| 多花蔷薇胚性细胞悬浮系原生质体分离及再生植株 |
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| 引用本文： | 陈颖,梁建丽,陈晓丽,郭艳超,田传卫,赵梁军.多花蔷薇胚性细胞悬浮系原生质体分离及再生植株[J].园艺学报,2008,35(10):1491-1496. |
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| 作者姓名： | 陈颖 梁建丽 陈晓丽 郭艳超 田传卫 赵梁军 |
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| 作者单位： | (中国农业大学农学与生物技术学院，北京 100094) |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 利用多花蔷薇‘无刺3号’假珠芽诱导的胚性愈伤细胞悬浮系分离得到原生质体，并培养获得再生植株。酶种类和浓度, 酶解时间及悬浮细胞继代时间对原生质体分离有重要影响，最佳酶液组成是2.0%纤维素酶，0.5%离析酶，5.0 mmol·L-1 MES, 0.5 mol·L-1甘露醇，0.5% CaCl2·2H2O。采用继代3 d的悬浮细胞系，酶解10 h，原生质体产量达到26.67×106g-1，原生质体活力为92.21%。采用液体浅层培养，肌醇比蔗糖更有利于纯化后的原生质体分裂和生长。愈伤组织形成增殖培养基为1/2MS + 2,4-D 1.0 mg·L-1+ EBR 0.2 mg·L-1 + 10 g·L-1 Ficoll + 500 mg·L-1谷氨酰胺 + 20 mg·L-1甘氨酸 + 20 mg·L-1天冬氨酸，分化培养基为1/2MS + 10 mg·L-1 TDZ + 500 mg·L-1谷氨酰胺+ 20 mg·L-1甘氨酸+ 20 mg·L-1天冬氨酸，后转入1/2MS培养基上获得完整植株。
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| 关 键 词： | 多花蔷薇 原生质体 细胞悬浮系 植株再生 |
| 收稿时间： | 2008-6-10 |
| 修稿时间： | 2008-9-20 |
Protoplast Isolation and Plant Regeneration from Somatic Embryogenic Cell Suspension Cultures in Rosa multiflora |
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| CHEN Ying,LIANG Jian-li,CHEN Xiao-li,GUO Yan-chao,TIAN Chuan-wei,ZHAO Liang-jun.Protoplast Isolation and Plant Regeneration from Somatic Embryogenic Cell Suspension Cultures in Rosa multiflora[J].Acta Horticulturae Sinica,2008,35(10):1491-1496. |
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| Authors: | CHEN Ying LIANG Jian-li CHEN Xiao-li GUO Yan-chao TIAN Chuan-wei ZHAO Liang-jun |
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| Institution: | (Agronomy and Biotechnology College, China Agriculture University, Beijing 100094, China) |
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| Abstract: | Protoplasts were successfully isolated from somatic embryogenic cell suspension induced by pseudobulbils, shoots regeneration were achieved. Enzyme sort and concentration, treatment time and the culture time of cell suspensions had influence on protoplasts isolation. Enzyme solution contained 2.0% cellulase, 0.5% macerozyme, 5.0 mmol·L-1 MES, 0.5 mol·L-1mannitol, 0.5% CaCl2·2H2O. Optimum enzyme treatment time was 10 hours and cell suspensions subcultured for 3 days were adopted. The yield and viability were 26.67×106 g-1 and 92.21%. Protoplasts were cultured on shallow liquid layers, where inositol was better than sucrose for promoting cells division and growth. Calli induced and proliferation medium was 1/2MS + 2, 4-D1.0 mg·L-1 + EBR0.2 mg·L-1 + 10 g·L-1 Ficoll + 500 mg·L-1 glutamine + 20 mg·L-1 glycine + 20 mg·L-1 aspartic acid. Differentiation medium was 1/2MS + 10 mg·L-1 TDZ + 500 mg·L-1 glutamine + 20mg·L-1 glycine + 20 mg·L-1 aspartic acid. Shoots regenerated on 1/2MS medium. |
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| Keywords: | Rosa multiflora Rosa multiflora protoplast cell suspension culture shoot regeneration |
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