Abstract
Hairy root transformation systems are increasingly recognized as effective tools for functional genomics and gene editing studies in various crop species. In this study, we employed the RUBY reporter gene as a visible marker to establish a rapid, simple, and efficient in vivo system for potato hairy root induction and transformation. Explants were wounded by transverse cuts and gently scraped across the surface of a culture plate containing Rhizobium rhizogenes harboring the desired construct, then directly planted into pots containing moist, sterile vermiculite. The 2-3-week-old shoots regenerated from tubers of the Atlantic potato cultivar were identified as sufficient explants for Rhizobium rhizogenes-mediated transformation, achieving transformation efficiencies of up to 100%. This procedure was further adapted for stem nodes and leaf explants and achieved high transformation efficiencies of 80% and 93%, respectively. In addition, the in vivo hairy root transformation system was successfully utilized to validate CRISPR/Cas9 activities by inducing targeted mutations of the StDL1 gene in potato. Heteroduplex analysis on PAGE revealed a high editing frequency (96%), and Sanger sequencing of selected lines confirmed diverse deletions at the target site. Overall, this in vivo hairy root transformation method enables rapid and robust assessment of genome editing efficiency and transgene expression, providing a valuable tool for functional genomics and genetic improvement in potato, with potential applications in other Solanaceae crops.
Abstract from the paper’s OpenAlex record; open copy licensed cc-by. Read at the source ↗
The text the model read, with the source record’s inline markup (italics, sub- and superscripts) rendered and its character entities decoded; quote offsets refer to the raw text.
Claims 6
Claim · model-written summary
Atlantic potato shoots gave transformation efficiencies up to 100%.
- Taxon
- potato
Solanum tuberosume0 - Material
- Atlantic potato cultivare0
- Organ / stage
- 2-3-week-old shootse0
- Interactor
- Rhizobium rhizogenes
Martinezella rhizogenese0other - Outcome
- transformation efficienciese0
- Value
- up to 100%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The 2-3-week-old shoots regenerated from tubers of the Atlantic potato cultivar were identified as sufficient explants for Rhizobium rhizogenes-mediated transformation, achieving transformation efficiencies of up to 100%.”
Claim · model-written summary
Stem nodes gave 80% transformation efficiency.
- Taxon
- potatoSolanum tuberosumt0
- Organ / stage
- stem nodese0
- Outcome
- transformation efficienciese0
- Value
- 80%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This procedure was further adapted for stem nodes and leaf explants and achieved high transformation efficiencies of 80% and 93%, respectively.”
t0“in potato”
Claim · model-written summary
Leaf explants gave 93% transformation efficiency.
- Taxon
- potatoSolanum tuberosumt0
- Organ / stage
- leaf explantse0
- Outcome
- transformation efficienciese0
- Value
- 93%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This procedure was further adapted for stem nodes and leaf explants and achieved high transformation efficiencies of 80% and 93%, respectively.”
t0“in potato”
Claim · model-written summary
The method induced targeted StDL1 mutations in potato.
- Taxon
- potatoSolanum tuberosume0
- Gene
- StDL1e0
- Outcome
- targeted mutationse0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the in vivo hairy root transformation system was successfully utilized to validate CRISPR/Cas9 activities by inducing targeted mutations of the StDL1 gene in potato.”
Claim · model-written summary
Heteroduplex analysis found a 96% editing frequency.
- Taxon
- potatoSolanum tuberosumt0
- Outcome
- editing frequencye0
- Value
- 96%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Heteroduplex analysis on PAGE revealed a high editing frequency (96%), and Sanger sequencing of selected lines confirmed diverse deletions at the target site.”
t0“in potato”
Claim · model-written summary
Sanger sequencing confirmed diverse target-site deletions.
- Taxon
- potatoSolanum tuberosumt0
- Outcome
- diverse deletionse0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Sanger sequencing of selected lines confirmed diverse deletions at the target site.”
t0“in potato”