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. 1988 Sep 1;107(3):907–919. doi: 10.1083/jcb.107.3.907

Biogenesis of thylakoid membranes is controlled by light intensity in the conditional chlorophyll b-deficient CD3 mutant of wheat

PMCID: PMC2115274  PMID: 3047153

Abstract

Biogenesis of thylakoid membranes in the conditional chlorophyll b- deficient CD3 mutant of wheat is dramatically altered by relatively small differences in the light intensity under which seedlings are grown. When the CD3 mutant is grown at 400 microE/m2 S (high light, about one-fifth full sunlight) plants are deficient in chlorophyll b (chlorophyll a/b ratio greater than 6.0) and lack or contain greatly reduced amounts of the chlorophyll a/b-binding complexes CPII/CPII (mobile or peripheral LHCII), CP29, CP24 and LHCI, as shown by mildly denaturing 'green gel' electrophoresis, by fully denaturing SDS-PAGE, and by Western blot analysis. High light CD3 chloroplasts display an unusual morphology characterized by large, sheet-like stromal thylakoids formed into parallel unstacked arrays and a limited number of small grana stacks displaced toward the edges of the arrays. Changes in the supramolecular organization of CD3 thylakoids, seen with freeze- fracture electron microscopy, include a reduction in the size of EFs particles, which correspond to photosystem II centers with variable amounts of attached LHCII, and a redistribution of EF particles from the stacked to the unstacked regions. When CD3 seedlings are grown at 150 microE/m2 S (low light) there is a substantial reversal of all of these effects. Thus, chlorophyll b and the chlorophyll a/b-binding proteins accumulate to near wild-type levels (chlorophyll a/b ratio = 3.5-4.5) and thylakoid morphology is more nearly wild type in appearance. Growth of the CD3 mutant in the presence of chloramphenicol stimulates the accumulation of chlorophyll b and its binding proteins (Duysen, M. E., T. P. Freeman, N. D. Williams, and L. L. Huckle. 1985. Plant Physiol. 78:531-536). We show that this partial rescue of the CD3 high light phenotype is accompanied by large changes in thylakoid structure. The CD3 mutant, which defines a new class of chlorophyll b- deficient phenotype, is discussed in the more general context of chlorophyll b deficiency.

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Selected References

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  1. Armond P. A., Staehelin L. A., Arntzen C. J. Spatial relationship of photosystem I, photosystem II, and the light-harvesting complex in chloroplast membranes. J Cell Biol. 1977 May;73(2):400–418. doi: 10.1083/jcb.73.2.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellemare G., Bartlett S. G., Chua N. H. Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the chlorina f2 mutant of barley. J Biol Chem. 1982 Jul 10;257(13):7762–7767. [PubMed] [Google Scholar]
  4. Boardman N. K., Highkin H. R. Studies on a barley mutant lacking chlorophyll b. I. Photochemical activity of isolated chloroplasts. Biochim Biophys Acta. 1966 Oct 10;126(2):189–199. doi: 10.1016/0926-6585(66)90054-9. [DOI] [PubMed] [Google Scholar]
  5. Camm E. L., Green B. R. Fractionation of Thylakoid Membranes with the Nonionic Detergent Octyl-beta-d-glucopyranoside: RESOLUTION OF CHLOROPHYLL-PROTEIN COMPLEX II INTO TWO CHLOROPHYLL-PROTEIN COMPLEXES. Plant Physiol. 1980 Sep;66(3):428–432. doi: 10.1104/pp.66.3.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carter D. P., Staehelin L. A. Proteolysis of chloroplast thylakoid membranes. II. Evidence for the involvement of the light-harvesting chlorophyll a/b-protein complex in thylakoid stacking and for effects of magnesium ions on photosystem II-light-harvesting complex aggregates in the absence of membrane stacking. Arch Biochem Biophys. 1980 Apr 1;200(2):374–386. doi: 10.1016/0003-9861(80)90367-7. [DOI] [PubMed] [Google Scholar]
  7. Day D. A., Ryrie I. J., Fuad N. Investigations of the role of the main light-harvesting chlorophyll-protein complex in thylakoid membranes. Reconstitution of depleted membranes from intermittent-light-grown plants with the isolated complex. J Cell Biol. 1984 Jan;98(1):163–172. doi: 10.1083/jcb.98.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. De Greef J., Butler W. L., Roth T. F. Greening of etiolated bean leaves in far red light. Plant Physiol. 1971 Apr;47(4):457–464. doi: 10.1104/pp.47.4.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dunahay T. G., Schuster G., Staehelin L. A. Phosphorylation of spinach chlorophyll-protein complexes. CPII, but not CP29, CP27, or CP24, is phosphorylated in vitro. FEBS Lett. 1987 May 4;215(1):25–30. doi: 10.1016/0014-5793(87)80107-2. [DOI] [PubMed] [Google Scholar]
  10. Dunahay T. G., Staehelin L. A. Isolation and Characterization of a New Minor Chlorophyll a/b-Protein Complex (CP24) from Spinach. Plant Physiol. 1986 Feb;80(2):429–434. doi: 10.1104/pp.80.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Duysen M. E., Freeman T. P., Williams N. D., Huckle L. L. Chloramphenicol stimulation of light harvesting chlorophyll protein complex accumulation in a chlorophyll B deficient wheat mutant. Plant Physiol. 1985 Jul;78(3):531–536. doi: 10.1104/pp.78.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Duysen M. E., Freeman T. P., Williams N. D., Olson L. L. Regulation of excitation energy in a wheat mutant deficient in light-harvesting pigment protein complex. Plant Physiol. 1984 Nov;76(3):561–566. doi: 10.1104/pp.76.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farchaus J., Dilley R. A. Purification and partial sequence of the Mr 10,000 phosphoprotein from spinach thylakoids. Arch Biochem Biophys. 1986 Jan;244(1):94–101. doi: 10.1016/0003-9861(86)90097-4. [DOI] [PubMed] [Google Scholar]
  14. GUNNING B. E. THE FINE STRUCTURE OF CHLOROPLAST STROMA FOLLOWING ALDEHYDE OSMIUM-TETROXIDE FIXATION. J Cell Biol. 1965 Jan;24:79–93. doi: 10.1083/jcb.24.1.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goodchild D. J., Highkin H. R., Boardman N. K. The fine structure of chloroplasts in a barley mutant lacking chlorophyll B. Exp Cell Res. 1966 Oct;43(3):684–688. doi: 10.1016/0014-4827(66)90045-0. [DOI] [PubMed] [Google Scholar]
  16. Greene B. A., Allred D. R., Morishige D. T., Staehelin L. A. Hierarchical Response of Light Harvesting Chlorophyll-Proteins in a Light-Sensitive Chlorophyll b-Deficient Mutant of Maize. Plant Physiol. 1988 Jun;87(2):357–364. doi: 10.1104/pp.87.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hallam T. J., Simpson A. W., O'Connor N., Rink T. J. Control and interrelation of aggregation and secretion; the roles of Ca2+, diacylglycerol and thromboxane with particular reference to ADP stimulation. Adv Exp Med Biol. 1985;192:145–162. doi: 10.1007/978-1-4615-9442-0_11. [DOI] [PubMed] [Google Scholar]
  18. Jensen K. H., Herrin D. L., Plumley F. G., Schmidt G. W. Biogenesis of photosystem II complexes: transcriptional, translational, and posttranslational regulation. J Cell Biol. 1986 Oct;103(4):1315–1325. doi: 10.1083/jcb.103.4.1315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kyle D. J., Staehelin L. A., Arntzen C. J. Lateral mobility of the light-harvesting complex in chloroplast membranes controls excitation energy distribution in higher plants. Arch Biochem Biophys. 1983 Apr 15;222(2):527–541. doi: 10.1016/0003-9861(83)90551-9. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Leto K. J., Keresztes A., Arntzen C. J. Nuclear Involvement in the Appearance of a Chloroplast-Encoded 32,000 Dalton Thylakoid Membrane Polypeptide Integral to the Photosystem II Complex. Plant Physiol. 1982 Jun;69(6):1450–1458. doi: 10.1104/pp.69.6.1450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mayfield S. P., Nelson T., Taylor W. C. The Fate of Chloroplast Proteins during Photooxidation in Carotenoid-Deficient Maize Leaves. Plant Physiol. 1986 Nov;82(3):760–764. doi: 10.1104/pp.82.3.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McDonnel A., Staehelin L. A. Adhesion between liposomes mediated by the chlorophyll a/b light-harvesting complex isolated from chloroplast membranes. J Cell Biol. 1980 Jan;84(1):40–56. doi: 10.1083/jcb.84.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miller K. R., Cushman R. A. A chloroplast membrane lacking photosystem II. Thylakoid stacking in the absence of the photosystem II particle. Biochim Biophys Acta. 1979 Jun 5;546(3):481–497. doi: 10.1016/0005-2728(79)90083-5. [DOI] [PubMed] [Google Scholar]
  25. Miller K. R., Miller G. J., McIntyre K. R. The light-harvesting chlorpohyll-protein complex of photosystem II. Its location in the photosynthetic membrane. J Cell Biol. 1976 Nov;71(2):624–638. doi: 10.1083/jcb.71.2.624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Miller K. R., Ohad I. Chloroplast membrane biogenesis in Chlamydomonas: correlation between the formation of membrane components and membrane structure. Cell Biol Int Rep. 1978 Nov;2(6):537–549. doi: 10.1016/0309-1651(78)90062-0. [DOI] [PubMed] [Google Scholar]
  27. Mullet J. E., Arntzen C. J. Simulation of grana stacking in a model membrane system. Mediation by a purified light-harvesting pigment-protein complex from chloroplasts. Biochim Biophys Acta. 1980 Jan 4;589(1):100–117. doi: 10.1016/0005-2728(80)90135-8. [DOI] [PubMed] [Google Scholar]
  28. Nakatani H. Y., Baliga V. A clover mutant lacking the chlorophyll a- and b-containing protein antenna complexes. Biochem Biophys Res Commun. 1985 Aug 30;131(1):182–189. doi: 10.1016/0006-291x(85)91787-5. [DOI] [PubMed] [Google Scholar]
  29. Palmer J. D. Comparative organization of chloroplast genomes. Annu Rev Genet. 1985;19:325–354. doi: 10.1146/annurev.ge.19.120185.001545. [DOI] [PubMed] [Google Scholar]
  30. Ryrie I. J., Anderson J. M., Goodchild D. J. The role of the light-harvesting chlorophyll a/b protein complex in chloroplast membrane stacking. Cation-induced aggregation of reconstituted proteoliposomes. Eur J Biochem. 1980 Jun;107(2):345–354. doi: 10.1111/j.1432-1033.1980.tb06035.x. [DOI] [PubMed] [Google Scholar]
  31. Ryrie I. J., Fuad N. Membrane adhesion in reconstituted proteoliposomes containing the light-harvesting chlorophyll a/b-protein complex: the role of charged surface groups. Arch Biochem Biophys. 1982 Apr 1;214(2):475–488. doi: 10.1016/0003-9861(82)90051-0. [DOI] [PubMed] [Google Scholar]
  32. Staehelin L. A., Arntzen C. J. Regulation of chloroplast membrane function: protein phosphorylation changes the spatial organization of membrane components. J Cell Biol. 1983 Nov;97(5 Pt 1):1327–1337. doi: 10.1083/jcb.97.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Staehelin L. A. Reversible particle movements associated with unstacking and restacking of chloroplast membranes in vitro. J Cell Biol. 1976 Oct;71(1):136–158. doi: 10.1083/jcb.71.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steinback K. E., Burke J. J., Arntzen C. J. Evidence for the role of surface-exposed segments of the light-harvesting complex in cation-mediated control of chloroplast structure and function. Arch Biochem Biophys. 1979 Jul;195(2):546–557. doi: 10.1016/0003-9861(79)90381-3. [DOI] [PubMed] [Google Scholar]

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