野生二倍体草莓子叶的再生体系建立

Establishment of Cytoledon Regeneration System of Yellow Wonder,a Wild Diploid Strawberry Variety

【目的】建立野生草莓(Fragaria vesca.)Yellow Wonder子叶的高效再生体系,为其遗传转化研究与应用奠定基础。【方法】以野生草莓Yellow Wonder子叶为材料,研究不同激素配比、不同暗培养时间及不同子叶部位的不定芽诱导情况;同时研究不同IBA浓度下不定根的生长情况。【结果】在无菌条件下,草莓种子在添加3%蔗糖的1/2 MS培养基中发芽率可达90%;最佳激素配比为3.4 mg/L 6-BA+0.3 mg/L IBA,出愈率和分化率分别为93.33%和83.33%;最佳暗培养时间为7 d;IBA浓度为0.2 mg/L时,生根率可达100%;诱导不定芽效果最佳的部位为子叶中部。【结论】建立野生草莓Yellow Wonder子叶的高效再生体系,即将Yellow Wonder草莓种子在萌发培养基(1/2 MS+3%蔗糖)中培养至发芽;将子叶中部切下,正面朝上置于子叶再生培养基(6-BA 3.4 mg/L+IBA 0.3 mg/L)上;在室温条件下暗处理7 d后,转入(25±2)℃长日照条件下培养40 d诱导出不定芽;不定芽转接到继代培养基(MS+6-BA 0.5 mg/L+IBA 0.1 mg/L)上持续生长,待芽苗伸长至1~2 cm后,再转接到生根培养基(MS+IBA 0.2 mg/L)中进行生根培养。

【Objective】The cytoledon high-efficiency regeneration system of Yellow Wonder,a wild diploid strawberry variety,is established to lay the foundation for research and application of its genetic transformation.【Method】The effect of different hormones ratios,dark culture time and cytoledon parts on adventitious bud induction of strawberry cytoledon and the effect of different IBA concentrations on growth of adventitious roots are studied.【Result】The germination rate of strawberry seeds cultured on 1/2 MS + 3% sucrose can reach 90%.The callus induction and differentiation rate of strawberry cytoledon is 93.33% and 83.33% under the optimal hormones ratio of 3.4 mg/L 6-BA +0.3 mg/L IBA respectively.The optimal dark culture time is 7 d.The rooting rate of adventitious buds can reach 100% when IBA concentration is 0.2 mg/L.The middle part of cytoledon is the optimal explant for induction of adventitious buds.【Conclusion】The cytoledon high-efficiency regeneration system of Yellow Wonder is successfully established.The cytoledon high-efficiency regeneration system includes culturing seeds on the germination medium(1/2 MS + 3% sucrose) for the germination,culturing the middle part of cytoledon right side up on cytoledon regeneration medium(6-BA 3.4 mg/L+IBA 0.3 mg/L)for 7 d dark culture at indoor temperature and then for 40 d long-day sunshine at(25±2)℃ for inducing adventitious buds.Culturing the adventitious buds on the subculture medium(MS+6-BA 0.5 mg/L+IBA 0.1 mg/L)for growth of adventitious buds and culturing the adventitious buds with 1~2 cm height on the rooting medium(MS+ 0.2 mg/L IBA)for rooting of adventitious buds.