洋桔梗小孢子培养初步研究

Preliminary Study on Microspore Culture of Eustoma grandiflorum

[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] To explore effects of pretreatment and culture medium types on microspore in vitro culture. [ Method ] With Eustoma grandiflorum as material, effects of pretreatment and culture medium types on microspore culture were studied. The appropriate induction culture medium, adventitious bud plant culture medium and optimum rooting culture medium for callus were obtained. [ Result] The result showed that treating microspore 4℃ for 24 hours benefited the growing of callus. Solid-liquid medium containing MS ＋ 3mg/L KT ＋ 2.5 mg/L 2.4-D was conducive to callus induction, medium containing MS ＋ 1 mg/L 6-BA ＋ 0. 2 mg/L NAA ＋ 30 g/L Sucrose ＋ 6.1 g/L Agar was beneficial to adventitious buds＇ induction, suitable medium for root growth contained MS ＋ 0.2 mg/L 6-BA ＋ 1.0 mg/L NAA ＋ l. 5mg/L IBA ＋ 30 g/L Sucrose ＋ 6.1 g/L Agar ＋ 0.3 g/L Active carbon. Result of the chromosome ploidy identification showed that the chromosome numbers were halved through microspore culture. [ Conclusion] The effects of pretreatment and culture medium types on microspore in vitro culture were studied, which will lay a foundation for establishment of Eustoma grandiflorum microspore culture system and haploid breeding.