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Transgene stacking in potato using the GAANTRY system.

Kent F McCue1, Ethan Gardner1, Ronald Chan1

  • 1Crop Improvement and Genetics, Western Regional Research Center, USDA-ARS, Albany, CA, USA.

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|July 27, 2019
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Summary
This summary is machine-generated.

The GAANTRY system efficiently generates high-quality transgenic potato plants. This gene stacking technology enables stable integration of multiple genes using Agrobacterium rhizogenes.

Keywords:
GAANTRYGene-stackingSite-specific recombinaseSolanum tuberosum

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Area of Science:

  • Plant biotechnology
  • Molecular biology
  • Agrobacterium-mediated transformation

Background:

  • Gene stacking is crucial for developing crops with multiple desirable traits.
  • Efficiently introducing large DNA constructs into plants remains a challenge.

Purpose of the Study:

  • To evaluate the GAANTRY (Gene Assembly in Agrobacterium by Nucleic acid Transfer using Recombinase technologY) system for generating transgenic potato plants.
  • To assess the efficiency and accuracy of introducing a 10-gene stack using Agrobacterium rhizogenes.

Main Methods:

  • Utilized the GAANTRY 10-stack strain of Agrobacterium rhizogenes for transformation.
  • Employed a tissue culture-based callus transformation method for Lenape potato plants.
  • Conducted molecular and phenotypic characterization of transgenic lines, including droplet digital PCR.

Main Results:

  • Achieved a 32% transformation efficiency in Lenape potato plants.
  • Six out of seven independent transgenic lines successfully integrated the entire 28.5 kb 10-stack T-DNA construct.
  • Transgenic plants predominantly contained one or two copies of the stacked transgenes with no backbone DNA contamination.

Conclusions:

  • The GAANTRY system is effective for generating high-quality transgenic potato plants with large, stacked gene constructs.
  • Demonstrated the utility of GAANTRY in Agrobacterium rhizogenes for efficient gene stacking in potato.
  • Confirmed the stability and integrity of the integrated multi-gene constructs.