Articles
Effectiveness of pollination and in vitro germination of Laelia anceps subsp. dawsonii seeds using cryopreserved pollen
Article number
1421_4
Pages
27 – 34
Language
English
Abstract
Laelia anceps subsp. dawsonii, a beautiful endemic orchid native to Mexico, has been in danger of extinction.
The application of a biotechnological method such as pollen cryopreservation may represent a valuable strategy to guarantee the long-term germplasm preservation of this species and be a useful tool for orchid breeding programs.
This work reports successful pollen cryopreservation and then evaluates capsule formation, in vitro seed germination and subsequent development of seedlings resulted from pollination by cryopreserved pollen in Laelia anceps subsp. dawsonii. Using cryopreserved and non-cryopreserved pollen one flower per plant was pollinated from a total of 10 individuals.
Capsule formation and size of formed capsules 154 days after pollination, as well as the effect of different sucrose (7.5, 15.0, 22.5 and 30.0 g L‑1) concentrations in the semi-solid MS medium on seed germination were assessed after 117 d of culture.
Capsule formation induced by using fresh and cryopreserved pollen resulted in 100 and 60%, respectively.
Fresh pollen produced larger capsules (4.3 cm length × 2.2 cm wide) than cryopreserved (3.6 cm length × 1.8 cm wide), however, cryo-derived seeds showed higher percentages of germination at the same culture time.
The in vitro germination ranged from 37 to 66% for seeds derived of capsules produced using fresh pollen.
The highest percentage (66%) was obtained after culture at the lowest (7.5 g L‑1) sucrose concentration tested.
Germination of seeds produced by cryopreserved pollen ranged from 44 to 92%. The significantly highest percentage (92%) was obtained using 15 g L‑1 sucrose in the culture medium.
Greater concentrations of sucrose did not improve germination.
These are the first results obtained on the cryopreservation of pollen from Laelia anceps spp. and the subsequent asymbiotic seed germination from cryo-derived capsules.
The application of a biotechnological method such as pollen cryopreservation may represent a valuable strategy to guarantee the long-term germplasm preservation of this species and be a useful tool for orchid breeding programs.
This work reports successful pollen cryopreservation and then evaluates capsule formation, in vitro seed germination and subsequent development of seedlings resulted from pollination by cryopreserved pollen in Laelia anceps subsp. dawsonii. Using cryopreserved and non-cryopreserved pollen one flower per plant was pollinated from a total of 10 individuals.
Capsule formation and size of formed capsules 154 days after pollination, as well as the effect of different sucrose (7.5, 15.0, 22.5 and 30.0 g L‑1) concentrations in the semi-solid MS medium on seed germination were assessed after 117 d of culture.
Capsule formation induced by using fresh and cryopreserved pollen resulted in 100 and 60%, respectively.
Fresh pollen produced larger capsules (4.3 cm length × 2.2 cm wide) than cryopreserved (3.6 cm length × 1.8 cm wide), however, cryo-derived seeds showed higher percentages of germination at the same culture time.
The in vitro germination ranged from 37 to 66% for seeds derived of capsules produced using fresh pollen.
The highest percentage (66%) was obtained after culture at the lowest (7.5 g L‑1) sucrose concentration tested.
Germination of seeds produced by cryopreserved pollen ranged from 44 to 92%. The significantly highest percentage (92%) was obtained using 15 g L‑1 sucrose in the culture medium.
Greater concentrations of sucrose did not improve germination.
These are the first results obtained on the cryopreservation of pollen from Laelia anceps spp. and the subsequent asymbiotic seed germination from cryo-derived capsules.
Authors
F. Hernández-Ramírez, A. Martínez-Lagunes, J.A. Alejandre-Rosas, C.A. Cruz-Cruz, M.T. González-Arnao
Keywords
endemic orchid, liquid nitrogen, flowers, capsules, sucrose concentration, asymbiotic germination
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