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RISS 인기검색어
- 검색키워드
- 저자 : Isabelle Sylvestre (검색결과 4 건)
- 무료
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- 유료
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Barraco, Giuseppe,Sylvestre, Isabelle,Iapichino, Giovanni,Engelmann, Florent 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.2
In this study, we investigated the possibility of using the droplet-vitrification technique for cryopreserving nodal segments of in vitro plantlets of the endangered plant species Lithodora rosmarinifolia. Among the three vitrification solutions tested, only solutions B1, containing (w/v) 50 % glycerol and 50 % sucrose, and B3, containing 40 % glycerol and 40 % sucrose, were able to induce cryotolerance in nodal explants, resulting in intermediate survival and recovery after cryopreservation. A three-step vitrification protocol, including an additional dehydration treatment with half-strength vitrification solution for 30 min before the treatment with full-strength vitrification solution, did not lead to any improvement in survival and recovery compared with the two-step protocol. The optimal protocol was the following: preculture of nodal segments in liquid medium with 0.3 M sucrose for 16 h and 0.7 M sucrose for 5 h, treatment for 20 min in loading solution containing 1.9 M glycerol + 0.5 M sucrose, dehydration with vitrification solution B1 (glycerol 50.0 %, sucrose 50.0 %, w/v) for 60 min at room temperature, rapid cooling in minute droplets of vitrification solution, and rapid rewarming by immersion of nodal segments for 20 min in unloading solution containing 1.2 M sucrose. Under these conditions, 33 % recovery of cryopreserved nodal explants was achieved. Regrowth of cryopreserved samples was rapid and direct. These results indicate that long-term storage of L. rosmarinifolia by means of cryopreservation of nodal segments is possible, thereby contributing to securing the diversity of this rare and endangered plant species.
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Giuseppe Barraco,Isabelle Sylvestre,Giovanni Iapichino,Florent Engelmann 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.2
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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Jung-Yoon Yi,Isabelle Sylvestre,Myriam Colin,Mohammad Salma,Sok-Young Lee,Haeng-Hoon Kim,Hong-Jae Park,Florent Engelmann 한국원예학회 2012 원예과학기술지 Vol.30 No.1
An efficient protocol for cryopreservation of madder hairy root cultures has been developed using droplet-vitrification. In previous study, combining loading solution C4 (35% PVS3) and vitrification solution B5 (80% PVS3) was the most effective method. In this study, we tried three types of vitrification solution, B5, A3 (90% PVS2, on ice), and A5 (70% PVS2, on ice). Combining loading solution C4 and vitrification solution A5 (on ice) showed the best regeneration rate in this study. Histological changes of the cells within the hairy root of madder were also observed in different steps. The cells from the hairy roots of the control treatment were full and intact with different size of vacuoles and obvious cell nucleus having a dark nucleolus. After the stage of preparing for cryopreservation (after preculturing, loading, followed by dehydration solution A5 or B5), intercellular spaces had become distinct, and within cells, the cytoplasms had become denser and week plasmolyses had appeared. The cell plasmolyses were much more apparent and we measured the degree of plasmolysis by calculating, the area of cell/the area of cytoplasm. The value of plasmolysis degree was the highest in the combination of preculture, loading solution C4, and dehydration solution A5, 1.97. Because the highest regeneration rates appeared in the treatment of A5 for 20 min, we could assume that the optimal degree of plasmolysis for cryopreservation might be around 1.97. The changes in cell structure during cryopreservation might be a useful basis for the development of a proper long-term preservation method for madder germplasms.
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Yi, Jung-Yoon,Sylvestre, Isabelle,Colin, Myriam,Salma, Mohammad,Lee, Sok-Young,Kim, Haeng-Hoon,Park, Hong-Jae,Engelmann, Florent Korean Society of Horticultural Science 2012 원예과학기술지 Vol.30 No.1
An efficient protocol for cryopreservation of madder hairy root cultures has been developed using droplet-vitrification. In previous study, combining loading solution C4 (35% PVS3) and vitrification solution B5 (80% PVS3) was the most effective method. In this study, we tried three types of vitrification solution, B5, A3 (90% PVS2, on ice), and A5 (70% PVS2, on ice). Combining loading solution C4 and vitrification solution A5 (on ice) showed the best regeneration rate in this study. Histological changes of the cells within the hairy root of madder were also observed in different steps. The cells from the hairy roots of the control treatment were full and intact with different size of vacuoles and obvious cell nucleus having a dark nucleolus. After the stage of preparing for cryopreservation (after preculturing, loading, followed by dehydration solution A5 or B5), intercellular spaces had become distinct, and within cells, the cytoplasms had become denser and week plasmolyses had appeared. The cell plasmolyses were much more apparent and we measured the degree of plasmolysis by calculating, the area of cell/the area of cytoplasm. The value of plasmolysis degree was the highest in the combination of preculture, loading solution C4, and dehydration solution A5, 1.97. Because the highest regeneration rates appeared in the treatment of A5 for 20 min, we could assume that the optimal degree of plasmolysis for cryopreservation might be around 1.97. The changes in cell structure during cryopreservation might be a useful basis for the development of a proper long-term preservation method for madder germplasms.
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