Quantifying succulence: a rapid, physiologically meaningful metric of plant water storage

Quantification of succulence should ideally convey information about physiological function and yet also be straightforward to measure. While important aspects of succulence and its physiological consequences may be quantified using parameters derived from pressure-volume (P-V) curves, this technique applied to succulent tissues is difficult, time consuming and generally not suitable for large comparative datasets. We performed P-V curves on leaves of 25 taxa from across Caryophyllales and compared the results with direct measures of saturated water content (SWC(meas) ), the ratio of water mass at full saturation to tissue dry mass, for the same taxa. SWC(meas) was significantly related to relative capacitance, the most physiologically relevant parameter describing tissue succulence. We developed a linear model describing SWC(meas) as a function of relative capacitance and leaf volume, which is also supported when accounting for the phylogenetic relationships among taxa. These results indicate that SWC(meas) is a suitable proxy for tissue succulence, and that both cellular properties and variation in gross morphology contribute towards a plant's relative water storage capacity. Quantifying SWC(meas) across many taxa showing variation in tissue succulence will provide a new avenue for exploring the evolutionary dynamics of this important ecological adaptation.     

Relationship between specific leaf area,leaf thickness,leaf water content and <Emphasis Type="Italic">SPAD-502</Emphasis> readings in six Amazonian tree species

The aim of this work was to assess the effect of leaf thickness, leaf succulence (L_S), specific leaf area (SLA), specific leaf mass (W_s) and leaf water content (LWC) on chlorophyll (Chl) meter values in six Amazonian tree species (Carapa guianensis, Ceiba pentandra, Cynometra spruceana, Pithecolobium inaequale, Scleronema micranthum and Swietenia macrophylla). We also tested the accuracy of a general calibration equation to convert Minolta Chl meter (SPAD-502) readings into absolute Chl content. On average, SPAD values (x) increased with fresh leaf thickness (FLT [μm] = 153.9 + 0.98 x, r ² = 0.06**), dry leaf thickness (DLT [μm] = 49.50 + 1.28 x, r ² = 0.16**), specific leaf mass (W_s [g (DM) m⁻²] = 6.73 + 1.31 x, r ² = 0.43**), and leaf succulence (L_S [g(FM)] m⁻² = 94.2 + 1.58 x, r ² = 0.19**). However, a negative relationship was found between SPAD values and either specific leaf area [SLA (m² kg⁻¹) = 35.1 − 0.37 x, r ² = 0.38**] or the leaf water content (LWC [%]= 80.0 − 0.42 x, r ² = 0.58**). Leaf Chl contents predicted by the general calibration equation significantly differed (p<0.01) from those estimated by species-specific calibration equations. We conclude that to improve the accuracy of the SPAD-502 leaf thickness and LWC should be taken into account when calibration equations are to be obtained to convert SPAD values into absolute Chl content.     

Sporogenic and vegetative tissues of Saccharina latissima (Laminariales,Phaeophyceae) exhibit distinctive sensitivity to experimentally enhanced ultraviolet radiation : photosynthetically active radiation ratio

Fertile Saccharina latissima sporophytes, collected in the Kongsfjorden, Ny‐Ålesund, Spitsbergen, Norway (78°56.87′ N, 11°51.64′ E) were investigated in relation to its sensitivity to experimentally enhanced ultraviolet radiation : photosynthetically active radiation (UVR : PAR) ratios. Irradiance of UVR were 4.30 W m^?2 of UV‐A (320–400 nm) and 0.40 W m^?2 of UV‐B (280–320 nm), and PAR (400–700 nm) was ～4.30 W m^?2 (=20 µmol photons m^?2 s^?1). Excised soral (sporogenic) and non‐soral (vegetative) tissues were separately irradiated for 16 h at 7°C. Transmission electron microscopy showed abundant occurrence of physodes, electron dense particles (～300–600 nm) in the sorus. Paraphysis cells, with partly crystalline content, large mitochondria and abundant golgi bodies were towering over the sporangia. In soral tissue, cells were not visibly altered by the PAR + UVR irradiation. The chloroplasts, flagella and nucleus of unreleased meiospores inside the sporangial parent cells were visibly intact. Severe changes in the chloroplast structure of vegetative tissue occurred after PAR + UVR irradiation. These changes included wrinkling and dilatation of the thylakoid membranes, and appearance of electron translucent areas inside the chloroplasts. In vegetative cells exposed to PAR + UVR, the total amount of physodes, was slightly higher as in cells exposed to PAR only. Initial values of optimum quantum yield of photosystem II (F_v/F_m) were 0.743 ± 0.04 in non‐soral and 0.633 ± 0.04 in soral tissue. Vegetative tissue was observed to be more sensitive to radiant exposure of PAR and PAR + UVR compared to reproductive tissue. Under PAR, a 20% reduction in Fv/Fm was observed in non‐soral compared to no reduction in soral tissue, whereas under PAR + UVR, 60% and 33% reduction in Fv/Fm was observed in non‐soral and soral tissues, respectively. This can be attributed to the corresponding three times higher antiradical power (ARP) capacity in soral compared to non‐soral tissue.     

Enzyme activities in an artificial stroma medium

When spinach leaf tissue was subjected to evaporative dehydration, photosynthetic capacity at very high (5%) CO₂ concentration and saturating irradiance (300 W·m^-2), decreased in parallel to the relative water content (RWC). A 50% inhibition was observed at 60–40% RWC. In order to examine whether the inhibition was caused by increased solute concentrations in chloroplasts or cytoplasm, an artificial stroma medium (ASM) was set up containing all major osmotically relevant solutes measured in isolated intact spinach chloroplasts. Subsequently, the response of enzyme activities to normal and to increased concentrations of ASM was examined. Inhibition of enzymes by a concerted increase of all solutes was well correlated to the in-vivo response of photosynthesis to dehydration (60% inhibition at double-strength ASM). Inhibitory solutes were mainly divalent inorganic anions, such as sulfate and phosphate. Inhibition of ribulose-1,5-bisphosphate carboxylase by these ions as studied in more detail. Inhibition of the enzyme by sulfate and phosphate was competitive with respect to ribulose-1,5-bisphosphate, but not with respect to CO₂. The K_I for sulfate was 2.1 mmol·l^-1 and for phosphate 0.57 mmol·l^-1. Sugars and amino acids at the concentrations found in spinach chloroplasts did not prevent inhibition of enzymes by anions. The results indicate that increased anion concentrations in cells and organelles are responsible for primary, quickly reversible effects of moderate dehydration on plant tissues.Abbreviations ASM artificial stroma medium - RuBP ribulose 1,5-bisphosphate - RuBPCase ribulose-1,5-bisphosphate-carboxylase/oxygenase - RWC relative water content     

Oxygen-evolution patterns from spinach Photosystem II preparations

Patterns of O₂ evolution resulting from sequences of short flashes are reported for Photosystem (PS) II preparations isolated from spinach and containing an active, O₂-evolving system. The results can be interpreted in terms of the S-state model developed to explain the process of photosynthetic water splitting in chloroplasts and algae. The PS II samples display damped, oscillating patterns of O₂ evolution with a period of four flashes. Unlike chloroplasts, the flash yields of the preparations decay with increasing flash number due to the limited plastoquinone acceptor pool on the reducing side of PS II. The optimal pH for O₂ evolution in this system (pH 5.5–6.5) is more acidic than in chloroplasts (pH 6.5–8.0). The O₂-evolution, inactivation half-time of dark-adapted preparations was 91 min (on the rate electrode) at room temperature. Dark-inactivation half-times of 14 h were observed if the samples were aged off the electrode at room temperature. Under our conditions (experimental conditions can influence flash-sequence results), deactivation of S₃ was first order with a half-time of 105 s while that of S₂ was biphasic. The half-times for the first-order rapid phase were 17 s (one preflash) and 23 s (two preflashes). The longer S₂ phase deactivated very slowly (the minimum half-time observed was 265 s). These results indicate that deactivation from S₃ → S₂ → S₁, thought to be the dominant pathway in chloroplasts, is not the case for PS II preparations. Finally, it was demonstrated that the ratio of S₁ to S₀ can be set by previously developed techniques, that S₀ is formed mostly from activated S₃ (S₄), and that both S₀ and S₁ are stable in the dark.     

REGULATION OF NaCl IN JAUMEA CARNOSA (ASTERACEAE), A SALT MARSH SPECIES,AND ITS EFFECT ON LEAF SUCCULENCE

A salt marsh species, Jaumea carnosa, was used in hydroponic experiments to test the effects of increasing NaCl concentrations on leaf succulence and plant accumulations of K, Ca, Mg, Na and Cl. A nested experimental design was used with four salinity levels. Plants were grown in full Hoagland's solution plus different amounts of NaCl (0.0–1.2 osmoles). Leaf succulence was measured as percent water content as well as vertical elongation of mesophyll cells. There were no corresponding increases in leaf succulence with increasing concentrations of NaCl in the root zone. Plants receiving aerosol spray (40 mg/dm²/day) did not show significant increases in leaf succulence. Leaf succulence was significantly increased when the plants were removed from the NaCl solutions and placed in non-salinized Hoagland's solution. Osmotic concentrations of cell sap in leaf tissues showed significant increases as NaCl concentrations increased in the root zone. The concentrations of K, Ca and Mg were higher in plants grown without NaCl than in those grown with NaCl. The accumulations of K in the root tissues were always higher than those of the shoot tissues. Although there was a two-fold difference in NaCl concentrations at the highest levels, the concentrations of Na in the shoot tissues were relatively similar. The results of the Cl analyses of shoot tissues showed a similar pattern of regulation of uptake. This regulation of salt uptake may be important in preventing injury by limiting accumulations of salt in plant tissues when growing in soils of high osmotic potentials.     

Electron transfer and proton pumping under excitation of dark-adapted chloroplasts with flashes of light

Proton release inside thylakoids, which is linked to the action of the water-oxidizing enzyme system, was investigated spectrophotometrically with the dye neutral red under conditions when the external phase was buffered. Under excitation of dark-adapted chloroplasts with four short laser flashes in series, the pattern of proton release as a function of the flash number was recorded and interpreted in the light of the generally accepted scheme for consecutive transitions of the water-oxidizing enzyme system: S₀ → S₁, S₁ → S₂, S₂ → S₃, S₃ → S₄, S₀. It was found that the proton yield after the first flash varied in a reproducible manner, being dependent upon the dark pretreatment given. In terms of the proton-electron reaction during these transitions, the pattern was as follows. In strictly dark-adapted chloroplasts (frozen chloroplasts thawed in darkness and kept for at least 7 min in the dark after dilution), it was fitted well by a stoichiometry of 1:0:1:2. In a less stringently dark-adapted preparation (as above but thawed under light), it was fitted by 0:1:1:2. Mechanistically this is not yet understood. However, it is a first step towards resolving controversy over this pattern among different laboratories. Under conditions where the 1:0:1:2 stoichiometry was observed, proton release was time resolved. Components with half-rise times of 500 and 1000 μs could be correlated with the S₂ → S₃ and S₃ → S₄ transitions, respectively. Proton release during the S₀ → S₁ transition is more rapid, but is less well attributable to the transitions due to error proliferation. A distinct component with a half-rise time of only 100 μs was observed after the second flash. Since it did not fit into the expected kinetics (based on literature data) for the S_i → S_i+1 transitions, we propose that it reflects proton release from a site which is closer to the reaction center of Photosystem (PS) II than the water-splitting enzyme system. This is supported by the observation of rapid proton release under conditions where water oxidation is blocked. Related experiments on the pattern of proton uptake at the reducing side of PS II indicated that protons act as specific counterions for semiquinone anions without binding to them.     

S-Adenosylmethionine metabolism in HL-60 cells: effect of cell cycle and differentiation

The effect of the cell cycle and differentiation on S-adenosylmethionine (SAM) metabolism in HL-60 cells has been investigated. Synthesis and pool sizes of SAM and S-adenosylhomocysteine (SAH) were cell-cycle-independent (SAM, 315, μM; SAH, 4.6 μM). The SAM-synthase (ATP: l-methionine S-adenosyltransferase) of HL-60 cells has a K_m for methionine of 12.8±2.0 μM and thus appears to be of the intermediate K_m type found in other malignant tissues. The enzyme does not show cell-cycle regulation. Treatment of cells with DMSO resulted in a rapid and marked decrease of SAM and SAH levels without affecting pool turnover or the SAM/SAH ratio. A decrease in SAM concentration could also be observed in a variant cell line resistant to differentiation with DMSO. DMSO inhibited SAM-synthase in cell-free extracts. This inhibition was noncompetitive with respect to l-methionine. Inhibition of SAM-synthase by cycloleucine lowered SAM levels in intact cells, but resulted in differentiation of only a minor percentage of cells. These data indicate that changes in SAM and SAH levels in HL-60 cells seem to be a consequence rather than a cause of differentiation.     

Changes in morphological and anatomical structure of cabbage (Brassica oleracea L.) outer leaves and in ultrastructure of their chloroplasts caused by an in vitro excess of nickel

Morphological, anatomical and ultrastructural changes, the quantity and quality of stomata, and the chlorophyll (Chl) content in primary outer leaves of cabbage plants cv. S?awa from Enkhouizen were examined. The plants were grown in agar with basic MS medium containing added nickel (as NiSO₄×7 H₂O) in concentrations of 0, 5, 10, and 20 g m^-3 (Ni₀, Ni₅, Ni₁₀, Ni₂₀). Reduction of leaf blade area, of succulence and of leaf density, and growth of specific leaf area were noticed in plants treated with all concentrations of Ni. In Ni-treated plants the total number of stomata and open stomata decreased, and the number of defective stomata in both adaxial and abaxial side of leaves was higher. In all Ni-treated samples the volume of spongy and palisade mesophyll cells was smaller in comparison to control, and it was decreasing when the Ni concentration was increasing; at the same time, the number of mesophyll cells on the same area of cross sections of leaves was increasing. In comparison to control, the intercellular spaces of mesophyll tissue decreased in Ni₁₀ and Ni₂₀ plants and increased in Ni₅ plants. In Ni5 plants the number of chloroplasts in mesophyll cells was higher than in the Ni₀ control. Reduction of grana size and increase of number of non-appressed lamellae, which often had central arrangement, were observed. In the Ni₁₀ and Ni₂₀ plants, the number and size of chloroplasts decreased, and their internal membranes (especially grana) were reduced and swollen. In Ni₅ plants the concentration of Chl in leaves was slightly higher than in the control; in Ni₁₀ and Ni₂₀ plants it was lower than in the control.

     

Changes in morphological and anatomical structure of cabbage (Brassica oleracea L.) outer leaves and in ultrastructure of their chloroplasts caused by an in vitro excess of nickel

Morphological, anatomical and ultrastructural changes, the quantity and quality of stomata, and the chlorophyll (Chl) content in primary outer leaves of cabbage plants cv. Sława from Enkhouizen were examined. The plants were grown in agar with basic MS medium containing added nickel (as NiSO₄×7 H₂O) in concentrations of 0, 5, 10, and 20 g m^-3 (Ni₀, Ni₅, Ni₁₀, Ni₂₀). Reduction of leaf blade area, of succulence and of leaf density, and growth of specific leaf area were noticed in plants treated with all concentrations of Ni. In Ni-treated plants the total number of stomata and open stomata decreased, and the number of defective stomata in both adaxial and abaxial side of leaves was higher. In all Ni-treated samples the volume of spongy and palisade mesophyll cells was smaller in comparison to control, and it was decreasing when the Ni concentration was increasing; at the same time, the number of mesophyll cells on the same area of cross sections of leaves was increasing. In comparison to control, the intercellular spaces of mesophyll tissue decreased in Ni₁₀ and Ni₂₀ plants and increased in Ni₅ plants. In Ni5 plants the number of chloroplasts in mesophyll cells was higher than in the Ni₀ control. Reduction of grana size and increase of number of non-appressed lamellae, which often had central arrangement, were observed. In the Ni₁₀ and Ni₂₀ plants, the number and size of chloroplasts decreased, and their internal membranes (especially grana) were reduced and swollen. In Ni₅ plants the concentration of Chl in leaves was slightly higher than in the control; in Ni₁₀ and Ni₂₀ plants it was lower than in the control. This revised version was published online in June 2006 with corrections to the Cover Date.     

Anatomical variation of mesophyll conductance under potassium deficiency has a vital role in determining leaf photosynthesis

Leaves exposed to potassium (K) deficiency usually present decreased mesophyll conductance (g_m) and photosynthesis (A). The relative contributions of leaf anatomical traits in determining g_m have been quantified; however, anatomical variabilities related to low g_m under K starvation remain imperfectly known. A one‐dimensional model was used to quantify anatomical controls of the entire CO₂ diffusion pathway resistance within a leaf on two Brassica napus L. cultivars in response to K deficiency. Leaf photosynthesis of both cultivars was significantly decreased under K deficiency in parallel with down‐regulated g_m. The mesophyll conductance limitation contributed to more than one‐half of A decline. The decreased internal air space in K‐starved leaves was associated with the increase of gas‐phase resistance. Potassium deficiency reduced liquid‐phase conductance by decreasing the exposed surface area of chloroplasts per unit leaf area (S_c/S), and enlarging the resistance of the cytoplasm that can be interpreted by the increasing distance of chloroplast from cell wall, and between adjacent chloroplasts. Additionally, the discrepancies of A between two cultivars were in part because of g_m variations, ascribing to an altered S_c/S. These results emphasize the important role of K on the regulation of g_m by enhancing S_c/S and reducing cytoplasm resistance.     

CHLOROPLAST STRUCTURE AND FUNCTION IN CULTURED TOBACCO TISSUE

A comparative study was made of the ability of cultured pith tissue, leaves of buds induced from callus, and mature leaf tissue of Nicotiana tabacum L. ‘Maryland Mammoth’ to fix carbon, as determined by light-induced C¹⁴O₂ incorporation. Photosynthetic ability was then correlated with the fine structure of chloroplasts from these tissues. The light to dark incorporation ratio for C¹⁴O₂ was at least 3 times as great in the leaf tissue as in growing cultured tissue. The chlorophyll content of the leaf tissue was 10 times as great. The carbon fixation pattern of all the tissues, as determined by radioautographs of chromatogramed extracts, was qualitatively the same. The rate of sucrose synthesis differed greatly, since 20% of the total radioactivity of the extracts from mature leaf tissue appeared in sucrose, while only 1.0% was found in sucrose from callus extracts. The incorporation of C¹⁴O₂ into sugars was inhibited in all the tissue by DCMU (3,4-dichlorophenyl,1, 1-dimethylurea). Cultured tissue past the log phase of growth was intermediate between the younger cultured tissue and the leaf tissue in its chlorophyll content and ability to incorporate C¹⁴O₂ in the light. Proplastids from dark-grown callus possessed stroma lamellae, but prolamellar bodies were not observed. The chloroplasts from growing callus were partially differentiated in comparison with chloroplasts from mature leaf tissue, since each granum had only 4-7 lamellae. Chloroplasts from callus past the log phase of growth were indistinguishable from those in mature leaves. This study establishes that the differentiation of chloroplasts in cultured tissue is a function of the growth rate of the tissue. The growth rate and degree of differentiation of the tissue can be regulated, so a well-defined system is available for the experimental study of chloroplast differentiation.     

Leaf anatomy and photosynthetic acclimation in <Emphasis Type="Italic">Valeriana jatamansi</Emphasis> L. grown under high and low irradiance

Relationship of leaf anatomy with photosynthetic acclimation of Valeriana jatamansi was studied under full irradiance [FI, 1 600 mol(PPFD) m^–2 s^–1] and net-shade [NS, 650 mol(PPFD) m^–2 s^–1]. FI plants had thicker leaves with higher respiration rate (R _D), nitrogen content per unit leaf area, chlorophyll a/b ratio, high leaf mass per leaf area unit (LMA), and surface area of mesophyll cell (S _mes) and chloroplasts (S _c) facing intercellular space than NS plants. The difference between leaf thickness of FI and NS leaves was about 28 % but difference in photon-saturated rate of photosynthesis per unit leaf area (P _Nmax) was 50 %. This indicates that P _Nmax can increase to a larger extent than the leaf thickness with increasing irradiance in V. jatamansi. Anatomical studies showed that the mesophyll cells of FI plants had no open spaces along the mesophyll cell walls (higher S _c), but in NS plants wide open spaces along the mesophyll cell wall (lower S _c) were found. Positive correlation between S _c and P _Nmax explained the higher P _Nmax in FI plants. Increase in mesophyll thickness increased the availability of space along the mesophyll cell wall for chloroplasts (increased S _c) and hence P _Nmax was higher in FI plants. Thus this Himalayan species can acclimate to full sunlight by altering leaf anatomy and therefore may be cultivated in open fields.     

Leaf responses to drought stress in Mediterranean accessions of Solanum lycopersicum: anatomical adaptations in relation to gas exchange parameters

In a previous study, important acclimation to water stress was observed in the Ramellet tomato cultivar (TR) from the Balearic Islands, related to an increase in the water‐use efficiency through modifications in both stomatal (g_s) and mesophyll conductances (g_m). In the present work, the comparison of physiological and morphological traits between TR accessions grown with and without water stress confirmed that variability in the photosynthetic capacity was mostly explained by differences in the diffusion of CO₂ through stomata and leaf mesophyll. Maximization of g_m under both treatments was mainly achieved through adjustments in the mesophyll thickness and porosity and the surface area of chloroplasts exposed to intercellular airspace (S_c). In addition, the lower g_m/S_c ratio for a given porosity in drought‐acclimated plants suggests that the decrease in g_m was due to an increased cell wall thickness. Stomatal conductance was also affected by drought‐associated changes in the morphological properties of stomata, in an accession and treatment‐dependent manner. The results confirm the presence of advantageous physiological traits in the response to drought stress in Mediterranean accessions of tomato, and relate them to particular changes in the leaf anatomical properties, suggesting specific adaptive processes operating at the leaf anatomical level.     

Preliminary Studies on the Photosynthetic Structures of Trifolium alpinum L. as Related to Productivity

Trifolium alpinum L. is a high-quality alpine forage plant growingspontaneously from 1900 to 2800 m above sea level and is widelydistributed in Piedmont and the Valle d'Aosta (Italy), whereit can reach population frequencies of 90 per cent. Yields weredetermined on forage harvested in the Valle dell'Orco (Piedmont)and were comparable to cultivated clovers from higher latitudes;yields decreased progressively as the elevation increased. Thechemical and nutritional characteristics of the forage, thoughcomparable to clovers cultivated in the Po valley (Italy), were,however, more constant. The structure of the leaf lamina asrelated to elevation was investigated using light microscopy,TEM and SEM. This is complemented by data on chlorophyll concentration,succulence, specific leaf weight and area. At all elevationsT. alpinum lacks, apart from bundle sheath cell chloroplastsin a centrifugal arrangement, the structural characteristicsof C₄ plants. The chlorophyll a:b ratio (less than four) istypical of a C₂ plant. Succulence indices (S and S_m) were verylow, making CAM pathway photosynthesis unlikely. Unusual anddifficult to interpret structures included: small functionalchloroplasts in both the epidermises, stomata present almostexclusively in the upper epidermis and mitochondria enveloped(or enclosed) by chloroplasts. It was observed that, as theelevation increases, populations are selected which are well-adaptedfor gas exchange (increase in specific leaf area, stomatal densityand intercellular spaces) and characterized by a decrease inthe grana thylacoid:integrana thylacoid ratio (consistent withthe increase in the chlorophyll a:b ratio), the per cent water,S_m and the specific leaf weight. Trifolium alpinum L., alpine trefoil, leaf structures, photosynthesis, yield, elevation, C₂, C₄     

Hydraulic acclimation in a Mediterranean oak subjected to permanent throughfall exclusion results in increased stem hydraulic capacitance

Stem water storage capacity and hydraulic capacitance (C_S) play a crucial role in tree survival under drought-stress. To investigate whether C_S adjusts to increasing water deficit, variation in stem water content (StWC) was monitored in vivo for 2 years and related to periodical measurements of tree water potential in Mediterranean Quercus ilex trees subjected either to permanent throughfall exclusion (TE) or to control conditions. Seasonal reductions in StWC were larger in TE trees relative to control ones, resulting in greater seasonal C_S (154 and 80 kg m⁻³ MPa⁻¹, respectively), but only during the first phase of the desorption curve, when predawn water potential was above −1.1 MPa. Below this point, C_S decreased substantially and did not differ between treatments (<20 kg m⁻³ MPa⁻¹). The allometric relationship between tree diameter and sapwood area, measured via electrical resistivity tomography, was not affected by TE. Our results suggest that (a) C_S response to water deficit in the drought-tolerant Q. ilex might be more important to optimize carbon gain during well-hydrated periods than to prevent drought-induced embolism formation during severe drought stress, and (b) enhanced C_S during early summer does not result from proportional increases in sapwood volume, but mostly from increased elastic water.     

Effect of water stress on functioning and structure ofCicer arietinum L. nodules

Chickpea (Cicer arietinum L. cv. 235) plants were grown in sand culture at moisture equal to 45–50% of sand saturation capacity under greenhouse conditions. 60 d after sowing, pots were divided into four lots, leaving one as control and sand moisture content of others was brought to 25–30% (S₁), 12–15% (S₂) and 5–6% (S₃) of sand saturation capacity, by withholding the water supply and then maintaining the required levels gravimetrically till the harvest. Relative water content of leaves and nodule water content were measured as indices of water stress. With increase in the severity and duration of water stress nitrogenase activity and nitrogen and leghemoglobin content of the nodules decreased and the ratio of leghemoglobin components I and II were changed. Nodules developed under limited water availability showed decreased branching, breakdown of the endodermis, greater compactness and decreased vacuolation of cells in the central symbiotic tissue as compared to the control.     

Structural and functional traits of Quercus ilex in response to water availability

Water potential and morpho-anatomical parameters were measured, during the course of 1 year, on leaves of Quercus ilex trees growing in two coastal stands in Tuscany (Central Italy) with different conditions of water availability: Colognole (CL, mesic site) and Cala Violina (CV, xeric site). Morpho-anatomical measurements included: general leaf features and sclerophylly indices (surface area, thickness, mass per area and density), leaf moisture indices (water content, relative water content, succulence) and histochemical analysis (detection and localization of cutine and tannins in the leaves and starch reserves in the twigs). During the warmest and driest period (August) pre-dawn water potential (ψ_pd) in Holm-oak leaves reached −2.7 MPa at CV and −0.6 MPa at CL. Leaf surface was lower (−34%) and total leaf thickness (+10%), as well as spongy-palisade parenchyma ratio (+20%) were higher at CV. The sclerophylly parameters (leaf mass per area and leaf tissue density) were higher at CV than at CL (+24% leaf mass per area and +19% leaf tissue density). Among the moisture parameters, water content was higher at CL (+8%) and succulence was higher at CV (+13%). No differences in relative water content were observed between the two sites. All the parameters considered were substantially stable during the study period, with the exception of relative water content at CL, that fluctuated within the year. Histochemical analysis revealed a greater thickness of the upper cuticular layer at CV, whereas there were no differences in tannin distribution and content between the two sites. Differences in starch storage were detected in branchlets: it was abundant in CV but very scarce at CL. The strategies of Quercus ilex to cope with water stress were discussed at morpho-structural level.     

Differential efficiency of photosynthetic oxygen evolution in flashing light in triazine-resistant and triazine-susceptible biotypes of Senecio vulgaris L.

Photosynthetic oxygen evolution in response to flashing light was studied in triazine-susceptible and triazine-resistant biotypes of Senecio vulgaris L. Studies were conducted to determine if the modification of the herbicide-binding site which confers s-triazine resistance also affects the oxygen-evolving system. Oxygen evolution was measured using a Joliot-type oxygen-specific electrode on broken, stroma-free chloroplasts of both biotypes. We observed abnormal patterns of oxygen evolution in resistant chloroplasts. The S′₁ → S₂ transition is slower while the S₂ decay is faster. The S′₂ → S₃ transition, in contrast, is slightly faster in resistant chloroplasts, while the decay of the S₃ state is the same as in susceptible chloroplasts. These altered kinetics may be due to altered Q → B (B^?) electron flow in resistant chloroplasts. These results are also consistent with the hypothesis that back-reactions from the reducing (acceptor) side of Photosystem II to the oxidizing (donor) side occur with greater frequency in resistant than susceptible chloroplasts. These events are responsible for lower oxygen yield and increased ‘misses’ and ‘double hits,’ resulting in abnormal yield patterns and lower quantum yield of CO₂ fixation in resistant chloroplasts compared to the susceptible ones.