Articles
Dormant bud cryopreservation of a variety of woody ornamentals and regeneration via in vitro meristem cultures
Article number
1421_14
Pages
103 – 110
Language
English
Abstract
A collection of woody landscaping plant material was cryopreserved from a cool greenhouse in Luke Laukaa, Finland in mid-winter in January 2017. The dormant bud stem material was cut to 3 cm segments, placed in 2-mL cryovials, and directly frozen in two steps.
The first step was performed in a liquid nitrogen driven controlled rate freezer to -30°C over 6 h (hold 5 min, -0.1°C min‑1, hold 30 min). Secondly, the cryovials were immersed in liquid nitrogen and the cryobox was placed in the gas phase liquid nitrogen tank.
The material was thawed overnight at +4°C in January 2020. Twig sections were surface sterilized, meristems with a few leaf primordia were then excised and grown on WPM media (20 mg L‑1 sucrose, 1.5 mg L‑1 zeatin) in test tubes.
Regrowth was observed after 6-10 weeks.
All plant species studied, regenerated into new microplantlets, except Spiraea betulifolia and some Rosa cultivars.
The regrowth levels varied for Dasiphora fruticosa 50-70%, Diervilla sp. 63-75%, Lonicera sp. 8-83%, Spiraea sp. 0-80% and Viburnum sp. 36-100%. Some species cultures, as in Spiraea, could be subcultured within a few weeks.
The method proved to be feasible to rapidly cryopreserve dormant material and regenerate it in vitro for a variety of woody ornamentals.
The first step was performed in a liquid nitrogen driven controlled rate freezer to -30°C over 6 h (hold 5 min, -0.1°C min‑1, hold 30 min). Secondly, the cryovials were immersed in liquid nitrogen and the cryobox was placed in the gas phase liquid nitrogen tank.
The material was thawed overnight at +4°C in January 2020. Twig sections were surface sterilized, meristems with a few leaf primordia were then excised and grown on WPM media (20 mg L‑1 sucrose, 1.5 mg L‑1 zeatin) in test tubes.
Regrowth was observed after 6-10 weeks.
All plant species studied, regenerated into new microplantlets, except Spiraea betulifolia and some Rosa cultivars.
The regrowth levels varied for Dasiphora fruticosa 50-70%, Diervilla sp. 63-75%, Lonicera sp. 8-83%, Spiraea sp. 0-80% and Viburnum sp. 36-100%. Some species cultures, as in Spiraea, could be subcultured within a few weeks.
The method proved to be feasible to rapidly cryopreserve dormant material and regenerate it in vitro for a variety of woody ornamentals.
Authors
S. Tuohimetsä, A. Nukari
Keywords
germplasm, micropropagation, plant genetic resources, two-step freezing
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