应用DNA荧光染料研究木本植物红萼花的减数分裂。

D Beardsell, R B Knox, E Williams
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引用次数: 2

摘要

荧光DNA探针DAPI和Hoechst 33258在小孢子细胞减数分裂和小孢子有丝分裂的各个阶段对木本灌木红萼花(Thryptomene calycina)小染色体的成像优于传统的乙酰胭脂红染色。由于背景荧光减弱,Hoechst 33258略优于DAPI。与dna特异性探针的结合需要含有氯仿的固定剂来去除自身荧光物质,在处理期间用乙酸预处理,pH至少为6。DAPI或Hoechst 33258处理后核仁无荧光。荧光染料对小孢子有丝分裂中期I、中期II和中期的单倍体数量进行了测定。
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Use of DNA fluorochromes for studying meiosis in the woody species Thryptomene calycina.

The fluorescent DNA probes DAPI and Hoechst 33258 produce superior images to the traditional acetocarmine stain of the small chromosomes of the woody shrub Thryptomene calycina at all stages of microsporocyte meiosis and microspore mitosis. Hoechst 33258 was slightly superior to DAPI because of reduced background fluorescence. Binding with the DNA-specific probes required a fixative containing chloroform to remove autofluorescent materials, a pretreatment with acetic acid and a pH of least 6 during treatment. The nucleoli did not fluoresce after treatment with DAPI or Hoechst 33258. Superior resolution of chromosomes after treatment with the fluorochromes enabled easy determination of the haploid number at metaphase I, metaphase II and at metaphase of the microspore mitosis.

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