Skip to main content

Main menu

  • Home
  • Content
    • Current Issue
    • Archive
    • Preview Papers
  • About
    • Editorial Board and Staff
    • About the Journal
    • Terms & Privacy
  • More
    • Alerts
    • Contact Us
  • Submit a Manuscript
    • Instructions for Authors
    • Submit a Manuscript
  • Other Publications
    • Plant Physiology
    • The Plant Cell
    • Plant Direct
    • The Arabidopsis Book
    • Teaching Tools in Plant Biology
    • ASPB
    • Plantae

User menu

  • My alerts
  • Log in

Search

  • Advanced search
Plant Cell
  • Other Publications
    • Plant Physiology
    • The Plant Cell
    • Plant Direct
    • The Arabidopsis Book
    • Teaching Tools in Plant Biology
    • ASPB
    • Plantae
  • My alerts
  • Log in
Plant Cell

Advanced Search

  • Home
  • Content
    • Current Issue
    • Archive
    • Preview Papers
  • About
    • Editorial Board and Staff
    • About the Journal
    • Terms & Privacy
  • More
    • Alerts
    • Contact Us
  • Submit a Manuscript
    • Instructions for Authors
    • Submit a Manuscript
  • Follow PlantCell on Twitter
  • Visit PlantCell on Facebook
  • Visit Plantae
Research ArticleResearch Article
You have accessRestricted Access

Phytochrome Regulation and Differential Expression of Gibberellin 3β-Hydroxylase Genes in Germinating Arabidopsis Seeds

Shinjiro Yamaguchi, Maria W. Smith, Robert G. S. Brown, Yuji Kamiya, Tai-ping Sun
Shinjiro Yamaguchi
aFrontier Research Program, Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan
bDevelopmental, Cell, and Molecular Biology Group, Department of Botany, Box 91000, Duke University, Durham, North Carolina 27708-1000
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: shinjiro@acpub.duke.edu
Maria W. Smith
aFrontier Research Program, Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert G. S. Brown
aFrontier Research Program, Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yuji Kamiya
aFrontier Research Program, Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tai-ping Sun
bDevelopmental, Cell, and Molecular Biology Group, Department of Botany, Box 91000, Duke University, Durham, North Carolina 27708-1000
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site

Published December 1998. DOI: https://doi.org/10.1105/tpc.10.12.2115

  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Sequence Alignment of GA 3β-Hydroxylases.

    An alignment is shown for the deduced amino acid sequences of Arabidopsis GA4 (Chiang et al., 1995, 1997) and GA4H (GenBank accession number AF070937), pea LE (Lester et al., 1997; Martin et al., 1997), and GA 2β,3β-hydroxylase from pumpkin (Cm; Lange et al., 1997). Identical amino acid residues conserved between GA4H and at least one other protein are indicated in reverse type, and similar residues are in gray boxes. Gaps introduced to optimize the alignment are indicated as dots, and sequence truncations are indicated by dashes. The boxes were drawn using the BOXSHADE web site (http://ulrec3.unil.ch/software/BOX_form.html).

    • Download figure
    • Open in new tab
    • Download powerpoint
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    In Vitro Functional Analysis of the Recombinant GA4H Protein.

    (A) Diagram showing conversion of GA9 and GA20 to GA4 and GA1 by 3β-hydroxylation.

    (B) Functional analysis of the GA4H protein. Lysates of E. coli containing the MBP (as a control) or MBP–GA4H were incubated with 14C-GA9. The reaction mixture was separated on a silica gel by thin-layer chromatography. The positions of authentic GA4 and GA9 are indicated by arrowheads.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Developmental Regulation of GA4 and GA4H Expression.

    Autoradiography of RNA gel blots containing 25 μg of total RNAs isolated from different tissues, as labeled. Germinating seeds under continuous white light were harvested 12 hr (h) after imbibition. Young seedlings (harvested at 48, 72, and 96 hr after imbibition) and 14- and 35-day (d)-old plants were grown under 16-hr-light and 8-hr-dark cycles. Aerial tissues (rosette) were harvested from 14-day-old plants. Cauline leaves (leaf), main stems (stem), flower clusters (flower), and siliques (siliques 1, 2, and 3) were harvested from 35-day-old plants. Silique 1 has embryos at globular and heart-shaped stages. Embryos at torpedo to upturn U stages are contained in silique 2. Silique 3 has mature green seeds. The membrane was hybridized with the radiolabeled GA4 or GA4H antisense RNA probe and then reprobed with the the radiolabeled 18S rDNA probe as a loading control.

    • Download figure
    • Open in new tab
    • Download powerpoint
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    R Light–Induced Expression of the GA4 and GA4H Genes and Germination in Wild-Type Arabidopsis Seeds.

    (A) Diagram showing different light treatments. The seeds were imbibed in the dark and then irradiated with an FR light pulse 1 hr after imbibition. The seeds were then either irradiated with an R light pulse (stippled box) 24 hr after the FR light pulse (R in [B]) or incubated without the R light pulse in the dark (D in [B]). The triangle indicates the starting time of imbibition. The vertical arrow indicates the beginning of the R light pulse, which was set as 0 hr in the experiments shown in (B).

    (B) Autoradiography of RNA blots containing 12.5 μg of total RNAs prepared from germinating wild-type seeds under different light conditions as described in (A). The time after the R light pulse is indicated above the blot. The membrane was hybridized with the GA4 or GA4H antisense RNA probe and then reprobed with the radiolabeled 18S rDNA probe as a loading control.

    (C) Germination frequency and the levels of GA4 and GA4H mRNAs after the R light pulse. The highest mRNA levels for each gene were set as 100.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    R Light–Induced Expression of the GA4 and GA4H Genes in Imbibed ga1-3 Seeds.

    (A) Autoradiography of RNA blots containing 12.5 μg of total RNAs from imbibed ga1-3 seeds after light treatments, as diagrammed in Figure 4A. The membranes were hybridized with the GA4 or GA4H antisense RNA probe and then reprobed with the radiolabeled 18S rDNA probe as a loading control. Abbreviations are as given in Figure 4A.

    (B) A diagram showing relative levels of cross-hybridization of the GA4H probe with the GA4 mRNAs (black bars) and the net GA4H mRNA (open bars) in each lane in (A). The highest level of the hybridization signal was set as 100.

    (C) GA4 and GA4H mRNA levels after the R light pulse. The highest mRNA levels for each gene were set as 100.

    • Download figure
    • Open in new tab
    • Download powerpoint
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Effects of the phyB-1 Mutation on GA4 and GA4H Expression.

    (A) Diagram showing different light treatments. The seeds were imbibed in the dark and then irradiated with an FR light pulse 1 hr after the start of imbibition. The seeds were then either irradiated with an R light pulse 2 hr after the FR light pulse (R in [B]) or incubated without an R light pulse in the dark (D in [B]). Symbols are as given in the legend to Figure 4A.

    (B) Autoradiography of RNA gel blots containing 12.5 μg of total RNAs prepared from germinating wild-type (WT) or phyB-1 seeds under the light conditions given in (A). The membrane was hybridized with the GA4 or GA4H antisense RNA probe and then reprobed with a radiolabeled 18S rDNA probe as a loading control.

    • Download figure
    • Open in new tab
    • Download powerpoint
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 7.

    Photoreversibility of GA4 and GA4H Expression.

    (A) Diagram showing different light treatments. The seeds were imbibed in the dark and then irradiated with an FR light pulse 1 hr after imbibition. The seeds were then incubated in the dark (D), irradiated with an R light pulse (R), or treated with a second FR light pulse immediately after the R light pulse (R+FR). Symbols are as given in the legend to Figure 4A.

    (B) Autoradiography of RNA blots containing 12.5 μg of total RNAs prepared from germinating wild-type seeds under the light conditions given in (A). The membrane was hybridized with the GA4 or GA4H antisense RNA probe and then reprobed with a radiolabeled 18S rDNA probe as a loading control.

  • Figure 8.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 8.

    Effects of the ga1-3 Mutant Background and GA4 Application on GA4 and GA4H Expression in Imbibed Seeds.

    Shown is autoradiography of RNA gel blots containing 12.5 μg of total RNA from imbibed wild-type (WT) or ga1-3 seeds.

    (A) RNA was isolated from seeds at 4 hr after the light treatments, as shown in Figure 4A.

    (B) The ga1-3 seeds were imbibed in water (−GA4) or in 100 μM GA4 under continuous white light. Germination percentage is shown below the blots.

    The membranes were hybridized with the GA4 or GA4H antisense RNA probe and then reprobed with a radiolabeled 18S rDNA probe as a loading control.

  • Figure 9.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 9.

    Proposed Model for the Regulation of GA4 and GA4H Expression.

    The last two steps of the GA biosynthetic pathway are shown in the dashed box. Arrows indicate positive regulation. The feedback inhibition is shown by the T-bar.

PreviousNext
Back to top

Table of Contents

Print
Download PDF
Email Article

Thank you for your interest in spreading the word on Plant Cell.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Phytochrome Regulation and Differential Expression of Gibberellin 3β-Hydroxylase Genes in Germinating Arabidopsis Seeds
(Your Name) has sent you a message from Plant Cell
(Your Name) thought you would like to see the Plant Cell web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Phytochrome Regulation and Differential Expression of Gibberellin 3β-Hydroxylase Genes in Germinating Arabidopsis Seeds
Shinjiro Yamaguchi, Maria W. Smith, Robert G. S. Brown, Yuji Kamiya, Tai-ping Sun
The Plant Cell Dec 1998, 10 (12) 2115-2126; DOI: 10.1105/tpc.10.12.2115

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Request Permissions
Share
Phytochrome Regulation and Differential Expression of Gibberellin 3β-Hydroxylase Genes in Germinating Arabidopsis Seeds
Shinjiro Yamaguchi, Maria W. Smith, Robert G. S. Brown, Yuji Kamiya, Tai-ping Sun
The Plant Cell Dec 1998, 10 (12) 2115-2126; DOI: 10.1105/tpc.10.12.2115
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • METHODS
    • ACKNOWLEDGMENTS
    • Footnotes
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

In this issue

The Plant Cell Online: 10 (12)
The Plant Cell
Vol. 10, Issue 12
Dec 1998
  • Table of Contents
  • About the Cover
  • Index by author
View this article with LENS

More in this TOC Section

  • Chloroplast Chaperonin-Mediated Targeting of a Thylakoid Membrane Protein
  • Ectopic Expression of the Transcriptional Regulator silky3 Causes Pleiotropic Meristem and Sex Determination Defects in Maize Inflorescences
  • SAUR17 and SAUR50 Differentially Regulate PP2C-D1 during Apical Hook Development and Cotyledon Opening in Arabidopsis
Show more RESEARCH ARTICLES

Similar Articles

Our Content

  • Home
  • Current Issue
  • Plant Cell Preview
  • Archive
  • Teaching Tools in Plant Biology
  • Plant Physiology
  • Plant Direct
  • Plantae
  • ASPB

For Authors

  • Instructions
  • Submit a Manuscript
  • Editorial Board and Staff
  • Policies
  • Recognizing our Authors

For Reviewers

  • Instructions
  • Peer Review Reports
  • Journal Miles
  • Transfer of reviews to Plant Direct
  • Policies

Other Services

  • Permissions
  • Librarian resources
  • Advertise in our journals
  • Alerts
  • RSS Feeds
  • Contact Us

Copyright © 2021 by The American Society of Plant Biologists

Powered by HighWire