Interconversions of Chlorophylls a and b Synthesized from Exogenous Chlorophyllides a and b in Etiolated and Post-Etiolated Rye Seedlings

A comparative study of reciprocal conversions of chlorophylls a and b (Chl aand Chl b) in etiolated and post-etiolated rye seedlings (Secale cereale L.) was performed. The production of these pigments was initiated by infiltration of exogenous chlorophyllides a and b (Chlide a and b). It was shown that Chlide b, when infiltrated into etiolated rye seedlings, was esterified, producing Chl b. A major portion of Chl b (more than 80%) was transformed into Chl aduring long-term seedling dark exposure. The high rate of Chl b conversion into Chl a in the pool of pigments of exogenous origin was also observed during the lag-phase when there was no chlorophyll formation from endogenous precursors. The infiltration of Chlide a resulted in Chl a formation. The efficiency of its conversion into Chl b was low (about 1%) in the etiolated seedlings but increased during their greening. In the post-etiolated seedlings infiltrated with Chlide b, which were preliminary illuminated for 6–12 h, the Chl /Chl a ratio was almost similar in the pools of pigments synthesized from both exogenous and endogenous precursors. The rates of direct and reverse reactions responsible for the interconversion of Chl aand Chl b depended on the stage of the formation of the photosynthetic apparatus during greening of etiolated seedlings, when the particular structural components are formed in a definite sequence.     

Chlorophylls <Emphasis Type="Italic">d</Emphasis> and <Emphasis Type="Italic">f</Emphasis> and their role in primary photosynthetic processes of cyanobacteria

The finding of unique Chl d- and Chl f-containing cyanobacteria in the last decade was a discovery in the area of biology of oxygenic photosynthetic organisms. Chl b, Chl c, and Chl f are considered to be accessory pigments found in antennae systems of photosynthetic organisms. They absorb energy and transfer it to the photosynthetic reaction center (RC), but do not participate in electron transport by the photosynthetic electron transport chain. However, Chl d as well as Chl a can operate not only in the light-harvesting complex, but also in the photosynthetic RC. The long-wavelength (Q_y) Chl d and Chl f absorption band is shifted to longer wavelength (to 750 nm) compared to Chl a, which suggests the possibility for oxygenic photosynthesis in this spectral range. Such expansion of the photosynthetically active light range is important for the survival of cyanobacteria when the intensity of light not exceeding 700 nm is attenuated due to absorption by Chl a and other pigments. At the same time, energy storage efficiency in photosystem 2 for cyanobacteria containing Chl d and Chl f is not lower than that of cyanobacteria containing Chl a. Despite great interest in these unique chlorophylls, many questions related to functioning of such pigments in primary photosynthetic processes are still not elucidated. This review describes the latest advances in the field of Chl d and Chl f research and their role in primary photosynthetic processes of cyanobacteria.     

Assembly of the photosynthetic apparatus in embryos from Fucus serratus L

The assembly of the photosynthetic apparatus was studied during the first six days of development of Fucus serratus L. embryos. HPLC analysis revealed that oospheres and zygotes contain the same photosynthetic pigments (i.e., chlorophyll a, chlorophyll c, fucoxanthin, violaxanthin, and β-carotene) as fully developed thalli. Total pigment amount increased after fertilization, mainly due to an active synthesis of Chl a and fucoxanthin. Spectral modifications revealing the progressive integration of Chl a and Chl c in the photosynthetic units are described. In particular, a distinct emission at 705 nm, reflecting the accumulation of LHC I, was clearly detected. The emission bands at 705 nm and 725 nm were characterized by 77 K excitation fluorescence measurements. Their spectra differed by the presence of a large band at approximately 550 nm due to fucoxanthin in the excitation spectrum of F705 nm. Room temperature variable fluorescence was first observed 30 h after fertilization indicating a functional Photosystem II electron transfer at this developmental stage. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of gamma radiation on mutant induction of <Emphasis Type="Italic">Fagopyrum dibotrys</Emphasis> Hara

Agronomic traits, photosynthetic pigments, gas exchange, and chlorophyll (Chl) fluorescence parameters of red stem buckwheat (Fagopyrum dibotrys Hara) mutants induced by γ-radiation were compared with green control at seedling stage. Plant height, number of first-class branches, and rhizome biomass were inhibited significantly (p<0.01). Chl a, Chl b, and Chl a+b contents decreased with elevated dose of γ-rays, while increasing carotenoid content indicated that buckwheat was capable of adjusting to the radiation damage. Decrease in net photosynthetic rate was the result of both stomatal and non-stomatal limitations. Fluorescence parameters, such as F₀, F_m, F_v/F_m, F_v/F₀, Φ_PS2, electron transport rate, and photochemical quenching declined significantly (p<0.01) as compared with control due to photoinhibition, while non-photochemical quenching increased to enhance thermal dissipation. Lower parameters implied that leaf tissue was damaged significantly by high dose of γ-radiation and therefore leaf senescence was accelerated.     

Light and temperature, but not UV radiation, affect chlorophylls and carotenoids in Norway spruce needles (Picea abies (L.) Karst.)

Concentrations of chlorophyll a/freshweight (Chl a FW) and photosynthetic pigments/chlorophyll a were studied during one growing season in the current year's (CYN) and last year's needles (LYN) from Norway spruce (Picea abies (L.) Karst.) grown under natural or close‐to‐natural climate. Climate regimes differed in photosynthetic active radiation (PAR), temperature (T) and UV‐B radiation. Pigments were not affected by UV‐B but most of the differences between climate regimes, and also seasonal variations within climate regimes, could be related to PAR and T. Generally, two types of response to climate were observed: firstly, pigments reacted primarily to PAR without marked sensitivity to T and exhibited slow response times (> 30 d), and, secondly, pigments were affected by the combined action of PAR and T and responded faster than 20 d. The Chl a FW and chlorophyll b/chloprophyll a ratio exhibited slow‐type response in CYN and fast‐type response in LYN. Higher amplitudes in CYN than in LYN were observed for the latter two parameters, which are known to be associated with levels of pigment–protein complexes. It is suggested that slow response in CYN ensures that the high investments in proteins in these needles occur only in response to longer‐lasting climate episodes.     

PIGMENT AND PHOTOSYNTHETIC RESPONSES OF OSCILLATORIA AGARDHII (CYANOPHYTA) TO PHOTON FLUX DENSITY AND SPECTRAL QUALITY1

The effects of photon flux density (PFD) and spectral quality on biomass, pigment content and composition, and the photosynthetic activity of Oscillatoria agardhii Gomont were investigated in steady-state populations. For alterations of PFD, chemostat populations were exposed to 50, 130 and 230 μmol photons·m^?2·s^?1 of photosynthetic active radiation (PAR). Decreases in biomass, chlorophyll a (Chl a) and c-phycocyanin (CPC) contents, and CPC: Chl a and CPC: carotenoid content was not altered. Increases in the relative abundances of myxoxanthophyll and zeaxanthin and deceases in the relative abundances of echinenone and β-carotene within the carotenoid pigments coincided with increasing PFD. Increases in Chl a-specific photosynthetic rates and maxima and decreases in biomass-specific photosynthetic rates and maxima with increasing PFD were attributed to increased light harvesting by carotenoids per unit Chl a and reduction in total pigment content, respectively. Responses to spectral quality were tested by exposing chemostat populations to a gradient of spectral transmissions at 50 μmol photons·m^?2·s^?1 PAR. Biomass differences among populations were likely attributable to the distinct absorption of the PAR spectrum by Chl a, CPC, and carotenoids. Although pigment contents were not altered by spectral quality, relative abundances of zeaxanthin and echinenone in the carotenoid pigments increased in populations exposed to high-wavelength PAR. The population adapted to green light possessed a greater photosynthetic maximum than populations adapted to other spectral qualities.     

Changes in contents of photosynthetic pigments and ribulose-1,5-bisphosphate carboxylase activity during the development of globular somatic embryo into the plantlet of Siberian ginseng

During the development of the globular embryos via heart-shaped, torpedo-shaped, and cotyledonary embryos into plantlets, contents of chlorophyll (Chl) a and b and carotenoids, and activity of ribulose-1,5-bisphosphate carboxylase (RuBPC, EC 4.1.1.39) were investigated. In the solid cultivation (SC) the contents of Chl a, Chl b, Chl a/b, and total pigments increased up to plantlet stage. In the liquid cultivation (LC), contents of Chl a, Chl b, and total pigments increased till the torpedo-shaped stage, but decreased with the further development up to plantlets stage. During SC, RuBPC activity increased up to the torpedo-shaped embryo stage, but in the LC RuPBC activity increased continuously with the progress in the developmental stages. The correlations between Chl a and RuBPC activity on the SC and LC were negative, r = −0.26 and −0.56, respectively.     

铝胁迫对木荷幼苗光合特性的影响及添加盐基阳离子和磷的调节作用

为探讨铝(Al)胁迫对木荷(Schima superba)幼苗光合特征的影响,采用营养液水培的方法,对铝(Al)胁迫下木荷幼苗的光合响应及盐基阳离子(BC)和磷(P)的调节作用进行了研究。结果表明,在低浓度Al(0.25 mmol L–1)处理下,木荷幼苗的光合色素(Chl a、Chl b、Car)含量、光合作用参数(P n、g s、WUE、C i/C a)以及光响应特征参数(P max、AQY、R d、LSP)均呈下降趋势,添加BC或同时添加BC和P均能缓解上述参数的降低。中、高浓度Al(0.75、1.50 mmol L–1)处理,除光合色素含量呈增加趋势外,光合作用参数、光响应特征参数均下降,且下降幅度随Al浓度的升高而增大,添加P比添加BC更能有效缓解Al胁迫对木荷幼苗的影响。这揭示了BC、P在缓解木荷Al胁迫的相对重要性。     

Genetic variation in leaf pigment,optical and photosynthetic function among diverse phenotypes of <Emphasis Type="Italic">Metrosideros polymorpha</Emphasis> grown in a common garden

Coordinated variation has been reported for leaf structure, composition and function, across and within species, and theoretically should occur across populations of a species that span an extensive environmental range. We focused on Hawaiian keystone tree species Metrosideros polymorpha, specifically, 13-year old trees grown (2–4 m tall) in a common garden (approximately 1 ha field with 2–3 m between trees) from seeds collected from 14 populations along an altitude–soil age gradient. We determined the genetic component of relationships among specific leaf area (SLA), the concentrations of nitrogen (N) and pigments (chlorophylls, carotenoids, and anthocyanins), and photosynthetic light-use efficiency. These traits showed strong ecotypic variation; SLA declined 35% with increasing source elevation, and area-based concentrations of N, Chl a + b and Car increased by 50, 109 and 96%, respectively. Concentrations expressed on a mass basis were not well related to source elevation. Pigment ratios expressed covariation that suggested an increased capacity for light harvesting at higher source elevation; Chl/N and Car/Chl increased with source elevation, whereas Chl a/b declined; Chl a/b was higher for populations on younger soil, suggesting optimization for low N supply. Parallel trends were found for the photosynthetic reactions; light-saturated quantum yield of photosystem II (Φ _PSII) and electron transport rate (ETR) increased with source elevation. Correlations of the concentrations of photosynthetic pigments, pigment ratios, and photosynthetic function across the ecotypes indicated a stoichiometric coordination of the components of the light-harvesting antennae and reaction centers. The constellation of coordinated morphological, biochemical and physiological properties was expressed in the leaf reflectance and transmittance properties in the visible and near-infrared wavelength region (400–950 nm), providing an integrated metric of leaf status among and between plant phenotypes.     

Impacts of seabird-derived nutrients on water quality and diatom assemblages from Cape Vera,Devon Island,Canadian High Arctic

Relationships between key phytoplankton attributes including Chl a-specific light absorption, pigment composition and concentration, photosynthesis, primary production and community structure were studied in two open shallow nutrient-poor coastal systems receiving similar amounts of sewage water. Both systems were significantly nitrogen limited. However, differences in wastewater treatment (primary vs secondary) and sewage dilution (50%) between the two systems caused a greater difference between systems than locally around the outflows. For both systems, water at the outlet had significantly lower water transparency caused by a 20% higher absorption by coloured dissolved organic matter. Nutrient concentrations were also elevated, gradually decreasing with distance north (governing current) of the outflows, causing higher abundance of nano-sized phytoplankton, higher content of carotenoid pigments, 20–50% higher Chl a-specific absorption coefficients and higher photosynthetic capacity. Although maximum rates of Chl a-normalised photosynthesis were strongly related to nitrate availability, no effects were found on the derived areal primary production or algal biomass suggesting that photosynthetic and optical parameters are more sensitive indicators of nutrient enrichment than biomass or productivity. Handling editor: Tasman Peter Crowe     

Seasonal variation in pigmentation of the dinoflagellate Peridinium gatunense (Dinophyceae) in Lake Kinneret, Israel

1. Pigment composition was measured in natural phytoplankton samples from Lake Kinneret, Israel. From March through June 1998, the dinoflagellate Peridinium gatunense Nygaard mostly contributed more than 95% of the algal biomass. Peak densities were found in April, close to the water surface, with >10⁹ cells m^?3, chlorophyll (Chl) a concentration of 380 mg m^?3 and areal Chl‐a density of >1300 mg m^?2. 2. Cellular concentrations of Chl‐a changed between 201 and 282 pg cell^?1, but did not show a defined temporal fluctuation. 3. The mass ratio of Chl‐c to Chl‐a changed from March to June between 0.16 and 0.22, and the peridinin to Chl‐a ratio changed from 0.25 to 0.41. Neither ratio showed a clear pattern of seasonal change. Conversely, there was a progressive increase in diadinoxanthin and β‐carotene ratios to Chl‐a through the season, parallel to the increase in photon flux impinging upon the lake surface. The diadinoxanthin to Chl‐a ratio changed from 0.11 to 0.28 and the β‐carotene to Chl‐a ratio varied from 0.03 to 0.08 from March through June. 4. Diatoxanthin was not detected in natural samples. However, it was present in experiments with P. gatunense cultures, when concentration of diatoxanthin increased rapidly, concurrent with a decrease in diadinoxanthin and β‐carotene concentrations, while Chl‐c and peridinin ratios to Chl‐a were almost stable with photon flux increase. 5. The seasonal variation in cellular pigmentation of P. gatunense in Lake Kinneret suggests that accumulation of photoprotective pigments is essential for optimisation of photosynthetic activity of this large dinoflagellate.     

Relative sensitivity of various spectral forms of photosynthetic pigments to leaf senescence in wheat (Triticum aestivum L.)

The change in the characteristics of the absorption spectrum of chloroplasts which were isolated from the mature and senescing primary wheat leaves, was examined at various wavelengths in which the photosynthetic pigments mostly absorb. Chlorophyll (Chl) a was observed to be relatively more sensitive to leaf senescence than Chl b and carotenoids. Furthermore, the various spectral in vivo forms of Chl a, did not degrade to a similar extent; the far red absorbing forms of Chl a including species that absorb maximally at 692 nm (Chl a-692), 700 nm (Chl a-700) and 708 nm (Chl a 708) were found to be extremely sensitive to senescence induced losses. Both attached and detached senscing primary wheat leaves exhibited nearly similar pattern in the loss of photosynthetic pigments which suggests that the loss in long wavelength absorbing forms of Chl a is a selective indicator of leaf senescence.     

Minor but key chlorophylls in Photosystem II

A ‘metal-free’ chlorophyll (Chl) a, pheophytin (Phe) a, functions as the primary electron acceptor in PS II. On the basis of Phe a/PS II = 2, Phe a content is postulated as an index for estimation of the stoichiometry of pigments and photosystems. We found Phe a in a Chl d-dominant cyanobacterium Acaryochloris marina, whereas Phe d was absent. The minimum Chl a:Phe a ratio was 2:2, indicating that the primary electron donor is Chl a, accessory is Chl d, and the primary electron acceptor is Phe a in PS II of A. marina. Chl d was artificially formed by the treatment of Chl a with papain in aqueous organic solvents. Further, we will raise a key question on the mechanisms of water oxidation in PS II.     

A fluorometric method for the differentiation of algal populations in vivo and in situ

Fingerprints of excitation spectra of chlorophyll (Chl) fluorescence can be used to differentiate `spectral groups' of microalgae in vivo and in situ in, for example, vertical profiles within a few seconds. The investigated spectral groups of algae (green group, Chlorophyta; blue, Cyanobacteria; brown, Heterokontophyta, Haptophyta, Dinophyta; mixed, Cryptophyta) are each characterised by a specific composition of photosynthetic antenna pigments and, consequently, by a specific excitation spectrum of the Chl fluorescence. Particularly relevant are Chl a, Chl c, phycocyanobilin, phycoerythrobilin, fucoxanthin and peridinin. A laboratory-based instrument and a submersible instrument were constructed containing light-emitting diodes to excite Chl fluorescence in five distinct wavelength ranges. Norm spectra were determined for the four spectral algal groups (several species per group). Using these norm spectra and the actual five-point excitation spectrum of a water sample, a separate estimate of the respective Chl concentration is rapidly obtained for each algal group. The results of dilution experiments are presented. In vivo and in situ measurements are compared with results obtained by HPLC analysis. Depth profiles of the distribution of spectral algal groups taken over a time period of few seconds are shown. The method for algae differentiation described here opens up new research areas, monitoring and supervision tasks related to photosynthetic primary production in aquatic environments. This revised version was published online in June 2006 with corrections to the Cover Date.     

Pigment exchange of Photosystem II reaction center by chlorophyll d

Pigment exchanges among photosystem reaction centers (RCs) are useful for the identification and functional analysis of chromophores in photosynthetic organisms. Pigment replacement within the spinach Photosystem II RC was performed with Chl d derived from the oxygenic alga Acaryochloris marina, using a protocol similar to that reported previously [Gall et al. (1998) FEBS Lett 434: 88–92] based on the incubation of reaction centers with an excess of other pigments. In this study, we analyzed Chl d-modified monomeric RC which was separated from Chl d-modified dimeric RC by size-exclusion chromatography. Based on the assumption of a constant ratio of two Pheo a molecules per RC, the number of Chl a molecules in Chl d-modified monomeric RCs was found to decrease from six to four. The absorption spectrum of the Chl d-modified monomeric RC at room temperature showed a large peak at 699.5 nm originating from Chl d and a small peak at 672.5 nm orignating from Chl a. Photoaccumulation of the Pheo a⁻ in Chl d-modified monomeric RC, in the presence of sodium dithionate and methyl viologen, did not differ significantly from that in control RC, showing that the Chl d-modified monomeric RC retains its charge separation activity and photochemically active Pheo a.     

How do changes in temperature during growth affect leaf pigment composition and photosynthesis in Zea mays genotypes differing in sensitivity to low temperature?

The changes in photosynthetic activity and composition of pigments induced by changes in temperature were examined in the third leaf of three chilling-tolerant and three chilling-sensitive genotypes of maize (Zea mays L.). The plants were grown under a controlled environment at a photon flux density of 550 mol m^-2 s^-1, a 12 h photoperiod and at a suboptimal temperature of 14/12 C (day/night) until the full expansion of the third leaf. After this treatment, the chilling-tolerant genotypes, when compared with the sensitive ones, displayed a higher photosynthetic activity, a higher content of chlorophyll (Chl) a+b, a higher Chl a/b ratio, a larger total carotenoid pool size as well as a different carotenoid composition. When temperature was subsequently increased to 24/22 C for 3 d the composition of the pigments changed, but the chilling-sensitive genotypes, while adjusting their lower Chl a/b ratio and their different carotenoid composition, were unable to adjust their lower content of chlorophyll, their smaller total carotenoid pool size or their lower photosynthetic performance. Moreover, while the chilling-tolerant genotypes converted the most part of zeaxanthin to violaxanthin in the xanthophyll cycle, the chilling-sensitive genotypes retained high amounts of zeaxanthin. The changes in pigment composition that occurred over the 3 d at 24/22 °C were largely conserved when the plants were returned to 14/12 °C, but photosynthetic activity decreased and zeaxanthin accumulated again. The results suggest that the capability of the chilling-tolerant genotypes, when compared with the sensitive ones, to retain high amounts of pigments and to form a competent photosynthetic apparatus at low temperature is the basis for their more vigorous growth in cool climates.     

Organisation of Photosynthetic Apparatus of Triticale in Relation to Productivity

Chlorophyll (Chl) content, photochemical activity of chloroplasts as well as photosynthetic and crop productivity were studied in different winter hexaploid Triticale (xTriticosecale Witt.) lines and their F₁ hybrids. Heterosis enhanced Chl content, photosynthetic potential, photosynthetic productivity, and grain yield only in several F₁ hybrids of Triticale. Indication in some genotypes of close correlations among morphological structure, Chl content, photochemical activity of chloroplasts, photosynthetic potential, and plant productivity may be used in breeding practice of Triticale. This revised version was published online in August 2006 with corrections to the Cover Date.     

Relative binding affinities of chlorophylls in peridinin–chlorophyll–protein reconstituted with heterochlorophyllous mixtures

Peridinin–chlorophyll–protein (PCP), containing differently absorbing chlorophyll derivatives, are good models with which to study energy transfer among monomeric chlorophylls (Chls) by both bulk and single-molecule spectroscopy. They can be obtained by reconstituting the N-terminal domain of the protein (N-PCP) with peridinin and chlorophyll mixtures. Upon dimerization of these “half-mers”, homo- and heterochlorophyllous complexes are generated, that correspond structurally to monomeric protomers of native PCP from Amphidinium carterae. Heterochlorophyllous complexes contain two different Chls in the two halves of the complete structure. Here, we report reconstitution of N-PCP with binary mixtures of Chl a, Chl b, and [3-acetyl]-Chl a. The ratios of the pigments were varied in the reconstitution mixture, and relative binding constants were determined from quantification of these pigments in the reconstituted PCPs. We find higher affinities for both Chl b and [3-acetyl]-Chl a than for the native pigment, Chl a.     

The Heterogeneity of Structural and Functional Photosynthetic Characteristics of Mesophyll Chloroplasts in Various Parts of Mature or Senescing Leaf Blade of Two Maize (Zea Mays L.) Genotypes

Differences in ultrastructural parameters of mesophyll cell (MC) chloroplasts, contents of photosynthetic pigments, and photochemical activities of isolated MC chloroplasts were studied in the basal, middle, and apical part of mature or senescing leaf blade of two maize genotypes. A distinct heterogeneity of leaf blade was observed both for structural and functional characteristics of chloroplasts. In both mature and senescing leaves the shape of MC chloroplasts changed from flat one in basal part of leaf to nearly spherical one in leaf apex. The volume density of granal thylakoids decreased from leaf base to apex in both types of leaves examined, while the amount of intergranal thylakoids increased in mature leaves but decreased in senescing leaves. The most striking heterogeneity was found for the quantity of plastoglobuli, which strongly increased with the increasing distance from leaf base. The differences in chloroplast ultrastructure were accompanied by differences in other photosynthetic characteristics. The Hill reaction activity and activity of photosystem 1 of isolated MC chloroplasts decreased from leaf base to apex in mature leaves. Apical part of senescing leaf blade was characterised by low contents of chlorophyll (Chl) a and Chl b, whereas in mature leaves, the content of Chls as well as the content of total carotenoids (Car) slightly increased from basal to apical leaf part. This was reflected also in the ratio Chl (a+b)/total Car; the ratio of Chl a/b did not significantly differ between individual parts of leaf blade. Both genotypes examined differed in the character of developmental gradient observed along whole length of leaf blade.     

Photochemical and Thermal Phases of Chlorophyll a Fluorescence

The measurement of variable chlorophyll (Chl) a fluorescence is widely used as a convenient and versatile tool in photosynthesis research. In many applications empirical correlations and simplified models of Chl a fluorescence are used with success. Nevertheless, variable Chl a fluorescence provides only indirect and complex image of processes occurring within photosynthetic membranes and such simplifications have only limited validity. In this review we elucidate some controversial and still unresolved questions about the origin and interpretation of the variable Chl a fluorescence induction and the proper use of variable Chl a fluorescence for studies of photochemical events in photosystem 2 (PS2). Although the major part of variable Chl a fluorescence reflects the photochemical closure of the PS2 reaction centers (RCs) and can be considered as a function of the redox state of the primary acceptor Q_A, up to 50 % of the change in the Chl a fluorescence yield can be of secondary, nonphotochemical origin. We review the possible sources of the inherent heterogeneity in the origin of variable Chl a fluorescence. We also comment on the practical implications this bears for the use of variable Chl a fluorescence. This revised version was published online in August 2006 with corrections to the Cover Date.