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园艺学报 ›› 2011, Vol. 38 ›› Issue (09): 1781-1790.

• 新技术与新方法 • 上一篇    下一篇

流式细胞术细胞核分离缓冲液的改良及大、小果型荔枝幼果和果皮细胞分裂活性比较

李彩琴,王泽槐,徐咏珊,张劲霭,李建国*   

  1. (华南农业大学中国荔枝研究中心,广州 510642)
  • 收稿日期:2011-01-10 修回日期:2011-08-12 出版日期:2011-09-25 发布日期:2011-09-25
  • 通讯作者: 李建国

Improvement of the Cell Nuclei Suspensions for Flow Cytometry and Comparison on the Cell Division Activity of Litchi Fruitlet and Pericarp Which Having Different Final Fruit Size

LI Cai-qin,WANG Ze-huai,XU Yong-shan,ZHANG Jin-ai,and LI Jian-guo*   

  1. (China Litchi Research Center,South China Agricultural University,Guangzhou 510642,China)
  • Received:2011-01-10 Revised:2011-08-12 Online:2011-09-25 Published:2011-09-25
  • Contact: LI Jian-guo

摘要: 以荔枝幼果为材料,比较5种常用细胞核分离缓冲液对制备样品DNA分辨率效果的影响,发现在Otto缓冲液基础上进行适当改进(增加还原剂种类和含量,把RNaseA直接加入缓冲液,降低离心力),可获得最佳的提取效果,分辨率高,细胞G1/G0峰产生的变异系数低。通过比较具有相似遗传背景的‘妃子笑’早花大果和晚花小果以及具有不同遗传背景的大果(‘四两果’)和小果(‘陈紫’)型荔枝品种之间果实发育前期的幼果和果皮的细胞分裂活性发现,大果的细胞分裂活性高于小果,且持续时间长,说明果实大小差异与果实发育前期细胞分裂活性有关。

关键词: 荔枝, 细胞分裂活性, 流式细胞术, 果实大小, 核悬液

Abstract: The effects of the five commonly used buffers on DNA nuclei isolation from fruitlet of litchi were evaluated. The results showed that improved method on Otto buffers(adding the content and type of reducing agent into Otto buffers,putting RNase A directly into the buffer and reducing the centrifugal force)could get the highest resolution and the lowest coefficient of variation G1/G0 peak of DNA nucleus,which was suitable for detecting the cell division activity of litchi fruitlet and pericarp. By comparing with cell division activities of fruitlet and pericarp in the first stage of fruit development between larged-fruit and smalled-fruit from the early and late flowering flowers of‘Feizixiao’litchi with the similar genetic background,and from‘Siliangguo’and‘Chenzi’litchi with the different genetic background respectively,it was found that the cell division activity of larged-fruit was higher and lasted longer compared with the smalled fruit,indicating that their differences in terms of the final fruit size was related to the cell division activity in early fruit development.

Key words: litchi, cell division activity, flow cytometry, fruit size, nuclei suspension

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