二倍体马铃薯试管苗的培养及对叶肉原生质体融合的影响

Culture of Diploid Potato Plantlets in vitro and Effect on Mesophyll Protoplast Fusion

以35份二倍体马铃薯试管苗为试验材料,对二倍体马铃薯试管苗培养的影响因素进行研究,目的是使试管苗的叶片增大,叶面积增加,并有利于进行叶肉原生质体的融合。结果表明:含有AgNO3的培养基(MS+0.5 mg/L NAA+1 mg/L AgNO3)适合22份供试二倍体马铃薯试管苗的培养;添加AgNO3比不添加AgNO3的试管苗的叶片明显增大,叶面积显著增加,添加2%蔗糖和2 mg/L AgNO3的培养基的试管苗叶面积最大;以二倍体野生种S.chacoense和二倍体材料DY4-5-10为亲本进行电融合,以添加2%蔗糖和1 mg/L AgNO3的培养基培养其试管苗,获得的融合产物可再生出完整植株,该培养基适合用于分离原生质体的二倍体马铃薯试管苗的培养。

Thirty-five diploid potato materials were used to study the factors influencing the culture of diploid potato plantlets in vitro in order to increase their blade size and leaf area, and benefit the fusion of mesophyl protoplast. Twenty-two diploid potato plantlets in vitro could be cultivated in the medium MS＋0.5 mg/L NAA＋1 mg/L AgNO3. The blade size was increased obviously in the medium containing AgNO3 than without it, and the leaf area was increased, significantly. The maximum leaf area was got in the medium added with 2 mg/L AgNO3 and 2%sucrose. Plants were regenerated from the electrofusion product using mesophyl protoplast isolated from diploid wild species S. chacoense and a diploid clone DY4-5-10, which were cultivated in the medium added with 1 mg/L AgNO3 and 2%sucrose. So the medium is suitable for the culture of diploid potato plantlets in vitro used for the isolation of protoplasts.