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United States Department of Agriculture

Agricultural Research Service

Research Project: MANAGEMENT OF TEMPERATE FRUIT NUT AND SPECIALTY CROP GENETIC RESOURCES

Location: National Clonal Germplasm Repository (Corvallis, Oregon)

Title: The antioxidant melatonin boosts recovery of cryopreserved shoot tips

Authors
item Uchendu, Esther -
item Shukla, M. -
item Reed, Barbara
item Saxena, P. -

Submitted to: Symposium Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 2012
Publication Date: N/A

Interpretive Summary: Many useful plant species found in Canada are of conservation concern. In vitro storage and cryopreservation (liquid nitrogen storage) techniques guarantee safety of these species and have potential applications which may result in sustainable agriculture. Shoot tips of in vitro-grown plantlets of American elm, St John’s Wort, tobacco and dormant winter buds of American elm were successfully cryopreserved in liquid nitrogen at -196 °C under controlled environment conditions following antioxidant treatment with two protocols. The antioxidant melatonin enhanced regrowth. Under optimized conditions for cryopreservation of these species, the two protocols resulted in 80 to 100% regrowth for all plant types. Shoot tips of dormant winter buds of the American elm consistently produced nearly 100% regrowth with both techniques. This study indicates that addition of the antioxidant melatonin during cryopreservation produced excellent recovery of these species and is a very viable option for long-term storage .

Technical Abstract: Many useful plant species found in Canada are of conservation concern. In vitro storage and cryopreservation techniques guarantee safety of these species and have potential applications which may result in sustainable agriculture. Shoot tips of in vitro-grown plantlets of American elm, St John’s Wort, tobacco and dormant winter buds of American elm were successfully cryopreserved in liquid nitrogen at -196 °C under controlled environment conditions following antioxidant treatment and cold acclimation with either vitrification or encapsulation-vitrification protocols. Explants of St John’s Wort and tobacco had optimal regrowth following cryopreservation, when treated with the Plant Vitrification Solution #2 (PVS2) for 20 min on ice and those of American elm for 10 min on ice. Melatonin (0.1 µM), added to both preculture and regrowth medium significantly enhanced regrowth compared to the control (p<0.05). Under optimized conditions for cryopreservation of these species, the two protocols resulted in 80 to 100% regrowth for all plant types. Shoot tips of dormant winter buds of the American elm consistently produced nearly 100% regrowth with both techniques. This study indicates that addition of the antioxidant melatonin during cryopreservation produced excellent recovery of these species and is a very viable option for long-term storage of these economic species.

Last Modified: 4/16/2014
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