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[用荧光染料Hoechst 33342对朱顶红发育早期花粉染色质状态的研究]

[Investigation of the pollen chromatin state of Hippeastrum hibridum at the early stages of development with fluorescent dye Hoechst 33342].

作者信息

Mel'nikova E V, Gordon R Ia, Roshchina V V

机构信息

Institute of Cell Biophysic RAS, Pushclino, Moscow District.

出版信息

Tsitologiia. 2002;44(4):400-3.

Abstract

The work was carried out on the pollen of Hippeastrum hibridum, whose size (ca 55 mcm) permits to follow both the pollen tube formation and development of the vegetative nucleus and generative cell within the first two hours before the growing point arising. Using fluorescent dye Hoechst 33342 chromatin state alterations accompanying changes in the pollen physiological state were investigated. The maximum fluorescent intensity was observed in 30 min of staining and reflected the maximum chromatin functional activity. Histograms of pollen distribution according to fluorescent intensity differ considerably with doses of irradiation (500, 1000, 2000, 3000 and 4000 Gy). Besides, a germinative index of fluorescence intensity was calculated. A comparative analysis of these data has shown that the lowest decrease in fluorescent intensity observed at 500, 3000 and 4000 Gy, was accompanied by different germinative indices. At 500 Gy, the index was 2.5 times lower than in the control, but at 4000 G no germination was not noted. The process of pollen grain development is supposed to be more intensive and faster at 500 Gy, than in the control. At 4000 Gy, a decrease in the functional pollen activity is accompanied by decrease or inhibition of chromatin functional activity.

摘要

该研究以朱顶红的花粉为材料,其大小(约55微米)使得在生长点出现前的头两个小时内,既能追踪花粉管的形成,又能观察营养核和生殖细胞的发育。使用荧光染料Hoechst 33342研究了伴随花粉生理状态变化的染色质状态改变。染色30分钟时观察到最大荧光强度,这反映了染色质的最大功能活性。根据荧光强度绘制的花粉分布图因辐照剂量(500、1000、2000、3000和4000戈瑞)的不同而有很大差异。此外,还计算了荧光强度的发芽指数。对这些数据的比较分析表明,在500、3000和4000戈瑞时观察到的荧光强度最低降幅,伴随着不同的发芽指数。在500戈瑞时,该指数比对照低2.5倍,但在4000戈瑞时未观察到发芽现象。500戈瑞时花粉粒的发育过程被认为比对照更强烈、更快。在4000戈瑞时,功能性花粉活性的降低伴随着染色质功能活性的降低或抑制。

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