1. Plant Biology
Download icon

Building customizable auto-luminescent luciferase-based reporters in plants

  1. Arjun Khakhar
  2. Colby G Starker
  3. James C Chamness
  4. Nayoung Lee
  5. Sydney Stokke
  6. Cecily Wang
  7. Ryan Swanson
  8. Furva Rizvi
  9. Takato Imaizumi
  10. Daniel F Voytas  Is a corresponding author
  1. University of Minnesota, United States
  2. University of Washington, United States
Tools and Resources
  • Cited 7
  • Views 6,459
  • Annotations
Cite this article as: eLife 2020;9:e52786 doi: 10.7554/eLife.52786

Abstract

Bioluminescence is a powerful biological signal that scientists have repurposed as a reporter for gene expression in plants and animals. However, there are downsides associated with the need to provide a substrate to these reporters, including its high cost and non-uniform tissue penetration. In this work we reconstitute a fungal bioluminescence pathway (FBP) in planta using a composable toolbox of parts. We demonstrate that the FBP can create luminescence across various tissues in a broad range of plants without external substrate addition. We also show how our toolbox can be used to deploy the FBP in planta to build auto-luminescent reporters for the study of gene-expression and hormone fluxes. A low-cost imaging platform for gene expression profiling is also described. These experiments lay the groundwork for future construction of programmable auto-luminescent plant traits, such as light driven plant-pollinator interactions or light emitting plant-based sensors.

Data availability

All the data collected for this study is depicted in the figures included in the manuscript. All the raw data used to make the figures has been uploaded.

Article and author information

Author details

  1. Arjun Khakhar

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Colby G Starker

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6774-7227
  3. James C Chamness

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Nayoung Lee

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Sydney Stokke

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Cecily Wang

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Ryan Swanson

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Furva Rizvi

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Takato Imaizumi

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Daniel F Voytas

    Genetics, Cell Biology, and Development, University of Minnesota, Saint Paul, United States
    For correspondence
    voytas@umn.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4944-1224

Funding

University of Minnesota (Grand Challenges Postdoctoral Fellowship)

  • Arjun Khakhar

National Institutes of Health (R01GM079712)

  • Nayoung Lee

Rural Development Administration (PJ013386)

  • Nayoung Lee

U.S. Department of Energy

  • Colby G Starker
  • James C Chamness

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Richard Amasino, University of Wisconsin Madison, United States

Publication history

  1. Received: October 16, 2019
  2. Accepted: March 24, 2020
  3. Accepted Manuscript published: March 25, 2020 (version 1)
  4. Version of Record published: April 17, 2020 (version 2)
  5. Version of Record updated: July 14, 2020 (version 3)

Copyright

© 2020, Khakhar et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 6,459
    Page views
  • 945
    Downloads
  • 7
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Genetics and Genomics
    2. Plant Biology
    Ranjith K Papareddy et al.
    Research Article

    DNA methylation has evolved to silence mutagenic transposable elements (TEs) while typically avoiding the targeting of endogenous genes. Mechanisms that prevent DNA methyltransferases from ectopically methylating genes are expected to be of prime importance during periods of dynamic cell cycle activities including plant embryogenesis. However, virtually nothing is known regarding how DNA methyltransferase activities are precisely regulated during embryogenesis to prevent the induction of potentially deleterious and mitotically stable genic epimutations. Here, we report that microRNA-mediated repression of CHROMOMETHYLASE 3 (CMT3) and the chromatin features that CMT3 prefers help prevent ectopic methylation of thousands of genes during embryogenesis that can persist for weeks afterwards. Our results are also consistent with CMT3-induced ectopic methylation of promoters or bodies of genes undergoing transcriptional activation reducing their expression. Therefore, the repression of CMT3 prevents epigenetic collateral damage on endogenous genes. We also provide a model that may help reconcile conflicting viewpoints regarding the functions of gene-body methylation that occurs in nearly all flowering plants.