摘 要

本研究以紫荆（Cercis chinensis）种子为材料，探究了解除休眠过程中紫荆种子内源抑制物活性的变化情况，并采用气相色谱-质谱联用分析法（GC-MS）对紫荆种子胚、胚乳和种皮中内源抑制物质的种类进行鉴定，旨在进一步探索内源抑制物在紫荆种子中存在的部位及内源抑制物与紫荆种子休眠的关系。主要结论如下：

（1）通过有机溶剂萃取的方法对紫荆种子的胚、胚乳和种皮甲醇粗提液进行初步分离，得到了石油醚相、乙醚相、乙酸乙酯相、甲醇相和水相，并进行了生物测定。生物测定结果显示，各分离相提取物抑制作用从大到小依次为：甲醇相gt;乙醚相gt;水相gt;乙酸乙酯相gt;石油醚相。

（2）紫荆种子胚、胚乳和种皮浸提液对白菜籽发芽率均有抑制作用，各部分提取物白菜种子发芽率及发芽指数的抑制作用从大到小依次为：胚乳gt;种皮gt;胚。

（3）GC-MS分析，紫荆种子有机相提取液中分离鉴定出多种内源抑制物，主要为：有机酸类物质、酚类、酯类、醇类等。种子中具有抑制作用的物质主要有：邻苯三酚、2,6-二叔丁基对甲酚、亚油酸、棕榈酸、硬脂酸和十八碳烯酸等。在生物抑制活性最强的甲醇和乙醚相中含有邻苯三酚、2,6-二叔丁基对甲酚2种抑制活性很强的酚类物质。

关键词：紫荆；解除休眠；内源抑制物；气质联用（GC-MS）；

Study on Determination of Endogenous Inhibitors in Seedings of Ziziphus Seeds

Abstract

In this study, the seeds of Cercis chinensis were used as material to explore the changes of the endogenous inhibitor activity of Cercis chinensis seedlings during the dormancy process, and GC-MS analysis was used to determine the embryo and endosperm of Cercis chinensis seedlings. The identification of the endogenous inhibitors in the seed coat was used to further explore the sites of endogenous inhibitors in the seeds of Cercis chinensis and the relationship between the endogenous inhibitors and the dormancy of the seeds of Cercis chinensis. The main conclusions are as follows:

（1）The crude methanol extracts of embryos, endosperms and seed coats of Ziziphus jujuba seed were separated by organic solvent extraction. The petroleum ether phase, the ether phase, the ethyl acetate phase, the methanol phase and the aqueous phase were obtained, and the bioassay was carried out. The results of bioassay showed that the inhibitory effect of the extracts of each phase was as follows: methanol phasegt;ether phasegt;aqueous phasegt;ethyl acetate phasegt;petroleum ether phase.

（2）Germination rate of seeds, endosperm and seed coat of Zizipia purpurea seedlings all inhibited the germination rate of Chinese cabbage seeds. The inhibitory effect of seed germination rate and germination index of the extracts of each part of Chinese cabbage was descended from the following order: endospermgt; seed coatgt; embryo.

（3）By GC-MS analysis, a variety of endogenous inhibitors were isolated and identified from organic extracts of Zijing seeds, mainly including organic acids, phenols, esters, and alcohols. Substances with inhibitory effects in seeds are mainly pyrogallol, 2,6-di-tert-butyl-p-cresol, linoleic acid, palmitic acid, stearic acid, and octadecenoic acid. In the methanol and ether phases with the strongest biosuppressive activity, pyrogallol and 2,6-di-tert-butyl-p-cresol were two phenolic substances with strong inhibitory activity.

Key words: Cercis chinensis; release of dormancy; endogenous inhibitors; GC-MS;

目 录

1 绪 论 - 1 -

1.1研究目的及意义 - 1 -

1.2种子休眠原因研究进展 - 1 -

1.2.1种壳休眠 - 2 -

1.2.2胚休眠 - 3 -

1.2.3综合休眠 - 4 -

1.3 研究内容 - 4 -

2 材料与方法 - 5 -

2.1实验材料 - 5 -

2.2试验方法 - 5 -

2.2.1 紫荆种子内源抑制物的提取 - 5 -

2.2.2 紫荆种子内源抑制物的初步分离 - 5 -

2.2.3 紫荆种子分离物的生物活性测定 - 6 -

2.2.4 紫荆种子胚、胚乳和种皮各分离相GC-MS鉴定 - 7 -

3 结果与分析 - 8 -

3.1 紫荆种子胚、胚乳和种皮中各分离相的生物测定 - 8 -

3.1.1 胚、胚乳和种皮中各有机相对白菜籽发芽的影响 - 8 -

3.2 紫荆种子种皮中提取的有机化合物和相对含量 - 9 -

3.2.1 紫荆种子胚的石油醚相提取物 - 9 -

3.2.2 紫荆种子胚的乙醚相提取物 - 10 -

3.2.3 紫荆种子胚的乙酸乙酯相提取物 - 12 -

3.2.4 紫荆种子胚的甲醇相提取物 - 13 -

3.3 紫荆种子胚乳中提取的有机化合物和相对含量 - 15 -

3.3.1 紫荆种子胚乳的石油醚相提取物 - 15 -

3.3.2 紫荆种子胚乳的乙醚相提取物 - 16 -

3.3.3 紫荆种子胚乳的乙酸乙酯相提取物 - 17 -

3.3.4 紫荆种子胚乳的甲醇相提取物 - 18 -

3.4 紫荆种子种皮中提取的有机化合物和相对含量 - 20 -

3.4.1 紫荆种子种皮的石油醚相提取物 - 20 -

3.4.2 紫荆种子种皮的乙醚相提取物 - 22 -

3.4.3 紫荆种子种皮的乙酸乙酯相提取物 - 23 -

3.4.4 紫荆种子种皮的甲醇相提取物 - 25 -

4 讨论 - 27 -

4.1 紫荆种子各部位中抑制物质的生物测定 - 27 -

4.2 紫荆种子中抑制物的种类 - 28 -

致 谢 - 29 -

参考文献 - 30 -

1 绪 论

1.1研究目的及意义

紫荆(Cercis chinensis）是苏木科（Caesalpiniaceae）紫荆属（Cercis）落叶乔木或灌木，又名满条红、苏芳花等^[1]。紫荆原产于我国东南部，北至河北，南至广东、广西，西至云南、四川，西北至陕西，东至浙江、江苏和山东等省区^[2]。紫荆早春开花，紫红色的花着生在老枝上，是春季较好的观花植物，亦为优良的蜜源植物^[3]；紫荆的花、果、树皮、根等均可入药，花可治风湿筋骨痛，果可治疗咳嗽，树皮有清热解毒，活血行气，消肿止痛之功效，可治产后血气痛、疗疮肿毒、喉痹^[4]。此外，紫荆木材结构较细，可供建筑、家具等用。因此对于紫荆种子休眠的研究就显得尤为重要。

紫荆是集观赏价值，药用价值和经济价值于一身的优良树种，随着繁育技术的提高以及生产生活的需要，其应用也将越来越广泛。