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Title of Journal: Appl Biochem Biotechnol

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Abbravation: Applied Biochemistry and Biotechnology

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Springer US

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DOI

10.1016/0040-1951(87)90039-4

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1559-0291

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Effects of Ascorbic Acid on PVS2 Cryopreservation

Authors: Jessica Jeyanthi James Antony Chan Lai Keng Maziah Mahmood Sreeramanan Subramaniam
Publish Date: 2013/07/07
Volume: 171, Issue: 2, Pages: 315-329
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Abstract

Regrowth of the cryopreserved protocormlike bodies PLBs of Dendrobium Bobby Messina was assessed based on the plant vitrification solution 2 PVS2 optimisation conditions The optimized protocol obtained based on TTC spectrophotometrical analysis and growth recovery were 3–4 mm of PLBs size precultured in 02 M sucrose for 1 day treated with a mixture of 2 M glycerol and 04 M sucrose supplemented with halfstrength liquid MS media at 25 °C for 20 min and subsequently dehydrated with PVS2 at 0 °C for 20 min prior to storage in liquid nitrogen Following rapid warming in a water bath at 40 °C for 90 s PLBs were treated with unloading solution containing halfstrength liquid MS media supplemented with 12 M sucrose Subsequently the PLBs were cultured on halfstrength semisolid MS media supplemented with 2  w/v sucrose without any growth regulators and resulted in 40  growth recovery In addition ascorbic acid treatment was used to evaluate the regeneration process of cryopreserved PLBs However growth recovery rates of noncryopreserved and cryopreserved PLBs were 30 and 10  when 06 mM ascorbic acid was added Scanning electron microscopy analysis indicates that there are not much damages observed on both cryopreserved and noncryopreserved PLBs in comparison to PLBs stock culture

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