Application of confocal microscopy to the assessment of pollen development stage in Keteleeria fortunei
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摘要: 油杉为我国特有易危树种.为探究油杉花粉发育过程,其花粉样品经荧光染料曙红和Hoechst33342双染并以冬青油透明后,在激光扫描共聚焦显微镜下观察.结果表明单核小孢子经过连续4次有丝分裂后形成5-细胞型花粉粒,成熟花粉由2个退化的原叶细胞、1个精原细胞(体细胞)、1个不育细胞(柄细胞)和1个管细胞组成.应用激光扫描共聚焦显微术结合荧光染色技术能更清晰地显示出油杉花粉发育过程中的核相变化,观察效果远较常规的醋酸洋红染色法理想.此技术可用于其他花粉发育阶段的鉴定.
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关键词:
- 激光扫描共聚焦显微镜 /
- 油杉 /
- 花粉 /
- 有丝分裂
Abstract: Keteleeria fortunei (Murr.) Carr. is a vulnerable endemic species to China. Using laser scanning confocal microscope(LSCM) technique, this study examined the whole development stages of pollen in K. fortunei. Pollen samples were treated with double fluorescence staining with Eosin and Hoechst33342, followed by methyl salicylate clearing. The results reveal that pollen development consists of several distinct stages. Each uninucleate microspore undergoes four times asymmetric mitotic division (the microspore mitosis) to give rise to a mature pollen grain containing two degenerated prothallial cells, a body cell, a stalk cell, and a vegetative cell. The application of confocal microscopy in conjugation with fluorescence staining produces superior images to the traditional acetocarmine staining of pollen and highlights the nuclei at all stages of pollen development. This technique represents an efficient tool for the assessment of nuclear status of pollen grains. -
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