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    板栗胚珠培养研究初报

    Ovule culture in vitro of Castanea mollissima Bl

    • 摘要: 以板栗胚珠为外植体,筛选了板栗胚珠培养适宜的外植体采摘时期、生长调节剂组合、培养基中碳源种类、培养基中糖浓度等指标,并深入探讨了板栗胚珠培养中存在的急需解决的技术问题,为板栗体细胞胚胎高频发生体系的建立奠定了基础.结果表明:①在板栗胚珠离体培养中,适宜的外植体采摘时间为板栗第1次盛花期后的30~44 d;②单独使用0.5或1.0 mg/L的2,4-D、6-BA、TDZ均不能诱导板栗胚珠的体细胞胚胎发生;③不同种类的细胞分裂素与2,4-D配合使用时,其种类和浓度直接影响出胚率,6-BA与2,4-D配合使用的效果明显优于KT、TDZ、ZT分别与2,4-D的组合,6.0 mg/L的2,4-D和0.5 mg/L的6-BA配合使用,有利于诱导胚珠的体细胞胚胎发生;④板栗胚珠培养中,胚性愈伤组织的发生与2,4-D浓度、细胞分裂素种类、细胞分裂素浓度都显著相关,4.0 mg/L的2,4-D和1.0 mg/L的TDZ配合使用,利于诱导胚性愈伤组织的发生;⑤使用白砂糖作为培养基中的碳源最好,其效果明显优于蔗糖、葡萄糖、麦芽糖;⑥培养基中白砂糖浓度为10~40 g/L时,都能诱导板栗胚珠的体细胞胚胎发生,但外植体死亡率呈现出随糖浓度增加而升高的趋势.

       

      Abstract: The ovules of Castanea mollissima Bl.were used as explants in this experiment to test the main active factors when cultured in vitro.These factors included hormone combinations,the collecting date to get ovules as well as the types and concentrations of carbohydrate source in the basic medium.The rush techniques used to overcome the difficulties existing in ovule culture in vitro of C.mollissima were also discussed in this paper.Results of the experiment indicated that:1) The optimal period to collect the bars for getting fitting ovules was about 30 to 44 days after the first full-blown florescence of C.mollissima in Huairou Zone of Beijing.2) When maintained C.mollissima's ovules in vitro,adding 2,4-D,6-BA and TDZ individually at the concentration level of 0.5 mg/L or 1.0 mg/L to WPM medium can not induce embryogenesis.3) While using cytokinins together with 2,4-D to basic medium,the variety and concentration of cytokinin in the medium affected embryogensis frequency greatly.The effects of adding 6-BA and 2,4-D simultaneously to the medium were pretty well than adding any one of KT,TDZ or ZT together with 2,4-D to the medium.And the analytic result also showed that adding 6.0 mg/L 2,4-D accompany with 0.5 mg/L 6-BA to the medium was effective to induce somatic embryos of C.mollissima.4) The concentration of 2,4-D contained in the medium,the sort and the concentration of cytokinins contained in the medium were all significant correlated with the frequency of embryonic callus induction.Adding 4.0 mg/L 2,4-D and 1.0 mg/L TDZ to the medium simultaneously was efficiently to induce embryonic callus in ovule culture in vitro of C.mollissima.5) White granulated sugar bought from food store was the best ingredient acting as the carbohydrate source when culturing ovules of C.mollissima in vitro and it was better than sucrose,glucose or maltose.6) 10-40 g/L of white granulated sugar contained in the medium all can induce embryogenesis of C.mollissima,but the death rate of the explants was rising with increasing the concentration of sugar.

       

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