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| 利用溴化钠和溴化氢代替溴素染色橡胶树乳管的染色方法研究 |
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郑硕恒1,2, 杜晓愚3, 史敏晶3, 杨署光3, 吴绍华3, 田维敏1, 张华2*, 张世鑫1*
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1. 中国科学院西双版纳热带植物园,云南 勐腊 666303；2. 西藏农牧大学 资源与环境学院,西藏 林芝 860000；3. 中国热带农业科学院橡胶研究所,农业农村部橡胶树生物学与遗传资源利用重点实验室/海南省热带作物栽培生理学重点实验室,海口571101
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| 摘要: |
| 传统的碘-溴染色（I?-Br?）显示乳管细胞的方法，虽然准确且高效，但是I?-Br?染色剂的主成分溴素（Br?）是一种危险化学品，被严格管控，使得I?-Br?染色方法的应用受到极大限制。为了寻求效果相似且更安全的染色试剂，该文采用溴化钠（NaBr）和溴化氢（HBr）代替Br?配制不同染色剂，对橡胶树萌条树皮和叶柄进行染色，使用光镜技术对树皮和叶柄横切片进行观察，以筛选出染色最佳的染色剂配方。结果表明：（1）NaBr可以代替Br?配制染色剂对乳管进行染色，其中0.5 mol?L-1 NaBr + 0.2 mol?L-1 I?的乙酸溶液的染色效果较好，并且使用稀释的冰乙酸作为溶剂能够使NaBr溶解更充分，染色效果更好。（2）HBr可以代替Br?配制染色剂对乳管进行染色且较低浓度的HBr就能使乳管染色，染色效果较好的配方为6.25 mmol?L-1 HBr + 0.2 mol?L-1 I?的乙酸溶液。该研究结果认为使用更安全实惠的NaBr和HBr可以替代Br?配制染色剂，用于橡胶树乳管细胞的形态结构研究。该文提供的橡胶树乳管细胞染色的新试剂，可为产胶植物的乳管生物学研究提供技术支持。 |
| 关键词: 巴西橡胶树，乳管，光镜技术，溴化钠，溴化氢 |
| DOI：10.11931/guihaia.gxzw202503016 |
| 分类号: |
| Fund project:云南省重大科技专项计划项目（202402AE09001901）；国家自然科学基金（31800577）；中国科学院先导B项目（E4XDB21B01）。 |
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| Staining method of laticifers in Hevea brasiliensis bark and petiole using sodium bromide (NaBr) and hydrogen bromide (HBr) instead of bromine (Br?) |
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ZHENG Shuoheng1,2, DU Xiaoyu3, SHI Minjing3, YANG Shuguang3, WU Shaohua3, TIAN Weimin1, ZHANG Hua2*, ZHANG Shixin1*
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1. Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla 666303, Yunnan, China; 2. College of Resources and Environment, Xizang Agricultural and Animal Husbandry University, Nyingchi 860000, Xizang, China; 3. Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture and Rural Affairs / Hainan Key Laboratory for Cultivation and Physiology of Tropical Crops, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
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| Abstract: |
| The iodine-bromine (I?-Br?) staining method is accurate and efficient for revealing the microstructure of laticifer. However, the main component of I?-Br? staining agent, bromine (Br?), has strong pungent odor and highly corrosive. It is classified as a precursor to toxic chemicals and a hazardous chemical, subject to strict control, which greatly limits the application of the I?-Br? staining method. To seek a similar but safer staining reagent, this study used sodium bromide (NaBr) and hydrogen bromide (HBr) to replace Br?, to prepare different staining agents for staining the bark and petioles of rubber tree shoots. Light microscopy was used to observe the cross-sections of the bark and petioles to screen for the best staining agent formula. The results were as follows: (1) NaBr could replace Br? to prepare staining agents for laticifer, and the staining effect of 0.5 mol?L-1 NaBr + 0.2 mol?L-1 I? in acetic acid solution was better. Moreover, using diluted glacial acetic acid as the solvent could make NaBr dissolve more thoroughly and improve the staining effect. (2) HBr could also replace Br? to prepare staining agents for laticifer, and a lower concentration of HBr could stain laticifer. The best staining agent formula was 6.25 mmol?L-1 HBr + 0.2 mol?L-1 I? in acetic acid solution. The results indicated that the safer and more economical NaBr and HBr could be used to replace Br? to prepare staining agents for the study of the morphology and structure of laticifer in rubber tree. The new staining reagents provided in this paper can provide technical support for the study of laticifer in rubber-producing plants. |
| Key words: Hevea brasiliensis, laticifer, optical microscopy technique, sodium bromide, hydrogen bromide |
