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Autonomous replication sequences from the Amaranthus palmeri eccDNA replicon enable replication in yeast.

William T Molin1, Allison Yaguchi2, Mark Blenner2

  • 1Crop Production Systems Research Unit, USDA-ARS, Stoneville, MS, 38776, USA.

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|July 12, 2020
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Summary

Autonomous replication sequences (ARS) from glyphosate-resistant Amaranthus palmeri

Keywords:
Autonomous replicationDNA bendingEACSeccDNA replicon

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Area of Science:

  • Plant molecular biology
  • Genetics
  • Molecular biology

Background:

  • Extrachromosomal circular DNA (eccDNA) plays a role in plant adaptation and evolution.
  • Glyphosate resistance in Amaranthus palmeri is associated with specific eccDNA elements.

Purpose of the Study:

  • To investigate if autonomous replication sequences (ARS) within the Amaranthus palmeri eccDNA can function in a heterologous yeast system.
  • To characterize the functional elements responsible for replication within the eccDNA replicon.

Main Methods:

  • Sequence analysis of the Amaranthus palmeri eccDNA replicon.
  • Functional validation of identified ARS elements in an ARS-less yeast plasmid system.

Main Results:

  • A region with distinct A+T/G+C content and bending, characteristic of ARS, was identified.
  • An extended autonomous replication sequence (EACS) was found near DNA unwinding elements (DUEs).
  • This eccDNA region successfully drove autonomous replication of an ARS-less yeast plasmid.

Conclusions:

  • The identified ARS elements from Amaranthus palmeri eccDNA are functional in a yeast replication system.
  • This finding provides insights into the replication mechanisms of plant eccDNA.
  • The EACS and DUE sequences are key components for autonomous replication in this system.