Growth parameters and resistance against Drechslera teres of spring barley (Hordeum vulgare L. cv. Scarlett) grown at elevated ozone and carbon dioxide concentrations.

Spring barley ( Hordeum vulgare L. cv. Scarlett) was grown at two CO2 levels (400 vs. 700 ppm) combined with two ozone regimes (ambient vs. double ambient) in climate chambers for four weeks, beginning at seedling emergence. Elevated CO2 concentration significantly increased aboveground biomass, root biomass, and tiller number, whereas double ambient ozone significantly decreased these parameters. These ozone-induced reductions in growth parameters were strongly overridden by 700 ppm CO2. The elevated CO2 level increased C : N ratio of the leaf tissue and leaf starch content but decreased leaf protein levels. Exposure to double ambient ozone did not affect protein content and C : N ratio but dramatically increased leaf starch levels at 700 ppm CO2. Resistance against Drechslera teres (Sacc.) Shoemaker was increased in leaves grown at double ambient ozone but was less obvious at 700 ppm than at 400 ppm CO2. Constitutive activities of beta-1,3-glucanase and chitinase were significantly higher in leaves grown at double ambient ozone compared to ambient ozone levels. The sum of methanol-soluble and alkali-released cell wall-bound aromatic metabolites (i.e., C-glycosylflavones and several structurally unidentified metabolites) and lignin contents did not show any treatment-dependent differences.

Gene induction of stilbene biosynthesis in Scots pine in response to ozone treatment, wounding, and fungal infection.

The S-adenosyl-L-methionine:pinosylvin-O-methyltransferase (PMT) gene was sequenced from Scots pine (Pinus sylvestris). The open reading frame is arranged in two exons spaced by one 102-bp intron. Promoter regulatory elements such as two "CAAT" boxes and one "TATA" box were identified. Several cis-regulatory elements were recognized: stress-responsive elements (Myb-responsive elements) as well as G, H, and GC boxes. Moreover, elicitor-responsive elements (W boxes) and a sequence resembling the simian virus 40 enhancer core were found. In phloem and needles of control trees, the transcripts of stilbene synthase (STS) and PMT were hardly detectable. Increased ozone fumigation up to 0.3 microL L(-1) enhanced the transcript level of STS and PMT in needles but not in healthy phloem. Wounding, e.g. mock inoculation, of stem-phloem was characterized by a transient increase in STS and PMT transcripts, which was more pronounced in the case of fungal inoculation. Combination of fungal-challenge or mock treatment with ozone resulted in a positive interaction at 0.3 microL L(-1). Scots pine stilbene formation appeared to be induced via STS and PMT gene expression upon ozone and fungal stress as well as wounding. The broad stress-responsiveness is in agreement with the range of various cis-acting elements detected in the STS and PMT promoters.

Effects of ozone, acid mist and soil characteristics on clonal Norway spruce (Picea abies (L.) Karst.)--an introduction to the joint 14 month tree exposure experiment in closed chambers.

This paper introduces a series of publications referring to a single 14-month laboratory study testing the hypothesis that the recent decline of Norway spruce (Picea abies (L.) Karst.) at higher elevations of the Bavarian Forest and comparable forests in medium-range mountains and in the calcareous Alps is caused by an interaction of elevated ozone concentrations, acid mist and site-specific soil (nutritional) characteristics. The effect of climatic extremes, a further important factor, was not included as an experimental variable but was considered by testing of the frost resistance of the experimental plants. Results of these individual studies are presented and discussed in the following 14 papers. Plants from six pre-selected clones of 3-year-old Norway spruce (Picea abies (L.) Karst.) were planted in April 1985 in an acidic soil from the Bavarian Forest, or a calcareous soil from the Bavarian Alps. After a transition period, plants were transferred, in July 1986, into four large environmental chambers and exposed for 14 months to an artificial climate and air pollutant regime based on long-term monitoring in the Inner Bavarian Forest. The climatic exposure protocol followed realistic seasonal and diurnal cycles (summer maximum temperature, 26 degrees C; total mean temperature, 9.8 degrees C; winter minimum, -14 degrees C; mean relative humidity, 70%; maximum irradiance, 500 W m(-2); daylength summer maximum, 17 h; winter minimum, 8 h). Plants were fumigated with ozone, generated from pure oxygen (control: annual mean of 50 microg m(-3); pollution treatment: annual mean of 100 microg m(-3) with 68 episodes of 130-360 microg m(-3) lasting 4-24 h), and background concentrations of SO(2) (22 microg m(-3)) and NO(2) (20 microg m(-3)); windspeed was set at a constant 0.6 m s(-1). Plants were additionally exposed to prolonged episodes of misting at pH 5.6 (control) and pH 3.0 (treatment). Simulation of the target climatic and fumigation conditions was highly reliable and reproducible (temperature +/-0.5 degrees C; rh+/-10%; ozone+/-10 microg m(-3);SO(2) and NO(2)+/-15 microg m(-3)).

Clone and soil effects on the growth of young Norway spruce during 14 months exposure to ozone plus acid mist.

Five clones of 3-year old Norway spruce (Picea abies [L.] Karst), planted in a soil from the Bavarian Forest (pH 4.4) or a soil from the Calcareous Bavarian Alps (pH 6.9), were exposed for two successive vegetation periods, in closed environmental chambers, to a pollution treatment consisting of acidic mist (pH 3.0) plus ozone levels of 100 microg m(-3) with episodes of 130-360 microg m(-3); control trees were exposed to mist of pH 5.6 and ozone levels of 50 microg m(-3). Climatic and pollution protocols followed the diurnal and seasonal pattern characteristic for the Inner Bavarian Forest in Southern Germany, an area affected by the new-type forest decline. Biometric parameters were strongly related to clone and soil. Pollution treatment had a limited effect on only a few growth parameters. The stem diameter growth increment of two clones was reduced by pollution treatment in both soils, a third clone was affected in the acidic soil only. Two other clones were not affected at all. Stem volume increment of three clones, calculated as D(2)H, was reduced by pollution treatment in the neutral soil, a fourth clone was affected in the acidic soil only. Bud break was either delayed (two clones) or accelerated (two other clones) by treatment. Depending on soil and clone, needle yellowing was observed in previous years' needles in both treatment and control trees exposed to increased light intensities. The 'spotted' yellowing was not identical to symptoms found in forest decline areas and was most likely a consequence of nutrient deficiencies during the vegetation period preceding the experiment. The results of this experiment are discussed with regard to field observations and forest productivity. The complex pattern of growth responses resulting from interactions between air pollution, soil and genetic factors is considered to reflect different susceptibilities of trees to air pollutants.

Effects of low-level long-term ozone fumigation and acid mist on photosynthesis and stomata of clonal Norway spruce (Picea abies (L.) Karst.).

Four clones of 3-year-old Norway spruce (Picea abies (L.) Karst.), grown on two soils, were from July 1986 to September 1987 exposed to ozone fumigation (50 microg m(-3) as a control, 100 microg m(-3) plus peaks between 130 and 360 microg m(-3) as treatment) and acid mist of pH 3.0 (versus mist pH 5.6 in the control). Climatic conditions, identical for both control and treatment, followed a diurnal and seasonal pattern characteristic of medium high altitudes of the Bavarian Forest, an area affected by the new-type forest decline. Gas-exchange measurements were carried out on the plants from December 1986 until the end of the 14-month's exposure using a series of climate-controlled minicuvettes. ANOVA of the four clones investigated towards the end of the experiment gave hints of a treatment-related depression of the photosynthetic capacity of the previous year's needles (age-class 1986). Within this age-class only one of the clones (11) showed a significant treatment effect, indicating an age-class dependence and a genetic influence of the treatment-related depression of the photosynthetic capacity. The current year's flush was not impaired through the ozone and acid mist exposure. Analysis also revealed clear effects of soil, clone and needle age on photosynthetic parameters.

Effects of ozone, acid mist and soil characteristics on clonal Norway spruce (Picea abies (L.) Karst.)--overall results and conclusions of the joint 14 month tree exposure experiment in closed chambers.

This paper summarizes and evaluates the main findings of 14 preceding papers related to the joint 14-month tree-exposure experiment carried out by the 'Munich Working Party on Air Pollution' at the GSF, Munich, FRG, from July 1986 to September 1987. The experiment tested the hypothesis that an interaction of ozone/acid mist/soil/extreme climatic conditions is the cause of decline of Norway spruce (Picea abies (L.) Karst.) at higher altitudes of the Inner Bavarian Forest. The main findings of the individual studies are presented and their implications for the hypothesis are discussed. Clear effects of soil and genetic factors (differences between clones), for example on growth and frost resistance were found. Treatment with O(3)/acid mist was shown to have effects on plant biochemistry, physiology, histology/ cytology, and growth. The wide scattering of these effects, and the lack of a consistent pattern of response across all clones does not permits a firm conclusion on the validity of the experimental hypothesis. These effects were not confounded by the nutrient stresses imposed during the initial exposure period and were not found to be cumulative during repeated treatments, as was proposed by the hypothesis. It is concluded that the experimental evidence does not indicate that ozone/acid mist are major factors to explain the Norway spruce decline on acidic sites at higher altitudes of the Inner Bavarian Forest and probably similar forest areas.

Concentrations of lead, cadmium, mercury and other elements in seeds of Lupinus mutabilis and of other legumes.

Several species of legumes and varieties of lupins were examined for their heavy metal content in the seeds. Atomic absorption spectrometry was the main analytical tool. Validity and reproducibility of the results was checked by analyzing the same materials at two different laboratories performing the same technique. Additionally, inductively coupled plasma emission spectroscopy was applied. In grains of Lupinus mutabilis which constitute a traditional food in Andean populations, the following contents of heavy metals were determined: 0.10-0.25 microgram/g Cd; 0.5-1.6 microgram/g Pb; 0.10-0.15 microgram/g Hg. Seeds of other legumes contained 0.05-0.35 microgram/g Cd; 0.1-0.2 microgram/g Pb; 0.01-0.04 microgram/g Hg. The high lead content of lupins is easily reduced to one tenth by traditional extraction with boiling water. The mercury content, too, is decreased by this technique. The high manganese content of 1 300-1,400 micrograms/g of Lupinus albus compared to other legumes (25-37 micrograms/g) seems to be characteristic for this species and may be of nutritional significance. The comparison of the heavy metal contents of legume seeds of different origin and variety indicates a complex pattern of environmental and genetic factors that contribute to the specific metal contents of individual harvests. On the level of varieties the environmental factors (climate, soil, geology, agricultural techniques) seem to exhibit more important influences on the specific accumulation of heavy metals than genetic factors. In contrast, on the level of species or genera, the accumulation of heavy metals seems to be dominated by genetic factors rather than by environmental influences.

Monthly deposition of cadmium in rural and industrial areas of Germany (Bayern, Pfalz, Ruhr district) and its influences upon an agricultural model system.

Monthly depositions of cadmium were collected by a modified Bergerhoff method and measured by AAS during a 3-year period in rural areas of the Pfalz and in an industrial area of the Ruhr district. Another one year period included measurements in rural areas of southern Bavaria and on a Dutch island. The log-normally distributed deposition rates of cadmium at the rural areas in southern Germany amounted to only 20% of those of the industrial district. The depositions on the Dutch island were twice as high as the depositions on the rural areas of southern Germany. The monthly cadmium deposition rates show only little periodical fluctuation during the year and scatter around more or less constant median values of 25 and 120 micrograms . m-2 . month-1 at the rural and industrial areas, respectively. When open air mass cultures of algae were taken as an agricultural model, the organisms, depending on their growth rate, accumulated 0.4-4.0 ppm of cadmium (dry matter based). The course of the cadmium accumulation reflects the deposition rate of the area where the algae were grown. No growth depression of the algae due to cadmium can be observed under the given deposition rates.

Effects of blue and red light on photosynthetic (14)CO 2 uptake, and distribution of (14)C in free and proteinbound amino acids in fern gametophytes [Dryopteris filix-mas (L) SCHOTT.

Morphogenesis and metabolism of the early gametophytes (= sporelings) of the common male fern are controlled by light. The "normal" two-dimensional development of the gametophytes in white or blue light is correlated with an increase in protein content; inred light alone, on the other hand, the sporelings remain filamentous, and the protein content is markedly lower (cf. MOHR, 1965). The problem has been whether blue light increases the rate of protein synthesis or decreases the rate of protein degradation. This problem was solved in the present paper by the use of (14)CO2. Blue light promotes specifically the rate of protein synthesis as indicated by the increase of (14)C incorporation into protein-bound amino acids under blue light as compared with red light.Using (14)CO2 we have analyzed the kinetics of free amino acid synthesis (Fig. 4) and protein synthesis (Fig. 5) under steady state conditions of photosynthetic CO2 incorporation in blue or red light (Fig. 3). Under our conditions the rate of photosynthesis is about 1.5 times higher under blue light than under red light (Fig. 3, Table 1).The facts that the total pool sizes of the free amino acids are smaller in blue than in red light (V. DEIMLING and MOHR, 1967; Table 2) and that, on the other hand, the (14)C-contents of the thoroughly labelled amino acid pools are virtually identical in blue and red (Table 3) indicate (a) that the pool sizes of these labelled amino acids may be equal in both light qualities and (b) that there is a compartmentation of free amino acid pools in the fern sporeling. This problem will be dealt with more in detail in a forthcoming paper on the behaviour of alanine in the fern sporeling (PAYER, 1969).Protein synthesis is obviously much stronger under blue light than under red light. The detailed kinetics (Fig. 5b) indicate the involvement of two sorts of proteins: a relatively small part with high turnover which is rapidly labelled with a small but significant difference in red and blue, and a larger part with a slower turnover, the synthesis of which is strongly favored by blue light. - The first sort could be enzyme protein; the latter sort might be structural protein of the chloroplasts. These organelles increase dramaticly in size under the influence of blue light (BERGFELD, 1963). The amino acid composition of the protein, however, does not show any qualitative difference in gametophytes grown in blue or red light (V. DEIMLING and MOHR, 1967, Table 4).

[Compartmentation of free alanine and its consequences for the quantitative consideration of protein synthesis: Experiments with fern gametophytes [Dryopteris filix-mas (L.) SCHOTT] in blue and red light]. / Untersuchungen zur Kompartimentierung der freien Aminosäure Alanin in den Farnvorkeimen von Dryopteris filix-mas (L.) Schott im Rotlicht und im Blaulicht.

In fern gametophytes (= sporelings) there is a strong correlation between the degree of blue light mediated photomorphogenesis and the protein content of the organism (cf. MOHR, 1963). In a previous paper (PAYER et al., 1969) we have shown that blue light specifically increases the rate of protein synthesis in the fern sporelings over the rate which is maintained under red light. - In the present paper blue light mediated protein synthesis has been dealt with further using one representative amino acid, alanine, which was labelled with (14)C from (14)CO2 under steady state conditions of photosynthetic (14)C incorporation under blue or red light.Synthesis of free alanine is proportional to the rate of photosynthesis (Table 1). For a number of reasons we conclude that alanine is derived directly from primary photosynthetic products. Since the pool size of the thoroughly (14)C-labelled pool of free alanine is much less than the actual, pool size of this amino acid, (Table 1), and since the specific activity of the isolated (14)C-alanine is much below the value we can expect on the basis of the specific activity of the (14)CO2 applied we conclude that there are separate pools of free alanine; "active" (with respect to protein synthesis) and "inactive" pools which do not mingle. Taking into account this possibility of compartmentation of pools of free amino acids we have calculated in the case of (14)C-alanine the rate of protein synthesis for two extreme instances (Table 2). A comparison of the theoretical values with the actual data indicates that indeed protein synthesis is fed from "active" pools of amino acids while the "inactive" pools are possibly located in the vacuoles. The total pool of alanine is much larger in red grown than in blue grown sporelings while the active pools seem to have the same size under both conditions. The cells of the red grown sporelings have much larger vacuoles than the cells of the blue grown sporelings.The rate of protein synthesis is under our conditions 1.8 times higher in blue light than in red light. The rate of turnover of the total protein is 0.29% per hour in the blue and 0.23% in the red light. The absolute turnover of protein is 1.5 times higher in blue light than in red light. It is concluded that the blue light mediated increase of protein synthesis is very real. Blue light must act specifically at the level of polypeptide synthesis.

[A specific effect of blue light on the incorporation of photosynthetically assimilated (14)C into the protein of fern sporelings [dryopteris filix-mas (L.) Schott]]. / Ein spezifischer Einfluß von Blaulicht auf den Einhau von photosynthetisch assimiliertem (14)C in das Protein von Farnvorkeimen [Dryopteris filix-mas (L.) Schott].

Morphogenesis and metabolism of the early gametophytes (=sporelings) of the common male fern are controlled by light. The "normal" two-dimensional development of the gametophytes takes place only in white or blue light; in red light alone, on the other hand, the sporelings remain filamentous even under conditions of equal photosynthetic rate.The problem has been whether blue light exerts its morphogenic influence through differential gene activation. In other words: does blue light mediate the synthesis of "morphogenic enzymes" which are required for "normal" morphogenesis. In an earlier paper (DRUMM and MOHR, 1967) we have shown that blue light increases the rate of RNA synthesis within an hour whereas the first indication of a morphogenic change due to blue light is only discernible about 3 hours after the onset of blue light (Figs. 1,2). Furthermore we have shown (MOHR, 1965) that Actinomycin D specifically inhibits the blue light mediated morphogenic alterations, and BERGFELD (1967) has shown that blue light will rapidly lead to changes in nuclei and nucleoli in the fern sporelings. In the present paper it has been shown that blue light does increase the rate of protein synthesis about an hour after the transfer of the sporelings from the red into the blue light of equal quantum flux density (350 pE·cm(-2)·s(-1)).The rate of protein synthesis was measured in shortterm experiments (40min) using (14)CO2. The photosynthetic rate was the same in red and blue; it was not influenced by the transfer(Fig. 3). Likewise the rate of (14)C incorporation into the pool of free amino acids was not significantly different in red and blue light (Fig.4). On the other hand, the rate of incorporation of (14)C into the protein increased rapidly after the transfer of the sporelings from the red into the blue light (Fig. 5). The same phenomenon (no influence of blue light on the specific activity of the free amino acid; a strong promotive influence on the specific activity of the protein-bound amino acid) was observed in the case of alanine which was investigated in detail (Figs. 6, 7). Since the increase of the protein content of the sporelings is not significant during the first six hours after transfer to blue light (Fig. 8) the protein induced by blue light and directly related to morphogenesis can only be a very small fraction of the total protein of the sporeling.The data strongly support the hypothesis (OHLENROTH and MOHR, 1964), that the morphogenic effect of blue light on the fern sporelings is due to the induction of "morphogenic enzymes" by blue light.