Articles
Efficiency of aluminum cryo-plates for cryopreservation of Dendrobium signatum Rchb. f. pollinia
Article number
1234_36
Pages
279 – 286
Language
English
Abstract
The efficiency of V cryo-plate and D cryo-plate methods for cryopreservation of pollinia of Dendrobium signatum Rchb. f., a Thai orchid was investigated.
Pollinia were collected from flowers and then placed on the aluminum cryo-plate containing 12 wells embedded with 3% sodium alginate gel.
In V cryo-plate method, cryo-plates with pollinia were immersed in 0.6 M sucrose + 2 M glycerol loading solution (LS) for 15 min, then dehydrated with PVS2 solution for 40 min at room temperature (29±2°C). In D cryo-plate method, cryo-plates with pollinia were treated with LS for 15 min, then dehydrated in a laminar air-flow cabinet for 3 h at room temperature (29±2°C). In both methods, cryo-plates were directly plunged into liquid nitrogen for 40 min, and rapidly warmed in 1.2 M sucrose for 15 min.
The cryopreserved and non-cryopreserved pollinia were used to pollinate flowers of the same species.
The results showed that cryopreserved pollinia retained fertilizing ability, and had similar pod formation as those pollinated with non-cryopreserved pollinia.
The pod formation from cryopreserved pollinia in V cryo-plate and D cryo-plate methods were 55.6 and 50%, respectively.
Seeds from non-cryopreserved and cryopreserved pollinia were successfully produced and germinated into plantlets with well-formed leaves and roots.
Multiple stems (4.8 stems plant-1) produced from one plantlet when cultured on modified VW agar medium (1949) supplemented with 100 g L-1 banana, 150 mL coconut water, 50 g L-1 potato, and 20 g L-1 sucrose for 120 d at 25±2°C. Cryopreserved pollinia using aluminum cryo-plate can be used successfully for pollination.
Pollinia were collected from flowers and then placed on the aluminum cryo-plate containing 12 wells embedded with 3% sodium alginate gel.
In V cryo-plate method, cryo-plates with pollinia were immersed in 0.6 M sucrose + 2 M glycerol loading solution (LS) for 15 min, then dehydrated with PVS2 solution for 40 min at room temperature (29±2°C). In D cryo-plate method, cryo-plates with pollinia were treated with LS for 15 min, then dehydrated in a laminar air-flow cabinet for 3 h at room temperature (29±2°C). In both methods, cryo-plates were directly plunged into liquid nitrogen for 40 min, and rapidly warmed in 1.2 M sucrose for 15 min.
The cryopreserved and non-cryopreserved pollinia were used to pollinate flowers of the same species.
The results showed that cryopreserved pollinia retained fertilizing ability, and had similar pod formation as those pollinated with non-cryopreserved pollinia.
The pod formation from cryopreserved pollinia in V cryo-plate and D cryo-plate methods were 55.6 and 50%, respectively.
Seeds from non-cryopreserved and cryopreserved pollinia were successfully produced and germinated into plantlets with well-formed leaves and roots.
Multiple stems (4.8 stems plant-1) produced from one plantlet when cultured on modified VW agar medium (1949) supplemented with 100 g L-1 banana, 150 mL coconut water, 50 g L-1 potato, and 20 g L-1 sucrose for 120 d at 25±2°C. Cryopreserved pollinia using aluminum cryo-plate can be used successfully for pollination.
Authors
N. Jitsopakul, P. Sangyojarn, P. Homchan, K. Thammasiri
Keywords
orchid, Dendrobium signatum, V cryo-plate, D cryo-plate, Vacin and Went medium, breeding
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