Abstract
Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific knock outediting of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, was were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop transgene-free potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.
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Claims 6
Claim · model-written summary
Up to 68% of regenerated potato plants had a mutation in at least one StPPO2 allele.
- Taxon
- Potato
Solanum tuberosumt0 - Gene
- StPPO2e1
- Outcome
- regenerated plants contained mutationse0
- Value
- 68%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Up to 68% of regenerated plants contained mutations in at least one allele of the target gene”
e1“to induce mutations in the StPPO2 gene”
t0“Potato Tubers”
Claim · model-written summary
Twenty-four percent of edited potato lines had mutations in all four alleles.
- Taxon
- PotatoSolanum tuberosumt0
- Gene
- StPPO2e1
- Outcome
- edited lines carried mutations in all four allelese0
- Value
- 24%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“24% of edited lines carried mutations in all four alleles”
e1“to induce mutations in the StPPO2 gene”
t0“Potato Tubers”
Claim · model-written summary
No off-target mutations were identified in other analyzed StPPO genes.
- Taxon
- PotatoSolanum tuberosumt0
- Gene
- StPPO genese0
- Outcome
- No off-target mutationse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“No off-target mutations were identified in other analyzed StPPO genes”
t0“Potato Tubers”
Claim · model-written summary
Four-allele StPPO2 mutations reduced potato tuber PPO activity by up to 69% against control.
- Taxon
- PotatoSolanum tuberosumt0
- Treatment
- Mutations induced in the four alleles of StPPO2 genee0
- Gene
- StPPO2e0
- Outcome
- tuber PPO activitye0
- Direction
- reductione0
- Value
- 69%e0
- Comparator
- controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control”
t0“Potato Tubers”
Claim · model-written summary
Four-allele StPPO2 mutations reduced potato tuber enzymatic browning by 73% against control.
- Taxon
- PotatoSolanum tuberosumt0
- Treatment
- Mutations induced in the four alleles of StPPO2 genee0
- Gene
- StPPO2e0
- Outcome
- enzymatic browninge0
- Direction
- reductione0
- Value
- 73%e0
- Comparator
- controle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control”
t0“Potato Tubers”
Claim · model-written summary
The authors conclude that editing one StPPO family member can develop transgene-free potato varieties with less tuber browning.
- Taxon
- potatoSolanum tuberosume0
- Gene
- StPPO gene familye0
- Outcome
- transgene-free potato varieties with reduced enzymatic browning in tuberse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the CRISPR/Cas9 system can be applied to develop transgene-free potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family”
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Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.