期刊
BMC RESEARCH NOTES
卷 12, 期 -, 页码 -出版社
SPRINGERNATURE
DOI: 10.1186/s13104-019-4493-8
关键词
GAANTRY; Gene-stacking; Site-specific recombinase; Solanum tuberosum
资金
- USDA Agricultural Research Service CRIS [2030-21220-002-00-D]
ObjectiveGAANTRY (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 examined the ability of the GAANTRY Agrobacterium rhizogenes ArPORT1 10-stack' strain to generate transgenic potato plants.ResultsThe 28.5 kilobase 10-stack T-DNA, was introduced into Lenape potato plants with a 32% transformation efficiency. Molecular and phenotypic characterization confirmed that six of the seven tested independent transgenic lines carried the entire desired construct, demonstrating that the GAANTRY 10-stack strain can be used can be used in a tissue culture-based callus transformation method to efficiently generate transgenic potato plants. Analysis using droplet digital PCR showed that most of the characterized events carry one or two copies of the 10-stack transgenes and that backbone' DNA from outside of the T-DNA was absent in the transgenic plants. These results demonstrate that the GAANTRY system efficiently generates high quality transgenic potato plants with a large construct of stacked transgenes.
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