Ein spezifischer Einfluß von Blaulicht auf den Einhau von photosynthetisch assimiliertem 14C in das Protein von Farnvorkeimen [Dryopteris filix-mas (L.) Schott]

Summary 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 ¹⁴CO₂. The photosynthetic rate was the same in red and blue; it was not influenced by the transfer(Fig. 3). Likewise the rate of ¹⁴C 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 ¹⁴C 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.     

Untersuchungen zur Kompartimentierung der freien Aminosäure Alanin in den Farnvorkeimen von Dryopteris filix-mas (L.) Schott im Rotlicht und im Blaulicht

Summary 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 ¹⁴C from ¹⁴CO₂ under steady state conditions of photosynthetic ¹⁴C 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 ¹⁴C-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 ¹⁴C-alanine is much below the value we can expect on the basis of the specific activity of the ¹⁴CO₂ 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 ¹⁴C-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.     

Eine Analyse der durch Blaulicht bewirkten Steigerung der Proteinsynthese bei Farnvorkeimen auf der Ebene der Aminosäuren

Zusammenfassung Bei der quantitativen Analyse der Protein-Aminosäuren von Farnvorkeimen (Dryopteris filix-mas) ergab sich, daß die Steigerung des Proteingehalts durch Blaulicht mit einer Zunahme aller untersuchten Aminosäuren einhergeht. Dabei bleibt der prozentuale Anteil der einzelnen Aminosäuren am Gesamtprotein immer annähernd gleich, abgesehen vom Prolin. — Offenbar kommt es unter dem Einfluß von Blaulicht hauptsächlich zu einer quantitativen Vermehrung und nur im geringen Maße zu einer qualitativen Veränderung der Proteinfraktion. — Da die pools der erfaßbaren freien Aminosäuren im Hellrot stets größer sind als im Blaulicht, ist der Schluß berechtigt, daß das Blaulicht seine Wirkung im Zusammenhang mit der Polypeptidsynthese und nicht über eine Steigerung der Aminosäuresynthese ausübt.

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An analysis of the blue light mediated increase of protein synthesis in fern gametophytes on the level of amino acids

Summary Morphogenesis and metabolism of the sporelings (= young gametophytes) of the common male fern Dryopteris filix-max are controlled by visible radiation. Short wavelengths visible radiation (= blue light) leads to an increase in protein synthesis and makes possible the formation of normal two-dimensional prothallia. Under long wavelengths visible radiation (= red light) the sporelings grow as cellular filaments, the protein contents of which are markedly lower than under blue light even under conditions of equal rate of dry matter accumulation in red and blue light (Fig. 2). — Quantitative amino acid analysis of the total protein of blue and red grown sporelings did not reveal any striking difference. The contents of all those amino acids which can be measured quantitatively after protein hydrolysis show about the same increase under the influence of blue light (Figs. 3, 4). Only in the case of proline are the differences between red grown and blue grown sporelings indicative of a qualitative change in the nature of the protein fraction (Fig. 3/h). —The pools of the free amino acids are always smaller in blue grown sporelings than in red grown ones (Figs. 6, 7). — The facts reported in this paper indicate that blue light leads mainly to a quantitative increase in the rate of protein synthesis. The influence of blue light on the qualitative nature of the protein fraction seems to be slight. On the other hand, the data on the pool sizes of the free amino acids support the conclusion that blue light controls the rate of protein synthesis at the stage of polypeptide synthesis and not through amino acid synthesis.

     

Die regulation der DNS-Synthese in Farngametophyten durch Licht

Zusammenfassung Bei Farnvorkeimen (Dryopteris filix-mas) steigert Blaulicht spezifisch DNS-Replikation und mitotische Aktivität, so daß sich die im Hellrot bzw. Blaulicht herangewachsenen Vorkeime, die gleichviel Trockensubstanz besitzen, bezüglich ihres DNS-Gehalts und ihrer Zellzahl wesentlich unterscheiden — Die Zelle scheint als Bezugssystem für RNS, Protein und entsprechende biochemische Größen ungeeignet zu sein. Der Gesamtkeimling ist das angemessene Bezugssystem.

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The regulation of DNA synthesis in fern gametophytes by light

Summary In young gametophytes (= sporelings) of the common male fern (Dryopteris filix-mas) short wavelength light (= blue light) specifically increases DNA replication and mitotic activity even if the sporelings have the same growth rate under blue and red light, as determined by dry matter increase. Cell number and DNA content of sporelings of the same age and the same dry matter content are much higher under blue than under red light (Figs. 1, 2). In the second part of the paper it is pointed out that the cell (or unit of DNA) may not be used as a system of reference for biochemical data such as protein or RNA content in the case of the fern sporelings (and possibly not in many other organisms either). The appropriate system of reference seems to be the entire multicellular system if precaution is taken that the systems grow with the same growth rate under the different experimental conditions (e.g. under blue and red light in the case of the fern sporelings).

     

Die Regulation der RNS-Synthese in Farngametophyten Durch Licht

Zusammenfassung In einer früheren Arbeit wurde gezeigt (Ohlenroth und Mohr, 1963), daß sich die Vorkeime von Dryopteris filix-mas im Blaulicht zu normalen zweidimensionalen Prothallien mit einem relativ hohen Proteingehalt entwickeln. Im Hellrot hingegen bilden sich Zellfäden (= Protonemen) aus, deren Proteingehalt bei gleicher Photosyntheseleistung wesentlich geringer ist.In der vorliegenden Arbeit wird gezeigt, daß auch der RNS-Gehalt im Blaulicht stets größer als im Hellrot ist. Die blaulichtabhängige RNS-Zunahme setzt zeitlich früher ein als die Proteinzunahme.In Umsetzexperimenten von Hellrot nach Blau manifestiert sich die morphogenetische Umsteuerung der Protonemen zeitlich eher als die blaulichtabhängige Steigerung des Proteingehalts. Kasemir und Mohr (1965) konnten zeigen, daß es sich bei dem Blaulichtprotein in erster Linie um eine Vermehrung des Strukturproteins der Chloroplasten hanhandelt. Die blaulichtabhängige RNS-Zunahme dagegen ist spätestens zum Zeitpunkt der morphologischen Umsteuerung faßbar. Dieses Ergebnis wird dadurch gestützt, daß Blaulicht im Vergleich zu Hellrot rasch einen gesteigerten Einbau von ¹⁴C, als ¹⁴C-Uridin (U) geboten, in die RNS-Fraktion verursacht. Das Blaulicht scheint in den Farnvorkeimen zwei verschiedene Vorgänge zu verursachen. 1. Steigerung einer autonomen Proteinsynthese in den Chloroplasten. 2. Auslösung oder Steigerung einer spezifischen Enzymsynthese im Cytoplasma. Die blaulichtabhängige Steigerung der RNS-Synthese scheint damit in Zusammenhang zu stehen. Die Daten der vorliegenden Arbeit werden als Indizien dafür angesehen, daß das Blaulicht seine morphogenetische Wirkung über eine differentielle Genaktivierung ausübt.

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The regulation of RNA synthesis in fern gametophytes by light

Summary Morphogenesis and metabolism of the gametophytes (= sporelings) of the common male fern Dryopteris filix-mas are controlled by visible radiation. Short wavelengths visible radiation (= blue light) leads to an increase in protein synthesis and makes possible the formation of normal two-dimensional prothallia. Under long wavelengths visible radiation (= red light) the sporelings grow as cellular filaments the protein contents of which are markedly lower than under blue irradiation even under conditions of equal rate of dry matter accumulation in red and blue (Ohlenroth and Mohr, 1963). — It is shown in the present paper that the RNA content of sporelings of the same age is always higher in blue light than in red light (Figs. 1, 3). The blue-dependent increase of RNA occurs faster than the blue-dependent increase of protein (Fig. 2). Furthermore the increase of protein per sporeling is much larger than the increase of RNA (Fig. 4). These facts are in agreement with the hypothesis that in some way or another blue light initiates differential gene activation.The blue light-dependent morphological changes which occur if we put red grown filamentous sporelings under blue light can be measured much faster than the blue light-dependent increase of the bulk protein (Figs. 5, 6). We have to conclude as we did in a previous paper (Kasemir and Mohr, 1965) that the blue light-dependent increase in the protein content of the sporelings might be mainly due to an increase of chloroplast protein. — The blue light-dependent increase of the RNA content occurs at least as fast as the morphological changes (Figs. 5, 6). This finding is supplemented by the observation (Fig. 8) that blue light markedly and rapidly stimulates the incorporation of ¹⁴C into RNA. The ¹⁴C was applied as ¹⁴C-uridine (U). — It seems that blue light causes an increase of protein synthesis in the chloroplasts as well as in the cytoplasm. Blue light-dependent RNA synthesis seems to be involved in this response. These data support the view that blue light might exert its morphogenetic control through differential gene activation.

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Die Arbeit wurde durch Sachbeihilfen der Deutschen Forschunggemeinschaft und der Stiftung Volkswagenwerk gefördert.     

Die Aufnahme von 14CO2 und die Verteilung des 14C auf freie Aminosäuren und auf Proteinaminosäuren im Hellrot und im Blaulicht. [Objekt: Farnvorkeime von Dryopteris filix-mas (L.) Schott]

Summary 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 ¹⁴CO₂. Blue light promotes specifically the rate of protein synthesis as indicated by the increase of ¹⁴C incorporation into protein-bound amino acids under blue light as compared with red light.Using ¹⁴CO₂ 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 ¹⁴C-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).

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Die Aufnahme von ¹⁴CO₂ und die Verteilung des ¹⁴C auf freie Aminosäuren und auf Proteinaminosäuren im Hellrot und im Blaulicht. [Objekt: Farnvorkeime von Dryopteris filix-mas (L.) Schott]

     

Photosynthese und Photomorphogenese bei Farnvorkeimen vonDryopteris filix-mas

Summary Under conditions of identical rate of photosynthesis (measured by dry weight increase under steady state conditions) growth and differentiation of the gametophytes of ferns (e.g.Dryopteris filix-mas) are completely different in red and blue light. In the blue light normal growth and morphogenesis take place and normal two or three-dimensional prothallia are formed (Fig. 3). In the red, however, the prothallia look very similar to those growing in complete darkness: they grow as one-dimensional filaments (Fig. 1).It has been shown in this paper that photosynthesis, which is important as a source of organic material and free energy, has no influence at all on morphogenesis. Morphogenesis, i.e. the formation of normal prothallia instead of filaments, is controlled by a photoreactive system which depends on blue light of suitable intensity and which is not related to photosynthesis as such. If no blue light is present no morphogenesis occurs in spite of high photosynthetic activity.In our opinion theprimary products of photosynthesis are the same in all wavelengths. But now the photomorphogenic light reaction which depends on blue light apparently directs the flow of metabolites. In this way even the same initial products of CO₂ fixation may lead subsequently to rather different photosynthetic products and consequently to the very great difference between prothallia growing with or without blue light.The addition of sucrose has practically no influence on growth and morphogenesis under our conditions. On the basis of our results we cannot agree with the general conclusions drawn byMiller andMiller (1961) who regard photosynthesis as a photomorphogenetic system in these gametophytes of ferns.Mit 6 Textabbildungen.Herrn Professor Dr.E. G. Pringsheim in Verehrung zum 80. Geburtstag.     

RNA and protein metabolism in the particulate fractions of the gametophytes of bracken fern during growth in red and blue light

Summary The metabolism of RNA and protein in the gametophytes of bracken fern (Pteridium aquilinum) is affected by the quality of light in which they are grown. When sporelings were grown as two-dimensional gametophytes in blue light, particulate fractions separated from the sporelings exhibited greater incorporation of uridine-³H and leucine-³H into RNA and protein, respectively, than those from sporelings grown as one-dimensional protonema in red light. After various periods of exposure of gametophytes to red or blue light in the presence of uridine-³H, the nuclei-rich fraction showed the highest specific activity in RNA, and irrespective of incubation time, blue light was more effective than red light. The possibility that enhanced synthesis of RNA in the nucleus in response to blue light is significantly related to the morphological growth pattern of the gametophytes, is discussed.     

Der Einfluss von Farblicht auf Wachstum und Zusammensetzung Pflanzlicher Gewebekulturen

Summary The growth of green cultures of callus tissue from Nicotiana tabacum var. Samsun is stimulated by light. To determine whether the increase in growth is caused by photosynthesis or by a blue light dependent increase of protein synthesis, a comparative study was made of the effect which blue and red light have on the growth and the composition of tobacco tissue. It is shown that the growth stimulation by light depends on the chlorophyll content of the tissues. Starting with chlorophyll-free tissue the cultures begin to grow faster in blue light only after they become visibly green. On the other hand, the growth of green tissue in red light decreases as soon as the chlorophyll content under this condition becomes less. There are no differences in the rate of growth of green tissues cultivated in blue and in red light of approximately the same flow of quanta; in both cases the cultures grow better than the controls in the dark. Furthermore there are no differences between the protein and carbohydrate content of tissues grown in blue or red light and in the dark. There is, however, a small but significant difference between the total nitrogen of green tissue and that of chlorophyll-free tissue which is due to a higher amount of soluble nitrogen in the green tissue. From these results it is concluded that the light dependent growth stimulation is caused by photosynthesis. As shown by a light dependent ¹⁴CO₂ incorporation in which sucrose is the main product, the green cells are able to fix CO₂ photosynthetically. However, the rate of photosynthesis in the tissue cultures is small and does not balance the respiration. It seems very unlikely, therefore, that the formation of carbohydrates by photosynthesis is responsible for the observed growth increase.     

Beeinflussung der Morphogenese,Substanzproduktion und Proteinzunahme vonAcetabularia mediterranea durch sichtbare Strahlung

Zusammenfassung Während in Blaulicht die Entwicklung vonAcetabularia mediterranea normal verläuft, wird in Rotlicht das Zellwachstum nach 2–3 Wochen fast vollständig eingestellt. Hutbildung konnte in Rotlicht nur dann beobachtet werden und auch nur unmittelbar nach der übertragung des Rotlichts, wenn mit Zellen im Stadium kurz vor der Hutbildung gearbeitet wurde.An Zellen, die nach Vorbehandlung mit Rotlicht aufgehört haben zu wachsen, kann Wachstum wieder induziert werden, wenn sie zusätzlich zum Rotlicht kurzfristig mit Blaulicht bestrahlt werden. Die Wachstumsgeschwindigkeit (Zunahme der Stiellänge) ist dabei zwischen 10 sec und 1 Std. Blaulicht täglich proportional dem Logarithmus der Bestrahlungsdauer. Die Hutbildung wird um so stärker gefördert, je mehr Blaulicht die Zellen zusätzlich erhalten.Da beiAcetabularia Wachstumsvorgänge mit einer Substanzzunahme gekoppelt sind, wird in Rotlicht die Substanzproduktion frühzeitig eingestellt, verläuft in Blaulicht dagegen normal. Entsprechend wird durch zusätzliche Blaulichtgaben auch die Substanzproduktion gefördert.Eine genauere Untersuchung der wirksamen Spektralbereiche ergab, daß nur Strahlung mit Wellenlängen < 545 nm einen fördernden Einfluß auf Zellwachstum und Substanzproduktion hat. Außerdem wird durch diesen Spektralbereich auch die Proteinvermehrung spezifisch gefördert, allerdings nur bei Dauerbestrahlung.Das Verhalten der Zellen in Rotlicht beruht nicht auf einem Mangel an morphogenetischen Substanzen.

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Summary In blue light cell parts ofAcetabularia mediterranea grow normally, they form stalks and finally caps. In red light, however, growth processes cease almost completely after 2–3 weeks. Cells in red light do not form caps, except those in the stage just before cap formation and then only immediatly after transfering to red light.Additional blue light to red light induces growth on cells pretreated with red light. Between 10 sec and 1 h of blue light per day, the growth rate (increase of stalk length) is proportional to the logarithm of the amount of additional blue light. Cap formation is also promoted by blue light.InAcetabularia, there is a close correlation between growth processes and increase of dry weight. Therefore, in blue light dry matter increases continuously but is limited in red light. Here again, additional breaks with blue light stimulate the formation of dry matter.A close examination of the different spectral regions shows that only radiation with wavelength < 545 nm promotes cell growth and formation of dry matter. In addition, there is also a specific promotion of protein synthesis in this spectral region, but only in continuous light.The reduction of growth processes in red light is not caused by a lack of morphogenetic substances.

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Frau R.Dittrich danke ich für die sorgfältige Betreuung der Kulturen und Mithilfe bei den Versuchen.

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Die Arbeit wurde durch eine Sachbeihilfe des Ministeriums für wissenschaftliche Forschung an Herrn Prof. J.Reinert unterstützt.     

Blue light-induced synthesis of ribulosebisphosphate carboxylase in cultured plant cells

Upon illumination with blue light (350–550 nm) of suspension cultured cells (Nicotiana tabacum var. Samsun) the transition of leucoplasts to functional chloroplasts is induced. During the subsequent greening period chlorophylls as well as membrane and enzyme proteins are synthesized. Thus the amount of ribulosebisphosphate carboxylase (EC. 4.1.1.39) being small in leucoplasts increases dramatically due to de novo synthesis. This change is also reflected in the level of translatable messenger RNA specific for the small subunit of ribulosebisphosphate carboxylase which accumulates only in blue-irradiated cells; its in vitro translation product isolated by immunoprecipitation corresponds mainly to the precursor protein (Mr 20 000) of the small subunit. In contrast, red light (600–700 nm) does not induce synthesis of ribulosebisphosphate carboxylase. According to these findings it is proposed that blue light exerts its influence on ribulosebisphosphate carboxylase in cultured tobacco cells at a level below translation.     

Morphogenetic plasticity of callus reinitiated from cell suspension cultures of the fern Platycerium coronarium

Cell suspension cultures were initiated from gametophyte-derived callus of the fern Platycerium coronarium. Two distinct types of callus masses, distinguished by their colouration, were obtained when the cells from suspension culture were plated on semisolid Murashige and Skoog (MS) medium containing 10 M kinetin. The two types of callus masses had distinct morphogenetic capacities despite their common origin. Morphogenesis into either gametophytes or sporophytes occurred when these callus masses were cultured on phytohormone-free MS medium depending on the type of callus used. The dark-green gametophytic callus showed a faster rate of growth and morphogenesis as compared to the pale-green sporophytic callus. Total chlorophyll content and autofluorescence and size of chloroplasts of the sporophytic callus and cell suspension cultures were lower than that of the gametophytic callus. Observations from confocal laser scanning microscopy were in agreement with the physiological parameters measured. The availability of cell cultures of the same ploidy level, but with two distinct pathways of development will be useful for comparative studies of developmental plasticity.     

Die Steigerung der durch Phytochrom bewirkten Anthocyansynthese des Senfkeimlings (Sinapis alba L.) durch Chloramphenicol

Zusammenfassung Chloramphenicol (CAP) steigert in einem gewissen Konzentrationsbereich (etwa 20–40 g/ml) die durch Dunkelrot (DR) ausgelöste Anthocyansynthese des Senfkeimlings. Die lag-Phase sowie der Zeitpunkt der Beendigung der Anthocyansynthese werden durch das Antibioticum nicht beeinflußt. Mit und ohne CAP beobachtet man bei allen untersuchten DR-Intensitäten eine konstante Akkumulationsrate an Anthocyan über einen Zeitraum von mindestens 24 Std. —Die Steigerung der Akkumulationsrate durch CAP (20 g/ml) liegt stets im Bereich von 25%, auch wenn die DR-Intensität auf die Hälfte oder ein Viertel gesenkt wird (Wirkung des CAP=25% · Wirkung des DR). Man kann daraus formal schließen, daß CAP und DR unabhängig voneinander die Anthocyansynthese beeinflussen. Molekular läßt sich der Sachverhalt folgendermaßen deuten: CAP hemmt bei einer Konzentration von 20 g/ml die Proteinsynthese der Plastiden. Dadurch wird der Phenylalanin-pool in den Kotyledonen erhöht. Da diese geringe CAP-Konzentration die cytoplasmatische Proteinsynthese noch nicht hemmt, kann die Anthocyansynthese ungestört ablaufen. Da Phenylalanin als eine Vorstufe der Flavonoidsynthese fungiert, führt das erhöhte Angebot an dieser Substanz zu einer Steigerung der Anthocyansynthese.

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The increase of phytochrome-mediated anthocyanin synthesis in the mustard seedling (Sinapis alba L.) by chloramphenicol

Summary Chloramphenicol (CAP) within a certain range of concentration (about 20–30g/ml) increases the rate of far-red mediated anthocyanin accumulation in the mustard seedling (Sinapis alba L.) (Fig. 1). The lag-phase after the onset of far-red and the time of termination of anthocyanin synthesis are not influenced by the presence of the antibiotic (Fig. 2). With and without CAP we observe a constant rate of anthocyanin accumulation over a period of at least 24 hours after the lag-phase at all far-red intensities investigated (Fig. 2). The percentage increase of the rate of anthocyanin accumulation which is due to CAP (20 g/ml) is independent of the far-red intensity applied and always amounts to about 25% (Table). Formally one can conclude that CAP and far-red seem to act as two independent factors in a multiplicative system. On the molecular level the observations can possibly be explained as follows: CAP at a concentration of 20 g/ml inhibits protein synthesis of the plastids. This inhibition leads to an increase of the pool of phenylalanine in the cotyledons. Because the small concentration of CAP does not interfere with protein synthesis (enzyme synthesis) in the cytoplasm, far-red mediated anthocyanin synthesis can proceed normally. Since phenylalanine acts as a precursor of flavonoids the increased pool of this substance will lead to an increase in the rate of anthocyanin accumulation.

     

In vitro culture of Vitis: the effects of light spectrum,manganese sulfate and potassium iodide on morphogenesis

The effects of blue and red light, manganese sulfate concentration (100 and 5 M), and potassium iodide (5 and 0 M) on shoot and root production from subcultured shoots of the Vitis hybrid Remaily Seedless were studied.Shoot production was greater in blue light. It was increased by lowering the manganese sulfate concentration. The response to manganese in blue light was greatest when there was no potassium iodide addition.Root production was decreased by red light and lower manganese concentration.The effects of manganese, iodide and light spectrum on morphogenesis are discussed in relation to their known effects on IAA metabolism.     

Ribonucleic Acid and Protein Changes in the Subcellular Components of the Gametophytes of Pteridium aquilinum during Growth in Red and Blue Light

The distribution of RNA and protein in the gametophytes of bracken fern, Pteridium aquilinum is affected by the quality of light in which they are grown. Two-dimensional gametophytes growing in blue light have a greater amount of RNA and protein than one-dimensional protonema growing in red light. Fractions rich in nuclei, chloroplasts, mitochondria, ribosomes and soluble supernatants obtained from blue-light grown gametophytes by differential centrifugation contain greater amounts of RNA and protein than corresponding fractions of red-light grown plants. Differences in RNA and protein content are detected in some of the fractions within 24 hours after start of the treatment.     

Light quality affects photosynthesis and leaf anatomy of birch plantlets in vitro

Cultures in vitro of Betula pendula Roth were subjected to light of different spectral qualities. Photosynthetic capacity was highest when the plantlets were exposed to blue light (max recorded photosynthesis, 82 mol CO₂ dm^–2 h^–1) and lowest when irradiated with light high in red and/or far-red wave lengths (max recorded photosynthesis, 40 mol CO₂ dm^–2 h^–1). Highest chlorophyll content (2.2 mg dm^–2 leaf area) was found in cultures irradiated with blue light, which also enhanced the leaf area. Morphometric analysis of light micrographs showed that the epidermal cell areas were largest in plantlets subjected to blue light and smallest in those subjected to red light. Morphometric analysis of electron micrographs of palisade cells, showed that the functional chloroplast area was largest in chloroplasts of leaves subjected to blue light and smallest in those exposed to red light. We suggest that light quality affects photosynthesis both through effects on the composition of the photosynthetic apparatus and on translocation of carbohydrates from chloroplasts.     

Gibberellic acid and the light inhibition of stem elongation

Summary The ability of gibberellic acid (GA₃) to prevent the light inhibition of stem elongation in peas was examined for several varieties under a wide range of irradiation conditions.A saturating dose of GA₃ largely prevented the inhibitory effect of red light on total stem height in Duke of Albany (tall), Alaska (medium) and Meteor (dwarf) although a small, but statistically significant, effect persisted in all varieties after 3 days of light. The growth of the second internode was, however, markedly inhibited by red light even with a saturating dose of GA₃. With gibberellin there was no difference between the effects of continuous red light and 15 minutes per day on height but the second internode was much shorter in the former treatment. The number of internodes present was the same in both cases and, therefore, the upper internodes in continuous light were as long or longer than in the 15-minute treatment. The number of internodes was only slightly fewer in darkness than in light so that, with GA₃, the effect of red light was transient and only the growth of the lower internodes was inhibited. Without GA₃ overall height was less in both red light treatments than in darkness for all three varieties.In blue light, on the other hand, there was no difference depending on whether height or internode length is considered, and even with a saturating dose of GA₃ the growth rate remained depressed in continuous blue light. There was, however, some interaction between blue light and GA₃.Red/far-red reversal experiments showed that in the varieties Alska and Duke of Albany the far-red stimulation of elongation persisted in the presence of a saturating dose of GA₃ while for the dwarf variety Meteor there was a significant interaction between far-red and GA₃.At least a quantitative difference was found between tall and dwarf peas in their response to light. Tall varieties showed a much greater effect of a prolonged exposure to blue and a smaller effect of a short exposure to red than dwarf varieties. Increasing the duration of exposure to red increasingly inhibited the growth of tall varieties. The medium variety Alaska grew to approximately the same height in continuous red and blue light.     

Structural and functional properties of the coleoptile chloroplast: Photosynthesis and photosensory transduction

Recent studies have shown that guard cell and coleoptile chloroplasts appear to be involved in blue light photoreception during blue light-dependent stomatal opening and phototropic bending. The guard cell chloroplast has been studied in detail but the coleoptile chloroplast is poorly understood. The present study was aimed at the characterization of the corn coleoptile chloroplast, and its comparison with mesophyll and guard cell chloroplasts. Coleoptile chloroplasts operated the xanthophyll cycle, and their zeaxanthin content tracked incident rates of solar radiation throughout the day. Zeaxanthin formation was very sensitive to low incident fluence rates, and saturated at around 800–1000 mol m^–2 s^–1. Zeaxanthin formation in corn mesophyll chloroplasts was insensitive to low fluence rates and saturated at around 1800 mol m^–2 s^–1. Quenching rates of chlorophyll a fluorescence transients from coleoptile chloroplasts induced by saturating fluence rates of actinic red light increased as a function of zeaxanthin content. This implies that zeaxanthin plays a photoprotective role in the coleoptile chloroplast. Addition of low fluence rates of blue light to saturating red light also increased quenching rates in a zeaxanthin-dependent fashion. This blue light response of the coleoptile chloroplast is analogous to that of the guard cell chloroplast, and implicates these organelles in the sensory transduction of blue light. On a chlorophyll basis, coleoptile chloroplasts had high rates of photosynthetic oxygen evolution and low rates of photosynthetic carbon fixation, as compared with mesophyll chloroplasts. In contrast with the uniform chloroplast distribution in the leaf, coleoptile chloroplasts were predominately found in the outer cell layers of the coleoptile cortex, and had large starch grains and a moderate amount of stacked grana and stroma lamellae. Several key properties of the coleoptile chloroplast were different from those of mesophyll chloroplasts and resembled those of guard cell chloroplasts. We propose that the common properties of guard cell and coleoptile chloroplasts define a functional pattern characteristic of chloroplasts specialized in photosensory transduction.Abbreviations Ant or A antheraxanthin - dv/dt fluorescence quenching rate - Fm maximum yield of fluorescence with all PS II reaction centers closed - Fo yield of instantaneous fluorescence with all PS II reaction centers open - Vio or V violaxanthin - Zea or Z zeaxanthin     

INTERACTION OF LIGHT QUALITY AND NUCLEASES IN THE GROWTH OF THE GAMETOPHYTES OF ASPLENIUM NIDUS

Appropriate concentrations of ribonuclease A and B selectively inhibited initiation of two-dimensional morphology in the gametophytes of Asplenium nidus, grown under a photoperiod of 5½ hr white light. Filamentous growth was promoted in such sporelings, the individual cells of which were significantly longer than corresponding cells of the control. Higher concentrations of enzymes were required to inhibit two-dimensional growth in the gametophytes grown in blue light. Concentrations of ribonuclease A or B which inhibited two-dimensional growth in white light promoted growth in length of the protonema in red light. Growth modifications in the sporelings induced by deoxyribonuclease in different light conditions were similar to those induced by the ribonucleases. The results lend further support to the postulated role of RNA in the regulation of two-dimensional growth in fern gametophytes.     

Regulation of Protein Synthesis during Photomorphogenesis of Gametophytes of the Fern Onoclea sensibilis

Gametophytes of the fern Onoclea sensibilis grow as filaments in the dark and in red light and become planar in blue light. Pulse-labeling 4-day-old gametophytes with [³⁵S]methionine at different times after transfer to dark, red, and blue light environments revealed higher rates of amino acid uptake and protein synthesis in blue light than in red light or in the dark. Characterization of the extant and newly synthesized soluble proteins by one- and two-dimensional gel electrophoresis showed that the patterns of protein accumulation and synthesis in gametophytes exposed to short periods of red or blue light were qualitatively indistinguishable from those of gametophytes maintained in the dark. However, some striking increases and decreases in the levels of certain polypeptides were noted and these changes were accentuated during continued growth of gametophytes in the different environments. The results show that photomorphogenesis of gametophytes of O. sensibilis is associated with quantitative rather than qualitative changes in the population of mRNAs available for translation.