冬凌草试管苗茎尖玻璃化法超低温保存及植株再生研究

对冬凌草Rabdosia rubescens(Hemsl.)Hara.]试管苗茎尖进行了玻璃化法超低温保存及植株再生研究。结果表明:1～2cm嫩梢在培养基MS+0.5mol/L蔗糖上培养3d,切取2～3mm茎尖,室温(20℃)下装载液(60%PVS2)过渡30min,0℃下玻璃化液(PVS2)处理45min,投入液氮保存1d后,40℃水浴化冻,1.2mol/L蔗糖的MS培养基洗涤20min,接种于6-BA0.8mg/L、IAA0.05mg/L的MS培养基上,暗处理1周后转到正常光下,成活率达到86%,再生植株生长和分化正常。

Study on Cryopreservation and Plant Regeneration of in Vitro Shoot-tips of Rabdosia Rubescens (Hemsl.) Hara. by Vitrification

The cryopreservation and plant regeneration of in vitro shoot-tips of Rabdosia rubescens(Hemsl.)Hara.by vitrification were studied.Shoot-tips,2～3 mm long,gotten from in vitro shoots(1～2 cm)were precultured on MS medium supplemented with 0.5 mol/L sucrose for 3 d,loaded with the 60 % PVS2 for 30 min at 20 ℃,and incubated in PVS2 for 45 min at 0 ℃,then directly plunged into liquid nitrogen,after rapid thawed in water at 40 ℃.The shoot-tips were rinsed in the MS medium supplemented with 1.2 mol/L sucrose for 20 min,plated on the MS regeneration medium supplemented with 0.8 mg/L 6-BA,0.05 mg/L IAA for one week in dark,then exposured to the light.In these conditions,the survival of shoot-tips was up to 86 % and grew and differenced normally.