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Leaf growth, photosynthesis and tissue water relations of greenhouse-grown Eucalyptus marginata seedlings in response to water deficits.

Leaf growth, rate of leaf photosynthesis and tissue water relations of shoots of Eucalyptus marginata Donn ex Sm. (jarrah) seedlings were studied during a soil drying and rewatering cycle in a greenhouse experiment. Rates of leaf growth and photosynthesis were sensitive to water deficits. The rate of leaf growth decreased linearly with predawn leaf water potential to reach zero at -1.5 MPa. Rate of leaf growth did not recover completely within the first three days after rewatering. Midday photosynthetic rates declined to 40% of those of well-watered seedlings at a predawn leaf water potential of -1.0 MPa and reached zero at -2.2 MPa. Photosynthetic rate recovered rapidly following rewatering and almost fully recovered by the second day after rewatering. All tissue water relations parameters, except the bulk modulus of elasticity, changed significantly as the soil dried and recovered completely by the third day after rewatering. Changes in osmotic pressure at full turgor of 0.4 MPa indicated considerable capacity for osmotic adjustment. However, because there was little osmotic adjustment until predawn leaf water potential fell below -1.5 MPa, this capacity would not have enhanced seedling growth, although it may have increased seedling survival. The sensitivity of photosynthesis and relative water content to water deficits suggests that greenhouse-grown E. marginata seedlings behave like mesophytic plants, even though E. marginata seedlings naturally grow in a drought-prone environment.

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Effects of leaf nutrient status on photosynthetic capacity in loblolly pine (Pinus taeda L.) seedlings grown in elevated atmospheric CO(2).

We measured needle photosynthesis of loblolly pine seedlings grown in a factorial experiment with two CO(2) partial pressures (35 and 65 Pa) and three nutrient treatments (7 mM NH(4)NO(3) + 1 mM PO(4); 7 mM NH(4)NO(3) + 0.2 mM PO(4); 1 mM NH(4)NO(3) + 1 mM PO(4)). The data were used to parameterize a physiologically based photosynthetic model that included limitations imposed by ribulose-1,5-bisphosphate carboxylase/oxygenase activity, electron transport capacity and inorganic phosphate availability. With nonlimiting nutrients, seedlings grown at 65 Pa CO(2) had significantly higher net photosynthesis and lower stomatal conductance than seedlings grown at 35 Pa CO(2). Nutrient limitations by either N or P significantly reduced photosynthetic capacity. When either N or P was limiting, there was no effect of growth CO(2) partial pressure on photosynthesis, but stomatal conductance was significantly lower for seedlings grown at 65 Pa CO(2). Modeled biochemical parameters suggest that, in all cases, photosynthesis was co-limited by carboxylation, electron transport and phosphate regeneration. Acclimation to growth in elevated CO(2) involved a reduction in leaf N content. In the low-N and low-P treatments, modeled parameters indicated that the biochemical processes of photosynthesis were down regulated to the point that there was no effect of increasing CO(2) partial pressure. The capacity to regenerate phosphate was reduced in both low nutrient treatments, but was only reduced by elevated CO(2) when seedlings were grown under low soil P conditions. Increased photosynthetic water use efficiency and nutrient use efficiency in response to CO(2) enrichment occurred in all three nutrient treatments and have important implications for whole-plant water and nutrient balance. These data support the contention that soil nutrient status in forest ecosystems will be a critical influence on tree seedling response to increasing atmospheric CO(2) partial pressures.

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Effect of elevated carbon dioxide concentration and root restriction on net photosynthesis, water relations and foliar carbohydrate status of loblolly pine seedlings.

To determine the effects of CO(2)-enriched air and root restriction on photosynthetic capacity, we measured net photosynthetic rates of 1-year-old loblolly pine seedlings grown in 0.6-, 3.8- or 18.9-liter pots in ambient (360 micro mol mol(-1)) or 2x ambient CO(2) (720 micro mol mol(-1)) concentration for 23 weeks. We also measured needle carbohydrate concentration and water relations to determine whether feedback inhibition or water stress was responsible for any decreases in net photosynthesis. Across all treatments, carbon dioxide enrichment increased net photosynthesis by approximately 60 to 70%. Net photosynthetic rates of seedlings in the smallest pots decreased over time with the reduction occurring first in the ambient CO(2) treatment and then in the 2x ambient CO(2) treatment. Needle starch concentrations of seedlings grown in the smallest pots were two to three times greater in the 2x ambient CO(2) treatment than in the ambient CO(2) treatment, but decreased net photosynthesis was not associated with increased starch or sugar concentrations. The reduction in net photosynthesis of seedlings in small pots was correlated with decreased needle water potentials, indicating that seedlings in the small pots had restricted root systems and were unable to supply sufficient water to the shoots. We conclude that the decrease in net photosynthesis of seedlings in small pots was not the result of CO(2) enrichment or an accumulation of carbohydrates causing feedback inhibition, but was caused by water stress.

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Photosynthetic gas exchange and accumulation of phytotoxins in mangrove seedlings in response to soil physico-chemical characteristics associated with waterlogging.

We evaluated photosynthetic gas exchange and the accumulation of iron, manganese and sulfur in seedlings of five mangrove species (Aegiceras corniculatum (L.) Blanco, Avicennia marina (Forsk.) Vierh., Bruguiera gymnorrhiza (L.) Lamk., Hibiscus tiliaceus L. and Rhizophora stylosa Griff.) growing under anoxic soil conditions at low irradiance. Seedlings of the viviparous mangroves showed no significant responses to root anoxia. The presence of ferrous sulfate or manganous sulfate had a smaller effect on CO(2) assimilation, transpiration rate and stomatal behavior than the presence of sodium sulfide. Sodium sulfide inhibited photosynthetic gas exchange and caused complete stomatal closure in all species. Stomatal closure was probably the result of the damaging effect of sulfide ions on root cell membranes. Some leaf epinasty and wilting were also observed in response to the Na(2)S treatment in all species. A combination of FeSO(4), MnSO(4) and Na(2)S had a smaller effect on photosynthesis than Na(2)S alone, especially for Avicennia marina and Rhizophora stylosa seedlings, which maintained appreciable rates of CO(2) assimilation (2.49 and 3.84 &mgr;mol m(-2) s(-1), respectively) in the presence of all three phytotoxins. Roots of phytotoxin-treated seedlings of all species accumulated significant amounts of the corresponding toxin compared with roots of the control plants. The FeSO(4) or MnSO(4) treatments had no effect on foliar concentrations of iron or manganese, whereas the Na(2)S treatment resulted in an accumulation of S in the leaves of all species. Interactions between Fe(2+) and Mn(2+) and sulfide in the culture medium appeared to reduce their uptake by the seedlings. We conclude that high concentrations of sulfides in mangrove sediments may adversely affect both growth and survival of mangrove seedlings at low irradiances.

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Influence of ozone on cold acclimation in sugar maple seedlings.

During summer 1994, sugar maple (Acer saccharum Marsh.) seedlings were grown in open-top chambers supplied with air containing near ambient ozone concentration (control, low O(3)) or three times the ambient ozone concentration (high O(3)). The rate of CO(2) assimilation was significantly reduced by chronic exposure to a high concentration of ozone during the summer. During fall, seedlings were removed from the open-top chambers and acclimated to cold under natural conditions. In both species during cold acclimation, the starch concentration decreased, whereas the sucrose concentration increased. There was no treatment effect on the freezing tolerance of roots, even though roots in the high-O(3) treatment accumulated higher concentrations of the cryoprotective oligosaccharides raffinose and stachyose than control roots. Cold acclimation occurred earlier and stachyose concentration of stems was higher in high-O(3)-treated seedlings than in low-O(3)-treated seedlings. Cold acclimation was associated with an earlier accumulation of ABA in the xylem sap of high-O(3)-treated seedlings compared with low-O(3)-treated seedlings.

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Effects of aluminum on growth, development, and nutrient composition of honeylocust (Gleditsia triacanthos L.) seedlings.

Hydroponic experiments were conducted to determine the effects of aluminum (Al) on root development, shoot morphology and the nutrient composition of honeylocust seedlings (Gleditsia triacanthos L.). Seedlings were grown at pH 4 in a nutrient solution containing 0, 50, 150, 600 or 1500 microM Al. Within seven days, there were significant differences in root growth and root nutrient composition between control seedlings and seedlings grown in the presence of 150 or 600 microM Al. By day 14, significant reductions in leaf production and plant height were observed in seedlings treated with 1500 microM Al. At the lowest Al concentration, 50 microM, leaf size and expansion rates were significantly lower than in the controls. By the third week of the experiment, Ca and Mg concentrations in young leaves of the Al-treated seedlings were significantly lower than in leaves of control plants. Analysis of old leaf tissue, however, revealed no consistent pattern of nutrient concentration with Al treatment.

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Effects of anaerobic growth conditions on phosphorus tissue concentrations and absorption rates of southern pine seedlings.

A non-circulating, continuously flowing solution culture was used to examine the long- and short-term effects of anaerobic growth conditions on phosphorus uptake in 12-week-old seedlings of three pine species. Sand pine (Pinus clausa (Engelm.) Sarg.) and a drought-hardy loblolly pine (P. taeda L.) had the largest reductions in biomass after 8 weeks in anaerobic solution, whereas the more flood-tolerant pond pine (P. serotina Michx.) and wet-site loblolly pine seedlings were least affected. Anaerobic growth conditions reduced P concentrations in the shoot and increased P concentrations in the root. Short-term (32)P-uptake experiments were conducted with intact seedlings to determine absorption rates (influx) of inorganic phosphate (Pi) by aerobically and anaerobically grown pine seedlings. Influx of Pi was weakly correlated with shoot fresh weight, root fresh weight, root P and shoot P per gram fresh weight root, suggesting that internal P levels and biomass influenced Pi absorption. The highest Pi absorption rates were found in anaerobically grown seedlings under anaerobic (32)P-uptake conditions. Of these, sand pine had the highest absorption rate in 50 microM KH(2)PO(4) (0.96 micromol Pi g(-1) FW root h(-1)), and the wet-site loblolly pine exhibited the lowest rate (0.24 micromol Pi g(-1) FW root h(-1)). Aerobically grown seedlings had similar Pi absorption rates that were not significantly affected by O(2) concentration in the (32)P-uptake solution.

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Effects of drought preconditioning on thermotolerance of photosystem II and susceptibility of photosynthesis to heat stress in cedar seedlings.

Changes in photosystem II (PSII) thermotolerance during drought and recovery were studied under controlled conditions in three Mediterranean cedar species (Cedrus brevifolia Henry, C. libani Loudon and C. atlantica Manetti). The temperature at which the quantum yield of PSII photochemistry was reduced by 15% of its value at 25 degrees C was 3 to 4 degrees C higher in drought-treated plants than in well-watered plants. The drought-induced increase in PSII thermotolerance was already evident 8 days after water had been withheld from the seedlings, when net CO(2) assimilation was still at 80% of its initial value, and was visible for up to 12 days after re-watering. When seedlings of the three species were exposed to temperatures above 45 degrees C for 5 h, both maximal quantum yield of PSII photochemistry and net CO(2) assimilation rate were significantly reduced in unconditioned seedlings, whereas drought-preconditioned seedlings were almost unaffected by the heat treatment. Drought-preconditioned seedlings still exhibited a higher tolerance to heat stress than unconditioned seedlings 60 days after re-watering, although the transient, drought-induced osmotic adjustment had fully disappeared. Among species, C. atlantica was the most heat sensitive, whereas the heat treatment had no significant effect on the parameters measured in C. brevifolia.

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Physiological responses of black spruce layers and planted seedlings to nutrient addition.

We investigated effects of nutrient addition on several physiological characteristics of 60-cm-tall black spruce (Picea mariana Mill. B.S.P.) layers (i.e., rooted branches of overstory trees) and 20-cm-tall planted seedlings on a clear-cut, N-limited boreal site. After two growing seasons, current-year and one-year-old needles of fertilized trees (layers and seedlings combined) had higher net photosynthetic rates (A(n)) and maximum capacity of Rubisco for CO(2) fixation (V(max)) than unfertilized trees. One-year-old needles of fertilized trees had higher stomatal conductance (g(s)), higher water-use efficiency, and lower intercellular to ambient CO(2) ratio than unfertilized trees. Additionally, fertilized trees had higher predawn and midday shoot water potentials than unfertilized trees. Stomatal conductance of 1-year-old needles was 23% higher in seedlings than in layers, but there were no significant differences in g(s) of current-year needles between the regeneration types. For both needle age-classes, A(n) and V(max) of layers were 25 and 40% higher, respectively, than the corresponding values for seedlings. The higher values of A(n), V(max) and foliar N concentration of layers compared with seedlings after two growing seasons may be associated with the larger root systems of the layers compared with the transplanted seedlings.

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Comparison of the responses to flooding of seedlings and cuttings of Gmelina.

The effects of flooding on Gmelina arborea Roxb. seedlings and cuttings of the same parent stock were compared to determine their suitability as transplanting stock. Flooding caused reductions in stomatal conductance, xylem pressure potential and dry matter accumulation in both groups of plants. In seedlings, flooding induced formation of hypertrophied lenticels, stem hypertrophy and production of short, thick, adventitious roots in seedlings, whereas in cuttings, only thin roots and numerous smaller lenticels were induced. For 8 days after the flooding treatment ended, the flooded seedlings grew faster than control seedlings, whereas in cuttings, post-flooding growth was similar to that of control plants. It is suggested that seedlings may perform better than cuttings in very wet or saturated soil.

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Influence of drought stress and low irradiance on plant water relations and structural constituents in needles of Pinus ponderosa seedlings.

The influence of low light on tolerance to prolonged drought was tested on unshaded and shaded seedlings of ponderosa pine (Pinus ponderosa var. scopulorum Dougl. ex Laws.). Unshaded seedlings of P. ponderosa var. ponderosa were also drought stressed to compare varietal responses to drought. The maximum irradiance received by shaded seedlings was 10% of full light. Seedlings were progressively drought stressed until predawn water potentials (Psi(x)) were -5.0 MPa. Relative water content (RWC) and the reciprocal of Psi(x) were analyzed by means of an unusual application of the pressure-volume relationship for determination of RWC of the apoplast (RWC(a)), osmotic potential at full turgor (Psi(oft)), and ratio of fully turgid weight to dry weight. Major varietal differences in drought response were in RWC(a) and needle cellulose content. The shaded seedlings showed tissue damage at relative water contents < 60%, and were killed by water deficits from which unshaded seedlings recovered. Correspondingly, shaded plants had significantly higher cell volume/cell mass ratio, Psi(oft), less cellulose in needle tissue, and lower RWC(a) than unshaded plants. These differences suggest that low irradiance restricts drought adaptation in ponderosa pine.

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Temporal variations in cold sensitivity of root growth in cold-stored white spruce seedlings.

We examined effects of soil temperature on the number of roots produced by white spruce (Picea glauca (Moench.) Voss) seedlings during the first month of growth following 0-30 weeks of storage in darkness at -2 degrees C. After storage, seedlings were planted in pots and placed in a controlled-environment chamber with a constant air temperature of 11 degrees C and a 16-h photoperiod. Water baths were used to keep soil temperature at 3, 7 or 11 degrees C. The number of long roots (> 10 mm) produced was strongly dependent on soil temperature. At soil temperatures of 3 or 7 degrees C, the number of long roots produced was only 11 to 30% that at 11 degrees C. Seedlings that had been stored for 14 weeks and then planted in soil at 11 degrees C produced the greatest number of long roots. For seedlings planted in soil at 11 degrees C, the number of long roots increased with time of storage up to 14-18 weeks and then declined progressively with length of storage. No increase in number of long roots with length of storage up to 18 weeks was evident in seedlings planted in soil at 3 or 7 degrees C. The maximum number of short roots (5-10 mm) was observed in seedlings that had been stored for 17 weeks and then planted in soil at 7 or 11 degrees C.

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Arabidopsis seedling growth, storage lipid mobilization, and photosynthetic gene expression are regulated by carbon:nitrogen availability.

The objective of the current work was to establish the degree to which the effects of carbon and nitrogen availability on Arabidopsis seedling growth and development are due to these nutrients acting independently or together. Growth of seedlings on low (0.1 mM) nitrogen results in a significant reduction of seedling and cotyledon size, fresh weight, chlorophyll, and anthocyanin content but a slight increase in endogenous sugars. The addition of 100 mM sucrose (Suc) to the nitrogen-depleted growth media results in a further reduction in cotyledon size and chlorophyll content and an overall increase in anthocyanins and endogenous sugars. Storage lipid breakdown is almost completely blocked in seedlings grown on low nitrogen and 100 mM Suc and is significantly inhibited when seedlings are grown on either low nitrogen or high Suc. Carbohydrate repression of photosynthetic gene expression can only be observed under low nitrogen conditions. Low (0.1 mM) nitrogen in the absence of exogenous carbohydrate results in a significant decrease in chlorophyll a/b-binding protein and ribulose bisphosphate carboxylase small subunit gene transcript levels. Thus, carbon to nitrogen ratio rather than carbohydrate status alone appears to play the predominant role in regulating various aspects of seedling growth including storage reserve mobilization and photosynthetic gene expression.

Anthocyanins↗

Changes in Isozyme Profiles of Catalase, Peroxidase, and Glutathione Reductase during Acclimation to Chilling in Mesocotyls of Maize Seedlings.

The response of antioxidants to acclimation and chilling in various tissues of dark-grown maize (Zea mays L.) seedlings was examined in relation to chilling tolerance and protection from chilling-induced oxidative stress. Chilling caused an accumulation of H2O2 in both the coleoptile + leaf and the mesocotyl (but not roots), and acclimation prevented this accumulation. None of the antioxidant enzymes were significantly affected by acclimation or chilling in the coleoptile + leaf or root. However, elevated levels of glutathione in acclimated seedlings may contribute to an enhanced ability to scavenge H2O2 in the coleoptile + leaf. In the mesocotyl (visibly most susceptible to chilling), catalase3 was elevated in acclimated seedlings and may represent the first line of defense from mitochondria-generated H2O2. Nine of the most prominent peroxidase isozymes were induced by acclimation, two of which were located in the cell wall, suggesting a role in lignification. Lignin content was elevated in mesocotyls of acclimated seedlings, likely improving the mechanical strength of the mesocotyl. One cytosolic glutathione reductase isozyme was greatly decreased in acclimated seedlings, whereas two others were elevated, possibly resulting in improved effectiveness of the enzyme at low temperature. When taken together, these responses to acclimation illustrate the potential ways in which chilling tolerance may be improved in preemergent maize seedlings.

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The Fungicide Phosphonate Disrupts the Phosphate-Starvation Response in Brassica nigra Seedlings.

The development of Brassica nigra seedlings over 20 d of growth was disrupted by the fungicide phosphonate (Phi) in a manner inversely correlated with nutritional inorganic phosphate (Pi) levels. The growth of Pi-sufficient (1.25 mM Pi) seedlings was suppressed when 10, but not 5, mM Phi was added to the nutrient medium. In contrast, the fresh weights and root:shoot ratios of Pi-limited (0.15 mM) seedlings were significantly reduced at 1.5 mM Phi, and they progressively declined to about 40% of control values as medium Phi concentration was increased to 10 mM. Intracellular Pi levels generally decreased in Phi-treated seedlings, and Phi accumulated in leaves and roots to levels up to 6- and 16-fold that of Pi in Pi-sufficient and Pi-limited plants, respectively. Extractable activities of the Pi-starvation-inducible enzymes phosphoenolpyruvate phosphatase and inorganic pyrophosphate-dependent phosphofructokinase were unaltered in Pi-sufficient seedlings grown on 5 or 10 mM Phi. However, when Pi-limited seedlings were grown on 1.5 to 10 mM Phi (a) the induction of phosphoenolpyruvate phosphatase and inorganic pyrophosphate-dependent phosphofructokinase activities by Pi limitation was reduced by 40 to 90%, whereas (b) soluble protein concentrations and the activities of the ATP-dependent phosphofructokinase and pyruvate kinase were unaffacted. It is concluded that Phi specifically interrupts processes involved in regulation of the Pi-starvation response in B. nigra.

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Regulation of gibberellin 20-oxidase and gibberellin 3beta-hydroxylase transcript accumulation during De-etiolation of pea seedlings.

Gibberellin (GA) 20-oxidase (GA 20-ox) and GA 3beta-hydroxylase (GA 3beta-hy) are enzymes that catalyze the late steps in the formation of active GAs, and are potential control points in the regulation of GA biosynthesis by light. We have investigated the photoregulation of the GA 20-ox and GA 3beta-hy transcript levels in pea (Pisum sativum L.). The GA 20-ox transcript level was higher in light-grown seedlings than in etiolated seedlings, whereas GA 3beta-hy mRNA accumulation was higher in etiolated seedlings. However, transfer of etiolated seedlings to light led to a 5-fold increase in the expression of both transcripts 4 h after transfer. GA 20-ox mRNA accumulation is regulated by both phytochromes A and B. Transfer to light also resulted in a 6-fold decrease in GA(1) levels within 2 h. These results suggest that the light-induced drop in GA(1) level is not achieved through regulation of GA 20-ox and GA 3beta-hy mRNA accumulation. The application of exogenous GA(1) to apical buds of etiolated seedlings prior to light treatments inhibited the light-induced accumulation of both GA 20-ox and GA 3beta-hy mRNA, suggesting that negative feedback regulation is an important mechanism in the regulation of GA 20-ox and GA 3beta-hy mRNA accumulation during de-etiolation of pea seedlings.

Darkness↗

Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in Arabidopsis seedlings.

Previous studies have shown that salicylic acid (SA) is an essential component of the plant resistance to pathogens. We now show that SA plays a role in the plant response to adverse environmental conditions, such as salt and osmotic stresses. We have studied the responses of wild-type Arabidopsis and an SA-deficient transgenic line expressing a salicylate hydroxylase (NahG) gene to different abiotic stress conditions. Wild-type plants germinated under moderate light conditions in media supplemented with 100 mM NaCl or 270 mM mannitol showed extensive necrosis in the shoot. In contrast, NahG plants germinated under the same conditions remained green and developed true leaves. The lack of necrosis observed in NahG seedlings under the same conditions suggests that SA potentiates the generation of reactive oxygen species in photosynthetic tissues during salt and osmotic stresses. This hypothesis is supported by the following observations. First, the herbicide methyl viologen, a generator of superoxide radical during photosynthesis, produced a necrotic phenotype only in wild-type plants. Second, the presence of reactive oxygen-scavenging compounds in the germination media reversed the wild-type necrotic phenotype seen under salt and osmotic stress. Third, a greater increase in the oxidized state of the glutathione pool under NaCl stress was observed in wild-type seedlings compared with NahG seedlings. Fourth, greater oxidative damage occurred in wild-type seedlings compared with NahG seedlings under NaCl stress as measured by lipid peroxidation. Our data support a model for SA potentiating the stress response of the germinating Arabidopsis seedling.

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The effect soaking pea seeds with or without seedcoats has on seedling growth.

Pea seeds (Pisum sativum L. ;Alaska') with intact seedcoats (WC) and with seedcoats removed (WOC) were soaked in distilled water for 24 hours at 20 degrees . The water, containing the pea diffusate, was decanted after the second, fourth, sixth, eighth, twelfth, and twenty-fourth hour and analyzed for total nitrogen, alpha-amino nitrogen, carbohydrate, and total solute dry weight. The seeds were germinated at 20 degrees in a 16 hour photoperiod of 300 foot candles. Stem lengths and dry weights of roots, shoots and cotyledons were determined after 4, 11, and 18 days of growth. WOC seeds imbibed more water than WC seeds during the 24 hour imbibition period. Diffusates from WOC seeds always contained more solute than diffusates from WC seeds. Maltose, glucose, and fructose were not detected in the early diffusates from WOC seeds but were found in WC seed diffusates at all times. Seedlings from WC seeds had longer stems than those from WOC seeds. The dry weight of stems and roots of WC seedlings was greater than those from WOC seedlings. The dry weight of cotyledons from 18 day-old WC seedlings was less than from WOC seedlings. Water absorption by WC seeds was slower than by WOC seeds. Removal of the seedcoat allowed rapid imbibition resulting in seed injury presumably because of the loss of solutes which included monosaccharides, disaccharides, amino acids, and other nitrogen containing compounds. These results are consistent with the hypothesis that rapid imbibition disrupts membrane organization leading to reduction of seedling growth.

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