Chlorophyll a′/P-700 and pheophytin a/P-680 stoichiometries in higher plants and cyanobacteria determined by HPLC analysis

Kobayashi, Masami; Watanabe, Tadashi; Nakazato, Masataka; Ikegami, Isamu; Hiyama, Tetsuo; Matsunaga, Tadashi; Murata, Norio

doi:10.1016/0005-2728(88)90254-x

Cited by 88 publications

(60 citation statements)

References 36 publications

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“…5,6 Though weakly basic compound might cause epimerization of Chl a, the effect of desiccants on the Chl a alteration was not studied in detail. The effect of desiccants on the pigment composition of thylakoid membranes was examined with the present reversed-phase HPLC.…”

Section: Alteration Of Chl a During Pigment Extraction With Desiccantsmentioning

confidence: 99%

“…[4][5][6] Though the amount of Chl a′ within PS I has not been finally determined yet, one possibility for the function of Chl a′ is a constituent of P700, the primary electron donor of PS I. [4][5][6] Presence of the "prime-type" pigments in the reaction center complexes of heliobacteria 7 and green sulfur bacterium, Chlorobium tepidum, 8 the evolutional counterparts of PS I, 9 suggests that "prime-type" pigments play essential roles in the photochemical reaction. Recently, Chl d′ was also detected in PS I of a Chl d-containing prokaryote, Acaryochloris marina.…”

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confidence: 99%

“…5,6 Though normal-phase HPLC showed high resolution for epimeric mixtures of Chl a and Pheo a, 11 stoichiometric comparison of Chl a′ with PhQ required two HPLC runs under different conditions. 15 Establishment of HPLC conditions which can separate PhQ, Chl a′, and Chl a is highly desirable for precise determination of Chl a′ within PS I, because direct comparison of the amount of Chl a′ with that of PS I is possible on a single HPLC trace.…”

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confidence: 99%

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Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Nakamura

Watanabe

2001

ANAL. SCI.

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Reversed-phase HPLC conditions for separation of chlorophyll (Chl) a, Chl a′ (the C13 2 -epimer of Chl a), pheophytin (Pheo) a (the primary electron acceptor of photosystem (PS) II), and phylloquinone (PhQ) (the secondary electron acceptor of PS I), have been developed. Pigment extraction conditions were optimized in terms of pigment alteration and extraction efficiency. Pigment composition analysis of light-harvesting complex II, which would not contain Chl a′ nor Pheo a, showed the Chl a′/Chl a ratio of 3 -4 × 10 -4 and the Pheo a/Chl a ratio of 4 -5 × 10 -4 , showing that the conditions developed here were sufficiently inert for Chl analysis. Preliminary analysis of thylakoid membranes with this analytical system gave the PhQ/Chl a′ ratio of 0.58 ± 0.03 (n = 4), in line with the stoichiometry of one molecule of Chl a′ per PS I.

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Section: Alteration Of Chl a During Pigment Extraction With Desiccantsmentioning

confidence: 99%

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confidence: 99%

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Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Nakamura

Watanabe

2001

ANAL. SCI.

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“…Chl a and b and Cars contents were determined according to Lichtenthaler [ 10 ]. The Pheo a content was determined using the equations proposed in [ 11 ] for pigment quantification in photosystem II (PSII) reaction centre preparations and corrected according to HPLC-established Chl a/ Pheo a molar ratio for leaf-pigment extracts from various species [ 12 ]. This correction was applied besause we established that for leaf extracts this method [ 11 ] gives values for Chl a coinciding with the values determined by the method of Lichthentaler [ 10 ], but the Pheo a content was overestimated.…”

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confidence: 99%

Effects of 24-epibrassinolide Pre-treatment on UV-B-Induced Changes in the Pigment Content of Pea Leaves

Dobrikova¹,

Vladkova²,

Stanoeva³

et al. 2013

Proceeding BAS

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In the present work, the effects of 24-epibrassinolide (EBR) on the UV-Binduced changes in the pigment content of pea leaves were studied. Control (non-EBR-treated) and EBR-treated plants were irradiated with UV-B for 3 h and pigment analysis was performed after 24 and 48 h. The results show that EBR spraying of plants 48 h prior to UV-B exposure alleviates its detrimental effect on chlorophyll a and b (Chl a and Chl b) content in comparison with control pea leaves. An increase in carotenoids (Car) and UV-B absorbing compounds was also observed at low dose of UV-B radiation. For the first time, it is shown that UV-B damage effect on control leaves is accompanied by a significant (more than 50%) increase in their pheophytin a (Pheo a) content 48 h after the UV-B exposure and that the EBR pre-treatment prevents the increase of Pheo a content in UV-B irradiated leaves. In addition, it is demonstrated that EBR application modifies UV-B-induced alterations of energy distribution between the main pigment-protein complexes in pea thylakoid membranes.

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“…3 for Chl a'; see ref. 7 for Chl a). No impurity other than a trace amount (less than 0.1 %) of epimer was detected in normal-phase HPLC, indicating that pigment denaturation in the column was sufficiently prevented.…”

Section: Resultsmentioning

confidence: 99%

Integrity of Chlorophyllous Pigments in Silica Normal-Phase HPLC

et al. 1996

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KeywordsChlorophyll a, chlorophyll a', fast atom chromatography bombardment mass spectrometry, high-performance liquidWe have been utilizing silica normal-phase HPLC for the preparation and analysis of chlorophylls (Chls)1, and have detected two molecules of Chl a' (C132 epimer of Chl a) in close association with P700, the primary donor of photosystem (PS) I of higher plants and cyanobacteria.2-4 However, this claim has occasionally met serious skepticism based on previous warnings against the use of siliceous adsorbents for the chromatography of chlorophylls.5,6The questions raised so far concern the identity of each peak, denaturation and recovery in the column, and the integrity of the pigments during extraction.Though we have repeatedly addressed these questions"2, we now feel that it is appropriate to assure the assignments and to compare the performance of our normal-phase HPLC with that of reversed-phase HPLC, which is generally considered to be sufficiently inert with respect to the elution behaviors of chlorophyllous pigments. Experimental InstrumentationFour HPLC systemsl''-9, of which the operational conditions are summarized in Table 1, were employed in the present work.1H-NMR spectra were recorded on a 270 MHz NMR spectrometer (JEOL GSX-270) for acetone-d6 solutions with tetramethylsilane as an internal standard. The pulse angle was 45° (4.3 µs), the acquisition time 5.5 s, and 64 transients were accumulated. The measurements were carried out at 25° C. Resonance peaks were assigned by comparisons with those reported by Hynninen and Lotjonen.10 Positive-ion FAB-MS measurements were performed with a JEOL JMS-700 MStation. Pigments (25 nmol) were dissolved in chloroform (10 µl), and 1 sl of the solution was submitted to analysis using 3-nitrobenzyl alcohol as the matrix.ll-13 Ions were produced by

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Chlorophyll a′/P-700 and pheophytin a/P-680 stoichiometries in higher plants and cyanobacteria determined by HPLC analysis

Cited by 88 publications

References 36 publications

Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Separation and Determination of Minor Photosynthetic Pigments by Reversed-phase HPLC with Minimal Alteration of Chlorophylls

Effects of 24-epibrassinolide Pre-treatment on UV-B-Induced Changes in the Pigment Content of Pea Leaves

Integrity of Chlorophyllous Pigments in Silica Normal-Phase HPLC

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