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A simple and efficient cryopreservation of in vitro-grown shoot tips of `Troyer' citrange [Poncirus trifoliata (L.) Raf. × Citrus sinensis (L.) Osbeck.] by encapsulation-vitrification

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Abstract

A simple and efficient method was developed for cryopreservation ofin vitro-grown shoot tips of `Troyer' citrange byencapsulation-vitrification. Excised shoot tips were precultured withincreasing sucrose concentrations of 0.3, 0.5, 0.75 and 1 M for 4 days.Precultured shoot tips were encapsulated and simultaneously osmoprotectedwith a loading solution of 2 M glycerol and 1 M sucrose.Osmoprotected and encapsulated shoot tips were dehydrated with a highlyconcentrated vitrification solution prior to direct immersion in LN for 1 h.Optimal survival of cryopreserved shoot tips was obtained when preculturedshoot tips were osmoprotected for 60 min during encapsulation.Dehydration by exposure to modified PVS2 solution for 90 min at24 °C and for 180 to 210 min at 0 °C was found optimalfor survival of cryopreserved shoot tips. Preculture duration largelyinfluenced survival of cryopreserved shoot tips, with the best result obtainedafter 2 to 5 days of preculture with stable 1 M sucrose. With theoptimized parameters, 100% survival of cryopreserved shoot tips wasachieved. Morphologies of plants regenerated from cryopreserved shoottips were similar to those of the seedlings.

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Authors and Affiliations

  1. Department of Virology, Agricultural Research Organization, the Volcani Center, P.O.B. 6, Bet Dagan, 50250, Israel

    Qiaochun Wang, Özgur Batuman, Ping Li, Moshe Bar-Joseph & Ron Gafny

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  1. Qiaochun Wang
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  2. Özgur Batuman
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  3. Ping Li
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  4. Moshe Bar-Joseph
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  5. Ron Gafny
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Wang, Q., Batuman, Ö., Li, P. et al. A simple and efficient cryopreservation of in vitro-grown shoot tips of `Troyer' citrange [Poncirus trifoliata (L.) Raf. × Citrus sinensis (L.) Osbeck.] by encapsulation-vitrification. Euphytica 128, 135–142 (2002). https://doi.org/10.1023/A:1020683305690

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