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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: Peroxidation due to cryoprotectant treatment is a vital factor for cell survival in Arabidopsis cryopreservation

Authors
item Ren, Li -
item Jiang, Xiang-Ning -
item Gai, Ying -
item Wang, Wei-Ming -
item REED, BARBARA
item Shen, Xiao-Hui -

Submitted to: Plant Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: July 20, 2013
Publication Date: July 29, 2013
Citation: Ren, L., Jiang, X., Gai, Y., Wang, W., Reed, B.M., Shen, X. 2013. Peroxidation due to cryoprotectant treatment is a vital factor for cell survival in Arabidopsis cryopreservation. Plant Science. 212:37-47.

Interpretive Summary: Cryopreservation, storage in liquid nitrogen, is an effective and cost-effective tool for the long-term storage of plants, but damage to plant tissues and death of plants sometimes occurs. Determining the causes of this damage is vital to improving plant recovery. When germinating seeds were cryopreserved, the ability to recover was lost as germination progressed. Germination could be restored after cryopreservation in seeds germinated for 48-h , but after 72-h all the seeds died. Genes expressed at the two times were compared and 166 were analyzed. Among these, oxidative-stress genes played important roles in cryopreservation, and peroxidation was a key factor related to cell survival. Germinating seeds underwent more peroxidation at 72-h than at 48-h. We determined that membranes were injured during the process, but in the 48-h germinating seeds some of these oxidative-stress genes prevented the injury. Reducing injury from oxidative stress was the key factor to successful cryopreservation. This study established a model to test and evaluate the effect of antioxidants and cryoprotectants in plant cryopreservation.

Technical Abstract: Cryopreservation is a safe and cost-effective tool for the long-term storage of plant germplasm, but damage to plant tissues and death of plants sometimes occurs. Determining the causes of this damage is vital to improving plant recovery from cryopreservation. When Arabidopsis germinating seeds were cryopreserved, the ability to recover from cryogenic treatment was lost as germination progressed. Germination could be restored in 48-h germinating seeds after cryopreservation, whereas 72-h germinating seeds died. Why seedling age and survival are negatively correlated is an interesting issue. A comparative transcriptomics was performed to screen differentially expressed genes between 48- and 72-h germinating seeds after exposure to the cryoprotectant in the vitrification protocol. A total of 7146 transcript-derived fragments (TDFs) were detected, and 166 differentially expressed TDFs were analyzed. Among these, oxidative-stress response genes played important roles in cryopreservation, and peroxidation was a key factor related to cell survival. Germinating seeds underwent more peroxidation at 72-h than at 48-h. A comprehensive analysis indicated that peroxidation injured membrane systems leading to photophosphorylation and oxidative phosphorylation damage. In 48-h germinating seeds some genes prevented the initiation of apoptosis induced by oxidative stress, leading to higher survival. Differential strategies of oxidative stress response affected germinating seed survival, and reducing the injury of oxidative stress was the key factor to successful cryopreservation. This study provided a novel insight of genetic regulatory mechanisms in cryopreservation, and established an excellent model to test and evaluate the effect of exogenous antioxidants and conventional cryoprotectants in plant cryopreservation.

Last Modified: 8/19/2014
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