黄瓜未受精胚珠离体培养及单倍体植株再生

Studies on Haploid Plant Induction via in vitro Unfertilized Ovule Culture of Cucumber

以欧亚杂交类型黄瓜(Cucumis sativus L.)65G×228的F_1为材料,对未受精胚珠进行离体培养,通过6因素5水平的正交试验,研究预处理温度、诱导培养基、TDZ、KT和BA浓度对未受精胚珠雌核发育的影响。结果证明TDZ浓度对胚发生频率有极显著影响,是诱导黄瓜离体雌核发育的最主要因素,当其浓度为0.02 mg·L^(-1)时胚状体的诱导率最高。诱导培养基和KT浓度显著影响胚发生率,而预处理温度和BA浓度影响不显著。硝态氮(NO_3^-)和铵态氮(NH_4^+)含量比例均接近1:1的诱导培养基,较其他比值较大的培养基的胚发生率有显著提高。利用流式细胞仪进行倍性鉴定,再生植株中单倍体率为18.2%。

The unpollinated ovules of cucumber（Cucumis sativus L.） line 65G×228 were used to obtain haploid plants via in vitro gynogenesis.With the unfertilized ovules,a regeneration system of cucumber was established by analizing the effects of pretreatment temperature,induction medium,concentration of TDZ,KT and BA on haploid induction.The results indicated that 0.02 mg· L^（-1） TDZ was better for the initiation of gynogenesis.Induction medium and concentration of KT significantly affected the frequency of embryo induction.Further,the highest embryo regeneration frequency was achieved at IM3 medium supplemented with 0.75 mg·L^（-1） KT and 2.0 mg · L^（-1） BA.The embryos could grow root and develop into the complete plant on MS medium.Flow cytometry analysis showed that 18.2% of the regenerated plants were haploid.