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浙江大学学报（农业与生命科学版）
Journal of Zhejiang University: Agric. & Life Sci.  2010, Vol. 36 Issue (5): 497-502    DOI: 10.3785/j.issn.1008-9209.2010.05.004
Biological sciences & biotechnology     
Microfilament cytoskeletons in BY-2 protoplasts and broad bean mesophyll protoplasts and the effect of latrunculin B on their distribution
YE Lu-fei1,LIU Cheng-ke1,MENG Chun-mei1,2,HONG Jian1,2
1.Institute of Biotechnology,Zhejiang University,Hangzhou 310029,China; 2.Center of Analysis and Measurement,Zhejiang University,Hangzhou310029,China
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Abstract  Protoplasts of tobacco suspension cultured BY-2 cells and broad bean (Vicia faba) mesophyll cells were prepared by enzyme digestion . The microfilament cytoskeletons in protoplasts were labelled by TRITC-phalloidin staining . Laser scanning confocal microscopy (LSCM) revealed delicate microfilament networks which evenly distributed in the protoplasts . LSCM also recorded the dynamic process of microfilament depolymerization and distribution after respective treatment with 10 and 20μm ol.L-1 latrunculin B . This study of protoplast preparation,microfilament staining,latrunculin B treatment and LSCM sets the experimental foundations for further studies of microfilament cytoskeleton in plant cells .

Published: 13 September 2010
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YE Lu-fei1
LIU Cheng-ke1
MENG Chun-mei1
2
HONG Jian1
2
Cite this article:

YE Lu-fei1,LIU Cheng-ke1,MENG Chun-mei1,2,HONG Jian1,2. Microfilament cytoskeletons in BY-2 protoplasts and broad bean mesophyll protoplasts and the effect of latrunculin B on their distribution. Journal of Zhejiang University: Agric. & Life Sci., 2010, 36(5): 497-502.

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http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2010.05.004     OR     http://www.zjujournals.com/agr/Y2010/V36/I5/497


烟草 BY-2细胞和蚕豆叶肉细胞原生质体微丝骨架及latrunculin B处理对其分布的影响

用酶解法分别制备烟草BY-2悬浮培养细胞和蚕豆叶肉细胞的原生质体,应用激光共聚焦显微镜 观察经 TRITC-鬼笔环肽荧光染色的2种原生质体的微丝骨架空间分布,以及观察用10和20μm ol.L-1微 丝抑制剂 latrunculin B分别处理2种原生质体后对其分布的影响 。结果表明 :激光共聚焦显微镜清楚地显 示出2种细胞原生质体中微丝精细排列的均匀网络结构 ;共聚焦显微成像显示了随 latrunculin B处理时 间的延长微丝解聚和微丝骨架分布的动态过程 。建立原生质体制备、微丝骨架荧光染色、latrunculin B处理 及激光共聚焦显微观察的实验体系为进一步研究植物微丝骨架的功能奠定了基础 。
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