Abstract:

To study the characteristic features of the amyloplast, a uniquely differentiated plastid-type which synthesizes and accumulates reserve starch, in comparison with those of the chloroplast, these two types of plastids were isolated from white-wild and green-mutant protoplasts of cultured sycamore (Acer pseudoplatanus L.) cells, respectively. The intactness of the isolated amyloplast preparations was 70%. Electron microscopic ultrastructural analysis of both plastid types revealed unique structural features of the green-mutant chloroplasts, including well developed grana membranes and abundant ribosomal particles and plastoglobuli. After osmotic rupture of the isolated amyloplasts and chloroplasts, a clear separation of the envelope-membranes was achieved by discontinuous sucrose density gradient centrifugation. Although the visible absorption spectra of the envelope lipid components were indistinguishable between the amyloplasts and chloroplasts, the envelope-membrane polypeptide patterns were clearly distinct as judged by denaturing electrophoresis. By immunoblotting analysis using the specific antiserum raised against the pea chloroplast 29-kilodalton Pi-translocator, the amount of this carrier-protein (31-kilodalton) in the white-wild amyloplast envelope-membranes was estimated to be at least 10-fold less than in the green-mutant envelopes.

Open in

PUBMED | PMC | Google Scholar | Wikipedia

Relations:

Content

Citations

(10)

References

(16)

Affiliates

(2)

Isolation and Characterization of the Amyloplast Envelope-Membrane from Cultured White-Wild Cells of Sycamore (<em>Acer pseudoplatanus</em> L.) <sup><a href="#fn1" rid="fn1" class=" fn">1</a></sup>

Abstract

To study the characteristic features of the amyloplast, a uniquely differentiated plastid-type which synthesizes and accumulates reserve starch, in comparison with those of the chloroplast, these two types of plastids were isolated from white-wild and green-mutant protoplasts of cultured sycamore (Acer pseudoplatanus L.) cells, respectively. The intactness of the isolated amyloplast preparations was 70%. Electron microscopic ultrastructural analysis of both plastid types revealed unique structural features of the green-mutant chloroplasts, including well developed grana membranes and abundant ribosomal particles and plastoglobuli. After osmotic rupture of the isolated amyloplasts and chloroplasts, a clear separation of the envelope-membranes was achieved by discontinuous sucrose density gradient centrifugation. Although the visible absorption spectra of the envelope lipid components were indistinguishable between the amyloplasts and chloroplasts, the envelope-membrane polypeptide patterns were clearly distinct as judged by denaturing electrophoresis. By immunoblotting analysis using the specific antiserum raised against the pea chloroplast 29-kilodalton Pi-translocator, the amount of this carrier-protein (31-kilodalton) in the white-wild amyloplast envelope-membranes was estimated to be at least 10-fold less than in the green-mutant envelopes.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. [PubMed] [Google Scholar]
Flügge UI, Heldt HW. Identification of a protein involved in phosphate transport of chloroplasts. FEBS Lett. 1976 Oct 1;68(2):259–262. [PubMed] [Google Scholar]
Heldt HW. Adenine nucleotide translocation in spinach chloroplasts. FEBS Lett. 1969 Sep;5(1):11–14. [PubMed] [Google Scholar]
Heldt HW, Rapley L. Specific transport of inorganic phosphate, 3-phosphoglycerate and dihydroxyacetonephosphate, and of dicarboxylates across the inner membrane of spinach chloroplasts. FEBS Lett. 1970 Oct 5;10(3):143–148. [PubMed] [Google Scholar]
Huber SC, Akazawa T. A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells. Plant Physiol. 1986 Aug;81(4):1008–1013.[PMC free article] [PubMed] [Google Scholar]
Joyard J, Grossman A, Bartlett SG, Douce R, Chua NH. Characterization of envelope membrane polypeptides from spinach chloroplasts. J Biol Chem. 1982 Jan 25;257(2):1095–1101. [PubMed] [Google Scholar]
Macherel D, Kobayashi H, Akazawa T, Kawano S, Kuroiwa T. Amyloplast nucleoids in sycamore cells and presence in amyloplast DNA of homologous sequences to chloroplast genes. Biochem Biophys Res Commun. 1985 Nov 27;133(1):140–146. [PubMed] [Google Scholar]
Macherel D, Viale A, Akazawa T. Protein Phosphorylation in Amyloplasts Isolated from Suspension-Cultured Cells of Sycamore (Acer pseudoplatanus L.). Plant Physiol. 1986 Apr;80(4):1041–1044.[PMC free article] [PubMed] [Google Scholar]
Ngernprasirtsiri J, Macherel D, Kobayashi H, Akazawa T. Expression of Amyloplast and Chloroplast DNA in Suspension-Cultured Cells of Sycamore (Acer pseudoplatanus L.). Plant Physiol. 1988 Jan;86(1):137–142.[PMC free article] [PubMed] [Google Scholar]
Oakley BR, Kirsch DR, Morris NR. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. [PubMed] [Google Scholar]
Pain D, Blobel G. Protein import into chloroplasts requires a chloroplast ATPase. Proc Natl Acad Sci U S A. 1987 May;84(10):3288–3292.[PMC free article] [PubMed] [Google Scholar]
RACKER E. Spectrophotometric measurements of the enzymatic formation of fumaric and cis-aconitic acids. Biochim Biophys Acta. 1950 Jan;4(1-3):211–214. [PubMed] [Google Scholar]
Sandelius AS, Selstam E. Localization of Galactolipid Biosynthesis in Etioplasts Isolated from Dark-Grown Wheat (Triticum aestivum L.). Plant Physiol. 1984 Dec;76(4):1041–1046.[PMC free article] [PubMed] [Google Scholar]
Selstam E, Sandelius AS. A Comparison between Prolamellar Bodies and Prothylakoid Membranes of Etioplasts of Dark-Grown Wheat Concerning Lipid and Polypeptide Composition. Plant Physiol. 1984 Dec;76(4):1036–1040.[PMC free article] [PubMed] [Google Scholar]
Stitt M. Fructose 2,6-bisphosphate and plant carbohydrate metabolism. Plant Physiol. 1987 Jun;84(2):201–204.[PMC free article] [PubMed] [Google Scholar]
Strzalka K, Ngernprasirtsiri J, Watanabe A, Akazawa T. Sycamore amyloplasts can import and process precursors of nuclear encoded chloroplast proteins. Biochem Biophys Res Commun. 1987 Dec 16;149(2):799–806. [PubMed] [Google Scholar]

Research Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Chikusa, Nagoya 464, Japan

Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa, Nagoya 464, Japan

^{Recipient of a predoctoral student fellowship provided by the Hitachi Scholarship Foundation (Tokyo). Permanent address: Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.}

^{Recipient of a predoctoral student fellowship from the Japanese government (Mombusho). Permanent address: Department of Biochemistry, Kasetsart University, Bangkok, Thailand.}

^{Recipient of a postdoctoral fellowship provided by the Japan (JSPS)-France (Centre National de al Recherche Scientifique) Scientist Exchange Program (1986). Current address: CNRS/CENG, Grenoble-Cedex, France.}

^{Recipient of a fellowship from the Visiting Scientist Program (1986) provided by the Japanese government. Current address: Jagellonian University, Krakow, Poland.}

^{Supported by grants from the Ministry of Education, Science and Culture (Mombusho) of Japan. This is Paper No. 74 in the series “Structure and Function of Chloroplast Proteins,” and paper No. 73 is Ref. 32.}

Copyright notice

Abstract

To study the characteristic features of the amyloplast, a uniquely differentiated plastid-type which synthesizes and accumulates reserve starch, in comparison with those of the chloroplast, these two types of plastids were isolated from white-wild and green-mutant protoplasts of cultured sycamore (Acer pseudoplatanus L.) cells, respectively. The intactness of the isolated amyloplast preparations was 70%. Electron microscopic ultrastructural analysis of both plastid types revealed unique structural features of the green-mutant chloroplasts, including well developed grana membranes and abundant ribosomal particles and plastoglobuli. After osmotic rupture of the isolated amyloplasts and chloroplasts, a clear separation of the envelope-membranes was achieved by discontinuous sucrose density gradient centrifugation. Although the visible absorption spectra of the envelope lipid components were indistinguishable between the amyloplasts and chloroplasts, the envelope-membrane polypeptide patterns were clearly distinct as judged by denaturing electrophoresis. By immunoblotting analysis using the specific antiserum raised against the pea chloroplast 29-kilodalton Pi-translocator, the amount of this carrier-protein (31-kilodalton) in the white-wild amyloplast envelope-membranes was estimated to be at least 10-fold less than in the green-mutant envelopes.

Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.

Learn More