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[目的]探索预处理和培养基类型对小孢子离体培养的影响。[方法]以洋桔梗为材料进行小孢子培养,研究预处理和培养基类型对小孢子离体培养的影响,并筛选胚性愈伤组织适宜诱导培养基、不定芽植株培养基以及最佳生根培养基。[结果]利用4℃低温预处理24 h有利于小孢子愈伤组织的形成。愈伤组织适宜诱导培养基为:MS+3 mg/L KT+2.5 mg/L 2,4-D;不定芽植株培养基为MS+1 mg/L 6-BA+0.2 mg/L NAA;最佳生根培养基为:MS+0.2 mg/L 6-BA+1.0 mg/L NAA+1.5 mg/L IBA+0.3 g/L活性炭。经过染色体倍性鉴定,显示小孢子培养后染色体数目减半。[结论]该方法研究了预处理和培养基类型对小孢子离体培养的影响,为洋桔梗游离小孢子培养体系的建立以及单倍体育种奠定基础。
[Objective] The research aimed to explore the effects of pretreatment and medium types on microspore culture in vitro. [Method] The microspore culture of Eustoma series was used to study the effect of pretreatment and culture medium on microspore culture in vitro. The embryogenic callus induction medium, adventitious bud plant medium and the best rooting Medium. [Result] Pretreatment with 4 ℃ for 24 h was beneficial to the formation of microspore callus. The optimal medium for callus induction was MS + 3 mg / L KT + 2.5 mg / L 2,4-D and MS + 1 mg / L 6-BA + 0.2 mg / L NAA The best rooting medium was MS + 0.2 mg / L 6-BA + 1.0 mg / L NAA + 1.5 mg / L IBA + 0.3 g / L activated carbon. After chromosome ploidy identification, showed that after microspore culture chromosome number halved. [Conclusion] This method studied the effects of pretreatment and culture medium on the microspore culture in vitro, and laid the foundation for establishment of microspore culture system and haploid breeding.