Transgene stacking in potato using the GAANTRY system

Authors: McCue, Kent; Gardner, Ethan; Chan, Ronald; Thilmony, Roger; Thomson, James - Jim

Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/16/2019
Publication Date: 7/25/2019
Citation: McCue, K.F., Gardner, E.B., Chan, R., Thilmony, R.L., Thomson, J.G. 2019. Transgene stacking in potato using the GAANTRY system. BMC Research Notes. 12:457. https://doi.org/10.1186/s13104-019-4493-8.
DOI: https://doi.org/10.1186/s13104-019-4493-8

Interpretive Summary: This work reports use of the Agrobacterium rhizogenes ArPORT1 ‘GAANTRY’ strain for T-DNA transfer of a large 28.5 kb T-DNA cassette into potato. Seven independent lines were fully characterized. Six had complete intact T-DNA inserts. Four were single copy and two contained duplicate copies of the T-DNA. Research demonstrates the successful use of the ArPORT1 ‘GAANTRY’ strain in the crop species potato.

Technical Abstract: GAANTRY (Gene Assembly in Agrobacterium by Nucleic acid Transfer using Recombinase technologY) is a flexible and effective system for stably stacking multiple genes within an Agrobacterium virulence plasmid Transfer-DNA (T-DNA). We used the Agrobacterium rhizogenes ArPORT1 ’10-stack’ GAANTRY strain to generate transgenic potato (cv. Lenape) plants. The 28.5 kilobase 10-stack T-DNA, was introduced with a 32.6 % transformation efficiency. Six independent transgenic lines, containing the entire construct were molecularly and phenotypically characterized, demonstrating that the GAANTRY system can be used to generate transgenic Lenape potato plants with a large stacked T-DNA construct.