Articles
Expression analysis of Trx M1 and M-Grx genes in Dendrobium nobile protocorm-like body response to cryopreservation
Article number
1234_12
Pages
97 – 104
Language
English
Abstract
Reactive oxygen species (ROS)-induced oxidative stress is a vital factor that reduces cell viability during cryopreservation.
Thioredoxin (Trx) and glutaredoxin (Grx) participate in ROS scavenging and manage redox homeostasis in cells.
Our previous proteomic study found proteins Trx M1 and monothiol-Grx (M-Grx) in protocorm-like bodies (PLBs) of Dendrobium nobile Lindl.
LSQUOHamana Lake DreamRSQUO were differentially expressed during cryopreservation.
Therefore, this study was to clarify the molecular regulation of Trx M1 and M-Grx, and their relationship with oxidative stress during cryopreservation.
The mRNA expression of Trx M1 and M-Grx in Dendrobium nobile PLBs was analyzed using real-time quantitative PCR method in both the control vitrification procedure and an improved vitrification procedure by adding ascorbic acid (AsA) in unloading solution.
The results showed that both Trx M1 and M-Grx genes were up-regulated after loading, PVS2-treated and unloading, however significantly down-regulated after preculture and cooling-rewarming.
Notably, our previous study also found the obvious decrease of survival, along with the occurrence of oxidative damage in PLBs after preculture and cooling-rewarming, which was consistent with the change of Trx M1 and M-Grx gene expression.
The addition of AsA significantly increased the survival and M-Grx expression in cryopreserved PLBs.
In conclusion, Trx M1 and M-Grx genes play important roles in Dendrobium nobile PLB tolerance to ROS-induced oxidative stress during cryopreservation.
Adding AsA in unloading solution might relieve oxidative damage by enhancing M-Grx gene expression.
Thioredoxin (Trx) and glutaredoxin (Grx) participate in ROS scavenging and manage redox homeostasis in cells.
Our previous proteomic study found proteins Trx M1 and monothiol-Grx (M-Grx) in protocorm-like bodies (PLBs) of Dendrobium nobile Lindl.
LSQUOHamana Lake DreamRSQUO were differentially expressed during cryopreservation.
Therefore, this study was to clarify the molecular regulation of Trx M1 and M-Grx, and their relationship with oxidative stress during cryopreservation.
The mRNA expression of Trx M1 and M-Grx in Dendrobium nobile PLBs was analyzed using real-time quantitative PCR method in both the control vitrification procedure and an improved vitrification procedure by adding ascorbic acid (AsA) in unloading solution.
The results showed that both Trx M1 and M-Grx genes were up-regulated after loading, PVS2-treated and unloading, however significantly down-regulated after preculture and cooling-rewarming.
Notably, our previous study also found the obvious decrease of survival, along with the occurrence of oxidative damage in PLBs after preculture and cooling-rewarming, which was consistent with the change of Trx M1 and M-Grx gene expression.
The addition of AsA significantly increased the survival and M-Grx expression in cryopreserved PLBs.
In conclusion, Trx M1 and M-Grx genes play important roles in Dendrobium nobile PLB tolerance to ROS-induced oxidative stress during cryopreservation.
Adding AsA in unloading solution might relieve oxidative damage by enhancing M-Grx gene expression.
Authors
W. Di, X.R. Jiang, M.X. Jia, J. Xu, R.F. Ren, Z.D. Li, B.L. Li, Y. Liu
Keywords
Dendrobium, vitrification, thioredoxin, glutaredoxin, gene expression, oxidative damage
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