Effects of nitrogen dioxide and nitrate nutrition on growth and nitrate assimilation in bean leaves

The influence of nutrient nitrate level (0-20 millimolar) on the effects of NO₂ (0-0.5 parts per million) on growth, K, photosynthetic pigment, N contents, and the activities of enzymes of N assimilation was studied in bean (Phaseolus vulgaris L. cv Kinghorn Wax) leaves. Exposing 7-day old bean seedlings for 5 days continuously to 0.02 to 0.5 parts per million NO₂ increased plant height, fresh weight, chlorophyll, carotenoid, organic N and nitrate contents, and nitrate reductase and glutamate synthase activities in the leaves of seedlings supplied with no external N. At 20 millimolar nitrate, most of the parameters examined were inhibited except for organic N and nitrate contents and glutamate synthase activity which increased in most cases. Generally, with an increase in NO₂ concentration, the stimulatory effect declined and/or the inhibitory effect increased. A 3-hour exposure of 12-day-old bean seedlings to 0.1 to 2.0 parts per million NO₂ increased nitrate content and nitrate reductase activity at each nutrient nitrate level except for a slight inhibition of enzyme activity during exposure to 2.0 parts per million NO₂ at 20 millimolar nitrate. The experiments demonstrated that the effect of NO₂ is strongly influenced by nutrient N level and that NO₂ is assimilated into organic nitrogenous compounds to serve as a source of N, only to a limited extent.     

Sodium Stimulates Growth of Amaranthus tricolor L. Plants through Enhanced Nitrate Assimilation

Effects of Na application on the capacity of NO₃⁻ assimilation were studied in Na-deficient Amaranthus tricolor L. cv Tricolor plants. On day 30 after germination, Na-deficient A. tricolor plants received either 0.5 millimolar NaCl or KCl. The level of nitrate reductase activity doubled within 24 hours by the addition of Na and the enhanced level was maintained thereafter. When the plants were exposed to 2 millimolar ¹⁵NO₃⁻, total ¹⁵N taken up by the plants was greater in the Na-treated plants than in the K-treated plants within 24 hours of the Na treatment. Incorporation of ¹⁵N into the 80% ethanol-insoluble nitrogen fraction of the Na-treated plants in the light period was about 260% of those of the K-treated plants indicating greater capacity of NO₃⁻ assimilation in the Na-treated plants. From these results, it was demonstrated that Na application to the Na-deficient A. tricolor plants promoted NO₃⁻ reduction and its subsequent assimilation into protein, resulting in growth enhancement.     

Development of Photochemical Activity and the Appearance of the High Potential Form of Cytochrome b-559 in Greening Barley Seedlings

The development of photochemical activity during the greening of dark-grown barley seedlings (Hordeum vulgare L. cv. Svalöfs Bonus) was studied in relation to the formation of the high potential form of cytochrome b-559 (cytochrome b-559_HP). Photosynthetic oxygen evolution from leaves was detected at 30 minutes of illumination. The rate of oxygen evolution per gram fresh weight of leaf was as high at 2 to 2.5 hours of greening as at 24 hours or in fully greened leaves. On a chlorophyll basis, the photosynthetic rate at 90 minutes of greening was 80-fold greater than the rate at 45 hours. It is concluded that the majority of photosynthetic units are functional at an early stage of greening, and that chlorophyll synthesis during greening serves to increase the size of the units.     

Effects of light quality on the chloroplastic ultrastructure and photosynthetic characteristics of cucumber seedlings

To understand how light quality influences plant photosynthesis, we investigated chloroplastic ultrastructure, chlorophyll fluorescence and photosynthetic parameters, Rubisco and chlorophyll content and photosynthesis-related genes expression in cucumber seedlings exposed to different light qualities: white, red, blue, yellow and green lights with the same photosynthetic photon flux density of 100 μmol m^?2 s^?1. The results revealed that plant growth, CO₂ assimilation rate and chlorophyll content were significantly reduced in the seedlings grown under red, blue, yellow and green lights as compared with those grown under white light, but each monochromatic light played its special role in regulating plant morphogenesis and photosynthesis. Seedling leaves were thickened and slightly curled; Rubisco biosynthesis, expression of the rca, rbcS and rbcL, the maximal photochemical efficiency of PSII (Fv/Fm) and quantum yield of PSII electron transport (Ф_PSII) were all increased in seedlings grown under blue light as compared with those grown under white light. Furthermore, the photosynthetic rate of seedlings grown under blue light was significantly increased, and leaf number and chlorophyll content of seedlings grown under red light were increased as compared with those exposed to other monochromatic lights. On the contrary, the seedlings grown under yellow and green lights were dwarf with the new leaves etiolated. Moreover, photosynthesis, Rubisco biosynthesis and relative gene expression were greatly decreased in seedlings grown under yellow and green light, but chloroplast structural features were less influenced. Interestingly, the Fv/Fm, Ф_PSII value and chlorophyll content of the seedlings grown under green light were much higher than those grown under yellow light.     

Effects of strontium on photosynthetic characteristics of oilseed rape seedlings

The threat of environmental strontium pollution led to an increased interest to elucidating the mechanisms of this metal toxicity in the organism. To investigate strontium effects on vital photosynthesis characteristics, three-leaf stage oilseed rape seedlings (Brassica napus L., cv. Mianyou No.15), raised in the hydroponic culture, were provided with a nutrient solution containing 0, 10, 20, and 40 mM SrCl₂. Strontium uptake and distribution in oilseed rape plants and its effect on various aspects of photosynthesis were investigated after 0, 7, 14, and 21 days of strontium treatment. Oilseed rape seedlings demonstrated a strong ability of strontium accumulation. Strontium absorbed by roots was primarily transferred to leaves and accumulated there. The leaf photosynthetic oxygen evolution rate, chlorophyll content, and Rubisco (EC 4.1.3.9) and phosphoenolpyruvate carboxylase (PEPCase; EC 4.1.1.31) activities declined progressively with increasing concentration of applied strontium and also with increasing the duration of exposure time. These results indicate that strontium accumulated in leaves damaged various processes of photosynthesis, such as energy absorption, energy transfer, and photosynthetic carbon assimilation.     

Arbuscular Mycorrhizal Fungi Alter Fractal Dimension Characteristics of Robinia pseudoacacia L. Seedlings Through Regulating Plant Growth,Leaf Water Status,Photosynthesis, and Nutrient Concentration Under Drought Stress

The influence of arbuscular mycorrhizal fungi (AMF), Funneliformis mosseae and Rhizophagus intraradices, on plant growth, leaf water status, chlorophyll concentration, photosynthesis, nutrient concentration, and fractal dimension (FD) characteristics of black locust (Robinia pseudoacacia L.) seedlings was studied in pot culture under well-watered, moderate drought stress, and severe drought stress treatments. Mycorrhizal seedlings had higher dry biomass, leaf relative water content (RWC), and water use efficiency (WUE) compared with non-mycorrhizal seedlings. Under all treatments, AMF colonization notably enhanced net photosynthetic rate, stomatal conductance, and transpiration rate, but decreased intercellular CO₂ concentration. Leaf chlorophyll a and total chlorophyll concentrations were higher in AM seedlings than those in non-AM seedlings although there was no significant difference between AMF species. AMF colonization improved leaf C, N, and P concentrations, but decreased C:N, C:P, and N:P ratios. Mycorrhizal seedlings had a larger FD value than non-mycorrhizal seedlings. The FD value was positively and significantly correlated to the plant growth parameters, photosynthesis, RWC, WUE, and nutrient concentration but negatively correlated to leaf/stem ratio, C:N and C:P ratios, and intercellular CO₂ concentration. We conclude that AMF lead to an improvement of growth performance of black locust seedlings under all growth conditions, including drought stress via improving leaf water status, chlorophyll concentration, photosynthesis, and nutrient uptake. Moreover, FD technology proved to be a powerful non-destructive method to characterize the effect of AMF on the physiology of host plants during drought stress.     

Effects of nitrogen dioxide and nitrate nutrition on nodulation, nitrogenase activity, growth, and nitrogen content of bean

The influence of nutrient nitrate level (0-20 millimolar) on the effects of NO₂ (0-0.5 parts per million) on nodulation and in vivo acetylene reduction activity of the roots and on growth and nitrate and Kjeldahl N concentration in shoots was studied in bean (Phaseolus vulgaris L. cv Kinghorn Wax) plants. Exposing 8-day old seedlings for 6 hours each day, for 15 days, to 0.02 to 0.5 parts per million NO₂ decreased total nodule weight at 0 and 1 millimolar nitrate, and nitrogenase (acetylene reduction) activity at all concentrations of nitrate. The pollutant had little effect on root fresh or dry weights. Shoot growth was inhibited by NO₂. The NO₂ exposure increased nitrate concentration in roots only at 20 millimolar nutrient nitrate. Exposure to NO₂ markedly increased Kjeldahl N concentration in roots but generally decreased that in shoots. The experiments demonstrated that nutrient N level and NO₂ concentration act jointly in affecting nodulation and N fixing capability, plant growth and composition, and root/shoot relationships of bean plants.     

Essentiality of Boron for Dinitrogen Fixation in Anabaena sp. PCC 7119

The relationship between the requirement for boron and the form of N supplied in nutrient media to cyanobacterium Anabaena sp. PCC 7119 was investigated. When cells were grown in a medium which contained nitrate or ammonium-N, boron deficiency in the nutrient media did not inhibit growth or change cell composition. However, when cells were dependent on N₂ fixation, the lack of boron inhibited growth (i.e. growth ceased after 96 hours under these conditions). Additionally, boron-deficient cells showed a significant decrease in their content of phycobiliproteins and chlorophyll and accumulated carbohydrates within 24 hours of removing boron from the nutrient media. Inhibition of photosynthetic O₂ evolution accompanied the decrease in photosynthetic pigments. Boron deficiency symptoms were relieved when either boron or combined N was added to boron-deficient cultures. The degree of recovery depended upon the age of the cultures. Assays of nitrogenase activity showed that, after 2 hours of growth, nitrogenase activity of boron-deficient cells was inhibited by 40%. After 24 hours a total inactivation of nitrogenase activity was observed in boron-deficient cells. These results strongly suggest an involvement of boron in N₂ fixation in cyanobacteria.     

Influence of Age and Light on the Distribution and Development of Nitrate Reductase in Greening Barley Leaves

The relation between leaf age and the induction of nitrate reductase activity by continuous and intermittent light was studied with barley seedlings (Hordeum vulgare L. cv. Club Mariout). In general, nitrate reductase activity declined as the period of growth in darkness was extended beyond 5 days. Maximum activity was found near the leaf tip while activity was lowest in the morphologically youngest tissue near the base of the lamina. Increased activity was observed after continuous illumination of dark-grown seedlings for 24 hours. The increase in activity in response to light was greatly reduced when the dark pretreatment period was extended beyond 8 days. The amount of nitrate reductase activity present in the different sections of the leaf was closely related to the amount of polyribosomes present. The pattern of chlorophyll accumulation closely parallelled that of increases in nitrate reductase activity. The initial lag in the induction of nitrate reductase activity was removed by a 10-minute light treatment 6 hours before placing dark-grown barley seedlings in light. The enzyme was also induced under flashing light with various dark intervals. These induction curves closely resembled those of chlorophyll accumulation under the same conditions. The development of photosynthetic CO₂ fixation follows the same induction pattern in this system. Our results suggest that photosynthetic products may be required for the induction of significant levels of nitrate reductase activity in leaves of dark-grown seedlings, although other light effects may not be discounted.     

Effects of ethephon on aging and photosynthetic activity in isolated chloroplasts

Chloroplasts, isolated from the primary leaves of 7-day-old seedlings, were incubated in vitro at 25°C with 2-chloroethylphosphonic acid (ethephon) under light (0.16 milliwatts per square centimeter) and dark conditions. Ethephon at 1 micromolar (0.1445 ppm), 0.1 and 1 millimolar, or 5 microliters ethylene promoted the deterioration of chloroplasts, increased proteolysis, and reduced the chlorophyll content and PSI and PSII during 72 hours under both light and dark conditions. The decline in PSI and PSII occurred prior to a measurable loss of chlorophyll. The loss of photosynthetic activity affected by ethephon was initiated prior to 12 hours of incubation. After 24 hours in light, 0.1 millimolar (1.445 ppm) epthephon significantly reduced PSI and PSII and promoted the total free amino acid liberation in isolated chloroplasts. In darkness the rate of loss of PSI activity was about 50% of that in light. After 24 hours, in light at 1 millimolar epthephon, PSII activity was 55% of the control, yet nearly 90% of the chlorophyll remained, which indicates that the loss of thylakoid integrity was promoted by ethephon. Ethylene injected in the chloroplast medium at 5 microliters (0.22 micromolar per milliliter) reduced PSI by nearly 50% of the initial in 12 hours. In leaf sections floated in 5 microliters per milliliter suspension medium, a 36% loss of chlorophyll of the control in 36 hours was observed. Cycloheximide at 0.5 millimolar masked the effect of 1 millimolar ethephon and maintained the initial chlorophyll content during the 72 hour period.     

Photosynthesis and ion content of leaves and isolated chloroplasts of salt-stressed spinach

Spinach (Spinacia oleracea) plants were subjected to salt stress by adding NaCl to the nutrient solution in increments of 25 millimolar per day to a final concentration of 200 millimolar. Plants were harvested 3 weeks after starting NaCl treatment. Fresh and dry weight of both shoots and roots was decreased more than 50% compared to control plants but the salt-stressed plants appeared healthy and were still actively growing. The salt-stressed plants had much thicker leaves. The salt-treated plants osmotically adjusted to maintain leaf turgor. Leaf K⁺ was decreased but Na⁺ and Cl⁻ were greatly increased.

The potential photosynthetic capacity of the leaves was measured at saturating CO₂ to overcome any stomatal limitation. Photosynthesis of salt-stressed plants varied only by about 10% from the controls when expressed on a leaf area or chlorophyll basis. The yield of variable chlorophyll a fluorescence from leaves was not affected by salt stress. Stomatal conductance decreased 70% in response to salt treatment.

Uncoupled rates of electron transport by isolated intact chloroplasts and by thylakoids were only 10 to 20% below those for control plants. CO₂-dependent O₂ evolution was decreased by 20% in chloroplasts isolated from salt-stressed plants. The concentration of K⁺ in the chloroplast decreased by 50% in the salt-stressed plants, Na⁺ increased by 70%, and Cl⁻ increased by less than 20% despite large increases in leaf Na⁺ and Cl⁻.

It is concluded that, for spinach, salt stress does not result in any major decrease in the photosynthetic potential of the leaf. Actual photosynthesis by the plant may be reduced by other factors such as decreased stomatal conductance and decreased leaf area. Effective compartmentation of ions within the cell may prevent the accumulation of inhibitory levels of Na⁺ and Cl⁻ in the chloroplast.

     

Photosynthetic and stomatal responses of spinach leaves to salt stress

The gas exchange of spinach plants, salt-stressed by adding NaCl to the nutrient solution in increments of 25 millimolar per day to a final concentration of 200 millimolar, was studied 3 weeks after starting NaCl treatment. Photosynthesis became light saturated at 1100 to 1400 micromoles per square meter per second in salt-treated plants and at approximately 2000 micromoles per square meter per second in control plants. Photosynthetic capacity of the mesophyll measured as a function of intercellular partial pressure of CO₂ at the light intensity prevailing during growth and at light saturation were both decreased in the salttreated plants. The CO₂ compensation points and relative enhancements of photosynthesis at low O₂ were not affected by salinity. The lower photosynthetic rates in salt-treated leaves at 450 micromoles per square meter per second were associated with a 70% reduction in stomatal conductance and low intercellular CO₂ (219 microbars; cf. 285 microbars for controls). Increasing photon flux density to light saturation extended the linear portions of the CO₂ response curves, increased stomatal conductances, increased intercellular CO₂ in the salt-treated plants, but lowered it in controls, and accentuated differences in photosynthetic rate (area basis) between the treatments.

Leaves from salt-treated plants were thicker but contained about 73% of the chlorophyll per unit area of control plants. When photosynthetic rates were expressed on a chlorophyll basis there was no difference in initial slope of assimilation versus intercellular CO₂ between treatments. Photosynthetic rates (chlorophyll basis) at light saturation differed only by 20% which was also observed earlier with isolated, intact chloroplasts (Robinson et al. 1983 Plant Physiol 73: 238-242).

Measurement of carbon isotope ratio revealed less discrimination against ¹³C with salt treatment and confirmed the persistence of low intercellular partial pressures of CO₂ during plant growth. The development of a thicker leaf with less chlorophyll per unit area during salt treatment permitted stomatal conductance and intercellular partial pressure of CO₂ to decline without restricting photosynthesis and had the benefit of greatly increasing water use efficiency.

     

Isolation of mesophyll protoplasts from mature leaves of soybeans

A procedure based on a combined cellulase-Pectolyase Y-23 enzyme digestion and metrizamide-sorbitol gradient purification protocol was developed for isolating mesophyll protoplasts from mature leaves of soybean (Glycine max L. Merr.). Based on chlorophyll content, this procedure results in a 10 to 15% protoplast yield from fully expanded mature leaves and a 20 to 30% yield from young (expanding) leaves within 3 hours. Isolated protoplasts displayed high rates of HCO₃⁻-dependent photosynthesis; greater than 75 micromoles O₂ evolved per milligram chlorophyll per hour at 25°C. This photosynthetic rate is comparable to that of mesophyll cells isolated mechanically from the same leaves.     

Changes in photosynthesis caused by adaptation of maize seedlings to short-term drought

Detached leaf is in the state of increasing water deficit; it is a good experimental model for looking into the hardening effect of adaptation of eight-day-old maize (Zea mays L.) seedlings to short-term drought (five days without watering). The light stage of photosynthesis and photosynthetic CO₂/H₂O exchange in detached leaves were studied. Specific surface density of leaf tissue (SSDL), the content of chlorophylls a and b, proline, MDA as well as photosynthetic parameters: quantum yield of photosystem II fluorescence, assimilation of CO₂, and transpiration at room temperature and light saturation (density of PAR quantum flux of 2000 μmol/(m² s)) at normal and half atmospheric CO₂ concentration were determined. The leaves of seedlings exposed to short-term drought differed from control material by a greater SSDL and higher content of proline. The hardening effect of the stress agent on the dark stage of photosynthesis was detected; it was expressed in the maintenance of the higher photosynthetic CO₂ assimilation against control material due to the elevation of stomatal conductance for CO₂ diffusing into the leaf. Judging from the lack of differences in the MDA content, short-term drought did not injure photosynthetic membranes. In detached leaves of experimental maize seedlings, photosynthesis was maintained on a higher level than in control material.     

不同种源花榈木苗期生长及生理特性比较

为了解不同种源花榈木在贵阳的生长特性和差异,该文通过对10个种源地花榈木进行育苗试验,测定其两年生实生苗的苗高、地径、生物量、叶片光合参数、光合色素、硝酸还原酶活性、硝态氮含量和根系活力,并进行差异性分析。结果表明:(1)10个种源花榈木净光合速率、气孔导度、胞间CO₂浓度、蒸腾速率和水分利用效率差异显著(P<0.05),表明不同种源花榈木光合特性及光能利用效率具有较大差异,浙江杭州和浙江永康花榈木是具有较高光合生长潜力的种源。(2)种源间的叶绿素含量、硝酸还原酶、硝态氮、根系活力存在显著差异,福建建瓯种源的叶绿素a、叶绿素b含量和叶绿素总量最高,能够将光合原初反应过程中积蓄的光能进行高效地传递,促进碳的同化; 贵州花溪种源硝酸还原酶活性最大,硝态氮含量最高,对氮元素的利用能力较强,能够促进植物蛋白质、氨基酸和叶绿素等的合成; 贵州望谟种源根系活力最大,吸收养分的能力强。(3)各种源间苗高、地径和生物量的分配存在显著差异,浙江杭州种源的植株枝叶繁茂、根系发达,生长表现好,安徽黄山种源的植株矮小,生长表现较差; 浙江杭州种源将生物量更多分配在根和叶,提高其根系吸收养分和叶片获取光能的能力,安徽黄山种源总体生物量积累最少,长势最差。(4)通过主成分分析法对各种源的花榈木适应性进行综合评价,结果显示浙江杭州种源>贵州黎平种源>浙江永康种源>贵州望谟种源>福建建瓯种源>贵州凯里种源>贵州石阡种源>贵州花溪种源>贵州平塘种源>安徽黄山种源。综上结果表明,浙江杭州、贵州黎平和浙江永康种源花榈木对贵阳地区立地环境具有较强的适应能力和生长潜力。     

Effects of Nitrate Application on Amaranthus powellii Wats. : I. Changes in Photosynthesis, Growth Rates, and Leaf Area

Physiological effects of different nitrate applications were studied using the C₄ plant, Amaranthus powellii Wats. Plants were grown in a controlled environment chamber and watered daily with nutrient solutions containing 45, 10, 5, or 1 millimolar nitrate. Chloride and sulfate were used to keep the cation and phosphate concentrations constant. Total leaf nitrogen concentration, chlorophyll concentration, specific leaf mass, leaf area, relative growth rate, relative leaf growth rate, unit leaf rate (increase of dry mass per unit leaf area per day), net photosynthetic rate, and incident quantum yield decreased with decreasing nitrate concentration. The per cent decrease of unit leaf rate was similar to the decrease of light-saturated net photosynthetic rate; however, the decrease in relative growth rate was less than that of unit leaf rate because leaf area ratio (leaf area per unit dry mass) increased with decreasing nitrate concentration. Essential mineral concentrations per unit leaf area were about equal among all treatments. Leaf expansion, determined by stomatal density, decreased except for the 1 millimolar treatment which showed relatively more cell expansion but less cell division. Decreased nitrate application was correlated with higher osmotic potentials and lower pressure potentials (determined by pressure-volume curves), whereas leaf water potentials were equal among treatments. Even though total leaf area and shoot mass decreased with decreasing applied nitrate, the increase of the leaf area ratio may be related to selection for the highest possible growth rate.     

Photosynthetic acclimation to low temperature by western red cedar seedlings

Imposition of low, but above freezing, temperatures resulted in a gradual increase in the cold hardiness of western red cedar seedlings. This was associated with a decrease in the maximum rates of photosynthetic CO₂ fixation and O₂ evolution, and changes in chlorophyll a fluorescence transients which indicated that photoinhibition had occurred. Maximum photosynthetic rates declined approximately 40% during cold hardening. The leaves changed colour from green to red-brown during the hardening process. The colour change was due to the synthesis of large amounts of the carotenoid rhodoxanthin. Lutein levels doubled, while chlorophyll declined slightly. Dehardening resulted in the rapid recovery of photosynthesis to control levels, the rapid disappearance of rhodoxanthin, and the return of lutein levels to control. It is suggested that rhodoxanthin accumulation at low temperature functions to decrease the light intensity reaching the photosynthetic apparatus. The combination of photoinhibition and rhodoxanthin synthesis probably serves to protect the photosynthetic capacity of the seedlings at low temperature.     

外源K+和水杨酸在缓解融雪剂对油松幼苗生长抑制中的效应与机理

随着融雪剂在国内外寒冷地区的广泛应用及其在城市使用量的逐年增加,融雪剂对城市生态环境的危害引起了广泛的重视。其中,融雪剂在城市道路土壤中的积累对植物生长的影响已日益凸现。以油松幼苗为材料,通过分析0.2%浓度融雪剂胁迫下外源钾(K+)和水杨酸(salicylic acid,SA)对油松幼苗各生长生理指标的影响,探讨外源K+和SA在缓解融雪剂对油松幼苗生长抑制中的机理与剂量效应关系。结果表明,0.2%浓度的融雪剂处理对油松生长有明显的抑制作用,而20 mmol/L KNO3和2 mmol/L SA能明显诱导过氧化物酶(peroxidase,POD)活性的增强,缓解膜脂过氧化作用,降低丙二醛(malondialdehyde,MDA)在叶片中的积累,维持细胞膜的稳定性。虽然外源K+和SA对油松幼苗叶片胞间CO2浓度(intracellular CO2concentrations,Ci)和气孔导度(stomatal conductance,Gs)的缓解作用并不显著,但其可通过提高叶绿素含量促进光合作用的进行,缓解融雪剂胁迫对油松幼苗生长的抑制,分别增加生物量24.9%和63.6%。可见,20 mmol/L KNO3和2 mmol/L SA处理能有效缓解融雪剂对油松幼苗的伤害,为城市化学融雪剂的污染防治提供科学依据。     

Adaptations of the Photosynthetic Mechanism of Sugar Maple (Acer saccharum) Seedlings Grown in Various Light Intensities

Sugar maple (Acer saccharum Marsh.) seedlings were grown in a nursery for three years in 13, 25, 45 and 100 per cent of full daylight. During the third year of growth, the rates of their apparent photosynthesis and respiration were measured periodically with an infra-red gas analyzer at various light intensities and normal CO₂ concentration. In addition, the rates of apparent photosynthesis of a single attached leaf of the same seedlings were measured at saturating light intensity, hut varying CO₂ concentrations. An increase in the light intensity in which seedlings were grown had no effect on their height or mean leaf area, hut resulted in thicker leaves, an increase in the total leaf area per seedling due to an increase in the number of leaves, an increase in the dry weight especially of roots and a decrease in the chlorophyll content of leaves. Throughout the growing season seedlings grown in full daylight, as compared with those grown in lower light intensities, had the lowest rates of apparent photosynthesis measured at standard conditions (21,600 lux light intensity and 300 ul/l of CO₂), when this was expressed per unit leaf area, hut the highest rates on a per seedling basis. Thus dry matter production attained at the end of the growing season correlated positively with the photosynthetic rate per seedling, but not per unit leaf area. The rates of apparent photosynthesis of seedlings grown at lower light intensities were more responsive to changes in light intensity or CO₂ concentration than those of seedlings grown in full daylight intensity.     

Reduced osmotic potential effects on photosynthesis : identification of stromal acidification as a mediating factor

Addition of sorbitol, which facilitated reductions in reaction medium osmotic potential from standard (0.33 molar sorbitol, −10 bars) isotonic conditions to a stress level of 0.67 molar sorbitol (−20 bars), inhibited the photosynthetic capacity of isolated spinach (Spinacia oleracea) chloroplasts. This inhibition, which ranged from 64 to 74% under otherwise standard reaction conditions, was dependent on reaction medium inorganic phosphate concentration, with the phosphate optimum for photosynthesis reduced to 0.05 millimolar at the low osmotic potential stress treatment from a value of 0.25 millimolar under control conditions.

Stromal alkalating agents such as NH₄Cl (0.75 millimolar) and KCl (35 millimolar) were also found to affect the degree of low osmotic potential inhibition of photosynthesis. Both agents doubled the rate of NaHCO₃-supported O₂ evolution under the stress treatment, while hardly affecting the control rate at optimal concentrations. These agents also reduced the length of the lag phase of photosynthetic O₂ evolution under the stress treatment to a much greater degree. The rate-enhancement effect of these agents under the stress treatment was reversed by sodium acetate, which is known to facilitate stromal acidification.

The reaction medium pH optimum for photosynthesis under the stress treatment was higher than under control conditions. In the presence of optimal NH₄Cl, this shift was no longer evident.

Internal pH measurements indicated that the stress treatment caused a 0.43 and 0.24 unit reduction in the stromal and intrathylakoid pH, respectively, under illumination. This osmotically induced acidification was not evident in the dark. The presence of 0.75 millimolar NH₄Cl partially reversed the osmotically induced reduction in the illuminated stromal pH. It was concluded that stromal acidification is a mediating mechanism of the most severe site of low osmotic potential inhibition of the photosynthetic process.