3种抗生素对尾巨桉愈伤组织诱导及植株再生的影响

摘要

【目的】研究潮霉素、卡那霉素和头孢霉素对尾巨桉愈伤组织诱导、不定芽分化及生根的影响,确立抑制农杆菌生长的头孢霉素质量浓度。【方法】以尾巨桉无菌苗茎段为外植体,先后接种于添加不同质量浓度抗生素的愈伤组织诱导培养基、不定芽诱导培养基以及生根培养基上,统计愈伤组织诱导率、不定芽诱导率和不定芽生根率,观察头孢霉素对农杆菌生长的影响。【结果】（1）培养基中的潮霉素、卡那霉素、头孢霉素的质量浓度分别为15,25,200mg/L时,尾巨桉外植体的愈伤组织诱导率分别为6.7%,12.1%和26.7%;（2）潮霉素、卡那霉素、头孢霉素的质量浓度分别为15,25,250mg/L时,不定芽分化受到明显抑制,不定芽诱导率分别仅有2.6%,10.2%和7.6%;（3）潮霉素、卡那霉素、头孢霉素的质量浓度分别为15,25,250mg/L时,不定芽生根困难,生根率分别为10.2%,16.2%和10.1%;当头孢霉素质量浓度≥200mg/L时,农杆菌生长被完全抑制。【结论】尾巨桉对不同抗生素的敏感性存在差异,在尾巨桉遗传转化过程中,抗性愈伤组织与不定芽诱导的潮霉素适宜质量浓度为10mg/L,卡那霉素为20mg/L;生根诱导适宜的潮霉素和卡那霉素的质量浓度分别为15和25mg/L;抑制农杆菌生长的头孢霉素较适宜的质量浓度为200mg/L。%【Objective】 The study was to discuss the effects of hygromycin,kanamycin,and cephalosporin on the callus induction,regeneration of adventitious buds and root induction of Eucalyptus urophylla×Eucalyptus grandis.The optimum mass concentrations of cephalosporin to inhibition Agrobacterium was determined.【Method】 The asepsis stem segments of E.urophylla×E.grandis sterile clones were cultured respectively on the callus induction culture media,adventitious buds induction culture media,and rooting culture media,which were added several kinds of mass concentration antibiotics.Callus induction rate,rate of adventitious bud induction and rooting of adventitious buds were measured,respectively,and the effect of cephalosporin on Agrobacterium was observed.【Result】（1） The rate of callus induction from the explants was 6.7%,12.1% and 26.7% respectively,while the mass concentration of hygromycin,kanamycin and cephalosporin in the medium was higher than 15,25 and 200 mg/L.（2） Regeneration of adventitious buds was remarkably inhibited and the rate of regeneration of adventitious buds was 2.6%,10.2% and 7.6%,respectively while the mass concentrations of hygromycin,kanamycin and cephalosporin were over 15,25 and 250 mg/L.（3）Mullberry shoots could not form healthy roots and the rate of rooting of adventitious buds was 10.2%,16.2% and 10.1%,respectively while the mass concentrations of hygromycin,kanamycin and cephalosporin were 15,25 and 250 mg/L.Cefotaxime strongly inhibited Agrobacterium while the mass concentration of cephalosporin was over 200 mg/L.【Conclusion】 The inhibitive effects upon E.urophylla×E.grandis vary with the different antibiotics.The optimum mass concentrations of hygromycin and kanamycin were 10,20 mg/L at the stage of screening resistant callus and adventitious buds,and were 15,25 mg/L at the stage of rooting of resistant buds.200 mg/L Cef was a better choice to inhibit Agrobacterium.