尾叶桉U6遗传转化再生体系的建立

以桉树品种尾叶桉U6叶盘为外植体,选用MS培养基为基本培养基,附加激素6-BA,IAA,IBA,NAAA,通过研究不同激素浓度组合对外植体愈伤诱导及不定芽分化的影响,建立了较好的遗传转化再生体系.结果表明,尾叶桉U6叶盘最适分化培养基为MS 6-BA 2.0 mg/L 2,4-D 1 mg/L IAA 0.2 mg/L 蔗糖30 g/L 琼脂粉5 g/L;小苗的最佳生根培养基为MS NAA 0.2 mg/L mA 0.5 m/L 蔗糖30 g/L 琼脂粉5 g,L.40mg/L卡那霉素可以抑制叶盘的分化;20 mg/L卡那霉素可以抑制再生植株的生根.2种抑菌抗生素中,头孢霉素对叶片再生影响较大;而250 mg/L的羧苄青霉素能有效地抑制农杆菌菌株EHA105的生长,却对尾叶桉叶盘的芽分化影响不大,为适宜的抑菌抗生素.3种农杆菌LBA4404,EHA105,GV3101的菌液浸染桉树叶盘,统计其愈伤组织GUS染色率,以EHA105对外植体的浸染能力最强,达83.3%.

Establishment of a Genetic Transformation System in Eucalyptus urophylla U6

Adventitious shoots were induced from leaf explants of "Eucalyptus urophylla U6". An efficient regeneration system was established based on the effects of hormone combinations of different concentrations on callus induction and regeneration of shoots from leaf explants. The results indicated that the optimal medium for leaf explants contained Murashige & Skoog (MS) basal medium supplemented with 2.0 mg/L 6-BA, 1.0 mg/L 24-D and 0.2mg/l IAA. The medium for rooting of shoots was optimized as MS basal medium supplemented with 0.2 mg/L NAA and 0.1 mg/L IBA. Kanamycin at a concentration of 40 mg/L in the medium gave effective inhibition to the regeneration of shoots from the leaf explants. In addition, kanamycin at a concentration of 20 mg/L in the medium inhibited obviously the regeneration of roots from the shoots. For the two anti-microbial antibiotics, cephamycin had a more obvious effect on leaf regeneration, whereas carbenicillin at a concentration of 250 mg/L inhibited the development of the agrobacterium strain EHA105 and affected bud differentiation slightly. Carbenicillin is hence considered to be an appropriate antimicrobial antibiotic.