二倍体马铃薯原生质体的游离与培养

Studies on the Protoplast Isolation and Culture of Diploid Potato

以3种不同基因型的二倍体马铃薯为材料,研究了材料来源和不同预处理及游离过程中渗透浓度对原生质体游离质量和后期发育的影响。结果表明,不同基因型二倍体马铃薯细胞酶解时的适宜渗透浓度不同,Dd-Ⅲ-5为0.35～0.40 mol/L,JK3为0.35 mol/L,而Dd-Ⅱ-10的适宜渗透浓度范围较广,在0.30～0.45mol/L内均可;不同预处理所游离的原生质体质量及后期发育存在差异,经预处理后的原生质体活性均高于85%,且分化能力强;同一基因型的叶肉和愈伤组织所游离的原生质体活性差异不大,在产量上叶肉细胞高于愈伤组织;愈伤组织分化时,分化培养基附加的3个激素组合中,NAA 1 mg/L+ZT 0.2 mg/L时愈伤组织全部褐化死亡,而NAA 1 mg/L+2,4-D 1 mg/L+GA30.2 mg/L和NAA 1 mg/L+BA 1 mg/L上的愈伤都分化,后者分化较早;基因型对原生质体后期发育影响较大,经培养后,仅Dd-Ⅲ-5得到完整植株。

The research on the effects of materials type, different prereatment and osmo-tic concentration on the protoplast yield and cell development in diploid potatos of three different genotype was conducted. The result showed that osmotic concentration of different genotype are different ,the suit- able osmotic concentration of Dd-Ⅲ-5 is 0.35-0.40 mol/L,JK3 is 0.35 mol/L, Dd-Ⅱ-10 have large extent osmotic concentration in 0.30-0.45 mol/L. The protoplast quality and the cell development from different pretreatment have contrast, viability of the protoplast pretreated is more than 85%, and the capability of cleavage is stronger. The protoplast viability have contrast between mesophy11 cell and calli, the yield of mesophyll cell is more than calli; Among differential medi-um, calli are browning in NAA 1 mg/L＋ZT 0.2 mg/L, calli formed the plant in NAA 1 mg/L＋2,4-D 1 mg/L＋ GA3 0.2 mg/L and NAA 1 mg/L＋BA 1 mg/L , and the latter is earlier. Genotype have dramatic effects on protoplast culture, through culture, the regenerated plants formed only in Dd-Ⅲ-5.