文献类型： 外文期刊

作者： Zhang, Jin-Mei ¹ ; Zhang, Xiao-Ning ¹ ; Lu, Xin-Xiong ¹ ; Xin, Xia ¹ ; Yin, Guang-Kun ¹ ; He, Juan-Juan ¹ ; Xu, You-Mi ¹ ;

作者机构： 1.Chinese Acad Agr Sci, Inst Crop Sci, Natl Genebank, Beijing 100081, Peoples R China

2.Huazhong Agr Univ, Coll Hort & Forestry Sci, Wuhan 430070, Peoples R China

3.JiangXi Acad Agr Sci, Inst Rice Res, Nanchang 330200, Peoples R China

关键词： Carnation;Dianthus caryophyllus L.;Droplet-vitrification;In vitro;Shoot tips;Ultrastructural investigation

期刊名称：ACTA PHYSIOLOGIAE PLANTARUM （ 影响因子：2.354； 五年影响因子：2.711 ）

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收录情况： SCI

摘要： 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 A degrees C, cryoprotecting with Plant vitrification solution No.2 (PVS2) for 60 min at 0 A degrees 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.