油茶胚囊观察体系优化及三维结构重建

高超1,2, 袁德义2,*
1贵州大学, 贵州省森林资源与环境研究中心, 贵阳550025; 2中南林业科技大学, 经济林培育与保护教育部重点实验室, 经济林育种与栽培国家林业局重点实验室, 长沙410004

通信作者:袁德义;E-mail: yuan-deyi@163.com

摘 要:

筛选并利用优化的染色和透明实验方法, 结合激光共聚焦扫描显微镜观察油茶成熟胚囊内部结构。结果表明, 采用1%曙红染色1 h并用水杨酸甲酯透明5 d是观察油茶成熟胚囊较好的实验处理方案, 胚囊内细胞轮廓清晰, 对所需观察的目标有很好的表现力。基于该优化体系, 构建了油茶胚囊的三维结构, 为油茶及山茶属植物胚囊发育、受精机理等方面的研究提供了更加快速、高效的观察方法。

关键词:油茶; 胚囊; 三维结构; 激光共聚焦扫描显微镜

收稿:2017-02-13   修定:2017-04-19

资助:贵州省森林培育及生态建设重点学科人才基地建设项目(黔人领发[2009]9号)、贵州省高层次创新型人才培养“十层次”人才项目(黔科合人才[2015]4003号)、提高马尾松用材林林分质量关键技术研究与示范(黔林科合[2010]03)和国家自然科学基金(31170639)。

Observation system optimization and three-dimentional reconstruction of embryo sac in Camellia oleifera

GAO Chao1,2, YUAN De-Yi2,*
1Institute for Forest Resources & Environment of Guizhou, Guizhou University, Guiyang 550025, China; 2Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees of Ministry of Education, the Key Laboratory of Non-Wood Forest Products of Forestry Ministry, Central South University of Forestry and Technology, Changsha 410004, China

Corresponding author: YUAN De-Yi; E-mail: yuan-deyi@163.com

Abstract:

The internal structure of mature embryo sac in Camellia oleifera was observed using confocal laser scanning microscope, based on choosing and utilizing the optimized staining and clearing experimental methods. The experiment methods, the samples were stained with 1% eosin solution for 1 h and treated with methyl salicylate transparent material for 5 d for embryo sac observation, was better than others. The embryo sac clearly showde cell configuration. According to the optimized experimental methods, the three-dimentional structure of C. oleifera embryo sac was reconstructed for the first time. The results provided an effective method for researching the development of embryo sac and the fertilization mechanism in C. oleifera.

Key words: Camellia oleifera; embryo sac; three-dimentional structure; confocal laser scanning microscope

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