摘要
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0.5 mol/L蔗糖的培养基上预培养1～2 d,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溶液后迅速投入液氮,保存24 h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 ℃ 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 ℃ 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 cultivars 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.
出处
《西南农业学报》
CSCD
北大核心
2011年第5期1875-1878,共4页
Southwest China Journal of Agricultural Sciences
关键词
菠萝
茎尖
超低温保存
玻璃化法
Pineapple
Shoot-tips
Cryopreservation
Vitrification
