番木瓜茎尖的玻璃化法超低温保存及其植株再生

园艺学报 ›› 2004, Vol. 31 ›› Issue (1): 29-33.

番木瓜茎尖的玻璃化法超低温保存及其植株再生

曾继吾^1,2; 易干军^1*;张秋明²

(¹ 广东省农业科学院果树研究所, 广州510640; ² 湖南农业大学园艺园林学院, 长沙410128)

收稿日期:2003-04-17 修回日期:2003-06-05 出版日期:2004-02-25 发布日期:2004-02-25

Cryopreservation of in Vitro Papaya Shoot2tips by Vitrification Technique andIts Regeneration

Zeng Jiwu^1,2; Yi Ganjun¹; Zhang Qiuming²

(¹ Fruit Research Institute, Guangdong Academy of Agricultural Sciences, Guangdong 510640, China ; ² College of Horticulture, Hunan Agricultural University, Changsha 410128, China)

Received:2003-04-17 Revised:2003-06-05 Online:2004-02-25 Published:2004-02-25

摘要/Abstract

摘要： 研究了番木瓜( Carica papaya L. ) 茎尖玻璃化法超低温保存的一些影响因素。结果表明: 无
菌试管苗在MS + BA 0.5 mg/L + NAA 0.1 mg/L + GA₃ 1.0 mg/L 的培养基上生长较好。番木瓜茎尖超低温保存较佳体系是: 3～5 cm的茎尖在含有5 %二甲基亚砜(DMSO) 培养基上预培养3 d , 解剖镜下剥取含1～2 个叶原基的茎尖(1.5～2.5 mm长) , 先在室温下于60 %的玻璃化溶液(PVS2) 中装载40～50 min , 再用PVS2于0 ℃下处理30 min , 换1 次PVS2 溶液后迅速投入液氮。保存24 h 后, 在40 ℃水浴中化冻, 用1.2 mol/L 的蔗糖培养液洗涤两次, 转入继代培养基上再培养, 成活率和再生率分别达53.7 %和52.6 %。再生植株生根后可移栽成活。

关键词: 番木瓜, 茎尖, 超低温保存, 玻璃化法

Abstract: Experiments were conducted about tissue culture systems and cryopreservation on papaya ( Carica papaya L.) . The cluster buds were subcultured on the proliferation medium containing 0.5 mg/ L BA , 0.1 mg/ L
NAA and 1.0 mg/ L GA3. The procedure of cryopreservation was studied using the shoot tips as materials. In vitro, the shoot tips of papaya about 3-5 cm in length were precultured for 3 days at 25 ℃on MS medium supplemented
with 5 %DMSO. Shoot tips about 1.5-2.5 mm in length with one or two leaf primordiumwere loaded in a solution containing 60 % PVS2 for 40-50 min at room temperature. The shoot2tips were sufficiently dehydrated with a highly concentrated vitrification solution (PVS2) for 40 min at 0 ℃. After changing the solution with fresh PVS2 , the shoots were immersed into liquid nitrogen directly and conservated for 24 hour. After rapidly thawing in a water
bath at 40 ℃, the shoot tips were washed twice with 1.2 mol/ L sucrose solution and transferred on the same MS medium as the previous one. Survival rates of shoot tips was 53.7 % , and regrowth rate reached 52.6 %. Almost
all the shoot tips formed roots on hormone2free solidified MS medium and were successfully transferred to soil in pots. The plantlets could normally root and survive after transplantion.

Key words: Papaya (Carica papaya L.), Shoot-tips, Cryopreservation, Vitrification

中图分类号:

S 667

曾继吾;易干军;张秋明. 番木瓜茎尖的玻璃化法超低温保存及其植株再生[J]. 园艺学报, 2004, 31(1): 29-33.

Zeng Jiwu;Yi Ganjun;Zhang Qiuming. Cryopreservation of in Vitro Papaya Shoot2tips by Vitrification Technique andIts Regeneration[J]. ACTA HORTICULTURAE SINICA, 2004, 31(1): 29-33.

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