCl and decreased by 47 and 44% in plants grown at 680 and 940 mol m⁻³ NaCl. The relative leaf expansion rate (RLER) was maximal at 170 mol m⁻³ NaCl (120 cm m⁻² d⁻¹) and decreased by 57 and 52% in plants grown at 680 and 940 mol m⁻³ NaCl, respectively. In the same manner as RGR and RLER, the leaf production (P) and leaf death (D) decreased in 81 and 67% when salinity increased from 170 to 940 mol m⁻³ NaCl, respectively. Since the decrease in P with salinity was more pronounced than the decrease in D, the net accumulation of leaves per plant decreased with salinity. Additionally, an evident increase in annual mortality rates (λ) and death probability was observed with salinity. Leaf half-life (t _0.5) was 425 days in plants grown at 0 mol m⁻³ NaCl, and decreased to 75 days at 940 mol m⁻³ NaCl. Thus, increasing salinity caused an increase in mortality rate whereas production of new leaves and leaf longevity decreased and, finally, the leaf area was reduced.     

Effects of external NaCl on the growth of Atriplex amnicola and the ion relations and carbohydrate status of the leaves

Abstract Atriplex amnicola, was grown in nutrient solution cultures with concentrations of NaCl up to 750 mol m^?3. The growth optimum was at 25–50 mol m^?3 NaCl and growth was 10–15% of that value at 750 mol m^?3 NaCl. Sodium chloride at 200 mol m^?3 and higher reduced the rate of leaf extension and increased the time taken for a leaf to reach its maximal length. Concentrations of Na⁺, K⁺ and Mg²⁺ in leaves of different ages were investigated for plants grown at 25, 200 and 400 mol m^?3 NaCl. Although leaves of plants grown at 200 and 400 mol m^?3 NaCl had high Na⁺ concentrations at young developmental stages, much of this Na⁺ was located in the salt bladders. Leaves excluding bladders had low Na⁺ concentrations when young, but very high in Na⁺ when old. In contrast to Na⁺, K⁺ concentrations were similar in bladders and leaves excluding bladders. Concentrations of K⁺ were higher in the rapidly expanding than in the old leaves. At 400 mol m^?3 NaCl, the K⁺:Na⁺ ratios of the leaves excluding bladders were 0.4–0.6 and 0.1 for rapidly expanding and oldest leaves, respectively. The Na⁺ content in moles per leaf, excluding bladders, increased linearly with the age of the leaves; concurrent increases in succulence were closely correlated with the Na ⁺ concentration in the leaves excluding the bladders. Soluble sugars and starch in leaves, stems and buds were determined at dusk and dawn. There was a pronounced diurnal fluctation in concentrations of carbohydrates. During the night, most plant parts showed large decreases in starch and sugar. Concentrations of carbohydrates in most plant organs were similar for plants grown at 25 and 400 mol m^?3 NaCl. One notable exception was buds at dusk, where sugar and starch concentrations were 30–35% less in plants grown at 400 mol m^?3 NaCl than in plants grown at 25 mol m^?3 NaCl. The data indicate that the growth of A. amnicola at 400 mol m^?3 NaCl is not limited by the availability of photosynthate in the plant as a whole. However, there could have been a growth limitation due to inadequate organic solutes for osmotic regulation.     

The role of nitrate in osmoregulation of Italian ryegrass

Summary The role of nitrate in osmotic control was studied with Italian ryegrass grown in a nutrient solution in a climate room. Quantum-flux density, osmotic potential of the nutrient solution and availability of nitrate and chloride were varied independently. Plants at high quantum flux density (650 mol m^–2 s^–1) had a lower osmotic potential, a higher carbohydrate concentration and a lower nitrate concentration than plants at low quantum flux density (310 mol m^–2 s^–1), the decrease in nitrate concentration was osmotically equivalent to the increase in carbohydrate concentration. When nitrate in the nutrient solution was partly replaced by chloride, the chloride taken up substituted an equivalent part of the nitrate in the plant. It is concluded that nitrate plays a role in osmoregulation of the plant and compensates for a shortage of other solutes.     

Seasonal dynamics of <Emphasis Type="Italic">Zostera noltii</Emphasis> Hornem. in two Mediterranean lagoons

The density, biomass and shoot morphology of two populations of Zostera noltii were monitored from January 1998 to July 1999 at two shallow Mediterranean lagoons of Biguglia and Urbino, which differ in hydro-morphological conditions and nutrient loading. Monitoring included the principal biological and foliar parameters (shoot density, aboveground and belowground biomass, length, width and number of leaves, LAI and coefficient A: percentage of leaves having lost their apex), the organic matter contents of the sediment and the environmental conditions (salinity, turbidity, temperature, nutrient concentrations and dissolved oxygen levels). The two populations of Z. noltii displayed seasonal changes in density (1600–19600 m²), aboveground biomass (11–153 g. DW. m⁻²), leaf length (33–255 mm), and leaf width (0.9–1.8 mm). Temperature and turbidity were significant environmental factors influencing the temporal changes observed in the Z. noltii meadows studied. Conversely, the belowground biomass, the number of leaves per shoot and the LAI did not undergo any seasonal changes. In the Biguglia lagoon, the functioning dynamics of the Z. noltii seagrass beds are determined by the catchment area and the inputs of nutrients derived from it, whereas in the Urbino lagoon the dynamics of the Z. noltiibeds depend on low levels of water turbidity.     

Analysis of Qualitative Contribution of Assimilatory and Non-Assimilatory De-Excitation Processes to Adaptation of Photosynthetic Apparatus of Barley Plants to High Irradiance

The adaptation of barley (Hordeum vulgare L. cv. Akcent) plants to low (LI, 50 µmol m^–2 s^–1) and high (HI, 1000 µmol m^–2 s^–1) growth irradiances was studied using the simultaneous measurements of the photosynthetic oxygen evolution and chlorophyll a (Chl a) fluorescence at room temperature. If measured under ambient CO₂ concentration, neither increase of the oxygen evolution rate (P) nor enhancement of non-radiative dissipation of the absorbed excitation energy within photosystem 2 (PS2) (determined as non-photochemical quenching of Chl a fluorescence, NPQ) were observed for HI plants compared with LI plants. Nevertheless, the HI plants exhibited a significantly higher proportion of Q_A in oxidised state (estimated from photochemical quenching of Chl a fluorescence, q_P), by 49–102 % at irradiances above 200 µmol m^–2 s^–1 and an about 1.5 fold increase of irradiance-saturated PS2 electron transport rate (ETR) as compared to LI plants. At high CO₂ concentration the degree of P stimulation was approximately three times higher for HI than for LI plants, and the irradiance-saturated P values at irradiances of 2 440 and 2 900 µmol m^–2 s^–1 were by 130 and 150 % higher for HI plants than for LI plants. We suggest that non-assimilatory electron transport dominates in the adaptation of the photosynthetic apparatus of barley grown at high irradiances under ambient CO₂ rather than an increased NPQ or an enhancement of irradiance-saturated photosynthesis.     

Hydraulic Properties of a Mangrove Avicennia Germinans as Affected by NaCl

Water transport was assessed in seedlings of the mangrove Avicennia germinans L. grown at 171 and 684 mol m^–3 NaCl. Leaf specific conductivity declined by 25 % at high salinity. This was related to low specific conductivity, because Huber values remained similar. Leaves of A. germinans featured low internal conductance to water transport. This was lowered further under high salinity. Water transport constrains imposed by whole shoot and leaf blade at high salinity were balanced by stomatal regulation of water loss, which possibly maintain stem water potentials above embolisms levels.     

Photosynthetic and leaf morphological characteristics in Leucaena leucocephala as affected by growth under different neutral shade levels

Morphological and physiological measurements on individual leaves of Leucaena leucocephala seedlings were used to study acclimation to neutral shading. The light-saturated photosynthetic rate (P_{n max}) ranged from 19.6 to 6.5 mol CO₂ m^–2 s^–1 as photosynthetic photon flux density (PPFD) during growth decreased from 27 to 1.6 mol m^–2 s^–1. Stomatal density varied from 144 mm^–2 in plants grown in high PPFD to 84 mm^–2 in plants grown in low PPFD. Average maximal stomatal conductance for H₂O was 1.1 in plants grown in high PPFD and 0.3 for plants grown in low PPFD. Plants grown in low PPFD had a greater total chlorophyll content than plants grown in high PPFD (7.2 vs 2.9 mg g^–1 on a unit fresh weight basis, and 4.3 vs 3.7 mg dm^–2 on a unit leaf area basis). Leaf area was largest when plants were grown under the intermediate PPFDs. Leaf density thickness was largest when plants were grown under the largest PPFDs. It is concluded that L. leucocephala shows extensive ability to acclimate to neutral shade, and could be considered a facultative shade plant.Abbreviations the initial slope of the photosynthesis vs PPFD curve - P_{n max} the light-saturated photosynthetic rate - PPFD photosynthetic photon flux density     

Content of Oxalate in Actinidia Deliciosa Plants Grown in Nutrient Solutions with Different Nitrogen Forms

Kiwifruit plants (Actinidia deliciosa cv. Hayward) were grown in Hoagland nutrient solution with calcium nitrate, potassium nitrate, ammonium nitrate or ammonium chloride as the nitrogen source. Plants grown in the solution with nitrate nitrogen displayed a higher oxalate content, greater shoot length and leaf area, and higher content of ascorbic acid and NO₃ ^– ions in the leaves. Plants grown in the solution with ammonium nitrate, and particularly with ammonium chloride, showed low oxalate content, low content of ascorbic acid and NO₃ ^–, high content of Cl^– and Na⁺, low shoot length and leaf area. Oxalate formation appeared to be connected with the assimulation of nitrate, more precisely with nitrate reduction, while ammonium nitrogen assimilation did not induce the synthesis of oxalic acid.     

Effects of calcium on growth and leaf ion concentrations of Gossypium hirsutum grown in saline hydroponic culture

The optimum Ca²⁺ concentration for growth of cotton (Gossypium hirsutum cv. Acala SJ-2) was in the range 1 to 15 mol m^–3 for plants growing in hydroponic culture with 100–150 mol m^–3 NaCl. Most saline (but not sodic) soils contain higher Ca²⁺ concentrations. CaCl₂ was inhibitory to the growth of cotton above 20–50 mol m^–3. Increasing concentrations of Ca²⁺ in the range 0–2 mol m^–2 drastically reduced Na⁺ accumulation in the leaves. As CaCl₂ concentrations were increased above the optimum for growth there was a further reduction in leaf Na⁺ accumulation, but this was more than offset by increased leaf Ca²⁺ and Cl^– concentrations. Leaf K⁺ concentrations were not much affected by changes in external CaCl₂ concentrations. The response of Mg²⁺ varied from an increase to a decrease with increasing external CaCl₂ and was influenced by nutritional status. There was no evidence that high Ca²⁺ caused a deficiency of Mg²⁺ in cotton. Except for Cl^–, whose concentrations tended to decrease initially and then increase as the CaCl₂ concentration increased, the anions were largely unaffected by changes in external CaCl₂.     

Effects of constitutive expression of nitrate reductase in transgenic Nicotiana plumbaginifolia L. in response to varying nitrogen supply

Transformed Nicotiana plumbaginifolia plants with constitutive expression of nitrate reductase (NR) activity were grown at different levels of nitrogen nutrition. The gradients in foliar NO ₃ ^– content and maximum extractable NR activity observed with leaf order on the shoot, from base to apex, were much decreased as a result of N-deficiency in both the transformed plants and wild type controls grown under identical conditions. Constitutive expression of NR did not influence the foliar protein and chlorophyll contents under any circumstances. A reciprocal relationship between the observed maximal extractable NR activity of the leaves and their NO ₃ ^– content was observed in plants grown in nitrogen replete conditions at low irradiance (170 mol photons·m^–2 ·s^–1). This relationship disappeared at higher irradiance (450 mol photons·m^–2·S^–1) because the maximal extractable NR activity in the leaves of the wild type plants in these conditions increased to a level that was similar to, or greater than that found in constitutive NR-expressors. Much more NO ₃ ^– accumulated in the leaves of plants grown at 450 mol photons·m^–2·s^–1 than in those grown at 170 mol photons·m^–2·s^–1 in N-replete conditions. The foliar NO ₃ ^– level and maximal NR activity decreased with the imposition of N-deficiency in all plant types such that after prolonged exposure to nitrogen depletion very little NO ₃ ^– was found in the leaves and NR activity had decreased to almost zero. The activity of NR decreased under conditions of nitrogen deficiency. This regulation is multifactoral since there is no regulation of NR gene expression by NO ₃ ^– in the constitutive NR-expressors. We conclude that the NR protein is specifically targetted for destruction under nitrogen deficiency. Consequently, constitutive expression of NR activity does not benefit the plant in terms of increased biomass production in conditions of limiting nitrogen.Abbreviations Chl chlorophyll - N nitrogen - NR NADH-nitrate reductase - WT wild type     

Growth characteristics and ion contents of non-selected and salt-selected callus lines of highbush blueberry (Vacdnium corymbosum) cultivars Blue Crop and Denise Blue

Non-selected and sodium chloride selected callus lines of Vacdnium corymbosum L.cv Blue Crop and cv. Denise Blue were grown on media supplemented with 0–100 mM NaCl. For both cultivars, fresh weight and dry weight yields were greater in selected lines on all levels of NaCl. Selected lines of Blue Crop displayed better growth than selected lines of Denise Blue at most concentrations of NaCl. Internal Na⁺ and Cl^– concentrations in selected and non-selected lines of both cultivars increased as external concentration was raised. However, selected lines of Blue Crop and Denise Blue accumulated more Na⁺ and Cl^– than non-selected lines. Selected lines of both cultivars maintained higher levels of K⁺ than non-selected lines on all external NaCl levels. Selected lines of Blue Crop had higher levels of Na⁺ and Cl^– than that of Denise Blue. The results suggest Na⁺ and Cl^– accumulation could be a mechanism allowing better growth in selected lines at moderate salinity levels (50–75 mM NaCl).     

Tolerance ofHolcus lanatus andAgrostis stolonifera to sodium chloride in soil solution and saline spray

Summary Inland and sea cliff populations of bothAgrostis stolonifera L. andHolcus lanatus L. were subjected to soil NaCl treatments, of 100 and 200 mol m⁻³ NaCl, and tolerance examined using plant dry weight data. A parallel experiment subjected them to salt spray treatments of 2.5%, 5% and 10% NaCl in distilled water, and tolerance assessed from leaf damage. Both populations of each species were equally sensitive to soil NaCl. When subjected to salt spray the sea cliff populations however showed marked resistance to leaf damage. Soil salinity resistance and salt spray resistance thus appear to be independent characteristics in these two species.     

Leaf Structure of Tobacco In Vitro Grown Plantlets as Affected by Saccharose and Irradiance

Tobacco plantlets were cultured in vitro under high (200 µmol m^–2 s^–1) or low (60 µmol m^–2 s^–1) irradiance with or without saccharose in the medium. Light microscopy and image analysis were used to evaluate the effect of these culture conditions on leaf anatomy. Addition of saccharose resulted in thicker leaves (all leaf layers) and larger mesophyll cells under both growth irradiances. Various irradiance affected leaf anatomy differently when plantlets had been cultivated in presence or absence of saccharose in the medium. While under high irradiance in presence of saccharose leaf thickness and number of chloroplasts per cell section were increased, plantlets grown under high irradiance in absence of saccharose had thinner leaves and less chloroplasts per cell section. The changes were more pronounced in palisade parenchyma layer.