玻璃化法超低温保存柑桔茎尖及植株再生

园艺学报 ›› 2001, Vol. 28 ›› Issue (4): 301-306.

玻璃化法超低温保存柑桔茎尖及植株再生

王子成；邓秀新^*

(华中农业大学作物遗传改良国家重点实验室, 武汉430070)

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-08-25 发布日期:2001-08-25

Cryopreservation of Citrus Shoot-tips by Vitrification and Regeneration

Wang Zicheng and Deng Xiuxin

(National Key Laboratory of Crops Genetic Improvement , Huazhong Agricultural University , Wuhan 430070)

Received:1900-01-01 Revised:1900-01-01 Online:2001-08-25 Published:2001-08-25

摘要/Abstract

摘要： 采用玻璃化法对柑桔茎尖的离体超低温保存进行了研究。约10 mm 长的柑桔茎尖于含5 %二甲基亚砜(DMSO) 的培养基上预培养3 d , 切取2～2. 5 mm 长的茎尖, 室温下60 %玻璃化溶液2 (PVS2) 装载20～30 min , 然后用PVS2 于0 ℃处理50～60 min , 换入新鲜的PVS2 , 迅速投入液氮中, 24 h 后在40 ℃水浴中迅速化冻, 再用1. 2 mol/L 蔗糖培养基洗涤2次, 接种于含BA 1. 0 mg/L 的MT培养基上, 26 ℃暗培养1 周后转于正常光下培养。枳壳茎尖超低温保存后用氯化三苯基四氮唑(TTC) 法检测, 成活率为100 % , 培养再生率达到90 % ,再生后的苗能正常生根, 与对照没有形态上的变异, 移栽可成活。

关键词: 柑桔, 茎尖, 超低温, 玻璃化法

Abstract: A procedure had been studied on cryopreservation of citrus shoot2tips in vitro by vitrification. Shoot-tips of citrus about 10 mm were precultured for 3 days in MT medium supplemented with 5 % DMSO. The excised shoot2tips , 2 - 2. 5 mm in length , were loaded with 60 % PVS2 for 20 - 30 min at room temperature , and were exposed to PVS2 at 0 ℃for 50 - 60 min. Followed by changing the solution with fresh PVS2 , the shoot2tips were immersed into LN directly , and kept for 24 h. After rapid thawing in a water bath at 40 ℃, the shoot2tips were washed with 1. 2 mol/ L sucrose solution for twice and transferred onto MT medium supplemented with BA 1. 0 mg/ L. The cultures were kept in dark for one week prior to exposure to the light . Survival rate of shoot2tips of Poncirus trifoliata was 100 % by TTC examination , and regeneration rate reached to 90 %. The plantlets rooted normally and survived after transplantion. No morphological difference between the regenerated
plantlets and the control was observed.

Key words: Citrus, Shoot-tips, Cryopreservation, Vitrification

中图分类号:

S 666
Q 813

王子成;邓秀新. 玻璃化法超低温保存柑桔茎尖及植株再生[J]. 园艺学报, 2001, 28(4): 301-306.

Wang Zicheng and Deng Xiuxin. Cryopreservation of Citrus Shoot-tips by Vitrification and Regeneration[J]. ACTA HORTICULTURAE SINICA, 2001, 28(4): 301-306.

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