铁线莲‘水晶喷泉’茎段组培再生体系的建立

Establishment of a Tissue Culture Regeneration System for Clematis Crystal Fountain'Fairy Blue'Segments

本研究以铁线莲品种‘水晶喷泉’(Clematis Crystal Fountain’Fairy Blue’)的带芽茎段为初始外植体,通过研究灭菌时间、激素浓度配比、基础培养基类型等因素对茎段腋芽的诱导、增殖、再生苗生根的影响,从而建立该品种从外植体—腋芽诱导—不定芽增殖—生根得到完整的植株再生体系。结果表明:最佳灭菌条件为5%次氯酸钠溶液灭菌10 min,污染率为22.2%,成活萌芽率为44.5%;带芽茎段诱导腋芽最适培养基为MS+6-BA 1.0 mg/L+NAA 0.05 mg/L+蔗糖30 g/L pH值为6.0~6.2,6-BA/NAA比值为20,诱导率为100%,此时苗生长良好且株高较高;腋芽增殖最适培养基为DKW+6-BA 2.0 mg/L+NAA 0.05 mg/L+蔗糖30 g/L pH值为6.0~6.2,6-BA/NAA比值为40,增殖系数为5.6;最佳生根培养基为DKW+IBA 1.0 mg/L+蔗糖30 g/L pH值为5.7,生根率可达94.4%,本研究可为铁线莲品种‘水晶喷泉’的组织培养、规模化种植提供相关资料和遗传转化体系建立提供了科学依据。

In this study,the budding stem segment of Clematis Crystal Fountain'Fairy Blue'was used as the initial explant.The effects of sterilization time,hormone concentration ratio,basic medium type and other factors on axillary bud induction,proliferation and rooting of regenerated seedlings were studied,so as to establish a complete plant regeneration system from explants-axillary bud induction-adventitious bud proliferation-rooting.The results showed that the optimal sterilization condition was 10 min for 5%sodium hypochlorite solution,the pollution rate was 22.2%,the incubation rate was 44.5%,and the optimal medium for inducing axillary buds with budding stem segments was MS+6-BA 1.0 mg/L+NAA 0.05 mg/L+Sucrose 30 g/L p H=6.0~6.2,6-BA/NAA ratio was 20,and the induction rate was 100%.Seedlings grow well and have higher plant height.The optimal medium for axillary bud proliferation was DKW+6-BA 2.0 mg/L+NAA 0.05 mg/L+Sucrose 30 g/L pH=6.0~6.2,6-BA/NAA ratio was40,with a proliferation coefficient of 5.6,and the optimal rooting medium was DKW+IBA 1.0 mg/L+Sucrose30 g/L pH=5.7,with a rooting rate of 87%.This study provides a scientific basis for tissue culture and large-scale planting of Clematis Crystal Fountain'Fairy Blue'.