Abstract
The embryogenic cell suspension culture of Gentiana cruciata, cryopreserved by the encapsulation/dehydration method, survived both short- (48 h) and long-term (1.5 years) cryostorage with more than 80% viability. To assess the influence of cryotreatments on the embryogenic potential, a proembryogenic mass was encapsulated and exposed to the following treatments: (1) osmotic dehydration (OD), (2) OD + air desiccation (AD) and (3) OD + AD + cryostorage (LN). The somatic embryogenesis efficiency increased ten times after osmotic dehydration. The AD and LN cryotreatments did not cause any significant alterations in somatic embryo production. We monitored the (epi)genetic stability of 288 regenerants derived from: non-cryotreated, short-term, and long-term cryostored tissue using metAFLP markers and ten primer combinations. Changes in the sequence and DNA methylation levels were studied by subjecting the DNA to digestion with two pairs of isoschisomer restriction enzymes (KpnI/MseI and Acc65I/MseI). Two new AFLP unique DNA fragments at the DNA sequence level, with no differences at the methylation level, were found between regenerants derived from cryopreserved tissue, compared with the non-cryotreated controls. The Acc65I/MseI methylation levels for the three groups of regenerants were not significantly different. Cluster analysis was capable of identifying a number of sub-clusters. Only one of the sub-clusters comprises almost all regenerants derived from non-cryotreated and short-term cryostored tissue. Plantlets derived from long-term cryostored tissue were grouped into separate clusters. The observed AFLP alterations did not appear to be associated with the use of cryopreservation, but were probably related to the process of in vitro culture.
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Claims 10
Claim · model-written summary
Cryopreserved embryogenic suspension culture of Gentiana cruciata kept more than 80% viability after short- and long-term cryostorage.
- Taxon
- Gentiana cruciata
Gentiana cruciatat0 - Organ / stage
- embryogenic cell suspension culturee0
- Treatment
- encapsulation/dehydration methode0
- Outcome
- viabilitye0
- Value
- more than 80%e0
- Qualifier
- short- (48 h) and long-term (1.5 years) cryostoragee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e0“The embryogenic cell suspension culture of Gentiana cruciata, cryopreserved by the encapsulation/dehydration method, survived both short- (48 h) and long-term (1.5 years) cryostorage with more than 80% viability”
Claim · model-written summary
Osmotic dehydration increased somatic embryogenesis efficiency ten times in the proembryogenic mass.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Organ / stage
- proembryogenic masse1
- Treatment
- osmotic dehydratione2
- Outcome
- somatic embryogenesis efficiencye2
- Direction
- increasede2
- Value
- ten timese2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e1“a proembryogenic mass was encapsulated and exposed to the following treatments: (1) osmotic dehydration (OD), (2) OD + air desiccation (AD) and (3) OD + AD + cryostorage (LN)”
e2“The somatic embryogenesis efficiency increased ten times after osmotic dehydration”
Claim · model-written summary
Air desiccation and cryostorage cryotreatments did not significantly alter somatic embryo production.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Treatment
- AD and LN cryotreatmentse3
- Outcome
- somatic embryo productione3
- Direction
- no_changeinferrede3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e3“The AD and LN cryotreatments did not cause any significant alterations in somatic embryo production”
Claim · model-written summary
Two new AFLP unique DNA fragments were found in regenerants from cryopreserved tissue compared with non-cryotreated controls.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Material
- regenerants derived from cryopreserved tissuee4
- Outcome
- AFLP unique DNA fragmentse4
- Value
- Two newe4
- Comparator
- non-cryotreated controlse4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e4“Two new AFLP unique DNA fragments at the DNA sequence level, with no differences at the methylation level, were found between regenerants derived from cryopreserved tissue, compared with the non-cryotreated controls”
Claim · model-written summary
Regenerants from cryopreserved tissue showed no differences at the methylation level.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Material
- regenerants derived from cryopreserved tissuee4
- Outcome
- differences at the methylation levele4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e4“Two new AFLP unique DNA fragments at the DNA sequence level, with no differences at the methylation level, were found between regenerants derived from cryopreserved tissue, compared with the non-cryotreated controls”
Claim · model-written summary
Acc65I/MseI methylation levels did not differ significantly among the three groups of regenerants.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Material
- three groups of regenerantse5
- Outcome
- Acc65I/MseI methylation levelse5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e5“The Acc65I/MseI methylation levels for the three groups of regenerants were not significantly different”
Claim · model-written summary
Cluster analysis identified a number of sub-clusters among the regenerants.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Outcome
- a number of sub-clusterse6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e6“Cluster analysis was capable of identifying a number of sub-clusters”
Claim · model-written summary
Only one sub-cluster comprised almost all regenerants from non-cryotreated and short-term cryostored tissue.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Material
- regenerants derived from non-cryotreated and short-term cryostored tissuee7
- Outcome
- Only one of the sub-clusters comprises almost all regenerantse7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e7“Only one of the sub-clusters comprises almost all regenerants derived from non-cryotreated and short-term cryostored tissue”
Claim · model-written summary
Plantlets from long-term cryostored tissue were grouped into separate clusters.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Material
- Plantlets derived from long-term cryostored tissuee1
- Outcome
- grouped into separate clusterse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e1“Plantlets derived from long-term cryostored tissue were grouped into separate clusters”
Claim · model-written summary
The observed AFLP alterations were probably not associated with cryopreservation but related to in vitro culture.
- Taxon
- Gentiana cruciataGentiana cruciatat0
- Treatment
- use of cryopreservatione1
- Outcome
- AFLP alterationse1
- Direction
- no_changeinferrede1
- Setting
- in vitro culturee1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“embryogenic capacity of Gentiana cruciata (L.) suspension culture”
e1“The observed AFLP alterations did not appear to be associated with the use of cryopreservation, but were probably related to the process of in vitro culture”