Effect of Irradiance, Sugars and Nitrogen on Leaf Size of In Vitro Grown Ceratonia Siliqua L.

Carob (Ceratonia siliqua L.) has compound pinnate leaves consisting of 4 – 6 pairs of leaflets. However, in conditions of in vitro culture only one pair of leaflets develops. With increasing irradiance from 9.3 to 74.1 µmol m^–2 s^–1, leaf area increased 5-fold. Sucrose also significantly increased leaf area and the maxima were at concentration 147 mM at high irradiance and 233.6 mM at low irradiance. Sucrose was superior to fructose, glucose and combination of both in increasing leaf area. Decreasing concentration of KNO₃ and NH₄NO₃ caused a 3-fold decline of leaf area.     

Corn (Zea mays L.) growth, leaf pigment concentration, photosynthesis and leaf hyperspectral reflectance properties as affected by nitrogen supply

Plant nitrogen (N)deficiency often limits crop productivity. Early detection of plant N deficiency is important for improving fertilizer N-use efficiency and crop yield. An experiment was conducted in sunlit, controlled environment chambers in the 2001 growing season to determine responses of corn (Zea mays L. cv. 33A14) growth and leaf hyperspectral reflectance properties to varying N supply. Four N treatments were: (1) half-strength Hoagland's nutrient solution applied throughout the experiment (control); (2) 20% of control N starting 15 days after emergence (DAE); (3) 0% N starting 15 DAE; and (4) 0% N starting 23 DAE (0% NL). Plant height, the number of leaves, and leaf lengths were examined for nine plants per treatment every 3–4 days. Leaf hyperspectral reflectance, concentrations of chlorophyll a, chlorophyll b,and carotenoids, leaf and canopy photosynthesis, leaf area, and leaf N concentration were also determined during the experiment. The various N treatments led to a wide range of N concentrations (11 – 48 g kg^–1 DW) in uppermost fully expanded leaves. Nitrogen deficiency suppressed plant growth rate and leaf photosynthesis. At final harvest (42 DAE), plant height, leaf area and shoot biomass were 64–66% of control values for the 20% N treatment, and 46-56% of control values for the 0% N treatment. Nitrogen deficit treatments of 20% N and 0% N (Treatment 3) could be distinguished by changes in leaf spectral reflectance in wavelengths of 552 and 710 nm 7 days after treatment. Leaf reflectance at these two wavebands was negatively correlated with either leaf N (r = –0.72 and –0.75^**) or chlorophyll (r = –0.60 and –0.72^**) concentrations. In addition, higher correlations were found between leaf N concentration and reflectance ratios. The identified N-specific spectral algorithms may be used for image interpretation and diagnosis of corn N status for site-specific N management.     

Amino acid contents and transport in oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) under different nitrogen conditions

Oilseed rape (Brassica napus L.) needs very high nitrogen fertilizer inputs. Significant amounts of this nitrogen are lost during early leaf shedding and are a source of environmental and economic concern. The objective of this study was to investigate whether the remobilization of leaf amino acids could be limiting for nitrogen use efficiency. Therefore, amino acid concentrations were analyzed in subcellular compartments of leaf mesophyll cells of plants grown under low (0.5 mM NO₃^–) and high (4 mM NO₃^–) nitrogen supply. With high nitrogen supply, young leaves showed an elevated amino acid content, mainly in vacuoles. In old leaves, however, subcellular concentrations were similar under high and low nitrogen conditions, showing that the excess nitrogen had been exported during leaf development. The phloem sap contained up to 650 mM amino acids, more than four times as much than the cytosol of mesophyll cells, indicating a very efficient phloem-loading process. Three amino acid permeases, BnAAP1, BnAAP2, and BnAAP6, were identified and characterized. BnAAP1 and BnAAP6 mediated uptake of neutral and acidic amino acids into Xenopus laevis oocytes at the actual apoplastic substrate concentrations. All three transporters were expressed in leaves and the expression was still detectable during leaf senescence, with BnAAP1 and BnAAP2 mRNA levels increasing from mature to old leaves. We conclude that phloem loading of amino acids is not limiting for nitrogen remobilization from senescing leaves in oilseed rape.     

Protective effect of exogenous polyamines against atrazine in pea plants

The effect of exogenously applied polyamines in reversing the effect of atrazine stress on pea seedlings (Pisum sativum L., cv. Koray) was investigated. The plants treated with combinations of atrazine (14 mM) + spermidine (1 mM) and atrazine (14 mM) + spermine (1 mM) possessed improved growth (30–35% increase of leaf area and 10–20% increase of fresh weight 10 days after treatment) and chlorophyll content (50–60% increase) in comparison with atrazine-(14 mM) treated plants. Spermine and spermidine also diminished the inhibitory effect of atrazine on gas exchange and photosystem II function. This fact supports the hypothesis of Yordanov and Goltsev (1990, Plant Physiol 4:42–51) that the interaction of polyamines with the thylakoid membrane surface led to their stacking, to separation of the photosystems, and to the association of light-harvesting complex II with the photosystem II core complex.     

Volatiles from different organs of Unxia camphorata L. f. growing wild in the Amazon

The essential oils obtained from fresh leaves, flowers, roots and stems of Unxia camphorata collected in the rainy and dry seasons in the state of Pará, were examined by GC and GC/MS. The highest oil yields were observed from the samples collected at the dry season (flowers: 0.6%, leaves: 0.6%, stems: 0.3%, roots: 0.4%). All parts of the plant were rich in α-phellandrene, with non-significant changes during the rainy and dry seasons (flowers: 61.9–53.7%, leaves: 18.3–17.4%, stems: 68.3–72.8%, roots: 51.9–48.3%). In the leaf oil, the monoterpenoid camphor was the largest component (rainy season: 28.5%, dry season: 28.8%). In the leaf oil, the high amount of α-phellandrene was followed by methylthymol (rainy season: 14.1%, dry season: 13.1%). Depending on the plant part studied, these oils can be characterized by α-phellandrene, camphor/α-phellandrene/camphene, or α-phellandrene/methylthymol type.     

Energy-dependent phases of the circadian clock and the clock-controlled leaf movement in Phaseolus coccineus L.

The energy requirements of the various phases of the circadian clock in the laminar pulvini cells of primary leaves of Phaseolus coccineus L. were investigated using 4-h pulses of NaCN (5 mM) and NaN₃ (1 mM). The induced phase shifts were calculated from the timing of the subjective night position during the third cycle after the treatment. Both inhibitors produce advances during phases which are correlated with the upward movement of the leaf (ca. 0–12 h after the maximum of the subjective night position) and during phases which are correlated with the downward movement of the leaf (ca. 20–28 h after the maximum of the subjective night position). Maximal advances are induced during the phase which is correlated with the maximum of the subjective night position (hour 0), whereas during phases which are correlated with the subjective day position (ca. 12–20 h after the maximum of the subjective night position) the inhibitors have no effect or induce only small advances. These results demonstrate that the part of the circadian cycle which, according to Bünning's tension-relaxation model of the circadian clock, is characterized by features of relaxation, represents a sequence of phases with decreasing energy requirement, whereas the tension part of the circadian cycle represents a sequence of phases with increasing energy requirement. The energy requirement for changing and maintaining the leaf positions was investigated by continuously offering NaCN, NaN₃, and dinitrophenol (DNP) to leaves with intact and half (flexor cut away) pulvini. The substances inhibit in both pulvini the upward movement or induce a downward movement, depending on the leaf position, when the transfer to the inhibitor solution takes place. These results give evidence that the movement of intact pulvini reflects the turgor (volume) state of the extensor cells and that the increase of turgor (volume) and high turgor (volume) state requires more energy than the decrease of turgor (volume) or low turgor (small volume) state. Therefore, the time course of the energy requirements of the circadian clock and the clock-controlled turgor (volume states or leaf movement) is out of phase during a circadian cycle. Consequently the reaction of the clock-controlled leaf movement to the reduced energy supply can mask the clock behavior in pulse and step experiments. The phase response curves towards CN^- and N ₃ ^- reflect the time course of the CN^--induced membrane depolarizations (the energy requirement of the electrogenic pump) in extensor cells of the pulvinus (Freudling et al. (1980), Plant Physiol. 65, 966–968), and both are out of phase with the time course of the energy requirement of the turgor. Consequently it is hypothesized that in Phaseolus advances are due to membrane depolarization and that at least in this organism electric properties of the plasmalemma are essentially involved in the mechanism of the circadian clock.Abbreviations LD light-dark cycle - LL continuous light - DNP dinitrophenol This paper is dedicated to Professor Erwin Bünning on the occasion of his 75th birthdayIn this paper zero corresponds to the second maximum of the subjective night position of the leaves after transfer to constant conditions. Zero to twelve hours corresponds approximately to the upward movement of the leaves, 12–20 h to the elevated (subjective day) position, and 20–28 h to the downward movement of the leaves. In other circadian systems Pittendrigh's CT (circadian time) convention is used. CT 00 is the time of dawn after a 12-h light/12-h dark cycle. Since in Phaseolus the plants are raised in a LD cycle different from 12:12 and since the phases at dawn differ considerably from leaf to leaf and are furthermore not precisely determinable (whereas the subjective night position of the leaves is a well-defined and recognizable phase) this convention is not followed in Phaseolus. Phase zero in Phaseolus corresponds to approximately CT 18 in other systems     

Characteristics of a photorespiratory mutant of barley (Hordeum vulgare L.) deficient in phosphogly collate phosphatase

A barley mutant RPr84/90 has been isolated by selecting for plants which grow poorly in natural air, but normally in air enriched to 0.8% CO₂. After 5 minutes of photosynthesis in air containing¹⁴CO₂ this mutant incorporated 26% of the¹⁴C carbon into phosphoglycollate, a compound not normally labelled in wild type (cv. Maris Mink) leaves.The activity of phosphoglycollate phosphatase (EC 3.1.1.18) was 1.2 nkat mg^–1 protein at 30°C in RPr 84/90 compared to 19.2 nkat mg^–1 protein in the wild-type leaves. Phosphoglycollate phosphatase activity was not detected after protein separation by electrophoresis of leaf extracts from the mutant on polyacrylamide gels; on linear 5% acrylamide gels three bands with enzyme activity were separated from extracts of wild type plants. Gradient gel electrophoresis followed by activity staining showed two bands in Maris Mink tracks of MW 86,000 and 96,000, but no bands in 84/90. This is the first report of isozymes of phosphoglycollate phosphatase in barley which were absent in the mutant extracts. Our results confirm an earlier report of isozymes of this phosphatase in Phaseolus vulgaris [18].The photosynthetic rate of RPr 84/90 in 1% O₂, 350 l CO₂ l^–1 was 9–12 mg CO₂ dm^–2 h^–1 at 20°C, whereas the wild-type rate was 27–29 mg CO₂ dm^–2 h^–1 at 20°C. In 21% O₂, 350 l CO₂ l^–1 the rate was 2–3 mg CO₂ dm^–2 h^–1 in the mutant and 20 mg CO₂ dm^–2 h^–1 in the wild type.Genetic analysis has shown that the mutation segregates as a single recessive nuclear gene.     

A culture unit system for the study of responses of mycorrhizal and non-mycorrhizal seedling to treatments

The time-dependence of Mn accumulation was confirmed in potato foliage (Solanum tuberosum. L.cv. Norland) grown in solution culture. Older leaves grown at 0.61 mM Mn had substantially higher Mn concentrations than younger leaves and stem samples. Levels of Mn in older leaves increased steadily from 4000 µg g^–1 at one week to 8–10,000 µg g^–1 at 6 weeks, but were relatively constant in the emerging leaves. Even foliage grown at low Mn levels (0.01 mM Mn) had 4 fold gradients in Mn concentration from younger (40 µg g^–1) to older leaves (180 µg g^–1).At 0.61 mM Mn, concentrations of 3–4000 µg g^–1 in the youngest fully-developed leaves did not bring about any decline in yield, and levels of up to 5000 µg g^–1 occurred in individual potato leaves before Mn toxicity symptoms were observed. Potato foliage grown at the high Mn had similar leaf numbers, but showed an increased stem length and smaller leaves than foliage grown at 0.01 mM Mn. In particular, the leaf area of the middle and lower leaf fractions were affected by the high Mn level.The ability of rapidly growing plants to withstand high concentrations of Mn is discussed in relation to the pattern of dry matter and Mn accumulation shown by potato foliage.     

Effects of Benzyladenine and Naphthalene Acetic Acid on Growth and Camptothecin Accumulation in Camptotheca acuminata Seedlings

The effects of the cytokinin benzyladenine (BA) and the auxin naphthalene acetic acid (NAA) on Camptotheca acuminata Decaisne growth and camptothecin (CPT) accumulation (leaf CPT concentration and total leaf CPT yield) were studied in a hydroponic culture system for three weeks. Increasing BA concentrations from 0 to 3 mg l^–1 in growth medium decreased plant height, stem weight, and leaf weight but increased root weight. High BA levels (1 and 3 mg l^–1) increased leaf CPT concentration (% of dry weight), whereas BA applications had no effect on total leaf CPT yield, the product of leaf CPT concentration and total leaf dry weight per seedling. There was a positive correlation between root weight and leaf CPT concentration under BA treatments. NAA supplementations (from 0.5 to 4 mg l^–1) to growth medium reduced plant height, leaf number, leaf length, specific leaf weight, plant weight, stem weight, and leaf weight compared with the NAA control. Meanwhile, there were no differences in plant height, leaf length, and specific leaf weight among the NAA supplementations. NAA applications had no effect on leaf CPT concentration and NAA applications decreased total leaf CPT yield. There were negative correlations between leaf number and leaf CPT concentration, leaf length and leaf CPT concentration under NAA treatments. Our results suggest that BA applications from 0.3 to 3 mg l^–1 are not helpful for achieving high total leaf CPT yield and NAA applications from 0.5 to 4 mg l^–1 decrease total leaf CPT yield.     

Microbial introductions to apple leaves: Influences of altered immigration on fungal community dynamics

Fungal immigration to apple leaves in the field was altered by the introduction of populations ofChaetomium globosum orAureobasidium pullulans to surface-disinfested leaves either immediately following, or 6 days after, disinfestation. Total numbers of fungal individuals and numbers of filamentous fungal and yeast individuals were estimated and compared over time for 4–7 weeks on control leaves (leaves disinfested but no populations applied), onAureobasidium-treated, and onChaetomium-treated leaves. Fungal communities developing on leaves during three experiments in two different time frames (experiment 1: July 9–August 27; experiments 2 and 3: July 29–August 27), and thus under different immigration regimes, were also compared. Survival of introduced populations was not related to the presence of prior fungal immigrants. Rates of increase in total numbers of fungi and numbers of filamentous fungi and yeasts per leaf varied among experiments, apparently in relation to differences in immigration and environmental history. Differences among leaves in immigration had a short-term (days) influence on community size. However, no long-term effects of altered immigration on phylloplane fungal community size were evident.     

Primary production and nitrogen allocation of field grown sugar maples in relation to nitrogen availability

Above ground net primary production (NPP), nitrogen (N) allocation, and retranslocation from senescing leaves were measured in 7 sugar-maple dominated sites having annual net N mineralization rates ranging from 26 to 94 kg · ha^–1 · yr^–1. The following responses were observed: (1) Green sun leaves on richer sites had higher N mass per unit leaf area than sun leaves on poorer sites; (2) Total canopy N varied much less than annual net mineralization, ranging from 81 to 111 kg · ha^–1; (3) This was due to the existence of a large and relatively constant pool of N which was retranslocated from senescing leaves for use the following year (54 to 80 kg · ha^–1); (4) The percentage of canopy N retranslocated by sugar maple was also relatively constant, but was slightly higher on the richer sites. Percent N in leaf litter did not change across the gradient; (5) Above ground NPP increased linearly in relation to N allocated above ground. Therefore, N use efficiency, expressed as above ground NPP divided by N allocated above ground was constant; (6) N use efficiency expressed as (NPP above ground/total N availability) was a curvilinear function of N availability; and (7) This pattern reflected a decreasing apparent allocation of N below ground with decreasing N availability.     

Natural and constrainment-induced factors influencing the breakdown of dogwood and oak leaves

Breakdown rates and microbial colonization patterns of dogwood and oak leaves were measured between November and June of 1987–88 and 1988–89. Leaves were placed in artificial streams loose (unconstrained), in bags, or in packs. Discharge was maintained at approximately 0.25 s^–1, and no shredders were present in the streams. Average microbial biomass as ATP, for all species and treatments, increased from near 0 mg g^–1 AFDW in November to over 8 mg g^–1 AFDW in June. Microbial respiration increased from about 0.01 µg glucose respired hr-g^–1 AFDW in November to about 0.03 µg hr-g^–1 AFDW in June. Microbial biomass and activity were significantly greater on dogwood leaves than on oak leaves. Dogwood and oak leaf breakdown rates were fastest when unconstrained, –0.0034 and –0.0027 degree-day^–1 respectively. Breakdown rates of dogwood leaves were faster in bags (–0.0025 degree-day^–1) than in packs (–0.0015 degree-day^–1) while rates of oak leaves were not significantly different between bags and packs (–0.0014 and –0.0018 degree-day^–1 respectively). Breakdown rates of dogwood and oak leaves obtained in this study were much slower than those obtained by other investigators either in the presence or absence of shredders. A comparison of results from this study with results from other studies revealed that dogwood leaves may be affected more by turbulence, while oak leaves may be influenced more by shredder activity.     

Recovery of photosynthesis in sunflower after a period of low leaf water potential

Photosynthesis was studied in sunflower plants subjected to 1 to 2 days of desiccation and then permitted to recover. The leaf water potential to which leaves returned after rewatering was dependent on the severity of desiccation and the evaporative conditions. Under moderately evaporative conditions, leaf water potential returned to predesiccation levels after 3 to 5 hours when desiccation was slight. Leaf water potentials remained below predesiccation levels for several days after rewatering when leaf water potentials decreased to −13 to −19 bars during desiccation. Leaf water potential showed no sign of recovery when leaf water potentials decreased to −20 bars or below during desiccation. The lack of full recovery of leaf water potential was attributable to increased resistance to water transport in the roots and stem. The resistance ultimately became large enough to result in death of the leaves because net water loss continued even after the soil had been rewatered.     

Ethylene-induced leaf abscission in cotton seedlings : the physiological bases for age-dependent differences in sensitivity

The speed of ethylene-induced leaf abscission in cotton (Gossypium hirsutum L. cv LG-102) seedlings is dependent on leaf position (i.e. physiological age). Fumigation of intact seedlings for 18 hours with 10 microliters per liter of ethylene resulted in 40% abscission of the still-expanding third true (3°) leaves but had no effect on the fully expanded first true (1°) leaves. After 42 hours of fumigation with 50 microliters per liter of ethylene, total abscission of the 3° leaves occurred while <50% abscission of the 1° leaves was observed. On a leaf basis, endogenous levels of free IAA in 1° leaves were approximately twice those of 3° leaves. Free IAA levels were reduced equally (approximately 55%) in both leaf types after 18 hours of ethylene (10 microliters per liter) treatment. Ethylene treatment of intact seedlings inhibited the basipetal movement of [¹⁴C]IAA in petiole segments isolated from both leaf types in a dose-dependent manner. The auxin transport inhibitor N-1-naphthylphthalamic acid increased the rate and extent of ethylene-induced leaf abscission at both leaf positions but did not alter the relative pattern of abscission. Abscission-zone explants prepared from 3° leaves abscised faster than 1° leaf explants when exposed to ethylene. Ethyleneinduced abscission of 3° explants was not appreciably inhibited by exogenous IAA while 1° explants exhibited a pronounced and protracted inhibition. The synthetic auxins 2,4-D and 1-naphthaleneacetic acid completely inhibited ethylene-induced abscission of both 1° and 3° explants for 40 hours. It is proposed that the differential abscission response of cotton seedling leaves is primarily a result of the limited abscission-inhibiting effects of IAA in the abscission zone of the younger leaves.     

The narrow sheath duplicate genes: sectors of dual aneuploidy reveal ancestrally conserved gene functions during maize leaf development

The narrow sheath mutant of maize displays a leaf and plant stature phenotype controlled by the duplicate factor mutations narrow sheath1 and narrow sheath2. Mutant leaves fail to develop a lateral domain that includes the leaf margins. Genetic data are presented to show that the narrow sheath mutations map to duplicated chromosomal regions, reflecting an ancestral duplication of the maize genome. Genetic and cytogenetic evidence indicates that the original mutation at narrow sheath2 is associated with a chromosomal inversion on the long arm of chromosome 4. Meristematic sectors of dual aneuploidy were generated, producing plants genetically mosaic for NARROW SHEATH function. These mosaic plants exhibited characteristic half-plant phenotypes, in which leaves from one side of the plant were of nonmutant morphology and leaves from the opposite side were of narrow sheath mutant phenotype. The data suggest that the narrow sheath duplicate genes may perform ancestrally conserved, redundant functions in the development of a lateral domain in the maize leaf.     

Insertion Profiles in Stomatal Density and Sizes in Nicotiana Tabacum L. Plantlets

Tobacco (Nicotiana tabacum L. cv. Samsun) plantlets were cultured in vitro on Murashige-Skoog medium photoautotrophically (without sucrose) or photomixotrophically (with 3 % sucrose) under two irradiances [70 or 230 µmol m^–2 s^–1]. Significant differences in stomatal density and sizes in leaves of different insertion levels (3^rd, 5^th and 7^th leaves from bottom) in photomixotrophic plantlets but not in photoautotrophic ones were found after 35 d of culture. Stomatal density was higher in upper leaves and on abaxial leaf side. Higher irradiance enhanced stomatal density in photoautotrophic plantlets. Stomatal sizes decreased with leaf insertion level but no significant differences between leaf sides were found. Abaxial stomata were more circular than the adaxial ones. In photomixotrophic plantlets stomata tended to be more elongated in the 3^rd and the 5^th leaves, whereas stomatal elongation in photoautotrophic plantlets was similar in all leaves.     

Ozone detoxification in the apoplasm and symplasm of spinach,broad bean and beech leaves at ambient and elevated concentrations of ozone in air

Spinach (Spinacia oleracea L.), broad bean (Vicia faba L.) and beech (Fagus sylvatica L.) plants were exposed to ozone at concentrations often measured in air during the summer months (120–300 g·m^–3) and antioxidants were determined in the leaf tissue and in the aqueous phase of the cell wall, the apoplasm. Concentrations of both reduced ascorbate (AA) and its oxidized form, dehydroascorbate (DHA), showed the tendency to increase transiently in the apoplasm of spinach leaves 6–24 h after starting fumigation with ozone. In beech leaves, apoplasmic AA and DHA increased 3–7 d after beginning of treatment. At the very high concentration of 1600 g O₃·m^–3, an increase of apoplasmic AA was already measured after 1 d in beech leaves. Apparently, spinach and beech leaves respond to oxidative stress by increasing AA transport into the apoplasm and by accelerating DHA export. In contrast to these observations, DHA accumulated during 3 d of fumigation with only 120 g O₃·m^–3 in the apoplasm of broad bean leaves, while AA contents did not increase. After termination of fumigation, the extracellular redox state of ascorbate normalized within 1 d. Glutathione could not be detected in the apoplasm of any of the three leaf species. Intracellular AA changed its redox state in response to exposure to elevated concentrations of ozone. After 4–6 weeks of fumigation with 200–300 g O₃·m^–3 an increase of intracellular DHA was measured in beech leaves. At the same time, chlorophyll contents decreased and characteristic symptoms of ozone damage could be observed. However, no significant change in the redox state of apoplasmic ascorbate could be detected in beech leaves. Evidently, detoxification of ozone by apoplasmic AA was insufficient to protect the leaf tissue. Fumigation with a high ozone concentration (1600 g·m^–3) caused an appreciable increase in the cellular contents of the oxidized forms of ascorbate and glutathione in beech leaves. Whereas in spinach leaves intracellular antioxidant contents and redox states were not altered during fumigation with 120–240 g O₃·m^–3, in broad bean leaves the intracellular DHA concentration increased and intracellular ascorbate became more oxidized after fumigation of the plants with 120 g O₃·m^–3. Apparently, broad bean leaves are more sensitive to ozone than beech and spinach leaves.Abbreviations AA ascorbate, reduced form - DHA ascorbate, oxidized form (dehydroascorbate) - FW fresh weight - GSH glutathione, reduced form - GSSG glutathione, oxidized form - IWF intercellular washing fluid - V_air intercellular air space volume of leaves - V_apo apoplasmic water volume of leaves This work was supported within the Sonderforschungsbereich 251 of the University of Würzburg.     

Do Polyamines Participate in the Long-Distance Translocation of Stress Signals in Plants?

Accumulation and ethylene-dependent translocation of free polyamines was studied in various organs, the phloem and xylem exudates of common ice plants (Mesembryanthemum crystallinum L.). Under normal conditions (23–25°C), spermidine predominated among polyamines. Cadaverine was found in old leaves, stems, and, in large quantities, in roots. The heat shock treatment (HS; 47°C, 2 h) of intact plant shoots induced intense evolution of ethylene from leaves but reduced the leaf content of polyamines. Under these conditions, the concentration of polyamines in roots, particularly of cadaverine, increased many times. The HS treatment of roots (40°C, 2 h) induced translocation of cadaverine to stems and putrescine to leaves. An enhanced polyamine content after HS treatment was also found in the xylem and phloem exudates. The exposure of detached leaves to ethylene led to a reduction in their putrescine and spermidine and accumulation of cadaverine, which implies the ethylene-dependent formation of cadaverine and a possible relation between the HS-induced translocation of this diamine to roots and the transient ethylene evolution by leaves. To validate this hypothesis, we compared the ethylene evolution rate and interorgan partitioning of cadaverine and other polyamines for two lines of Arabidopsis thaliana: the wild type (Col-0) and ein4 mutant with impaired ethylene reception. In plants grown in light at 20–21°C, the rate of ethylene evolution by rosetted leaves was higher in the mutant than in the wild type. The content of putrescine and spermidine was reduced in mutant leaves, whereas cadaverine concentration increased almost threefold compared with the wild type. In roots, cadaverine was found only in the wild type and not in the mutant line. Our data indicate the ethylene-dependent formation of cadaverine in leaves and possible involvement of cadaverine and ethylene in the long-distance translocation of stress (HS) signal in plants.     

Evaluation of DNA damage and mutagenicity induced by lead in tobacco plants

Tobacco (Nicotiana tabacum L. var. xanthi) seedlings were treated with aqueous solutions of lead nitrate (Pb²⁺) at concentrations ranging from 0.4 mM to 2.4 mM for 24 h and from 25 μM to 200 μM for 7 days. The DNA damage measured by the comet assay was high in the root nuclei, but in the leaf nuclei a slight but significant increase in DNA damage could be demonstrated only after a 7-day treatment with 200 μM Pb²⁺. In tobacco plants growing for 6 weeks in soil polluted with Pb²⁺ severe toxic effects, expressed by the decrease in leaf area, and a slight but significant increase in DNA damage were observed. The tobacco plants with increased levels of DNA damage were severely injured and showed stunted growth, distorted leaves and brown root tips. The frequency of somatic mutations in tobacco plants growing in the Pb²⁺-polluted soil did not significantly increase. Analytical studies by inductively coupled plasma optical emission spectrometry demonstrate that after a 24-h treatment of tobacco with 2.4 mM Pb²⁺, the accumulation of the heavy metal is 40-fold higher in the roots than in the above-ground biomass. Low Pb²⁺ accumulation in the above-ground parts may explain the lower levels or the absence of Pb²⁺-induced DNA damage in leaves.     

Phenotypic analysis and molecular cloning of discolored-1 (dsc1), a maize gene required for early kernel development

Recessive mutations in the maize dsc1 locus prevent normal kernel development. Solidification of the endosperm in homozygous dsc1– mutant kernels was undetectable 12 days after pollination, at which time the tissue was apparently completely solidified in wild-type kernels. At later times endosperm did solidify in homozygous dsc1– mutant kernels, but there was a marked reduction in the volume of the tissue. Embryo growth in homozygous dsc1– kernels was delayed compared to wild-type kernels, but proceeded to an apparently normal stage 1 in which the scutellum, coleoptile, and shoot apex were clearly defined. Embryo growth then ceased and the embryonic tissues degraded. Late in kernel development no tissue distinctions were obvious in dsc1– mutant embryos. Immature mutant embryos germinated when transplanted from kernels to tissue culture medium prior to embryonic degeneration, but only coleoptile proliferation was observed. The dsc1 gene was isolated by transposon tagging. Analysis of the two different dsc1– mutations confirmed that transposon insertion into the cloned genomic locus was responsible for the observed phenotype. Dsc1 mRNA was detected specifically in kernels 5–7 days after pollination. These data indicate Dsc1 function is required for progression of embryo development beyond a specific stage, and also is required for endosperm development.