Abstract
Cryopreservation of rare plant materials is an important approach for preserving germplasms and is a good added concept to tissue banking. The preservation of embryogenic cell suspensions is even more valuable as the tissues facilitate in producing millions of embryos, plantlets and generates transgenics en masse. Catharanthus roseus is a medicinally important plant that produces a variety of anticancerous phytocompounds and needs conservation of alkaloid producing cell lines. In this study, embryogenic tissue banking has been attempted in C. roseus by the two-step cryopreservation method combining cryoprotection and dehydration. Prior to plunging into liquid nitrogen (LN), the tissues were exposed to osmotic-and cryoprotective agents. Two osmotic agents (sugar and sorbitol) and three cryoprotective compounds, polyethylene glycol (PEG), dimethyl sulfoxide (DMSO) and glycerol were used at varying concentrations to protect cells from freezing damages. Both sucrose and sorbitol increased callus biomass post-cryopreservation; the influence of sucrose was however, more prominent. Embryogenic tissue treated in medium with 0.4 M sucrose for 2 days followed by 5% PEG for 2 h showed maximum viability before (83%) and after (55%) cryopreservation, high regrowth percentage (77%) and produced an average 9 cell colonies per Petri dish. Additionally, dehydration (1-5 h) was tested to reduce water content for improving viability and regrowth of cryopreserved embryogenic cells. Among the various tested cryoprotective conditions, the highest (72%) viability was observed following the combination of treatments with 0.4 M sucrose (2 days),10% PEG (2 h) and dehydration (2 h). Maximum regrowth percentage (88%) and 12 colonies/petri dish was noted in combination of 0.4 M sucrose + 5% PEG. The cryopreserved calli differentiated into somatic embryos (52.78-54.33 globular embryos/callus mass) in NAA (0.5 mg/l) and BAP (0.5-1.0 mg/l) added media. Plantlets were successfully regenerated from cryopreserved tissue and the 2C DNA was estimated through flow cytometry. The genome size of cryopreserved regenerant was 1.51 pg/2C, which is similar to field-grown Catharanthus plants. Vinblastine and vincristine levels were nearly the same in mother plant's and frozen (cryopreserved) leaf tissue. The post cryopreservation embryogenesis protocol may be used for continuous production of plants for future applications.
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Claims 15
Claim · model-written summary
Sucrose increased callus biomass after cryopreservation.
- Taxon
- Catharanthus roseus
Catharanthus roseust0 - Treatment
- sucrosee0
- Outcome
- callus biomasse0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both sucrose and sorbitol increased callus biomass post-cryopreservation; the influence of sucrose was however, more prominent.”
t0“Catharanthus roseus”
Claim · model-written summary
Sorbitol increased callus biomass after cryopreservation.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- sorbitole0
- Outcome
- callus biomasse0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both sucrose and sorbitol increased callus biomass post-cryopreservation; the influence of sucrose was however, more prominent.”
t0“Catharanthus roseus”
Claim · model-written summary
Sucrose had a more prominent effect on callus biomass than sorbitol.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- sucrosee0
- Outcome
- influence of sucrosee0
- Direction
- more prominente0
- Comparator
- sorbitole0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Both sucrose and sorbitol increased callus biomass post-cryopreservation; the influence of sucrose was however, more prominent.”
t0“Catharanthus roseus”
Claim · model-written summary
The 0.4 M sucrose and 5% PEG regimen gave 83% pre-freeze viability.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- 0.4 M sucrose for 2 days followed by 5% PEG for 2 he0
- Outcome
- viability beforee0
- Direction
- maximume0
- Value
- 83%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Embryogenic tissue treated in medium with 0.4 M sucrose for 2 days followed by 5% PEG for 2 h showed maximum viability before (83%) and after (55%) cryopreservation”
t0“Catharanthus roseus”
Claim · model-written summary
The 0.4 M sucrose and 5% PEG regimen gave 55% post-freeze viability.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- 0.4 M sucrose for 2 days followed by 5% PEG for 2 he0
- Outcome
- viabilitye0
- Direction
- maximume0
- Value
- 55%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Embryogenic tissue treated in medium with 0.4 M sucrose for 2 days followed by 5% PEG for 2 h showed maximum viability before (83%) and after (55%) cryopreservation”
t0“Catharanthus roseus”
Claim · model-written summary
The 0.4 M sucrose and 5% PEG regimen yielded 77% regrowth.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Outcome
- regrowth percentagee0
- Direction
- highe0
- Value
- 77%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“high regrowth percentage (77%) and produced an average 9 cell colonies per Petri dish.”
t0“Catharanthus roseus”
Claim · model-written summary
The same regimen produced nine cell colonies per dish.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Outcome
- cell coloniese0
- Value
- 9 cell colonies per Petri dishe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“high regrowth percentage (77%) and produced an average 9 cell colonies per Petri dish.”
t0“Catharanthus roseus”
Claim · model-written summary
Sucrose, 10% PEG and dehydration produced the highest viability of 72%.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- 0.4 M sucrose (2 days),10% PEG (2 h) and dehydration (2 h)e0
- Outcome
- viabilitye0
- Direction
- higheste0
- Value
- 72%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the highest (72%) viability was observed following the combination of treatments with 0.4 M sucrose (2 days),10% PEG (2 h) and dehydration (2 h).”
t0“Catharanthus roseus”
Claim · model-written summary
Sucrose plus 5% PEG gave maximum regrowth of 88%.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- 0.4 M sucrose + 5% PEGe0
- Outcome
- regrowth percentagee0
- Direction
- Maximume0
- Value
- 88%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Maximum regrowth percentage (88%) and 12 colonies/petri dish was noted in combination of 0.4 M sucrose + 5% PEG.”
t0“Catharanthus roseus”
Claim · model-written summary
Sucrose plus 5% PEG yielded 12 colonies per dish.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- 0.4 M sucrose + 5% PEGe0
- Outcome
- coloniese0
- Value
- 12 colonies/petri dishe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Maximum regrowth percentage (88%) and 12 colonies/petri dish was noted in combination of 0.4 M sucrose + 5% PEG.”
t0“Catharanthus roseus”
Claim · model-written summary
Cryopreserved calli formed 52.78-54.33 globular embryos per callus mass in NAA and BAP medium.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Treatment
- NAA (0.5 mg/l) and BAP (0.5-1.0 mg/l) added mediae0
- Outcome
- somatic embryose0
- Value
- 52.78-54.33 globular embryos/callus masse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The cryopreserved calli differentiated into somatic embryos (52.78-54.33 globular embryos/callus mass) in NAA (0.5 mg/l) and BAP (0.5-1.0 mg/l) added media.”
t0“Catharanthus roseus”
Claim · model-written summary
Plantlets were regenerated from cryopreserved tissue.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Outcome
- Plantlets were successfully regenerated from cryopreserved tissuee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Plantlets were successfully regenerated from cryopreserved tissue”
t0“Catharanthus roseus”
Claim · model-written summary
Regenerants had a genome size of 1.51 pg/2C, similar to field-grown plants.
- Taxon
- Catharanthus
_W2_IMG_3209.jpg?width=60)
Catharanthuse0 - Outcome
- genome sizee0
- Direction
- similare0
- Value
- 1.51 pg/2Ce0
- Comparator
- field-grown Catharanthus plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_W2_IMG_3209.jpg?width=500)
Verbatim
e0“The genome size of cryopreserved regenerant was 1.51 pg/2C, which is similar to field-grown Catharanthus plants.”
Claim · model-written summary
Vinblastine levels were nearly the same in mother and cryopreserved leaf tissue.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Organ / stage
- leaf tissuee0
- Outcome
- Vinblastinee0
- Direction
- nearly the samee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Vinblastine and vincristine levels were nearly the same in mother plant's and frozen (cryopreserved) leaf tissue.”
t0“Catharanthus roseus”
Claim · model-written summary
Vincristine levels were nearly the same in mother and cryopreserved leaf tissue.
- Taxon
- Catharanthus roseusCatharanthus roseust0
- Organ / stage
- leaf tissuee0
- Outcome
- vincristinee0
- Direction
- nearly the samee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Vinblastine and vincristine levels were nearly the same in mother plant's and frozen (cryopreserved) leaf tissue.”
t0“Catharanthus roseus”
The same questions in other papers
sucrose · growth · Catharanthus roseus (Madagascar periwinkle)Results agree
1 paper in the same direction as this paper
- positive
sucrose · molecular: metabolites · Catharanthus roseus (Madagascar periwinkle)Results agree
1 paper in the same direction as this paper
- positive
drought · growth · Catharanthus roseus (Madagascar periwinkle)Positive and negative results
1 paper in the same direction as this paper · 4 in the opposite direction
- negative
- negative
- negative
2 papers more in this group
See all 6 papers →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.