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目的了解激素对杜仲不同发育程度完整未成熟种胚的影响。方法以杜仲三个不同发育时期的完整未成熟种胚为实验材料(分别标记为I#:心形胚早期,胚长约0.5 mm;II#:鱼雷形胚,胚长约1~2mm;III#:胚长约5mm,约为成熟胚的1/2),以MS为基本培养基,添加不同激素及组合(包括2,4-D、NAA、6-BA、TDZ、GA3)进行离体培养,观察不同激素对完整幼胚的影响。结果实验结果显示,I#材料接种后15天几乎全部死亡,显示简单的无机盐培养基不能满足发育早期种胚离体培养的营养需求;III#材料接种后生长迅速,在附加细胞分裂素的培养基中多数萌发形成了幼苗;II#材料在培养30天及60天进行结果统计显示:1 2,4-D促进疏松愈伤组织形成,抑制不定芽形成;2细胞分裂素类,包括6-BA、TDZ,均能诱导形成不定芽,培养60天诱导率显著增加;在6-BA 2mg/L中不定芽发生率达91%;3在附加0.5mg/L 2,4-D的培养基中,在培养约90天后40个外植体中有3个形成体细胞胚。结论 II#鱼雷形胚对激素的反应与杜仲成熟胚或幼苗离体培养结果相近,该发育状态的幼胚可能适合于诱导胚状体。
Objective To understand the effects of hormones on intact embryos of different developmental stages of Eucommia ulmoides. Methods The intact immature embryos of three different developmental stages of Eucommia ulmoides were used as experimental materials (labeled as I #: embryo length about 0.5 mm at the early stage of heart embryo, II #: torpedo embryo with embryo length of 1 ~ 2 mm, III #: Embryo length of about 5mm, about half of the mature embryo), with MS as the basic medium, add different hormones and combinations (including 2,4-D, NAA, 6-BA, TDZ, GA3) Cultivation, observation of different hormones on the integrity of immature embryos. Results The experimental results showed that almost all of the I # materials died within 15 days after inoculation, indicating that the simple inorganic salt medium could not meet the nutritional requirements of the in vitro culture of early embryo development. The III # material grew rapidly after inoculation, The results showed that: 1 2, 4-D promoted the formation of loose callus, inhibited the formation of adventitious buds; 2 cytokinins, including 6 -BA and TDZ could induce the formation of adventitious buds, and the induction rate was significantly increased after 60 days of culture. The incidence of adventitious buds in 6-BA 2mg / L was 91% .3 In the culture supplemented with 0.5mg / L 2,4-D In the base, 3 out of 40 explants formed somatic embryos after about 90 days of culture. Conclusion The response of II # torpedo-shaped embryos to hormones is similar to that of mature embryos or seedlings of Eucommia ulmoides subtilis. The embryos of this developmental state may be suitable for inducing embryoid bodies.