https://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
https://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
https://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
https://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2015, Vol. 42 ›› Issue (6): 1167-1174.doi: 10.16420/j.issn.0513-353x.2015-0071

• 研究报告 • 上一篇    下一篇

‘富士’苹果花粉原生质体分离初探

张宁,李威,顾钊宇,陈秋菊,段续伟,杨清,李天忠   

  1. 中国农业大学农学与生物技术学院,北京 100193
  • 出版日期:2015-06-25 发布日期:2015-06-25
  • 基金资助:
    国家自然科学基金项目(31171941,31372035)

Preliminary Study on the Isolation of Mature Pollen Protoplasts in‘Fuji’Apple

ZHANG Ning, LI Wei, GU Zhao-Yu, CHEN Qiu-Ju, DUAN Xu-Wei, YANG Qing, LI Tian-Zhong   

  1. College of Agriculture and Biotechnology,China Agricultural University,Beijing 100193,China
  • Online:2015-06-25 Published:2015-06-25

摘要: 以‘富士’苹果(Malus × domestica Borkh.‘Fuji’)成熟花粉为试材,采用“萌发—酶解二步法”,初步探讨了影响原生质体分离的关键因子,获得了花粉原生质体分离的最佳条件:用萌发后45 min的花粉,转入含1%纤维素酶和1%离析酶的混合酶液中,以18%甘露醇调节渗透压,静置酶解6 h,花粉原生质体分离效率可达6.83%,用0.1%荧光增白剂检测表明脱壁完全,用0.01% FDA检测表明其具有生活力,可以作为后续细胞融合等的材料。

关键词: 苹果, 花粉, 原生质体, 分离

Abstract: Little information is available about the research on plant pollen protoplasts,especially the isolation of apple pollen protoplasts. In this study,the protoplasts from mature pollen of Malus × domestica Borkh.‘Fuji’by the method of‘germination-enzymatic treatment’were isolated. To screen the most effective condition of isolation,some key factors were analyzed. The results indicated that:The isolation percentage was up to 6.83% when the pollens germinating for 45 min and then treated with mixed enzyme solution including 1% Cellulase Onozuka R-10,1% Macerozyme R-10 and 18% Mannitol for 6 h. The cell walls of isolated protoplasts were degraded entirely according to the examination of 0.1% calcoflower white staining. 0.01% fluorescein diacetate(FDA)staining indicated the protoplasts were viable. Therefore,the proroplasts can be used for further research.

Key words: apple, pollen, protoplast, isolation

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