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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (2): 60-63.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

柚单染色体微分离制片技术研究

王平1,2,乔锋1,3,王发明1,4,柳燕挣1,4,唐琦1,4   

  • 收稿日期:2009-09-16 修回日期:2009-09-29 出版日期:2010-01-20 发布日期:2010-01-20
  • 基金资助:

    柚单染色体AFLP分子标记以及文库的构建;柚单染色体微分离和微克隆研究

Study on single chromosome microdissection technique for pomelo（Citrus grandis）

  • Received:2009-09-16 Revised:2009-09-29 Online:2010-01-20 Published:2010-01-20

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摘要：

以‘琯溪蜜柚’幼胚为材料，对单染色体微分离制片过程中酶解时间、冰水固缩时间和染色三因素的影响效果进行了研究。结果表明：用2%纤维素酶与0.5%果胶酶的混合液处理40～60 min，后低渗处理之后进行20～30 min冰水固缩处理和卡宝品红染色组合可以得到理想的制片效果。该方法制片背景干净，各单染色体充分分散，适合下一步的小型单染色体微分离研究

Abstract:

With immature embryos of Citrus grandis‘Guanximiyou’ as material, the effect of the three factors, enzyme treating time, pycnosis treating time in ice water and dyeing, were studied in the process of single chromosome microdissection technique. The resluts indicated that the ideal effects had been proved with enzymatic hydrolysis of mixing 2% cellulose enzyme and 0.5% pectic enzyme for 40～60 min, then pycnosis treatment in ice water for 20～30 min after post-hypotonic treatment, and carbol fuchsin solution dyeing. Using this technique, the chromosome images were clear with satisfying background, each chromosome separation and extension adequately, it is suitable for next research for single small chromosome microdissection.