Abstract
Abstract Carnations ( Dianthus caryophyllus L.) are amongst the three most commercially valuable cut flowers worldwide. However, traditional breeding methods are often time-consuming and labor-intensive. Although genome editing is used as an alternative method for creating new varieties, the high heterozygosity of carnations inhibits the ability to maintain varietal characteristics in null segregants except for target-derived traits. The use of homozygous lines is a possible solution. Therefore, this study aimed to establish regeneration, transformation, and genome editing methods using seed-carnation varieties. The effects of four auxins (indole-3-butyric acid, IBA; a-naphthaleneacetic acid, NAA; 2,4-dichlorophenoxyacetic acid, 2,4-D; and 3-indoleacetic acid, IAA) and five cytokinins (6-benzyladenine, BA; thidiazuron, TDZ; kinetin, KT; zeatin, ZT; and N 6 -2-isopentenyl adenine, 2IP) on callus and shoot induction were evaluated. The combination of 0.05 mg/l 2,4-D and 4 mg/l TDZ had the highest shoot formation rate at 28%. In addition, shoot hyperhydricity was reduced by increasing the size of culture vessels. Sucrose, agar, and AgNO 3 concentrations, as well as pH, were optimized to facilitate regeneration. Hygromycin at 12.5 mg/l was subsequently used as the selection agent after Agrobacterium -mediated transformation. Finally, the phytoene desaturase gene was knocked out using the CRISPR/Cas9 system. The obtained albino shoot had a one-base deletion or two-base insertion in the genome sequence. To our knowledge, this is the first study to establish a system for genome editing of callus-derived shoots from a homozygous seed-propagated carnation, which may contribute to the rapid breeding of the new varieties.
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Claims 4
Claim · model-written summary
The combination of 0.05 mg/l 2,4-D and 4 mg/l TDZ gave the highest shoot formation rate, at 28%.
- Taxon
- carnation
Dianthus caryophyllust0 - Treatment
- 0.05 mg/l 2,4-D and 4 mg/l TDZe1
- Outcome
- shoot formation ratee1
- Direction
- higheste1
- Value
- 28%e1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“The combination of 0.05 mg/l 2,4-D and 4 mg/l TDZ had the highest shoot formation rate at 28%”
t0“for a seed-propagated carnation”
Claim · model-written summary
Enlarging culture vessels reduced shoot hyperhydricity.
- Taxon
- carnationDianthus caryophyllust0
- Treatment
- increasing the size of culture vesselse2
- Outcome
- shoot hyperhydricitye2
- Direction
- reducede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“In addition, shoot hyperhydricity was reduced by increasing the size of culture vessels”
t0“for a seed-propagated carnation”
Claim · model-written summary
Sucrose, agar, AgNO3 concentrations and pH were optimized to facilitate regeneration.
- Taxon
- carnationDianthus caryophyllust0
- Treatment
- Sucrose, agar, and AgNO 3 concentrations, as well as pHe3
- Outcome
- regeneratione3
- Direction
- facilitatee3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e3“Sucrose, agar, and AgNO 3 concentrations, as well as pH, were optimized to facilitate regeneration”
t0“for a seed-propagated carnation”
Claim · model-written summary
CRISPR/Cas9 knockout of the phytoene desaturase gene gave albino shoots with a one-base deletion or two-base insertion in the genome sequence.
- Taxon
- carnationDianthus caryophyllust0
- Treatment
- CRISPR/Cas9e5
- Gene
- phytoene desaturase genee5
- Outcome
- one-base deletion or two-base insertion in the genome sequencee4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e4“The obtained albino shoot had a one-base deletion or two-base insertion in the genome sequence”
e5“Finally, the phytoene desaturase gene was knocked out using the CRISPR/Cas9 system”
t0“for a seed-propagated carnation”
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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.