阳春砂花粉的结构和生殖细胞的分离
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引用本文:林美珍,陈荣珠,赖钟雄,林艺华.阳春砂花粉的结构和生殖细胞的分离[J].西北植物学报,2019,39(11):1998~2002
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作者单位
林美珍1,陈荣珠1,赖钟雄2,林艺华1* (1 漳州卫生职业学院 药学系福建漳州 3630002 福建农林大学 园艺植物生物工程研究所福州 350002) 
基金项目:漳州市自然科学基金(ZZ2019J10);
中文摘要:阳春砂为自交植物,但花的雌蕊高于雄蕊,是研究单子叶植物纲生殖细胞分子生物学的典型材料。为了解决授粉难的问题,该研究采用植物组织化学方法,对阳春砂花粉的发育过程进行观察,以明确其花粉的结构特征;选择渗透压冲击法分离生殖细胞,以探讨阳春砂的授粉技术方法。结果表明:(1)阳春砂花粉发育的特殊性从四分体时期开始,在四分体时期缺少典型的胼胝质壁,而4个小孢子是由多糖性质的细胞壁分隔、包裹着;成熟的阳春砂花粉粒是二胞型花粉,且花粉细胞质中含有丰富的多糖淀粉和脂滴物质;成熟花粉外壁是由多糖物质构成,而非胡萝卜素和类胡萝卜素酯的孢粉素物质构成。(2)阳春砂花粉在2%甘露醇+5%丙酮的爆破液中,爆破率可达10.67%,并释放出花粉内含物,其中包含生殖细胞;在释放出的花粉细胞质中,生殖细胞可完整保留约30 min;用显微操作仪将游离的生殖细胞收集成一定数量的群体,置离心管后即可保存在液氮罐中,每天可收集上百个阳春砂生殖细胞。
中文关键词:阳春砂  花粉  生殖细胞
 
Pollen Structure and Isolation of Generative Cells from Amomum villosum
Abstract:Amomum villosum is a kind of self pollinated plant, whose pistil stands higher than the stamen and it is a typical material for studying generative cell of Monocotyledoneae on molecular biology. To solve the difficulty of A. villosum pollination, the researcher observed development process to clarify the structural characteristics of its pollen by phytohistochemistry method and isolated generative cells to explore its pollination technology by osmotic shock method. This research shows that: (1) the particularity of pollen development of A. villosum begins with tetrad stage and callosewall is absent. The four small spores are separated and enclosed by a polysaccharide cell wall during the tetrad period. Mature pollen is 2 celled and it stores abundant polysaccharide starches and lipids in cytoplasm of A. villosum. Pollen exine is composed of polysaccharide not sporopollen substances of carotene and carotenoid esters. (2) The pollen of A. villosum broken rate is up to 10.67% and release pollen inclusions that contain generative cells in 2% mannitol+5% acetone solution. The generative cells remain intact for about 30 minutes in the released pollen cytoplasm. The isolated hundreds generative cells of A. villosum were collected by micromanipulation and stored in liquid nitrogen tank after placed in centrifuge tube every day.
keywords:Amomum villosum Lour.  pollen  generative cell
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