Investigating the cryopreservation of nodalexplants of Lithodora rosmarinifolia (Ten.)Johnst., a rare, endemic Mediterraneanspecies

Giovanni Iapichino, Giuseppe Barraco, Florent Engelmann, Isabelle Sylvestre, Giuseppe Barraco

Risultato della ricerca: Article

2 Citazioni (Scopus)

Abstract

In this study, we investigated the possibility ofusing the droplet-vitrification technique for cryopreservingnodal segments of in vitro plantlets of the endangered plantspecies Lithodora rosmarinifolia. Among the three vitrificationsolutions tested, only solutions B1, containing (w/v)50 % glycerol and 50 % sucrose, and B3, containing 40 %glycerol and 40 % sucrose, were able to induce cryotolerancein nodal explants, resulting in intermediate survivaland recovery after cryopreservation. A three-step vitrificationprotocol, including an additional dehydration treatmentwith half-strength vitrification solution for 30 minbefore the treatment with full-strength vitrification solution,did not lead to any improvement in survival andrecovery compared with the two-step protocol. The optimalprotocol was the following: preculture of nodal segments inliquid medium with 0.3 M sucrose for 16 h and 0.7 Msucrose for 5 h, treatment for 20 min in loading solutioncontaining 1.9 M glycerol ? 0.5 M sucrose, dehydrationwith vitrification solution B1 (glycerol 50.0 %, sucrose50.0 %, w/v) for 60 min at room temperature, rapid coolingin minute droplets of vitrification solution, and rapidrewarming by immersion of nodal segments for 20 min inunloading solution containing 1.2 M sucrose. Under theseconditions, 33 % recovery of cryopreserved nodal explantswas achieved. Regrowth of cryopreserved samples wasrapid and direct. These results indicate that long-termstorage of L. rosmarinifolia by means of cryopreservationof nodal segments is possible, thereby contributing tosecuring the diversity of this rare and endangered plantspecies.
Lingua originaleEnglish
pagine (da-a)141-146
Numero di pagine6
RivistaPlant Biotechnology Reports
Volume7
Stato di pubblicazionePublished - 2013

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Vitrification
Cryopreservation
cryopreservation
vitrification
Sucrose
Glycerol
sucrose
glycerol
droplets
Immersion
Dehydration
regrowth
plantlets
ambient temperature
explants
Temperature
Therapeutics

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Plant Science

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Investigating the cryopreservation of nodalexplants of Lithodora rosmarinifolia (Ten.)Johnst., a rare, endemic Mediterraneanspecies. / Iapichino, Giovanni; Barraco, Giuseppe; Engelmann, Florent; Sylvestre, Isabelle; Barraco, Giuseppe.

In: Plant Biotechnology Reports, Vol. 7, 2013, pag. 141-146.

Risultato della ricerca: Article

Iapichino, Giovanni ; Barraco, Giuseppe ; Engelmann, Florent ; Sylvestre, Isabelle ; Barraco, Giuseppe. / Investigating the cryopreservation of nodalexplants of Lithodora rosmarinifolia (Ten.)Johnst., a rare, endemic Mediterraneanspecies. In: Plant Biotechnology Reports. 2013 ; Vol. 7. pagg. 141-146.
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title = "Investigating the cryopreservation of nodalexplants of Lithodora rosmarinifolia (Ten.)Johnst., a rare, endemic Mediterraneanspecies",
abstract = "In this study, we investigated the possibility ofusing the droplet-vitrification technique for cryopreservingnodal segments of in vitro plantlets of the endangered plantspecies Lithodora rosmarinifolia. Among the three vitrificationsolutions tested, only solutions B1, containing (w/v)50 {\%} glycerol and 50 {\%} sucrose, and B3, containing 40 {\%}glycerol and 40 {\%} sucrose, were able to induce cryotolerancein nodal explants, resulting in intermediate survivaland recovery after cryopreservation. A three-step vitrificationprotocol, including an additional dehydration treatmentwith half-strength vitrification solution for 30 minbefore the treatment with full-strength vitrification solution,did not lead to any improvement in survival andrecovery compared with the two-step protocol. The optimalprotocol was the following: preculture of nodal segments inliquid medium with 0.3 M sucrose for 16 h and 0.7 Msucrose for 5 h, treatment for 20 min in loading solutioncontaining 1.9 M glycerol ? 0.5 M sucrose, dehydrationwith vitrification solution B1 (glycerol 50.0 {\%}, sucrose50.0 {\%}, w/v) for 60 min at room temperature, rapid coolingin minute droplets of vitrification solution, and rapidrewarming by immersion of nodal segments for 20 min inunloading solution containing 1.2 M sucrose. Under theseconditions, 33 {\%} recovery of cryopreserved nodal explantswas achieved. Regrowth of cryopreserved samples wasrapid and direct. These results indicate that long-termstorage of L. rosmarinifolia by means of cryopreservationof nodal segments is possible, thereby contributing tosecuring the diversity of this rare and endangered plantspecies.",
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AU - Sylvestre, Isabelle

AU - Barraco, Giuseppe

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N2 - In this study, we investigated the possibility ofusing the droplet-vitrification technique for cryopreservingnodal segments of in vitro plantlets of the endangered plantspecies Lithodora rosmarinifolia. Among the three vitrificationsolutions tested, only solutions B1, containing (w/v)50 % glycerol and 50 % sucrose, and B3, containing 40 %glycerol and 40 % sucrose, were able to induce cryotolerancein nodal explants, resulting in intermediate survivaland recovery after cryopreservation. A three-step vitrificationprotocol, including an additional dehydration treatmentwith half-strength vitrification solution for 30 minbefore the treatment with full-strength vitrification solution,did not lead to any improvement in survival andrecovery compared with the two-step protocol. The optimalprotocol was the following: preculture of nodal segments inliquid medium with 0.3 M sucrose for 16 h and 0.7 Msucrose for 5 h, treatment for 20 min in loading solutioncontaining 1.9 M glycerol ? 0.5 M sucrose, dehydrationwith vitrification solution B1 (glycerol 50.0 %, sucrose50.0 %, w/v) for 60 min at room temperature, rapid coolingin minute droplets of vitrification solution, and rapidrewarming by immersion of nodal segments for 20 min inunloading solution containing 1.2 M sucrose. Under theseconditions, 33 % recovery of cryopreserved nodal explantswas achieved. Regrowth of cryopreserved samples wasrapid and direct. These results indicate that long-termstorage of L. rosmarinifolia by means of cryopreservationof nodal segments is possible, thereby contributing tosecuring the diversity of this rare and endangered plantspecies.

AB - In this study, we investigated the possibility ofusing the droplet-vitrification technique for cryopreservingnodal segments of in vitro plantlets of the endangered plantspecies Lithodora rosmarinifolia. Among the three vitrificationsolutions tested, only solutions B1, containing (w/v)50 % glycerol and 50 % sucrose, and B3, containing 40 %glycerol and 40 % sucrose, were able to induce cryotolerancein nodal explants, resulting in intermediate survivaland recovery after cryopreservation. A three-step vitrificationprotocol, including an additional dehydration treatmentwith half-strength vitrification solution for 30 minbefore the treatment with full-strength vitrification solution,did not lead to any improvement in survival andrecovery compared with the two-step protocol. The optimalprotocol was the following: preculture of nodal segments inliquid medium with 0.3 M sucrose for 16 h and 0.7 Msucrose for 5 h, treatment for 20 min in loading solutioncontaining 1.9 M glycerol ? 0.5 M sucrose, dehydrationwith vitrification solution B1 (glycerol 50.0 %, sucrose50.0 %, w/v) for 60 min at room temperature, rapid coolingin minute droplets of vitrification solution, and rapidrewarming by immersion of nodal segments for 20 min inunloading solution containing 1.2 M sucrose. Under theseconditions, 33 % recovery of cryopreserved nodal explantswas achieved. Regrowth of cryopreserved samples wasrapid and direct. These results indicate that long-termstorage of L. rosmarinifolia by means of cryopreservationof nodal segments is possible, thereby contributing tosecuring the diversity of this rare and endangered plantspecies.

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