柳杉优良无性系组培快繁体系研究

Study on rapid propagation system of superior clones in Cryptomeria fortunei

【目的】柳杉Cryptomeria fortunei外植体丛芽分化能力强而芽伸长不足。为探索分段培养过程中的柳杉不定芽诱导与增殖、伸长培养及生根诱导的最佳培养基,建立柳杉分段培养的高效组培快繁体系。【方法】以20年生柳杉优良无性系的当年生带芽茎段为外植体,采用正交试验设计分段培养的组培体系。柳杉外植体分段培养再生体系主要包括丛生芽诱导、伸长、生根及移栽等步骤。【结果】①在3种基本培养基(MS、DCR和WPM)中分别添加不同质量浓度6-苄氨基腺嘌呤(6-BA)、吲哚丁酸(IBA)及水解酪蛋白(CH),3个柳杉无性系均能诱导出不定芽,但诱导率存在显著差异,以WPM+1.00 mg·L^-1 6-BA+0.10 mg·L^-1IBA中不定芽诱导率(100%)和增殖系数(9.13)最高。②高质量浓度激素培养基与低质量浓度激素培养基交替分段培养方式对柳杉无性系进行继代增殖培养,效果良好,有效枝条增长率可达到456.87%。③柳杉无性系离体生根的最适宜培养基为DCR+0.10 mg·L^-1 IBA,且3个无性系生根率最高可达100%。④生根的无菌苗移栽至V(泥炭)∶V(蛭石)=1∶1的混合基质,经过15~20 d炼苗,成活率高达96.70%,繁殖潜能达3 865株·母株^-1·a^-1。【结论】柳杉分段培养的组培模式极大提高了柳杉组培再生效率,有助于柳杉优良品种的工业化育苗。图1表5参23。

[Objective]The induced clustered buds of Cryptomeria fortunei have strong differentiative capacity but insufficient elongation ability.In order to obtain the optimum medium of adventitious bud induction,proliferation,elongation culture and rooting induction,the rapid propagation system of C.fortunei was established.[Method]The annual shoot from 20-year-old C.fortunei were used as explants;the orthogonal test design was used to select the optimum medium.The propagation system of C.fortunei includes induction,proliferation,elongation,rooting and transplant.[Result](1)Adventitious buds from three clones could be induced on MS,DCR and WPM basic medium supplemented with different combinations of 6-BA,IBA and CH.But the proliferation ability of buds varied significantly among different mediums.When clones were cultured in WPM medium with 1.0 mg·L^-1 6-BA and 0.1 mg·L^-1 IBA,the induction rate and proliferation coefficient was the highest,which was 100%and 9.13,respectively.(2)When three clones of C.fortunei were subcultured alternately with high concentration hormone and low concentration hormone,the effective shoot growth rate reached 456.87%.(3)The most suitable medium for the C.fortunei rooting was DCR medium with0.1 mg·L^-1 IBA,under which the rooting rate ratio was 100%.(4)The rooted seedlings were transplanted to a mixed matrix with equal vermiculite and peat,after 15-20 days of seedling hardening,and the survival rate was 96.70%.Therefore,the potential propagation ability is 3865 seedlings per maternal plant per year.[Conclusion]Thus,an efficient propagation technology for C.fortunei was established,which laid a technical foundation for industrialization of improved varieties.[Ch,1 fig.5 tab.23 ref.]