Formation of Chlorophyll <i>b</i>, and the Fluorescence Properties and Photochemical Activities of Isolated Plastids from Greening Pea Seedlings

Thorne, S.W.; Boardman, N.K.

doi:10.1104/pp.47.2.252

Cited by 69 publications

(31 citation statements)

References 31 publications

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“…At the later stages of needle development the relative amplitude of the 685-/ 695-nm bands decreased with respect to that at 735 nm. The evolution of the fluorescence spectrum was very similar to that found with P. jeffreyi cotyledons cultured in similar conditions (Michel-Wolwertz, 1977;Schoefs et al, 1995a) and in greening angiosperm leaves (Thorne and Boardman, 1971;Schoefs and Franck, 1991;Schoefs et al, 1992). By analogy with previous studies done on etiolated, greening, and fully green leaves (Govindjee and Wasielewski, 1989;Schoefs et al, 1992Schoefs et al, , 1994Dreyfuss and Thornber, 1994), we attributed the different bands to nonphotoactive Pchlide a (630 nm), to photoactive Pchlide a (654 nm), to the internal PSII light-harvesting complexes (CP43 and CP47, 685 and 695 nm, respectively), and to the PSI light-harvesting complex (735 nm).…”

Section: Changes In the In Situ 77 K Fluorescence Spectrum Of Pchlidesupporting

confidence: 62%

Chlorophyll Synthesis in Dark-Grown Pine Primary Needles

Schoefs

Franck

1998

Plant Physiology

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The pigment content of dark-grown primary needles of Pinus jeffreyi L. and Pinus sylvestris L. was determined by highperformance liquid chromatography. The state of protochlorophyllide a and of chlorophylls during dark growth were analyzed by in situ 77 K fluorescence spectroscopy. Both measurements unambiguously demonstrated that pine primary needles are able to synthesize chlorophyll in the dark. Norflurazon strongly inhibited both carotenoid and chlorophyll synthesis. Needles of plants treated with this inhibitor had low chlorophyll content, contained only traces of xanthophylls, and accumulated carotenoid precursors. The first form of chlorophyll detected in young pine needles grown in darkness had an emission maximum at 678 nm. Chlorophyll-protein complexes with in situ spectroscopic properties similar to those of fully green needles (685, 695, and 735 nm) later accumulated in untreated plants, whereas in norflurazon-treated plants the photosystem I emission at 735 nm was completely lacking. To better characterize the light-dependent chlorophyll biosynthetic pathway in pine needles, the 77 K fluorescence properties of in situ protochlorophyllide a spectral forms were studied. Photoactive and nonphotoactive protochlorophyllide a forms with emission properties similar to those reported for dark-grown angiosperms were found, but excitation spectra were substantially red shifted. Because of their lower chlorophyll content, norflurazon-treated plants were used to study the protochlorophyllide a photoreduction process triggered by one light flash. The first stable chlorophyllide photoproduct was a chlorophyllide a form emitting at 688 nm as in angiosperms. Further chlorophyllide a shifts usually observed in angiosperms were not detected. The rapid regeneration of photoactive protochlorophyllide a from nonphotoactive protochlorophyllide after one flash was demonstrated.

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Section: Changes In the In Situ 77 K Fluorescence Spectrum Of Pchlidesupporting

confidence: 62%

Chlorophyll Synthesis in Dark-Grown Pine Primary Needles

Schoefs

Franck

1998

Plant Physiology

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“…The (19) and the build-up of chlorophyll heterogeneity during greening (4,5) suggests that the photosynthetic apparatus is assembled in steps. Therefore, it appears possible that these differences in the preservation of the inner plastid membranes might reflect light-induced differences in their chemical composition.…”

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Chloroplast Maintenance and Partial Differentiation in Vitro

Rebeiz¹,

Larson²,

Weier³

et al. 1973

Plant Physiol.

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Tissue homogenates, etioplasts, and developing chloroplasts were prepared from cucumber (Cumucis sativus L.) cotyledons in tris-sucrose. They were incubated aerobically in the dark or in the light at pH 7.7 in the presence or absence of a cofactor mixture containing coenzyme A, glutathione, potassium phosphate, methyl alcohol, magnesium, nicotinamide adenine dinucleotide, and adenosine triphosphate. These (1,6,8,9). These organelles appear to possess certain amounts of RNA and DNA as well as the machinery for the biosynthesis of lipid, protein, and nucleic acids (9). They seem capable of dividing in vitro (17) and of forming their own protochlorophyll and chlorophyll from simple substrates (14,15,22). The differentiation of etioplasts into chloroplasts during greening is accompanied by substantial synthesis and accumulation of chlorophyll (11), carotenoids (12, 13), insoluble proteins, and colorless lipids (10). Likewise, dividing chloroplasts must grow before reaching full size (17). An assessment of cellular control during chloroplast differentiation, growth, maintenance, and division is therefore indispensable in evaluating the degree of autonomy of this organelle. (14,15). Cotyledons to be preirradiated were harvested with full hypocotyl hook as previously described (7). They were placed in beakers with enough distilled H20 to keep them moist and were illuminated with 240 ft-c of white fluorescent light at 28 C for 2.5 or 4.5 hr (15).Chemicals. The commercial sources of chemicals were reported elsewhere (14, 15). Preparation of Unfortified and Fortified Crude Homogenates. "Unfortified" crude homogenates were prepared from 4.5-day-old cotyledons as follows. Four grams of etiolated or greening cotyledons were gently ground with mortar and pestle without sand in 6.0 ml of 0.5 M sucrose and 0.2 M tris-HCl, pH 8.0, at 0 to 5 C. The brei was filtered through four layers of cheesecloth. About 5 ml of unfortified crude homogenates were obtained. "Fortified" crude homogenates were prepared by grinding 4 g of etiolated or greening cotyledons in 6.0 ml of fortified tris-HCl and 0.

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“…The structural changes which occur in pea etioplasts following illumination of dark-grown seedlings were examined in this TIhe onset of photochemical oxygen evolution (Hill activity) in plastids isolated from 10-day-old pea seedlings is first observable at about 5 hr of greening (2,16). The Hill activity increases rapidly between 5 and 8 hr of greening, and this correlates with the formation of grana.…”

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“…Previously in this laboratory we have studied the development of photochemical activity in plastids from greening pea (16) and bean seedlings (1) in relation both to chlorophyll formation and the changes in fine structure of the plastids.…”

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Lipid Composition of Pea and Bean Leaves during Chloroplast Development

Roughan

Boardman

1972

Plant Physiol.

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The changes in composition of the complex lipids were followed during the greening of dark-grown pea (Pisum sativum) and bean (Phaseolus vulgaris) seedlings. No Whole leaves were used in the present work to avoid the possibility of enzymic degradation of glycerolipids during plastid isolation (8,15). Certain glycerolipids (e.g., the galactolipids) are confined mainly to the plastids (6), and therefore particular attention has been paid to changes in the galactolipids during greening. Since the completion of this work, Tremolieres and Lepage (17) have reported on the lipid composition of greening pea seedlings. However, their study was concerned more with long term changes (up to 96 hr), while the present work has concentrated on the early stages of the greening process, during which the dramatic changes in structure of the plastids occur (7). The onset of photosynthetic oxygen evolution is also observed during the first few hours of greening (1 6).The changes in fine structure which occur in higher plant plastids during chloroplast development have been studied extensively (7). The internal membranes of an etioplast of a dark-grown seedling are organized mainly into one or more paracrystalline three-dimensional tubular structures which are termed "prolamellar bodies." On illumination of the etiolated seedlings, the prolamellar bodies are dispersed into sheets of perforated membranes which give rise to the lamellae. Then follows a fusion and elaboration of the lamellae in certain regions to form grana. These electron microscope studies strongly suggest that there is a continuity of membrane structures during chloroplast development, rather than a breakdown of the membranes of the prolamellar bodies of the etioplast followed by a resynthesis of the membranes of the developing chloroplast.In the present study, we have examined the glycerolipid composition of greening pea and bean seedlings. Chloroplast thylakoids contain about 50% by weight of lipid, mainly glycerolipid (6), and therefore it seemed likely that changes in membrane structure and the acquisition of photochemical activity might be reflected in changes in the various chloroplast glycerolipids. On the other hand, a constancy of lipid composition in the early stages of greening would support the view 1Permanent address: Plant Physiology Division, D.S.I.R., Palmerston North, New Zealand. 31MATERIALS AND METHODS Peas (Pisum sativum L. cv. Greenfeast) and beans (Phaseolus vulgaris L. cv. Brown Beauty) were germinated in the dark at 25 C in trays containing vermiculite wetted with Hoagland's nutrient solution. After 10 to 11 days (for peas) and 11 to 14 days (beans), trays were transferred to continuous illumination and maintained at 25 C. White fluorescent tubes provided an average of 400 ft-c at leaf level. At appropriate intervals 1 g fresh weight of leaves was harvested from four trays, wrapped in aluminum foil, and steamed for 10 min (19).The killed leaves were then extracted with 20 ml of chloroform-methanol (2: 1 v/v) in a Tenbroeck tissue gr...

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Formation of Chlorophyll b, and the Fluorescence Properties and Photochemical Activities of Isolated Plastids from Greening Pea Seedlings

Cited by 69 publications

References 31 publications

Chlorophyll Synthesis in Dark-Grown Pine Primary Needles

Chlorophyll Synthesis in Dark-Grown Pine Primary Needles

Chloroplast Maintenance and Partial Differentiation in Vitro

Lipid Composition of Pea and Bean Leaves during Chloroplast Development

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