An Improved Simplified High-Sensitivity Quantification Method for Determining Brassinosteroids in Different Tissues of Rice and Arabidopsis
An Improved Simplified High-Sensitivity Quantification Method for Determining Brassinosteroids in Different Tissues of Rice and Arabidopsis
复制标题
一种改进的简化高灵敏度定量方法，用于测定水稻和拟南芥不同组织中的油菜素类固醇
DOI：
10.1104/pp.113.221952
复制
发表时间：
2013-08-01
期刊：
影响因子：
7.4
通讯作者：
Yan, Cunyu
中科院分区：
文献类型：
--
作者：
Xin, Peiyong;Yan, Jijun;Yan, Cunyu
油菜素类固醇的定量对于研究其在植物生长和发育中生理作用的分子机制至关重要。在此，我们提出了一种简单、材料和节省成本的高性能方法来测定模型植物中的内源油菜素类固醇（BR）。这种新方法可以通过离子交换固相萃取同时富集多种生物活性 BR，如油菜素内酯、蓖麻油酮、茶甾酮和斑黄甾醇，并基于同位素稀释结合内标法对这些 BR 进行高灵敏度定量。对于常规分析，植物材料的消耗量减少到之前报道的二十分之一，整个过程可以在1天内完成，而之前需要3至4天。通过对水稻（Oryza sativa）和拟南芥（Arabidopsis thaliana）不同生态型和突变体中的 BR 进行分析验证了该策略，并用新方法确定了不同模型植物组织中的 BR 分布。该方法使植物生理学家能够监测1克鲜重模型植物组织中BR的动态和分布，这将加快未来工作中利用这些模型植物进行BR分子机制研究的进程。
Quantification of brassinosteroids is essential and extremely important to study the molecular mechanisms of their physiological roles in plant growth and development. Herein, we present a simple, material and cost-saving high-performance method for determining endogenous brassinosteroids (BRs) in model plants. This new method enables simultaneous enrichment of a wide range of bioactive BRs such as brassinolide, castasterone, teasterone, and typhasterol with ion exchange solid-phase extraction and high-sensitivity quantitation of these BRs based on isotope dilution combined with internal standard approach. For routine analysis, the consumption of plant materials was reduced to one-twentieth of previously reported and the overall process could be completed within 1 day compared with previous 3 to 4 days. The strategy was validated by profiling BRs in different ecotypes and mutants of rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana), and the BR distributions in different model plants tissues were determined with the new method. The method allows plant physiologists to monitor the dynamics and distributions of BRs with 1 gram fresh weight of model plant tissues, which will speed up the process for the molecular mechanism research of BRs with these model plants in future work.