黄独脱毒苗种质离体保存及其遗传稳定性和病毒变化的初步研究

Preliminary Study on In vitro Conservation of Dioscorea bulbifera L.Virus-free

以黄独脱毒苗带芽茎段为外植体,以MS为基本培养基,采用正交实验和单因子实验方法研究了无机盐水平、蔗糖、甘露醇和植物生长抑制剂PP333对黄独脱毒苗离体保存的影响。结果表明,黄独脱毒苗带芽茎段在25±1℃下保存于附加2.0 mg.L-1KT0、.5 mg.L-1NAA3、0 g.L-1蔗糖、20 g.L-1甘露醇和4 mg.L-1 PP333的1/4MS培养基上,180 d后其成活率可达90%以上。离体保存后的脱毒苗经形态指标和生理生化指标以及RT-PCR分析测定没有发生遗传变异,也没有检测到PVY病毒。本实验结果为药用植物黄独脱毒苗的种质保存提供了一条简单有效的途径。

The effect of inorganic salts level,sucrose,mannitol and plant growth inhibitor PP333 on In vitro conservation of Dioscorea bulbifera L.virus-free plantlet stems with a bud in MS basic medium was studied using single-factor experiments and orthogonal experimental method.The results showed that the survival rate of D.bulbifera L.virus-free plantlet stems with a bud which cultured on 1/4MS medium containing 2.0mg·L^-1 KT,0.5mg·L^-1 NAA,30g·L^-1 sucrose,20g·L^-1 mannitol and 4mg·L^-1 PP333 and stored at 25±1℃ temperature was over 90%.After morphological indexes measurement,physiological and biochemical indexes determination and RT-PCR analysis,the genetic variation and PVY of In vitro conservation D.bulbifera L.virus-free plantlets were not detected.The results provided a simple and effective way for In vitro conservation of D.bulbifera L.virus-free plantlets germplasm.