茄子小孢子再生体系的优化

Optimization of Microspore Regeneration System in Eggplant

【目的】开展影响茄子(Solanum melongena)小孢子愈伤组织诱导、生长和分化的研究,为茄子小孢子再生体系的建立及应用提供技术支持。【方法】以二倍体茄子游离小孢子为材料,探讨培养方式、诱导条件、培养基配方等若干因素对茄子小孢子愈伤组织诱导、生长和分化的影响,初步建立茄子小孢子培养体系。【结果】黑暗静止培养条件下采用2~3次更换部分新鲜培养基利于愈伤组织诱导和生长;愈伤组织分化培养时,愈伤组织直接转接到固体培养基时其成活率与大小有关,>2 mm时其成活率可达82.67%以上,固液共培对愈伤组织大小要求不严格,添加1~2 mL N3液体培养基时有利于愈伤组织生长和成活,液体培养基>2 mL会降低愈伤组织分化培养成活率;在添加36 g/L甘露醇的MS固体培养基培养的愈伤组织转绿(成熟)率最高,达94.64%;ZT 1.5 mg/L+NAA 0.05 mg/L 激素组合有利于芽分化,分化率达57.78%。【结论】黑暗静止培养时,定时更换部分培养基利于愈伤组织诱导和生长,添加1~2 mL N3液体培养基固液共培养时愈伤组织成活率最高,生长快,固体培养基附加 36 g/L甘露醇和ZT 1.5 mg/L+NAA 0.05 mg/L 激素组合有利于愈伤组织分化。

【Objective】The technology of microspores callus induction,growth and differentiation of eggplant (Solanum melongena) were studied to provide technical support for the establishment and application of the microspore regeneration system of eggplant.【Method】With the free microspores of diploid eggplant as materials,the effects of induction conditions,culture methods and culture medium types on the callus induction,growth and differentiation of eggplant microspores were explored,and the microspore culture system of eggplant was established.【Result】Under the dark static culture conditions,and replacing with some fresh culture medium for 2-3 times were beneficial to callus induction and growth.Upon the differentiation culture of callus,the survival rate of callus was related to its size when the callus was directly transferred to solid medium.When the callus was more than 2 mm,the survival rate could reach 82.67%.The requirement of solid-liquid culture on callus size was not strict,adding 1-2 mL N3 liquid medium was conducive to the growth and survival of callus,while the survival rate of differentiation culture of callus would be reduced when the liquid medium was over 2 mL.The highest greening rate of callus(maturity)was 94.64% in MS solid medium with 36 g/L mannitol.The hormone combination of ZT 1.5 mg/L and NAA 0.05 mg/L was beneficial to bud differentiation,and the differentiation rate was 57.78%.【Conclusion】Under the dark still culture,changing part of medium regularly was beneficial to callus induction and growth.When 1-2 mL N3 liquid medium was added into solid medium,the survival rate of callus was the highest and the growth rate was fast.The hormone combination of ZT 1.5 mg/L + NAA 0.05 mg/L with 36 g/L mannitol on the solid medium was conducive to callus differentiation.