以菠萝为试材,建立试管无性系,进行玻璃化法超低温保存及植株再生研究,结果表明,约2.0 ～3.0 cm的菠萝茎尖于含有0.Smol/L蔗糖的培养基上预培养l～2d,剥取含1～2片叶原基(1.0～2.5 mm长)的茎尖,室温(25℃)下用LS装载液预处理50min,再用PVS2溶液于0℃下处理40 min,换入新鲜的PVS2溶液后迅速投入液氪,保存24h后在40℃水浴中迅速化冻90 s,用1.2 mol/L的蔗糖培养液洗涤2次,每次10 min,然后转入含0.3 mol/L蔗糖的MS培养基上,暗培养48 h后转入含BA 0.5 mg/L的MS培养基上,暗培养1周后转移到正常光下,4个菠萝品种(澳大利亚卡因、夏威夷、巴厘、台农16号)的成活率分别为96.5％、95.3％、78.6％、87.7％,再生率分别为93.2％、92.6％、76.7％、87.7％.再生植株生长和分化正常,其形态上与对照一致,生根后可移栽成活.%Taking pineapple as tested materials, cryopreservation of pineapple shoot-tips in vitro by vitrification and plant regeneration were studied, the result showed that the shoot-tips of pineapple in length of 2.0 -3.0 cm were precultured on MS medium supplemented with 0. 5 mol/L sucrose for 1 -2 days. Dissected shoot apices about 1.0 -2.5 mm in length with one or two primordium leaves were loaded with a mixture of 2 mol/L glycerol plus 0.4 mol/L sucrose for 50 min under room temperature, and treated with the solution of PVS2 at 0 "C for 40 min, and then plunged directly into liquid nitrogen and conserved for at least 1 h. After being rapidly thawed in a water bath at 40 "C for 90s, the shoot tips were rinsed with 1.2 mol/L sucrose solution for 20 min under room temperature. Then the shoot tips were plated on MS medium supplemented with 0.3 mg/L sucrose, after 48 h of culture in darkness, the cryopreserved materials were transferred onto MS medium supplemented with 0.5 mg/L BA and cultured in darkness for one week. The survival rates of four pineapple culn'vars were 95.5 % ,95. 3 % ,78.6 % and 87.7 % .respectively, regrowth rate were 93.2 % ,92.6 % ,76.7 % and 87.7 % .respectively. Almost all the shoot tips formed roots and were successfully transferred to soil in pots. The plantlets could normally root and survive after transplantation.
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