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United States Department of Agriculture

Agricultural Research Service

Research Project: Management of Temperate-Adapted Fruit, Nut, and Specialty Crop Genetic Resources and Associated Information

Location: National Clonal Germplasm Repository (Corvallis, Oregon)

Title: Antioxidants and anti-stress compounds improve the survival of cryopreserved Arabidopsis seedlings

Authors
item Ren, Li -
item Zhang, Di -
item Shen, X.H. -
item Reed, Barbara

Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 15, 2014
Publication Date: June 1, 2014
Citation: Ren, L., Zhang, D., Shen, X., Reed, B.M. 2014. Antioxidants and anti-stress compounds improve the survival of cryopreserved Arabidopsis seedlings. Acta Horticulturae. 1039:57-62.

Interpretive Summary: Cryopreservation is a safe and cost-effective tool for the long-term storage of plant germplasm. Successful cryopreservation depends on having a suitable cryoprotection protocol. This study was done to determine how antioxidants improve the survival of Arabidopsis seedlings after cryopreservation. Several antioxidants were added during a cryopreservation protocol to improve the survival ratio in seedlings that were allowed to germinate for 60-h and 72-h . Four of the compounds increased the survival by approximately two times in 60-h seedlings and for 72-h seedlings increased the survival from zero to as much as 14.3%. This confirms that antioxidants and anti-stress compounds used in cryopreservation protocols can significantly increase the regrowth of seedlings and this may be due to reducing or repairing the oxidative damage.

Technical Abstract: Cryopreservation is a safe and cost-effective tool for the long-term storage of plant germplasm. Successful cryopreservation depends on suitable cryoprotection protocol. In Arabidopsis seedlings cryopreservation, the growth ability could be partly restored in 60-h seedlings, whereas 72-h seedlings died after cryopreservation. In this study, glutathione (GSH), ascorbic acid (VC), lipoic acid (LA), melatonin (M), polyvinylpyrrolidone (PVP), abscisic acid (ABA), glycine betaine (GB), CaCl2, and KNO3 were added to Plant Vitrification Solution 2 (PVS2) to improve the survival ratio in 60-h and 72-h seedlings. Among of them, four compounds (GSH, VC, ABA and GB) increased the survival ratio approximately two times in 60-h seedlings cryopreservation, and FDA viability staining analysis also indicated that adding these four compounds made 60-h seedlings keep higher viability than control. Other exogenous compounds had neutral or negative impacts on 60-h seedlings regrowth. Moreover, in 72-h seedlings cryopreservation, GSH, GB, ABA and VC increased the survival ratio from 0 to up tp 14.3%. These four compounds were usually considered as antioxidants or anti-stress compounds. In general, vitrification-based cryopreservation improves survival of plant materials by increasing cell viscosity, also produces complex stresses especially oxidative stress. Antioxidants and anti-stress compounds in cryopreservation protocol can significantly increase the regrowth of seedlings may be due to reducing or repairing the oxidative damage.

Last Modified: 10/1/2014
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