植物激素分析平台在内源油菜素甾醇分析方法研究中取得新进展
Published:26 Dec.2017    Source:http://www.genetics.ac.cn/xwzx/kyjz/201712/t20171215_4918477.html

油菜素甾醇(brassinosteroids, BRs)是继生长素、赤霉素、细胞分裂素、脱落酸和乙烯之后发现的第六大类植物激素,参与调控植物细胞的伸长与分裂、维管束分化、花粉发育和育性、植株衰老以及植物抗逆反应等一系列重要的生理过程。由于其含量低、基质复杂、质谱离子化效率低等因素,内源性BRs的准确分析检测对于植物学家和分析化学家都是巨大的挑战,一直是制约其在生物学上深入研究的瓶颈问题。

 

植物激素分析平台作为专业性植物激素分析研究平台,长期致力于高效、高灵敏度BRs分析方法研究和技术开发,取得了一系列的成果。首先从BRs的结构与物理化学性质入手,设计了一种基于混合模式离子交换机理的高效样品前处理方法,结合化学衍生技术使BRs定量分析灵敏度大幅提高,所需植物材料用量从传统方法的几十克减少至1克(Plant Physiology, 2013, 162, 2056-2066)。在此基础上提出了基于多种质谱学新技术从植物组织中筛选和鉴定活性BRs的策略,希望成为传统活性筛选方法的补充乃至替代技术,并筛选到一种全新的活性BR化合物 6-deoxo-28-homoTY,为BRs生物合成和代谢路径研究提供了帮助(Frontiers in Plant Science, 2016, 7: 1786)。接下来,针对BRs的化学结构和特性,技术人员专门设计合成了一种对邻二醇化合物具有特异识别作用的硼亲和磁性纳米颗粒(boronate affinity magnetic nanoparticles, BAMNPs),可以用于去除内源BRs分析中对BRs信号有抑制作用的高丰度干扰物,进而提高BRs定量灵敏度,降低植物材料用量至200毫克以下(Analyst, 2013, 138, 1342-1345)。近期,平台技术研究人员又将BAMNPs用于植物组织中内源BRs的直接固相萃取,设计了分步氧化-水解洗脱策略,将BRs回收率提高了2-3倍,所需植物材料用量降至10毫克级,极大地方便了植物学家进行样品准备以及特殊材料的获取。该方法直接提取策略节约了大量的溶剂提取时间,使整个分析时间缩短至3小时以内,大幅提高了分析通量。该检测技术的建立进一步提升了植物激素平台在BRs分析领域水平,有利于为BRs相关生物学研究提供更高质量的技术支持。

 

该研究于2017年12月14日正式发表于Analytical and Bioanalytical Chemistry杂志(DOI: 10.1007/s00216-017-0777-9),文章题目为“Pursuing extreme sensitivity for determination of endogenous brassinosteroids through direct fishing from plant matrices and eliminating most interferences with boronate affinity magnetic nanoparticles”。植物激素分析平台辛培勇博士为论文的第一作者,褚金芳博士为通讯作者。该研究获得了国家自然科学基金、中国科学院关键技术人才项目及中国博士后基金的资助。

 

植物激素分析平台已经开发建立了所有已知种类植物激素的定量分析方法,涵盖生长素、脱落酸、水杨酸、茉莉酸、乙烯、细胞分裂素、赤霉素、油菜素甾醇、独脚金内酯以及小肽类植物激素。所建立的技术体系已经为中科院遗传发育所、北京大学、中科院植物所、中国农业大学、中国水稻所、中山大学、浙江大学、Iowa State University等国内外近百家科研单位提供了高水平的技术支持,部分合作研究成果发表于Nature、Nature Biotechnology、PNAS、Plant Cell等优秀期刊上,有力地支撑了国内外植物激素分子作用机理及分子育种相关的研究工作。

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