Internal conversion in the photosynthetic mechanism of blue-green algae

1. In Chroococcus a quantum of light absorbed by phycocyanin has 90 per cent the chance of doing photosynthesis that a quantum absorbed by chlorophyll has. 2. By a process analogous to internal conversion in radioactivity (but with the linear dimensions and the wave length 10⁴ times larger) there will be transferred from phycocyanin to chlorophyll See PDF for Equation (a number of the order of 100) quanta for every one emitted as fluorescent light by the phycocyanin in the Chroococcus cell. 3. The yield of fluorescent light in Chroococcus is between 1 and 2 per cent. 4. The transfer of energy by internal conversion can account for the photosynthesis by phycocyanin observed by Emerson and Lewis.     

Photosynthetic action spectra of marine algae

A polarographic oxygen determination, with tissue in direct contact with a stationary platinum electrode, has been used to measure the photosynthetic response of marine algae. These were exposed to monochromatic light, of equal energy, at some 35 points through the visible spectrum (derived from a monochromator). Ulva and Monostroma (green algae) show action spectra which correspond very closely to their absorption spectra. Coilodesme (a brown alga) shows almost as good correspondence, including the spectral region absorbed by the carotenoid, fucoxanthin. In green and brown algae, light absorbed by both chlorophyll and carotenoids seems photosynthetically effective, although some inactive absorption by carotenoids is indicated. Action spectra for a wide variety of red algae, however, show marked deviations from their corresponding absorption spectra. The photosynthetic rates are high in the spectral regions absorbed by the water-soluble "phycobilin" pigments (phycoerythrin and phycocyanin), while the light absorbed by chlorophyll and carotenoids is poorly utilized for oxygen production. In red algae containing chiefly phycoerythrin, the action spectrum closely resembles that of the water-extracted pigment, with peaks corresponding to its absorption maxima (495, 540, and 565 mµ). Such algae include Delesseria, Schizymenia, and Porphyrella. In the genus Porphyra, there is a series P. nereocystis, P. naiadum, and P. perforata, with increasingly more phycocyanin and less phycoerythrin: the action spectra reflect this, with increasing activity in the orange-red region (600 to 640 mµ) where phycocyanin absorbs. In all these red algae, photosynthesis is almost minimal at 435 mµ and 675 mµ, where chlorophyll shows maximum absorption. Although the chlorophylls (and carotenoids) are present in quantities comparable to the green algae, their function is apparently not that of a primary light absorber; this role is taken over by the phycobilins. In this respect the red algae (Rhodophyta) appear unique among photosynthetic plants.     

The fluorescence spectra of red algae and the transfer of energy from phycoerythrin to phycocyanin and chlorophyll

1. The fluorescence spectra of the alga Porphyridium have been recorded as energy distribution curves for eleven different incident wave lengths of monochromatic incident light between wave lengths 405 and 546 mµ. 2. In these spectra chlorophyll fluorescence predominates when the incident light is in the blue part of the spectrum which is strongly absorbed by chlorophyll. 3. For blue-green and green light the spectrum excited in Porphyridium contains in addition to chlorophyll fluorescence, the fluorescence bands characteristic of phycoerythrin and of phycocyanin. 4. From these spectra the approximate curves for the fluorescence of the individual pigments phycoerythrin, phycocyanin, and chlorophyll in the living material have been derived and the relative intensity of each of them has been obtained for each of the eleven incident wave lengths. 5. The effectiveness spectrum for the excitation of the fluorescence of these three pigments in vivo has been plotted. 6. From comparisons of the effectiveness spectrum for the excitation of each of these pigments it appears that both phycocyanin and chlorophyll receive energy from light which is absorbed by phycoerythrin. 7. It is suggested that phycocyanin may be an intermediate in the resonance transfer of energy from phycoerythrin to chlorophyll. 8. Since phycoerythrin and phycocyanin transfer energy to chlorophyll, it appears probable that chlorophyll plays a specific chemical role in photosynthesis in addition to acting as a light absorber.     

THE EFFECTIVENESS OF THE SPECTRUM IN CHLOROPHYLL FORMATION

1. Although the carotenoid pigments are present in large concentration in the plastids of etiolated Avena seedlings as compared with protochlorophyll, the pigment precursor of chlorophyll, it is possible to show that the carotenoids do not act as filters of the light incident on the plant in the blue region of the spectrum where they absorb heavily. This suggests that the carotenoids are located behind the protochlorophyll molecules in the plastids. 2. Since the carotenoids do not screen and light is necessary for chlorophyll formation, an effectiveness spectrum of protochlorophyll can be obtained which is the reciprocal of the light energy necessary to produce a constant amount of chlorophyll with different wavelengths. The relative effectiveness of sixteen spectral regions in forming chlorophyll was determined. 3. From the effectiveness spectrum, one can conclude that protochlorophyll is a blue-green pigment with major peaks of absorption at 445 mµ, and 645 mµ, and with smaller peaks at 575 and 545 mµ. The blue peak is sharp, narrow, and high, the red peak being broader and shorter. This differs from previous findings where the use of rougher methods indicated that red light was more effective than blue and did not give the position of the peaks of absorption or their relative heights. 4. The protochlorophyll curve is similar to but not identical with chlorophyll. The ratio of the peaks of absorption in the blue as compared to the red is very similar to chlorophyll a, but the position of the peaks resembles chlorophyll b. 5. There is an excellent correspondence between the absorption properties of this "active" protochlorophyll and what is known of the absorption of a chemically known pigment studied in impure extracts of seed coats of the Cucurbitaceae. Conclusive proof of the identity of the two substances awaits chemical purification, but the evidence here favors the view that the pumpkin seed substance, which is chemically chlorophyll a minus two hydrogens, is identical with the precursor of chlorophyll formation found in etiolated plants.     

Relation between pigment content and photosynthetic characteristics in a blue-green algae

1. The blue-green alga Anacystis nidulans was cultured under steady state conditions at 25 and 39°C. and under several different light intensities to give five different types of cells. 2. Cells were submitted to pigment analysis based upon acetone extracts and aqueous extracts obtained by sonic disintegration. The different cell types show a threefold range of chlorophyll content and a fourfold range of phycocyanin content with only minor changes in the chlorophyll/phycocyanin ratio. Cells of highest pigment content were estimated to contain 2.8 per cent chlorophyll a and 24 per cent phycocyanin, the latter on a total chromoproteid basis. 3. Light intensity curves of photosynthesis were obtained for each of the cell types at 25 and at 39°C. The slopes of the light-limited regions of the curves are approximately linear functions of chlorophyll and phycocyanin contents. Maximum light-saturated rates of photosynthesis at 25 and 39° show no simple relation to pigment content.     

THE EFFECT OF ULTRAVIOLET LIGHT ON PHOTOSYNTHESIS

1. An unidentified unit in the mechanism of the photosynthesis of Chlorella pyrenoidosa is rendered inactive by the absorption of one quantum of ultraviolet light (2537 Å wave length). 2. The same irradiation has no effect on the normal respiration of Chlorella pyrenoidosa. Experiments have not yet been made on the respiration inhibitable by HCN. 3. No chemical change was detected in the chlorophyll extracted from irradiated cells.     

PHOTOSYNTHETIC LIGHT-HARVESTING FUNCTION OF VIOLAXANTHIN IN NANNOCHLOROPSIS SPP. (EUSTIGMATOPHYCEAE)1

Whole cell absorption spectra of the Eustigmatophycean algae Nannochloropsis salina Bourrelly and Nannochloropsis sp. reveal the presence of a distinct absorption peak at 490 nm. The lack of chlorophylls b and c in these species indicates that this peak must be attributed to carotenoid absorption. In vivo fluorescence excitation spectra for chlorophyll a emission show a corresponding maximum at 490 nm. This peak is more clearly resolved than carotenoid maxima in other algal classes due to the absence of accessory chlorophylls. The carotenoid composition of the two Nannochloropsis species shows that violaxanthin and vaucheriaxanthin are the main contributors to 490 nm absorption. Violaxanthin accounts for approximately 60% of the total carotenoid in both clones. We conclude that light absorption by violaxanthin, and possibly by vaucheriaxanthin, is coupled in energy transfer to chlorophyll a and that violaxanthin is the major light-harvesting pigment in the Eustigmatophyceae. This is the first report of the photosynthetic light-harvesting function of this carotenoid.     

PHOTOSYNTHETIC LIGHT-HARVESTING FUNCTION OF VIOLAXANTHIN IN NANNOCHLOROPSIS SPP. (EUSTIGMATOPHYCEAE)1

Whole cell absorption spectra of the Eustigmatophycean algae Nannochloropsis salina Bourrelly and Nannochloropsis sp. reveal the presence of a distinct absorption peak at 490 nm. The lack of chlorophylls b and c in these species indicates that this peak must be attributed to carotenoid absorption. In vivo fluorescence excitation spectra for chlorophyll a emission show a corresponding maximum at 490 nm. This peak is more clearly resolved than carotenoid maxima in other algal classes due to the absence of accessory chlorophylls. The carotenoid composition of the two Nannochloropsis species shows that violaxanthin and vaucheriaxanthin are the main contributors to 490 nm absorption. Violaxanthin accounts for approximately 60% of the total carotenoid in both clones. We conclude that light absorption by violaxanthin, and possibly by vaucheriaxanthin, is coupled in energy transfer to chlorophyll a and that violaxanthin is the major light-harvesting pigment in the Eustigmatophyceae. This is the first report of the photosynthetic light-harvesting function of this carotenoid.     

Incompetence of dark-synthesized phycocyanin in excitation transfer to photosystem II chlorophyll

Summary When cells of Anabaena variabilis, all the phycobilin pigments of which had been newly synthesized in the dark, were excited by light absorbed in phycocyanin, the fluorescence emission spectrum showed a peak corresponding to the emission from allophycocyanin, but no emission from chlorophyll. These cells were active in photosynthesis and, when excited by light absorbed by chlorophyll, the emitted fluorescence was characteristic of photosystem II chlorophyll. This indicates that dark synthesized phycocyanin is capable of excitation transfer to allophycocyanin but not to photosystem II chlorophyll.Abbreviation CMU 3-(p-chlorophenyl)-1,1-dimethylurea     

Photoinhibition of chloroplast reactions. I. Kinetics and action spectra

A study was made of photoinhibition of spinach chloroplast reactions. The kinetics and spectral characteristics of the photoinhibition over a range between 230 and 700 mμ have been examined. The decline of activity due to preillumination was independent of wavelength, and dependent upon the number of quanta applied, not upon the rate of application. The effectiveness spectra of photoinhibition indicate that active ultraviolet light is absorbed by a pigment which is not a normal light absorber for photosynthesis and acts with a high quantum efficiency (> 0.1) for photoinhibition.

Active visible light is absorbed by the pigments which sensitize photosynthesis (chlorophyll, carotenoids). A very low quantum efficiency (about 10⁻⁴) was observed for the photoinhibition with visible light.

The action spectrum of the photoinhibition of dye reduction by chloroplasts and lyophylized Anacystis cells indicated that the damage caused by visible light is due to quanta absorbed by photosystem II. However, since system I might not be involved in dye reduction, the spectra may reflect only damage to photosystem II.

     

Transfer of the Excitation Energy in Anacystis nidulans Grown to Obtain Different Pigment Ratios

The blue-green alga, Anacystis nidulans, was grown in lights of different colors and intensities, and its absorption and fluorescence properties were studied. Strong orange light, absorbed mainly by phycocyanin, causes reduction in the ratio of phycocyanin to chlorophyll a; strong red light, absorbed mainly by chlorophyll, causes an increase in this ratio. This confirms the earlier findings of Brody and Emerson (12) on Porphyridum, and of Jones and Myers (8) on Anacystis. Anacystis cultures grown in light of low intensity show, upon excitation of phycocyanin, emission peaks at 600 mmu and 680 mmu, due to the fluorescence of phycocyanin and chlorophyll a, respectively. Changes in the efficiency of energy transfer from phycocyanin to chlorophyll a are revealed by changes in the ratios of these two bands. A decrease in efficiency of energy transfer from phycocyanin to chlorophyll a seems to occur whenever the ratio of chlorophyll a to phycocyanin deviates from the normal. Algae grown in light of high intensity show, upon excitation of phycocyanin, only a fluorescence band at 660 mmu and no band at 680 mmu. This suggests reduced efficiency of energy transfer from phycocyanin to the strongly fluorescent form of chlorophyll a (chlorophyll a(2)) and perhaps increased transfer to the weakly fluorescent form of chlorophyll a (chlorophyll a(1)).     

The relationship between chlorophyll and the carotenoids in the algal flagellate,Euglena

1. We have obtained an action spectrum for chlorophyll formation in Euglena gracilis. This action spectrum is similar to the absorption spectrum of protochlorophyll. However, efforts to isolate and identify this pigment have been unsuccessful. 2. Porphyrins have been extracted from both the normal and dark-adapted Euglena and a chlorophyll-free mutant. 3. The "action" spectra for chlorophyll and carotenoid synthesis have been found to almost coincide, indicating that the same porphyrin-like molecule may influence the synthesis of both pigments. 4. It is indicated that two porphyrin-like systems are in operation simultaneously, one concerned with carotenoid "removal" and another involved in carotenoid and chlorophyll synthesis.     

CHLOROPLAST DEVELOPMENT IN OCHROMONAS DANICA

When dark-grown cells of Ochromonas danica are placed in the light, the amount of chlorophyll a per cell increases 82-fold; the content of carotenoid pigment, 24-fold. Concomitantly with this increase in chlorophyll and carotenoid pigment, the small proplastid of dark-grown cells develops into a large lamellate chloroplast. During the first 12 hours in the light, vesicles appear within the loose clusters of dense chloroplast granules, enlarge, align themselves into rows (plates in three dimensions), and fuse into discs. Double discs may form from the more or less simultaneous fusion of two adjacent plates of vesicles or by the addition of vesicles to an already formed single disc. Three-disc bands arise by the addition of a disc to an already formed two-disc band through the approach and fusion of more vesicles. After 24 hours in the light, most of the chloroplast bands contain three discs, but the chloroplasts are still small. After 48 hours in the light, almost all the cells contain full-sized chloroplasts with a full complement of three-disc bands. However, at this time the amount of chlorophyll a and carotenoid pigment is only one-half of maximum. During the next 3 days in the light, as the number of chlorophyll and carotenoid molecules per chloroplast approximately doubles, there is a compression of the discs in each band (from 180 to 130 A) and a precise alignment of their membranes. Changes also occur in the nucleus when dark-grown cells are placed in the light. There is an increase in the number of small nucleolar bodies, many of which lie directly against the nuclear envelope, and in a few cells a dense mass of granules is seen between the two membranes of the nuclear envelope.     

Photoacoustic spectroscopy of Anacystis nidulans. II. Characterization of pigment holochroms and thermal deactivation spectrum

The photoacoustic spectrum of Anacystis nidulans recorded at room temperature is qualitatively similar to low-temperature absorption or fluorescence excitation spectra. The bands of pigment holochroms are well resolved compared to room-temperature absorption spectra. The thermal deactivation spectrum obtained by extrapolating acoustic data for an infinitely thin sample indicates that the photosynthetic efficiency decreases from phycocyanin to chlorophyll a and carotenoids.     

Zeaxanthin Protects Plant Photosynthesis by Modulating Chlorophyll Triplet Yield in Specific Light-harvesting Antenna Subunits

Plants are particularly prone to photo-oxidative damage caused by excess light. Photoprotection is essential for photosynthesis to proceed in oxygenic environments either by scavenging harmful reactive intermediates or preventing their accumulation to avoid photoinhibition. Carotenoids play a key role in protecting photosynthesis from the toxic effect of over-excitation; under excess light conditions, plants accumulate a specific carotenoid, zeaxanthin, that was shown to increase photoprotection. In this work we genetically dissected different components of zeaxanthin-dependent photoprotection. By using time-resolved differential spectroscopy in vivo, we identified a zeaxanthin-dependent optical signal characterized by a red shift in the carotenoid peak of the triplet-minus-singlet spectrum of leaves and pigment-binding proteins. By fractionating thylakoids into their component pigment binding complexes, the signal was found to originate from the monomeric Lhcb4–6 antenna components of Photosystem II and the Lhca1–4 subunits of Photosystem I. By analyzing mutants based on their sensitivity to excess light, the red-shifted triplet-minus-singlet signal was tightly correlated with photoprotection in the chloroplasts, suggesting the signal implies an increased efficiency of zeaxanthin in controlling chlorophyll triplet formation. Fluorescence-detected magnetic resonance analysis showed a decrease in the amplitude of signals assigned to chlorophyll triplets belonging to the monomeric antenna complexes of Photosystem II upon zeaxanthin binding; however, the amplitude of carotenoid triplet signal does not increase correspondingly. Results show that the high light-induced binding of zeaxanthin to specific proteins plays a major role in enhancing photoprotection by modulating the yield of potentially dangerous chlorophyll-excited states in vivo and preventing the production of singlet oxygen.     

Adaptations in Pigment Composition and Photosynthesis by Far Red Radiation in Chlorella pyrenoidosa

Chlorella pyrenoidosa has been cultivated in radiation of wavelengths between 690–975 nm for several months. Absorption spectra and action spectra of photo-synthesis have been determined for far red and “white” light brown cultures, In vivo spectrophotometric analyses and action spectra showed that fur red growth Chlorella adapted to the extreme light conditions by an increase both in absorption and photosynthesis above 700 nm. It is proposed that som of the in vivo normal chlorophyll a forms were converted to a far red absorbing chlorophyll a form, giving the far red exposed suspension an increased photosynthetic activity between 700–740 nm. The analyses of far red grown Chlorella have also shown an increased photosynthesis in the blue part of the spectrum, presumably due to a decrease in photosynthetically inactive carotenoid content. By culturing Chlorella in a “white” light gradient between 0.5 × 10⁴ and 3.7 × 10⁴ erg cm^?2 s^?1, it has been demonstrated that light intensity did not influence pigment ratios between 500–750 nm. In the blue part, however, high light levels caused increased absorption because of increased carotenoid content. Some ecological aspects of this far red effect have also been discussed.     

Photosynthetic efficiency of marine plants

Multicellular marine plants were collected from their natural habitats and the quantum efficiency of their photosynthesis was determined in the laboratory in five narrow wave length bands in the visible spectrum. The results along with estimates of the relative absorption by the various plastid pigments show a fairly uniform efficiency of 0.08 molecules O₂ per absorbed quantum for (a) chlorophyll of one flowering plant, green algae, and brown algae, (b) fucoxanthol and other carotenoids of brown algae, and (c) the phycobilin pigments phycocyanin and phycoerythrin of red algae. The carotenoids of green algae are sometimes less efficient while those of red algae are largely or entirely inactive. Chlorophyll a of red algae is about one-half as efficient (_o2 = 0.04) as either the phycobilins, or the chlorophyll of most other plants. These results as well as those of high intensity and of fluorescence experiments are consistent with a mechanism in which about half the chlorophyll is inactive while the other half is fully active and is an intermediate in phycoerythrin- and phycocyanin-sensitized photosynthesis.     

Action Spectra for Two Effects of Light on Luminescence in Gonyaulax polyedra

The luminescence of the marine dinoflagellate Gonyaulax polyedra shows an endogenous diurnal rhythm. The effect of light during the phase of low luminescence capacity may be observed as an enhancement of luminescence during the subsequent bright phase. During the bright phase, however, illumination diminishes the capacity for luminescence. The action spectra for these two effects of light have been determined, and the major pigments of Gonyaulax have been examined. A consideration of the action spectrum and the pigment complement of Gonyaulax suggests that photosynthesis during the day is responsible, directly or indirectly, for the enhancement of luminescence during the following night. Photoinhibition of luminescence is in part attributable to light absorbed by the photosynthetic pigments. However, activity observed in the far red region of the spectrum beyond the absorption maximum of chlorophyll a suggests that an additional pigment, present in small amounts, may also act as sensitizer for photoinhibition.     

DIFFERENCES BETWEEN IN VIVO ABSORPTION AND FLUORESCENCE EXCITATION SPECTRA IN NATURAL SAMPLES OF PHYTOPLANKTON

The fluorescence excitation spectrum of live phytoplankton cells represents the portion of light absorbed that has been effectively transferred to chlorophyll a of photosystem II, whereas light absorbed by photoprotective pigments will not lead to fluorescence. Therefore, the in vivo fluorescence excitation spectrum of phytoplankton has been used as a proxy for the action spectrum of phytoplankton in computations of primary production in the ocean. The distribution of chlorophyll a between photosystems, as well as variations in the pathway of energy inside the photosynthetic membrane, can also influence the fluorescence excitation spectrum. In this study, we investigated the contribution of photoprotective pigments to the differences found between in vivo absorption and fluorescence excitation spectra of phytoplankton measured during two cruises: one from Las Islas Canarias to Nova Scotia and another in the Labrador Sea. A comparison of normalized fluorescence excitation and absorption spectra showed high variability in the difference between absorption and fluorescence in the blue region of the spectrum for samples from the two cruises. This difference was not entirely correlated with the concentration of photoprotective carotenoids. In this paper, results are interpreted in terms of differences in pigment composition and known patterns of energy distribution in the photosystems of different algal groups.     

真眼点藻类色素的提取与测定方法

分别采用甲醇、乙醇和丙酮3种有机溶剂提取7种真眼点藻的色素,比较3种有机溶剂提取色素的效果,测定3种有机溶剂色素提取液的吸收光谱,利用分光光度法计算藻的叶绿素a和类胡萝卜素的含量,并比较甲醇和乙醇色素提取液在A₄₇₀和A₆₆₆的最大吸收峰。结果表明:使用乙醇比甲醇和90%丙酮操作更简便、快捷并且毒害低。3种有机溶剂色素提取液的叶绿素a和类胡萝卜素的含量均无显著性差异(P>0.05),提取率基本一致。色素在3种有机溶剂中的吸收光谱相似,甲醇和乙醇的色素提取液在A₄₇₀和A₆₆₆的最大吸收峰并无显著性差异(P>0.05)。乙醇色素提取液可使用Lichtenthaler的公式计算色素含量。