Characterization of the Xanthophyll Cycle and Other Photosynthetic Pigment Changes Induced by Iron Deficiency in Sugar Beet (Beta vulgaris L.)

In this work we characterize the changes induced by iron deficiency in the pigment composition of sugar beet (Beta vulgaris L.) leaves. When sugar beet plants were grown hydroponically under limited iron supply, neoxanthin and β-carotene decreased concomitantly with chlorophyll a, whereas lutein and the carotenoids within the xanthophyll cycle were less affected. Iron deficiency caused major increases in the lutein/chlorophyll a and xanthophyll cycle pigments/chlorophyll a molar ratios. Xanthophyll cycle carotenoids in Fe-deficient plants underwent epoxidations and de-epoxidations in response to ambient light conditions. In dark adapted Fe-deficient plants most of the xanthophyll cycle pigment pool was in the epoxidated form violaxanthin. We show, both by HPLC and by in vivo 505 nanometers absorbance changes, that in Fe deficient plants and in response to light, the de-epoxidated forms antheraxanthin and zeaxanthin were rapidly formed at the expense of violaxanthin. Several hours after returning to dark, the xanthophyll cycle was shifted again toward violaxanthin. The ratio of variable to maximum chlorophyll fluorescence from intact leaves was decreased by iron deficiency. However, in iron deficient leaves this ratio was little affected by light conditions which displace the xanthophyll cycle toward epoxidation or de-epoxidation. This suggests that the functioning of the xanthophyll cycle is not necessarily linked to protection against excess light input.     

Photosystem II efficiency in low chlorophyll,iron-deficient leaves

Iron deficiency (iron chlorosis) is the major nutritional stress affecting fruit tree crops in calcareous soils in the Mediterranean area. This work reviews the changes in PS II efficiency in iron-deficient leaves. The iron deficiency-induced leaf yellowing is due to decreases in the leaf concentrations of photosynthetic pigments, chlorophylls and carotenoids. However, carotenoids, and more specifically lutein and the xanthophylls of the V+A+Z (Violaxanthin+ Antheraxanthin+Zeaxanthin) cycle are less affected than chlorophylls. Therefore, iron-chlorotic leaves grown in either growth chambers or field conditions have increases in the molar ratios lutein/chlorophyll a and (V+A+Z)/chlorophyll a. These pigment changes are associated to changes in leaf absorptance and reflectance. In the chlorotic leaves the amount of light absorbed per unit chlorophyll increases. The low chlorophyll, iron-deficient leaves showed no sustained decreases in PS II efficiency, measured after dark adaptation, except when the deficiency was very severe. This occurred when plants were grown in growth chambers or in field conditions. However, iron-deficient leaves showed decreases in the actual PS II efficiency at steady-state photosynthesis, due to decreases in photochemical quenching and intrinsic PS II efficiency. Iron-chlorotic leaves were protected not only by the decrease in leaf absorptance, but also by down-regulation mechanisms enhancing non-photochemical quenching and thermal dissipation of the light absorbed by PS II within the antenna pigment bed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Relationships between carotenoid composition and growth habit in British plant species

The pigment composition of leaves from a number of different plant species collected from field sites in the region of Sheffield, UK, have been compared using high-performance liquid chromatography. Expression of pigment content per unit leaf area was dominated by variation in the total leaf chlorophyll. Neither chlorophyll per unit area nor the chlorophyll a/b ratio were found to be correlated with the habitat from which the plants originated. When the amounts of different carotenoids were expressed relative to the total carotenoid pool, it was found that whilst neither total carotene (α- +β-carotene) nor neoxanthin correlated with ability to grow in shade, the leaf content of both lutein and the total xanthophyll cycle carotenoids (zeaxanthin, anther-axanthin and violaxanthin) did, with lutein content being high in shade species and xanthophyll cycle intermediates low. There was a strong negative correlation between the relative amounts of each of these groups of carotenoids. The ratio of lutein to xanthophyll cycle carotenoids was strongly correlated to an index of shade tolerance.     

Photosystem II efficiency and mechanisms of energy dissipation in iron-deficient,field-grown pear trees (Pyrus communis L.)

The dark-adapted Photosystem II efficiency of field-grown pear leaves, estimated by the variable to maximum chlorophyll fluorescence ratio, was little affected by moderate and severe iron deficiency. Only extremely iron-deficient leaves showed a decreased Photosystem II efficiency after dark adaptation. Midday depressions in Photosystem II efficiency were still found after short-term dark-adaptation in iron-deficient leaves, indicating that Photosystem II down-regulation occurred when the leaves were illuminated by excessive irradiance. The actual Photosystem II efficiency at steady-state photosynthesis was decreased by iron deficiency both early in the morning and at midday, due to closure of Photosystem II reaction centers and decreases of the intrinsic Photosystem II efficiency. Iron deficiency decreased the amount of light in excess of that which can be used in photosynthesis not only by decreasing absorptance, but also by increasing the relative amount of light dissipated thermally by the Photosystem II antenna. When compared to the controls, iron-deficient pear leaves dissipated thermally up to 20% more of the light absorbed by the Photosystem II, both early in the morning and at midday. At low light iron-deficient leaves with high violaxanthin cycle pigments to chlorophyll ratios had increases in pigment de-epoxidation, non-photochemical quenching and thermal dissipation. Our data suggest that pH could be the major factor controlling thermal energy dissipation, and that large (more than 10-fold) changes in the zeaxanthin plus antheraxanthin to chlorophyll molar ratio caused by iron deficiency were associated only to moderate increases in the extent of photoprotection.This revised version was published online in October 2005 with corrections to the Cover Date.     

Characterization of photosynthetic pigment composition, photosystem II photochemistry and thermal energy dissipation during leaf senescence of wheat plants grown in the field

Photosynthetic pigment composition and photosystem II (PSII) photochemistry were characterized during the flag leaf senescence of wheat plants grown in the field. During leaf senescence, neoxanthin and beta-carotene decreased concomitantly with chlorophyll, whereas lutein and xanthophyll cycle pigments were less affected, leading to increases in lutein/chlorophyll and xanthophyll cycle pigments/chlorophyll ratios. The chlorophyll a/b ratio also increased. With the progression of senescence, the maximal efficiency of PSII photochemistry decreased only slightly in the early morning (low light conditions), but substantially at midday (high light conditions). Actual PSII efficiency, photochemical quenching and the efficiency of excitation capture by open PSII centres decreased significantly both early in the morning and at midday and such decreases were much greater at midday than in the early morning. At the same time, non-photochemical quenching, zeaxanthin and antheraxanthin contents at the expense of violaxanthin increased both early in the morning and at midday, with a greater increase at midday. The results in the present study suggest that a down-regulation of PSII occurred in senescent leaves and that the xanthophyll cycle plays a role in the protection of PSII from photoinhibitory damage in senescent leaves by dissipating excess excitation energy, particularly when exposed to high light.     

Carotenoid composition in Zea mays developed at sub-optimal temperature and different light intensities

The content and composition of pigments were examined in the third leaf of Zea mays L. plants grown under controlled environment at near-optimal temperature (24°C) or sub-optimal temperature (14°C) at a light intensity of either 200 or 600 μmol m^?2 s^?1. Compared to leaves grown at 24°C, leaves grown at 14°C showed a large reduction in the chlorophyll (Chl) content, a marked decrease in the Chl a/b ratio, and a large increase in the ratio of total carotenoids/Chl a+b. Leaves grown at 14°C showed a much lower content of β-carotene than leaves grown at 24°C, while the content of the carotenoids of the xanthophyll cycle (violaxanthin [V] + antheraxanthin [A] + zeaxanthin [Z]) was markedly higher in the former leaves as compared to the latter leaves; neoxanthin and lutein were affected by the growth temperature to a much lesser extent. The xanthophylls/β-carotene ratio was about three times higher in leaves grown at 14°C as compared to leaves grown at 24°C. On a chlorophyll basis, the two types of leaves hardly differed in their level of β-carotene, while the levels of the xanthophylls (including lutein and neoxanthin) were higher in 14°C-grown leaves as compared to 24°C-grown leaves. In leaves grown at 14°C, 40 and 56% of the V+A+Z pool was in the form of zeaxanthin at low light intensity and high light intensity, respectively. Only trace amounts of zeaxanthin, if any, were present in leaves grown at 24°C. The changes in the pigment composition induced by growth at sub-optimal temperature were more pronounced at a light intensity of 600 as compared to 200 μmol m^?2 s^?1. In the given range, the light intensity slightly affected the composition of pigments in leaves grown at 24°C. The physiological significance of the modifications to the pigment composition induced by growth at sub-optimal temperature is discussed.     

Assessing leaf pigment content and activity with a reflectometer

This study explored reflectance indices sampled with a 'leaf reflectometer' as measures of pigment content for leaves of contrasting light history, developmental stage and functional type (herbaceous annual versus sclerophyllous evergreen). We employed three reflectance indices: a modified normalized difference vegetation index (NDVI), an index of chlorophyll content; the red/green reflectance ratio ( R _RED: R _GREEN), an index of anthocyanin content; and the change in photochemical reflectance index upon dark–light conversions (ΔPRI), an index of xanthophyll cycle pigment activity. In Helianthus annuus (sunflower), xanthophyll cycle pigment amounts were linearly related to growth light environment; leaves in full sun contained approximately twice the amount of xanthophyll cycle pigments as leaves in deep shade, and at midday a larger proportion of these pigments were in the photoprotective, de-epoxidized forms relative to shade leaves. Reflectance indices also revealed contrasting patterns of pigment development in leaves of contrasting structural types (annual versus evergreen). In H. annuus sun leaves, there was a remarkably rapid increase in amounts of both chlorophyll and xanthophyll cycle pigments along a leaf developmental sequence. This pattern contrasted with that of Quercus agrifolia (coast live oak, a sclerophyllous evergreen), which exhibited a gradual development of both chlorophyll and xanthophyll cycle pigments along with a pronounced peak of anthocyanin pigment content in newly expanding leaves. These temporal patterns of pigment development in Q. agrifolia leaves suggest that anthocyanins and xanthophyll cycle pigments serve complementary photoprotective roles during early leaf development. The results illustrate the use of reflectance indices for distinguishing divergent patterns of pigment activity in leaves of contrasting light history and functional type.     

Effects of iron on growth and reflectance spectrum of the bloom‐forming cyanobacterium Microcystis viridis

Iron is an important factor in algal blooms because it is involved in cyanobacterial pigment biosynthesis and therefore has the ability to influence the pigment status of algal cells. This role in pigment biosynthesis offers the opportunity for rapid monitoring of iron availability to cyanobacteria through spectral reflectance characterization. In the present study, the freshwater cyanobacterium Microcystis viridis was cultured with different levels of iron. Cell density, cellular content of iron and photosynthetic pigments, and spectral reflectivity of M. viridis were determined daily during the course of the culture experiment. The results showed that at the lowest iron concentration (0.01 μM) the growth of M. viridis was seriously limited, and the maximal cell density was only approximately 6.4% of the density observed with an iron concentration of 18 μM. Iron availability dramatically affected chlorophyll a, carotenoid and phycocyanin content, with the greatest impact on chlorophyll a. The iron‐induced changes in content and ratios of pigments were detectable through spectral reflectance. Eleven spectral indices previously developed for the estimation of concentrations and/or ratios of pigments and a newly proposed chlorophyll a/phycocyanin index were found to be suitable for generating sensitive regression models between cellular iron content and spectral parameters. The comprehensive application of key sensitive spectral indices and regression equations should help to support monitoring and diagnosis of iron availability to cyanobacteria via remote sensing.     

Pigment Content of Selected Planktonic Algae in Response to Simulated Natural Light Fluctuations and a Short Photoperiod

The lipophilic photosynthetic pigments in Limnothrix redekei, Planktothrix agardhii (cyanobacteria), Stephanodiscus minutulus, Synedra acus (diatoms), Scenedesmus acuminatus, and Scenedesmus armatus (chlorophycean) all isolated from an eutrophic lake were quantitatively determined by HPLC. The algae were grown semi-continuously under nutrient sufficient conditions at 20°C at a 12/12 h light/dark cycle with constant irradiance or with simulated natural light fluctuations as well as at a 6/18 h light/dark cycle with constant irradiance, all at the same daily light exposure. The zeaxanthin and the myxoxanthophyll contents of cyanobacteria were not influenced by fluctuating light, a short photoperiod or a different sampling time. The chlorophyll b/a ratio, the lutein/chlorophyll a ratio, and the neoxanthin content of chlorophycean as well as the chlorophyll c/a and the fucoxanthin/chlorophyll a ratio of diatoms were only slightly influenced by these factors. Therefore in some cases marker pigment contents and in other cases marker pigment/chlorophyll a ratios may be more useful for quantifying the relative importance of different taxonomic groups in natural phytoplankton. Simulated natural light fluctuations or the length of the photoperiod only slightly influenced the pigment content or the marker pigment/chlorophyll a ratio.     

The chloroplast pigments of the algal classes Eustigmatophyceae and Xanthophyceae. II. Xanthophyceae

The chloroplast pigments of Pleurochloris meiringensis Vischer, Mischococcus sphaerocephalus Vischer and Tribonema aequale Pascher have been analysed by a variety of chromatographic methods. There are significant differences between these xanthophycean algae and the eustigmatophycean algae formerly classified with them. In particular the former contain diadinoxanthin as the major xanthophyll whereas diadinoxanthin is absent from the latter and violaxanthin occurs as the major xanthophyll pigment. The other pigments of the Xanthophyceae sensu stricto include chlorophyll a, β-carotene, vaucheriaxanthin-ester, heteroxanthin, diatoxanthin, cryptoxanthin epoxide and a neoxanthin-like pigment.     

LIPOPHILIC PIGMENTS FROM CYANOBACTERIAL (BLUE-GREEN ALGAL) AND DIATOM MATS IN HAMELIN POOL,SHARK BAY,WESTERN AUSTRALIA1

Lipophilic pigments were examined in microbial mat communities dominated by cyanobacteria in the intertidal zone and by diatoms in the subtidal and sublittoral zones of Hamelin Pool, Shark Bay, Western Australia. These microbial mats have evolutionary significance because of their similarity to lithified stromatolites from the Proterozoic and Early Paleozoic eras. Fucoxanthin, diatoxanthin, diadinoxanthin, β-carotene, and chlorophylls a and c characterized the diatom mats, whereas cyanobacterial mats contained myxoxanthophyll zeaxanthin, echinenone, β-carotene, chlorophyll a and, in some cases, sheath pigment. The presence of bacteriochlorophyll a with in the mats suggest a close association of photosynthetic bacteria with diatoms and cyanobacteria. The high carotenoids: chlorophyll a ratios (0.84–2.44 wt/wt) in the diatom mats suggest that carotenoids served a photoprotective function in this high light environment. By contrast, cyanobacterial sheath pigment may have largely supplanted the photoprotective role of carotenoids in the intertidal mats.     

Effects of combined NaCl and CaCl2 salinity on photosynthetic parameters of barley grown in nutrient solution

The changes caused by NaCl− and CaCl₂-induced salinity on several leaf parameters have been measured in two cultivars of barley ( Hordeum vulgare L.) growing in a growth chamber in nutrient solution. Salinity was induced by adding to the nutrient solution equal weights of NaCl and CaCl₂, to obtain conductivities of 2, 6, 12, 19 and 26 dS m⁻¹. Salinity induced decreases in the leaf water potential and in the osmotic potential. Salinity did not induce significant changes in the relative photosynthetic pigment composition of barley leaves, the photosynthetic pigment stoichiometry for neoxanthin:violaxanthin cycle pigments:lutein:β-carotene:Chl b :Chl a being close to 3:6:14:12:25:100 (mol:mol). Salinity per se did not induce interconversions in the carotenoids within the violaxanthin cycle in most barley leaves. The PSII photochemistry of most barley leaves was unchanged by salinity. However, some apparently healthy leaves growing in high salinity exhibited sudden decreases in PSII photochemistry and increases in zeaxanthin (at the expense of violaxanthin), that preceded rapid leaf drying. Salinity induced significant changes in the slow part of the chlorophyll fluorescence induction curve from barley leaves.     

Photosynthetic UV-B Response of Beech (Fagus sylvatica L.) Saplings

Cloned saplings of beech (7-y-old) were exposed to enhanced UV-B irradiation (+25 %) continuously over three growing seasons (1999–2001). Analysis of CO₂ assimilation, variable chlorophyll (Chl) a fluorescence, and pigment composition was performed in late summer of the third growing season to evaluate the influence of long-term elevated UV-B irradiation. This influence was responsible for the stimulation of the net assimilation rate (P _N) over a range of irradiances. The increase in P _N was partially connected to increase of the area leaf mass, and thus to the increased leaf thickness. Even a higher degree of UV-B induced stimulation was observed at the level of photosystem 2 (PS2) photochemistry as judged from the irradiance response of electron transport rate and photochemical quenching of Chl a. The remarkably low irradiance-induced non-photochemical quenching of maximum Chl a fluorescence (NPQ) in the UV-B plants over the entire range of applied irradiances was attributed both to the reduced demand on non-radiative dissipation processes and to the considerably reduced contribution of the quenching localised in the inactivated PS2 reaction centres. Neither the content of Chls and total carotenoids expressed per leaf area nor the contents of lutein, neoxanthin, and the pool of xanthophyll cycle pigments (VAZ) were affected under the elevated UV-B. However, the contributions of antheraxanthin (A) and zeaxanthin (Z) to the entire VAZ pool in the dark-adapted UV-B treated plants were 1.61 and 2.14 times higher than in control leaves. Surprisingly, the retained A+Z in UV-B treated plants was not accompanied with long-term down-regulation of the PS2 photochemical efficiency, but it facilitated the non-radiative dissipation of excitation energy within light-harvesting complexes (LHC) of PS2. Thus, in the beech leaves the accumulation of A+Z, induced by other factors than excess irradiance itself, supports the resistance of PS2 against combined effects of high irradiance and elevated UV-B.     

CHLOROPLAST PIGMENTS OF CHLOROMONADOPHYCEAE

The chloroplast pigments of 2 species of Chloro-monadophyceae have been examined. Both Gonyo-stomum semen and Vacuolaria virescens possessed only one chlorophyll, chlorophyll a. Gonyostomum contained 4 carotenoids, but not fucoxanthin and peridinin. Vacuolaria also possessed 4 carotenoids which have been identified as β-carotene and the 3 xanthophylls commonly found in the Xanthophyceae (as represented by Tribonema aequale). These are antheraxanthin, lutein monoepoxide, and an hydroxy lutein monoepoxide-like xanthophyll. The taxonomic significance of these findings, the first for the Chloromonadophyceae, is discussed.     

Reversed-phase high-performance liquid chromatography of chlorophylls and carotenoids

Reversed-phase high-performance liquid chromatography with octadecyl- or octylsilylated silica gel as the stationary phase provides a powerful tool in the analysis of chloroplast pigments from higher plants and green algae. Chromatographic columns packed with 10 μm chemically bonded silica gel particles allow the simultaneous separation of chlorophylls a and b, chlorophyll isomers, pheophytins a and b, α-carotene, β-carotene, lutein, violaxanthin, lutein-5,6-epoxide, antheraxanthin, neoxanthin and several minor carotenoids from a single sample within a short analysis time. The quantitative analysis requires a minimum of 1–5 pmol for carotenoids and 5–10 pmol for chlorophylls. Pigment degradation products, formed on polar stationary phases, are not found in reversed-phase high-performance liquid chromatography due to the weak hydrophobic forces on which the separation mechanism is based. The production of altered pigments however, either induced by various treatments or generated during the isolation, can be monitored as the reversed-phase system is selective enough to separate cis-isomers and oxidation products from their parent compounds. The reproducibility of the individual retention time for each pigment is better than ±1.5% which facilitates the identification of unknown pigments. The method is applied to the analysis of the pigment composition of Chlorella fusca, spinach (Spinacia oleracea) chloroplasts, and to the rapid determination of the ratio of chlorophyll a to chlorophyll b.     

Iron Deficiency in the Blue-Green Alga Anacystis nidulans: Changes in Pigmentation and Photosynthesis

Phycocyanin-free photosynthetic lamellae (PSI-particles) were prepared from Anacystis nidulans, grown in complete and iron-deficient media. French press treatment and fractionated centrifugation were used. Absorption studies of the particles revealed an iron deficiency-induced shift of the main red chlorophyll a absorption peak from 679 to 673 nm as reported before for whole cells. The shift may reflect a changed distribution between different chlorophyll a forms. Action spectra for photo-oxidation of mammalian cytochrome c with photosynthetic lamellae revealed an iron deficiency-induced shift, corresponding to that found in the absorption spectra. As photo-oxidation of cytochrome c is mediated by PSI, it is believed that chlorophyll a also after the shift towards shorter wavelengths, is active in PSI. A decreased photosynthetic capacity of PSI, due to iron deficiency, was shown by time course studies of photosynthetic oxygen evolution, by photo-oxidation studies of P700 and mammalian cytochrome c, by photo-reduction studies of NADP and by combined studies of light-induced and chemical oxidation of P700. The ration chlorophyll a/700 was also determined for whole cells, lyophilized cells and PSI-particles. Iron deficiency caused an increased ratio in all studied fractions. The results of this work imply that energy is transferred with less efficiency within the photosynthetic units of PSI in iron-deficient A. nidulans than in iron-supplied algae.     

Research note: Irradiance of photosynthetically active radiation determines ultraviolet-susceptibility of photosynthesis in Ulva lactuca L.(Chlorophyta)

The respective ratio of photosynthetically active to ultraviolet radiation is of crucial importance to results obtained in ultraviolet (UV)‐research on photoautotrophic organisms. Specimens of the green macroalga Ulva lactuca L. were exposed to a constant irradiance of UV‐radiation at increasing irradiances of photosynthetically active radiation (PAR). The effects of experimental irradiance and spectral composition on photoinhibition of photosynthesis and its recovery were monitored by chlorophyll fluorescence measurements and the activity of the xanthophyll cycle was assessed by high performance liquid chromatography‐(HPLC) based pigment analysis. Results indicate a UV‐induced delay in recovery from PAR‐induced photoinhibition and a deceleration of violaxanthin conversion within the xanthophyll cycle due to the presence of UV‐radiation. Also the concentration of the protective pigment lutein increased considerably and could be indicative of the existence of an additional light‐protective mechanism, as, for example, the lutein‐epoxid cycle in Ulva. In total, results clearly show that the extent of UV‐induced inhibition of photosynthesis to be found in UV‐exposure experiments is highly dependent on the irradiance of background photosynthetically active radiation: with increasing irradiance of PAR the UV‐effects were diminished. Exemplified by the green algae Ulva lactuca this study demonstrates the crucial importance of the ratios of PAR:UV applied in UV‐research, particularly when conducting laboratory experiments in an ecological context.     

Photoacclimation impacts the molecular features of photosystem supercomplexes in the centric diatom Thalassiosira pseudonana

In diatoms, light-harvesting processes take place in a specific group of proteins, called fucoxanthin chlorophyll a/c proteins (FCP). This group includes many members and represents the major characteristic of the diatom photosynthetic apparatus, with specific pigments bound (chlorophyll c, fucoxanthin, diadino- and diatoxanthin besides chlorophyll a). In thylakoids, FCP and photosystems (PS) form multimeric supercomplexes.In this study, we compared the biochemical properties of PS supercomplexes isolated from Thalassiosira pseudonana cells grown under low light or high light conditions, respectively. High light acclimation changed the molecular features of the PS and their ratio in thylakoids. In PSII, no obvious changes in polypeptide composition were observed, whereas for PSI changes in one specific group of FCP proteins were detected. As reported before, the amount of xanthophyll cycle pigments and their de-epoxidation ratio was increased in PSI under HL. In PSII, however, no additional xanthophyll cycle pigments occurred, but the de-epoxidation ratio was increased as well. This comparison suggests how mechanisms of photoprotection might take place within and in the proximity of the PS, which gives new insights into the capacity of diatoms to adapt to different conditions and in different environments.     

A New Reversed Phase-HPLC Method Resolving All Major Higher Plant Photosynthetic Pigments

A new reversed phase-high performance liquid chromatography method has been developed to analyze the full complement of higher plant photosynthetic pigments (cis-neoxanthin, neoxanthin, violaxanthin, taraxanthin, anteraxanthin, lutein, zeaxanthin, cis-lutein, chlorophyll b, chlorophyll a, α- and β-carotene). The separation is carried out on a C₁₈ column in about 10 minutes, using a single high-pressure pump and three different mobile phases in three isocratic steps. This method introduces a major improvement in higher plant photosynthetic pigment analysis, resolving in only 10 minutes all photosynthetic pigments while achieving good separation of lutein from its isomer zeaxanthin. Zeaxanthin is involved in the xanthophyll cycle, which recently has been proposed to play a significant role in the protection of the photosynthetic apparatus from photoinhibitory conditions (Demmig et al. [1987] Plant Physiol 84: 218-224).     

The relative proportions of the chloroplast pigments as influenced by different levels of iron and potassium supply

Summary 1. Potato plants were grown in pot sand cultures at different levels of iron and potassium supply. Plants grown at the lowest level of iron developed iron deficiency chlorosis and potassium deficiency symptoms when maintained at the lowest level of potassium but not at the highest level.2. Rapid procedures were developed for the extraction and estimation of chloroplast pigments from small samples of lamina—usually less than 1 g of fresh material.3. The expression of pigment content on an area basis gave results which were in better agreement with visual observations than those expressed on either fresh or dry weight basis.4. Both iron and potassium additions increased the chloroplast pigment content. Leaves exhibiting iron deficiency contained reduced quantities of all pigments per unit area of lamina.5. A linear relationship existed between chlorophyll and carotene, chlorophyll and xanthophyll, and carotene and xanthophyll contents.6. The results suggested that laminae completely deficient in chlorophyll would contain no carotene but might still contain xanthophyll.7. The relative proportions of chlorophyll, carotene and xanthophyll do not remain constant under varying conditions of iron status. When iron is deficient the proportion of xanthophyll increased in relation to the chlorophyll and carotene contents.