Abstract
Embryogenic calli of Anemarrhena asphodel- oides Bunge were successfully cryopreserved by vitrifica- tion. Excised embryogenic calli from in vitro grown tillers were precultured in liquid basal tissue culture medium from Murashige and Skoog (MS medium) containing 2m g L -1 Kinetin (Kn), 0.1 mg L -1 a-naphthalene acetic acid (NAA) and 0.5 M sucrose for 2 days under continuous light at 25 ± 1C. Following preculture the calli were transferred to a 2 mL plastic cryotube. The calli was then loaded with a mixture of 2 M glycerol and 0.4 M sucrose for 30 min at 25 ± 1C and dehydrated with a highly concentrated vitrification solution (PVS2) for 40 min at 0C. After changing the solution with fresh PVS2, the calli were directly immersed in liquid nitrogen (LN). After rapid thawing in a water-bath at 35C for 5 min, the calli were then transferred onto solid MS medium supplemented with Kn 2 mg L -1 , NAA 0.1 mg L -1 , 3% (w/v) sucrose and 0.75% (w/v) agar. The cultures were kept in the dark for 5 days prior to exposure to the light (14 h light/dark cycle). After 30 days, the embryoids developed from embryogenic calli were transferred to fresh solid basal MS media sup- plemented with Kn 2 mg L -1 , NAA 0.1 mg L -1 , 3% (w/v) sucrose and 0.75% (w/v) agar (regeneration medium) and developed normal shoots. The regrowth rate of vitrified embryogenic calli reached over 60%. Cryopreservation did not affect the plant regeneration potential of A. asphodel- oides Bunge embryogenic calli through somatic embryo- genesis. Morphologies of plants regenerated from cryopreserved calli were similar to those of the seedlings. This efficient cryopreservation protocol would be useful for the ex-situ conservation of A. asphodeloides Bunge germ- plasm in gene banks at very low temperatures.
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Claims 6
Claim · model-written summary
Embryogenic calli were successfully cryopreserved by vitrification.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Outcome
- successfully cryopreservede0
- Value
- successfullye0
Verbatim
e0“Embryogenic calli of Anemarrhena asphodel- oides Bunge were successfully cryopreserved by vitrifica- tion”
t0“Anemarrhena asphodeloides Bunge”
Claim · model-written summary
Embryoids developed normal shoots after transfer to fresh medium.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Organ / stage
- embryoidse0
- Outcome
- developed normal shootse0
Not reportedDirection
Verbatim
e0“the embryoids developed from embryogenic calli were transferred to fresh solid basal MS media sup- plemented with Kn 2 mg L -1 , NAA 0.1 mg L -1 , 3% (w/v) sucrose and 0.75% (w/v) agar (regeneration medium) and developed normal shoots”
t0“Anemarrhena asphodeloides Bunge”
Claim · model-written summary
Regrowth rate of vitrified calli exceeded 60%.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Outcome
- regrowth ratee0
- Direction
- reachede0
- Value
- over 60%e0
Verbatim
e0“The regrowth rate of vitrified embryogenic calli reached over 60%”
t0“Anemarrhena asphodeloides Bunge”
Claim · model-written summary
Cryopreservation did not affect plant regeneration potential.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Treatment
- Cryopreservatione0
- Outcome
- plant regeneration potentiale0
- Direction
- did not affecte0
Not reportedComparator
Verbatim
e0“Cryopreservation did not affect the plant regeneration potential of A. asphodel- oides Bunge embryogenic calli through somatic embryo- genesis”
t0“Anemarrhena asphodeloides Bunge”
Claim · model-written summary
Regenerated plant morphology was similar to seedlings.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Outcome
- Morphologiese0
- Direction
- similare0
- Comparator
- seedlingse0
Verbatim
e0“Morphologies of plants regenerated from cryopreserved calli were similar to those of the seedlings”
t0“Anemarrhena asphodeloides Bunge”
Claim · model-written summary
The authors propose the protocol for ex situ germplasm conservation in gene banks.
- Taxon
- Anemarrhena asphodeloides BungeAnemarrhena asphodeloidest0
- Outcome
- ex-situ conservatione0
- Setting
- gene bankse0
Verbatim
e0“This efficient cryopreservation protocol would be useful for the ex-situ conservation of A. asphodeloides Bunge germ- plasm in gene banks at very low temperatures”
t0“Anemarrhena asphodeloides Bunge”