悬铃木遗传转化过程中选择压力的确定

Determination of the Selecting Pressure for Genetic Transformation of Platanus acerifolia Willd.

以二球悬铃木(Platanus acerifolia)的下胚轴和离体叶片为材料,研究了植物遗传转化过程中常用的2种选择剂卡那霉素(Kan)和除草剂(PPT)对其不定芽分化的抑制作用,同时试验了2种选择剂对试管苗生根的影响。结果表明,Kan和PPT对悬铃木下胚轴和离体叶片的不定芽分化及试管苗生根均表现出较强的抑制作用,其中12.5-20.0 mg/L Kan或2.0 mg/L PPT能完全抑制叶片外植体正常的不定芽分化,50 mg/L Kan或1.0 mg/L PPT可完全抑制下胚轴的不定芽分化;Kan和PPT抑制试管苗生根的选择浓度分别为50 mg/L和4.0 mg/L。试验确定了以悬铃木的下胚轴和叶片为转化受体时不同筛选阶段Kan和PPT的选择压力,为其基因工程改良奠定了基础。

Effects of two commonly used selecting agents in genetic transformation, kanamycin （Kan） and phosphinothricin （PPT）,on the differentiation of adventitious shoots from hypocotyl and in vitro-leaf explants and on rooting of in vitro shoots of Platanus acerifolia were investigated. The results indicated that both Kan and PPT showed a strong inhibition effect on the growth and differentiation of the hypocotyl and leaf explants of the species,with 12.5-20. 0 mg/L Kan or 2.0 mg/L PPT inhibiting shoot regeneration from leaves completely, and 50 mg/L Kan or 1.0 mg/L PPT inhibiting shoot regeneration from hypoeotyls. The concentrations of Kan and PPT to inhibit the rooting of in vitro - shoots were 50 mg/L and 4. 0 mg/L, respectively. In this paper, proper concentrations of Kan and PPT for selection in genetic transformation using hypocotyl or leaf as explants were determined, which established a foundation for the genetic imorovement of P. aceri folia via gene engineering.