Optimization of droplet-vitrification protocol for carnation genotypes and ultrastructural studies on shoot tips during cryopreservation

• Autorzy: Zhang J.-M. , Zhang X.-N. , Lu X.-X. , Xin X. , Yin G.-K. , He J.-J. , Xu Y.-M. , Chen X.-L.
• Języki publikacji: EN

Abstrakty

EN

This study was carried out to optimize a modified droplet-vitrification procedure for the cryopreservation of shoot tips from different carnation genotypes. The best procedure was developed by applying orthogonal tests to the experimental data and by further investigation of the effects on the regrowth percentage. It consisted in preculturing shoot tips in liquid Murashige and Skoog (MS) medium with 0.3 M sucrose for 2 days, pretreating them in liquid MS medium with 5 % Dimethyl sulfoxide +5 % glycerol + 0.3 M sucrose for 10 min, osmoprotecting in Loading solution for 20 min at 25°C, cryoprotecting with Plant vitrification solution No.2 (PVS2) for 60 min at 0°C, transferring in drops of fresh PVS2 over aluminum strips and finally storing them in Liquid nitrogen. With the application of the optimized protocol, four carnation cultivars (‘Master’, ‘Calibra’, ‘Lamour’ and ‘Ofcar’) achieved regrowth percentage after cryopreservation ranging from 41 to 73 %. Ultrastructural observations investigated by using transmission electron microscopy showed that the cells encountered the stress during cryopreservation and the main damages occurred during the dehydration step. For surviving cells, the most of the damaged cells could be repaired after recovery growth. This modified protocol will aid in the long-term conservation of carnation germplasm and the ultrastructural studies will benefit for understanding the damage and recovery of the cells during cryopreservation.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2014
• Tom: 36
• Numer: 12
• Opis fizyczny: p.3189-3198,fig.,ref.

Twórcy

autor

Zhang J.-M.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

autor

Zhang X.-N.

• College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, China
• Institute of Rice Research, JiangXi Academy of Agricultural Sciences, Nanchang, 330200, China

autor

Lu X.-X.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

autor

Xin X.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

autor

Yin G.-K.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

autor

He J.-J.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

autor

Xu Y.-M.

• College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, China

autor

Chen X.-L.

• National Genebank, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

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Identyfikatory

DOI

10.1007/s11738-014-1685-7