Quantitative fluorescence microscopy on dynamic changes of plastid nucleoids during wheat development

Miyamura, Shinichi; Nagata, Toshiyuki; Kuroiwa, Tsuneyoshi

doi:10.1007/bf01293188

Cited by 110 publications

(65 citation statements)

References 15 publications

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“…During the development of chloroplasts from proplastids in wheat (Miyamura et al, 1986) and Arabidopsis thaliana (Fujie et al, 1994), nucleoids first increase in size and then fragment and disperse. What changes at the level of cpDNA molecules accompany these cytological changes?…”

Section: The Chloroplast Nucleoid: a Revised Viewmentioning

confidence: 99%

Circular Chloroplast Chromosomes: The Grand Illusion

2004

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Section: The Chloroplast Nucleoid: a Revised Viewmentioning

confidence: 99%

Circular Chloroplast Chromosomes: The Grand Illusion

2004

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“…Amyloplast-nucleoids (nuclei) were isolated from BY-2 cells that had been cultured in the modified medium for 48 h. The cells were collected from 1.8 L of culture by sedimentation under gravity, and they were treated with enzyme solution (0. 6 Protoplasts were disrupted by repeated passages through layers of nylon mesh, the pore sizes of which was decreased stepwise from 300 to 20 Am as the disruption proceeded.…”

Section: Isolation Of Proplastid-nucleoids and Amyloplastnucleoidsmentioning

confidence: 99%

“…In addition, 0.2 M sucrose, 0.2 mm EDTA, 0.5 mM spermidine, 2.8 mnim 2-mercaptoethanol, and 0.16 mm PMSF were introduced from the TAN buffer. The reaction mixtures were incubated for 1 h at 260C, and incorporation of [5,6-3H]UTP into RNA was determined by scintillation counting after spotting the reaction mixtures onto DE-81 paper, with four washes in 5% (w/v) Na2HPO4, two in water, and two in ethanol. 32P-labeled transcripts, which were used as hybridization probes, were synthesized by the isolated plastidnucleoids as described above with the exception that [5,6-3H]-UTP was replaced by 0.1 Mm [a-32P]UTP.…”

Section: In Vitro Transcriptionmentioning

confidence: 99%

Conversion of Proplastids to Amyloplasts in Tobacco Cultured Cells Is Accompanied by Changes in the Transcriptional Activities of Plastid Genes

Sakai¹,

Kawano²,

Kuroiwa³

1992

Plant Physiol.

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When tobacco (Nicotiana tabacum L.) cultured cells at the stationary phase were transferred to culture medium that contained cytokinin (benzyladenine) instead of auxin (2,4-dichlorophenoxyacetic acid), proplastids in the BY-2 cells were converted to amyloplasts within 48 h. The data obtained from in vitro transcription assays using isolated plastid-nucleoids (nuclei) strongly suggested that amyloplast formation in BY-2 cells was accompanied by changes in the transcriptional activities of plastid genes.Plastids in higher plants have their own DNA, and they differentiate into various specialized organelles in an organspecific manner. It is of interest to determine how the expression of plastid genes is controlled during the process of plastid differentiation. The regulation of plastid gene expression during the biogenesis of chloroplasts has been extensively studied because of the importance of their principal function, photosynthesis. By contrast, the biogenesis of amyloplasts and the regulation of plastid gene expression during this process have not been examined in detail, even though amyloplasts are the sites of synthesis and storage of starch and are important constituents of the storage organs of higher plants. Studies of the processes and the mechanisms involved in the formation of amyloplasts are essential if we are to clarify the way in which plants turn on their storage functions.To study the process of amyloplast formation, we have established procedures for the induction of amyloplast formation in suspension-cultured tobacco (Nicotiana tabacum L.) cells (line BY-2). We have also analyzed the regulation of transcription of plastid genes in amyloplasts by an in vitro transcription assay using isolated plastid-nucleoids. The conditions for the culture of tobacco cell line BY-2, derived from Nicotiana tabacum L. cv Bright Yellow-2 (3), were described previously (14). For the induction of the transformation of proplastids to amyloplasts, auxin (2,4-D; 0.2 mg L-'), which was present in the conventional medium, was replaced by cytokinin (BA; 1 mg L-`), and 8.5 mL of a suspension of cells in stationary phase were transferred into 150 mL of the modified medium. The formation of amyloplasts was monitored by counting numbers of amyloplasts per cell under a light microscope. For measurement of the starch content of the cells, protoplasts prepared from 1 x 105 cells were treated with Sarkosyl (1%, w/v) and proteinase K (1 mg mLU-) at 300C for 1 h, and the hot-ethanol (80%, v/v) insoluble fraction was used for the extraction of starch. Starch was extracted with hot water and perchloric acid and quantified by the phenol-sulfuric acid method (1). Isolation of Proplastid-Nucleoids and AmyloplastNucleoidsProcedures for the isolation of proplastid-nucleoids (nuclei) were described previously (8). Amyloplast-nucleoids (nuclei) were isolated from BY-2 cells that had been cultured in the modified medium for 48 h. The cells were collected from 1.8 L of culture by sedimentation under gravity, and they were treated wi...

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“…Although the replication and transcription of cpDNA are likely to be regulated by the organization of plastid nucleoids, little is known about the relationship between the structure and function of nucleoids. In this regard, we (Sato et al, 1993 as well as others (Miyamura et al, 1986;Sodmergen et al, 1989) have pointed out that the morphology of plastid nucleoids is subject to dynamic changes during the development of plastids. The proplastid contains a single nucleoid, which is located at the center of the plastid.…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of the PEND Protein, a Novel bZIP Protein Present in the Envelope Membrane That Is the Site of Nucleoid Replication in Developing Plastids

Sato

Ohshima²,

Watanabe

et al. 1998

Plant Cell

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Plastid nucleoids are known to bind to the envelope membrane in developing chloroplasts. Here, plastid DNA is extensively replicated. We previously detected a DNA binding protein in the inner envelope membranes of developing plastids in pea and named it PEND (for plastid envelope DNA binding) protein. In this study, we report on the structure and molecular characterization of a cDNA for the PEND protein. As a result of screening cDNA libraries in gt11 with one of the target sequences of the PEND protein as a probe, we obtained a clone (PD2) for a novel DNA binding protein consisting of 633 amino acid residues. Analysis of the N-terminal sequence of the purified PEND protein indicated that the transit peptide is just 16 residues long. The PEND protein was detected specifically in the plastid envelope membrane of young unopened leaf buds by immunoblot analysis. The PEND protein consists of a basic region plus zipper region, an unprecedented sextuple repeat region, and a putative membrane-spanning region. The basic region with a zipper region seems to have diverged from that of other plant transcription factors. In addition, the PEND protein could be a distant homolog of the trans -Golgi network integral membrane proteins. The PEND protein is therefore a novel type of DNA binding protein that binds to the membrane as an intrinsic membrane protein.

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Quantitative fluorescence microscopy on dynamic changes of plastid nucleoids during wheat development

Cited by 110 publications

References 15 publications

Circular Chloroplast Chromosomes: The Grand Illusion

Circular Chloroplast Chromosomes: The Grand Illusion

Conversion of Proplastids to Amyloplasts in Tobacco Cultured Cells Is Accompanied by Changes in the Transcriptional Activities of Plastid Genes

Molecular Characterization of the PEND Protein, a Novel bZIP Protein Present in the Envelope Membrane That Is the Site of Nucleoid Replication in Developing Plastids

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