Development of mPing-based activation tags for crop insertional mutagenesis

Authors

Alexander Johnson, University of Georgia
Edward Mcassey, University of Georgia
Stephanie Diaz, University of South Carolina - Aiken
Jacob Reagin, University of South Carolina - Aiken
Priscilla S. Redd, University of South Carolina - Aiken
Daymond R. Parrilla, University of South Carolina - Aiken
Hanh T. Nguyen, University of Nebraska-LincolnFollow
Adrian Stec, University of Minnesota
Lauren A.L. McDaniel, University of Georgia
Thomas E. Clemente, University of Nebraska - LincolnFollow
Robert M. Stupar, University of Minnesota
Wayne A. Parrott, University of Georgia
C Nathan Hancock, University of South Carolina - Aiken

ORCID IDs

Alexander Johnson

Edward Mcassey

Stephanie Diaz

Jacob Reagin

Robert M. Stupar

Wayne A. Parrott

C. Nathan Hancock

Date of this Version

12-7-2020

Citation

Johnson A, Mcassey E, Diaz S, et al. Development of mPing-based activation tags for crop insertional mutagenesis. Plant Direct. 2021;00:1–12. https:// doi.org/10.1002/pld3.300

Comments

OPEN ACCESS

Abstract

Modern plant breeding increasingly relies on genomic information to guide crop improvement. Although some genes are characterized, additional tools are needed to effectively identify and characterize genes associated with crop traits. To address this need, the mPing element from rice was modified to serve as an activation tag to induce expression of nearby genes. Embedding promoter sequences in mPing resulted in a decrease in overall transposition rate; however, this effect was negated by using a hyperactive version of mPing called mmPing20. Transgenic soybean events carrying mPing-based activation tags and the appropriate transposase expression cassettes showed evidence of transposition. Expression analysis of a line that contained a heritable insertion of the mmPing20F activation tag indicated that the activation tag induced overexpression of the nearby soybean genes. This represents a significant advance in gene discovery technology as activation tags have the potential to induce more phenotypes than the original mPing element, improving the overall effectiveness of the mutagenesis system.