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为了优化史密斯桉愈伤组织诱导与分化系统,为史密斯桉的无性繁殖和转基因研究提供参考。通过建立无菌苗,研究不同品种、外植体部位、激素种类及其配比、不同质量浓度2,4-D与NAA对史密斯桉愈伤组织诱导的影响,激素种类及配比、基本培养基种类、6-BA质量浓度、NAA质量浓度、不同转种周期对史密斯桉愈伤组织分化成芽的影响以及激素种类及其质量浓度对史密斯桉不定芽分化成根的影响。结果表明,使用1.54 mol/L NaClO对史密斯桉种子消毒30~50 min建立的无菌苗效果较好;史密斯桉形成愈伤组织的能力在品种间有显著差异,品种9的愈伤形成率最高;不同外植体部位形成愈伤组织的能力从大到小依次为:子叶、真叶、下胚轴、来源于种子的幼茎、种子、老树幼茎、根;0.5~1.0 mg/L 2,4-D对史密斯桉愈伤组织的形成是必要的,1.0 mg/L 2,4-D+1.0 mg/L NAA组合对史密斯桉愈伤组织诱导效果最好,愈伤形成率最高;NAA与6-BA的质量比为1∶1时,有利于史密斯桉愈伤组织的形成,质量比为1∶5时有利于茎段的快速生长,质量比为1∶10时有利于不定芽的形成;单独使用0.5~1.0mg/L 2,4-D或2,4-D与NAA组合有利于愈伤组织的形成;单独使用0.5 mg/L的NAA或IBA有利于史密斯桉根的形成,其中NAA的效果较好;在MS培养基中苗高、苗的长势及腋芽萌发成苗的能力均较其他培养基有显著优势;转种20 d的愈伤分化成芽率相对较高。说明史密斯桉愈伤组织诱导与分化系统易于优化。
In order to optimize the callus induction and differentiation system of E. fortunei, it provided a reference for the research on asexual reproduction and transgenic study of E. fortunei. The effects of different concentrations of 2,4-D and NAA on callus induction, hormone type and proportion, basic culture 6-BA, NAA and different transplanting cycles on the callus differentiation of E. fortunei and their effects on the differentiation of Smith’s eucalyptus into adventitious buds. The results showed that the use of 1.54 mol / L NaClO disinfected 30 ~ 50 min of E. fortunei seed germination established a better effect; ability to form callus of E. eucalyptus significant differences in varieties, the highest rate of callus 9 varieties The ability of different explants to form callus in descending order was as follows: cotyledon, true leaf, hypocotyl, young stem from seed, seed, young stem and root of old tree; 0.5-1.0 mg / L 2,4-D was necessary for callus formation of E. fortunei. The combination of 1.0 mg / L 2,4-D + 1.0 mg / L NAA had the best induction effect on callus of E. fortunei with the highest callus formation rate. The mass ratio of NAA to 6-BA was 1: 1, which was in favor of the formation of callus of E. fortunei. The mass ratio of NAA to 6-BA was beneficial to the rapid growth of stem segments at the mass ratio of 1:10, . The combination of 0.5-1.0 mg / L 2,4-D or 2,4-D with NAA alone was beneficial to the formation of callus. The use of 0.5 mg / L of NAA or IBA alone was beneficial to the formation of S. eucalyptus , Of which NAA had a better effect. In MS medium, seedling height, seedling growth and axillary bud germination ability were significantly higher than those in other media. The callus transplanted for 20 days Into germination rate is relatively high. Explain the Smithii callus induction and differentiation system easy to optimize.