头孢霉素与卡那霉素对花生组培苗生根的影响

摘要

Resistance screening is an indispensable step in Agrobacterium mediated genetic transformation of peanut,but the use of antibiotics have significant influence on peanut seedling growth and rooting.In order to make clear the sensitive concentration of antibiotics on rooting of peanut seedlings,the peanut cultivation variety Fenghua 1 tissue culture seedlings were used as experimental materials,and the culture media with different concentrations of kanamycin or cefotaxime were used to detect the peanut seedling rooting rate and growth status.The results showed that 0~250 mg/L cefotaxime had little effect on peanut growth,while kanamycin had significant effect on peanut seedling rooting.0~10 mg/L kanamycin has no obvious effect on peanut seedling rooting and growth,but under the condition of 10 mg/L kanamycin,the seedlings had no branch roots,what''s more,15 mg/L kanamycin significantly inhibited the elongation of roots.When the rootless plantlets were transfered into MS medium after cutting off the bottom of the seedlings,the roots grew out again,whereas the rootless plantlets did not grow roots if not cutting off the bottom.After screening with 75 mg/L kanamycin,the rooting rate of the kanamycin-resistant tissue culture plantlets could reach 31%.This study could provide certain theoretical supports for roots induction during the late stage of peanut genetic transformation.%抗性筛选是农杆菌介导的花生遗传转化过程中不可或缺的步骤,但是抗生素的使用对花生组培苗的生长与生根有着显著影响.为了明确抗生素对花生组培苗生根的敏感质量浓度,本试验以花生栽培品种丰花1号组培苗为材料,在培养基中添加不同浓度的头孢霉素与卡那霉素,检测花生组培苗的生根率及植株生长状况并进行统计分析.结果表明:0~250 mg/L的头孢霉素对花生组培苗生根及植株生长基本无影响.卡那霉素对花生组培苗生根影响较为显著,0~10 mg/L卡那霉素对花生组培苗生根与生长基本无影响,10 mg/L卡那霉素条件下组培苗不长侧根,15 mg/L卡那霉素显著抑制根的伸长;切去不生根的组培苗底端后移入MS培养基又能重新生根,不切去底端的组培苗则不生根.经过75 mg/L卡那霉素筛选的抗性组培苗的生根率可达31%.本研究可为花生遗传转化后期根的诱导提供一定的理论支持.