棉 花 学 报    Cotton Science    2008，20(2)：94-98

Preparation of De-exined Pollen Grains and Observation of Pollen Grains Nuclear DNA in Gossypium hirsutum L.
YUE Jie-yu¹，WU Li-jun²，WU Yue-jin²，ZHANG Bao-long³，TANG Can-ming^1,**
(1.Agronomy College，Nanjing Agricultural University，Nanjing 210095，China;2.Institute of Plasma Physics ，Chinese Academy of Science，Hefei 230031，China；3.Genetics and Physiology Institute，Jiangsu Academy of Agriculture Science，Nanjing 210014，China)

Abstract：The objective of this study was demonstrating nuclei DNA in the pollen grains of upland cotton ( Gossypium hirsutum cv . Sumian 22 ). The procedures of detaching the exine and observing nuclei DNA of pollen grains were established. The pollen grains were fixed with 4% paraformaldehyde solution，oxidated with 10% sodium hypochlorite solution, heat shocked at 55 ℃ for 30 minutes，then the slide with pollens solution was pressed by hand. Finally, the exine of pollen grains could be detached. The pollen grains and de-exined pollen grains were stained with DAPI and observed with fluorescence microscope and laser scanning confocal microscope (Leica TCS SP2), respectively. The nucleus of cotton pollen grain could not be observed, but the nuclear DNA of de-exined pollen grain could be observed clearly. The inner wall fluorescence of pollen grain was not found. The three nuclei of the pollen grain of upland cotton cultivar Sumian 22 were observed in the experiment.
Key words：upland cotton；Gossypium hirsutum L.；pollen grain；de-exined pollen grain；DAPI
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