Abstract
Cannabis sativa L. cv. ‘Cheungsam’ is an industrial hemp plant of Republic of Korea origin, primarily cultivated for fiber and seed production. In vitro seed germination and tissue culture are valuable tools for developing various biotechnological techniques. In the present study, we aimed to develop a tissue culture process for hemp plants using Cheungsam as a model plant and examine the secondary metabolites produced from its callus. We also developed a method to prepare pathogen-free seedlings from field-derived seeds using hydrogen peroxide (H2O2) solution as a liquid germination medium. Treating seedlings with removed seed coat in 3% H2O2 significantly reduced the contamination rate. Callus formation and de novo organogenesis of shoots and roots from callus were successfully achieved using cotyledon and leaf tissues prepared from the pathogen-free seedlings. The most effective in vitro regeneration results were obtained using the Murashige and Skoog (MS) medium supplemented with certain targeted growth regulators. An optimal combination of 0.5 mg/L thidiazuron (TDZ) and 1.0 mg/L 1-naphthalene acetic acid proved highly effective for callus induction. The addition of 0.5 mg/L TDZ in the MS medium significantly stimulated shoot proliferation, while robust root development was best supported by MS medium supplemented with 2.5 mg/L indole-3-butyric acid for both cotyledon and leaf explants. Finally, gas chromatography–mass spectrometry (GC–MS) analysis of ethanol extract from Cheungsam leaf callus revealed the presence of different secondary metabolites, including 9-octadecenamide, methyl salicylate, dodecane, tetradecane, and phenol, 2,4-bis-(1,1-dimethylethyl). This study provides a comprehensive de novo regeneration protocol for Cheungsam plants and insight into the secondary metabolite profiles of its callus.
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Claims 11
Claim · model-written summary
A hydrogen-peroxide liquid germination method produced pathogen-free seedlings from field seeds.
- Taxon
- Cannabis sativa
Cannabis sativat0 - Treatment
- hydrogen peroxide (H2O2) solutione0
- Outcome
- pathogen-free seedlingse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We also developed a method to prepare pathogen-free seedlings from field-derived seeds using hydrogen peroxide (H2O2) solution as a liquid germination medium.”
t0“Cannabis sativa”
Claim · model-written summary
Treating seedlings without seed coats in 3% H2O2 significantly reduced contamination.
- Taxon
- Cannabis sativaCannabis sativat0
- Treatment
- 3% H2O2e0
- Outcome
- contamination ratee0
- Direction
- reducede0
- Qualifier
- removed seed coate0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Treating seedlings with removed seed coat in 3% H2O2 significantly reduced the contamination rate.”
t0“Cannabis sativa”
Claim · model-written summary
Cotyledon tissues from pathogen-free seedlings supported callus formation and shoot and root organogenesis.
- Taxon
- Cannabis sativaCannabis sativat0
- Organ / stage
- cotyledone0
- Outcome
- Callus formation and de novo organogenesis of shoots and rootse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Callus formation and de novo organogenesis of shoots and roots from callus were successfully achieved using cotyledon and leaf tissues prepared from the pathogen-free seedlings.”
t0“Cannabis sativa”
Claim · model-written summary
Leaf tissues from pathogen-free seedlings supported callus formation and shoot and root organogenesis.
- Taxon
- Cannabis sativaCannabis sativat0
- Organ / stage
- leaf tissuese0
- Outcome
- Callus formation and de novo organogenesis of shoots and rootse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Callus formation and de novo organogenesis of shoots and roots from callus were successfully achieved using cotyledon and leaf tissues prepared from the pathogen-free seedlings.”
t0“Cannabis sativa”
Claim · model-written summary
MS medium with targeted growth regulators gave the most effective regeneration.
- Taxon
- Cannabis sativaCannabis sativat0
- Treatment
- Murashige and Skoog (MS) medium supplemented with certain targeted growth regulatorse0
- Outcome
- in vitro regeneration resultse0
- Direction
- most effectivee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The most effective in vitro regeneration results were obtained using the Murashige and Skoog (MS) medium supplemented with certain targeted growth regulators.”
t0“Cannabis sativa”
Claim · model-written summary
0.5 mg/L TDZ with 1.0 mg/L 1-naphthalene acetic acid was optimal for callus induction.
- Taxon
- Cannabis sativaCannabis sativat0
- Treatment
- 0.5 mg/L thidiazuron (TDZ) and 1.0 mg/L 1-naphthalene acetic acide0
- Outcome
- callus inductione0
- Direction
- optimale0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“An optimal combination of 0.5 mg/L thidiazuron (TDZ) and 1.0 mg/L 1-naphthalene acetic acid proved highly effective for callus induction.”
t0“Cannabis sativa”
Claim · model-written summary
Adding 0.5 mg/L TDZ to MS medium significantly stimulated shoot proliferation.
- Taxon
- Cannabis sativaCannabis sativat0
- Treatment
- 0.5 mg/L TDZe0
- Outcome
- shoot proliferatione0
- Direction
- stimulatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The addition of 0.5 mg/L TDZ in the MS medium significantly stimulated shoot proliferation”
t0“Cannabis sativa”
Claim · model-written summary
MS medium with 2.5 mg/L indole-3-butyric acid best supported cotyledon-explant root development.
- Taxon
- Cannabis sativaCannabis sativat0
- Organ / stage
- cotyledone0
- Treatment
- MS medium supplemented with 2.5 mg/L indole-3-butyric acide0
- Outcome
- root developmente0
- Direction
- best supportede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“robust root development was best supported by MS medium supplemented with 2.5 mg/L indole-3-butyric acid for both cotyledon and leaf explants”
t0“Cannabis sativa”
Claim · model-written summary
MS medium with 2.5 mg/L indole-3-butyric acid best supported leaf-explant root development.
- Taxon
- Cannabis sativaCannabis sativat0
- Organ / stage
- leaf explantse0
- Treatment
- MS medium supplemented with 2.5 mg/L indole-3-butyric acide0
- Outcome
- root developmente0
- Direction
- best supportede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“robust root development was best supported by MS medium supplemented with 2.5 mg/L indole-3-butyric acid for both cotyledon and leaf explants”
t0“Cannabis sativa”
Claim · model-written summary
Cheungsam leaf callus contained 9-octadecenamide, methyl salicylate and dodecane.
- Taxon
- Cannabis sativaCannabis sativat0
- Material
- Cheungsame0
- Organ / stage
- leaf calluse0
- Outcome
- 9-octadecenamide, methyl salicylate, dodecanee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GC–MS) analysis of ethanol extract from Cheungsam leaf callus revealed the presence of different secondary metabolites, including 9-octadecenamide, methyl salicylate, dodecane, tetradecane, and phenol, 2,4-bis-(1,1-dimethylethyl).”
t0“Cannabis sativa”
Claim · model-written summary
Cheungsam leaf callus contained tetradecane and phenol, 2,4-bis-(1,1-dimethylethyl).
- Taxon
- Cannabis sativaCannabis sativat0
- Material
- Cheungsame0
- Organ / stage
- leaf calluse0
- Outcome
- tetradecane, and phenol, 2,4-bis-(1,1-dimethylethyl)e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GC–MS) analysis of ethanol extract from Cheungsam leaf callus revealed the presence of different secondary metabolites, including 9-octadecenamide, methyl salicylate, dodecane, tetradecane, and phenol, 2,4-bis-(1,1-dimethylethyl).”
t0“Cannabis sativa”
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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.