Junker Laura V, Ensminger Ingo
Department of Biology, Graduate Programs in Cell & Systems Biology and Ecology and Evolutionary Biology, University of Toronto, Mississauga Campus, Mississauga, ON, Canada.
Abteilung Waldnaturschutz, Forstliche Versuchs- und Forschungsanstalt Baden-Württemberg, 79100, Freiburg, Germany.
Physiol Plant. 2016 Dec;158(4):369-381. doi: 10.1111/ppl.12512. Epub 2016 Oct 14.
Rapid developments in remote-sensing of vegetation and high-throughput precision plant phenotyping promise a range of real-life applications using leaf optical properties for non-destructive assessment of plant performance. Use of leaf optical properties for assessing plant performance requires the ability to use photosynthetic pigments as proxies for physiological properties and the ability to detect these pigments fast, reliably and at low cost. We describe a simple and cost-effective protocol for the rapid analysis of chlorophylls, carotenoids and tocopherols using high-performance liquid chromatography (HPLC). Many existing methods are based on the expensive solvent acetonitrile, take a long time or do not include lutein epoxide and α-carotene. We aimed to develop an HPLC method which separates all major chlorophylls and carotenoids as well as lutein epoxide, α-carotene and α-tocopherol. Using a C -column and a mobile phase with a gradient of methanol, methyl-tert-butyl-ether (MTBE) and water, our method separates the above pigments and isoprenoids within 28 min. The broad applicability of our method is demonstrated using samples from various plant species and tissue types, e.g. leaves of Arabidopsis and avocado plants, several deciduous and conifer tree species, various crops, stems of parasitic dodder, fruit of tomato, roots of carrots and Chlorella algae. In comparison to previous methods, our method is very affordable, fast and versatile and can be used to analyze all major photosynthetic pigments that contribute to changes in leaf optical properties and which are of interest in most ecophysiological studies.
植被遥感和高通量精准植物表型分析的快速发展，有望实现一系列实际应用，即利用叶片光学特性对植物性能进行无损评估。利用叶片光学特性评估植物性能，需要能够将光合色素用作生理特性的替代指标，并具备快速、可靠且低成本检测这些色素的能力。我们描述了一种使用高效液相色谱法（HPLC）快速分析叶绿素、类胡萝卜素和生育酚的简单且经济高效的方法。许多现有方法基于昂贵的溶剂乙腈，耗时较长，或者不包括环氧叶黄素和α-胡萝卜素。我们旨在开发一种HPLC方法，能够分离所有主要的叶绿素和类胡萝卜素以及环氧叶黄素、α-胡萝卜素和α-生育酚。使用C柱和含有甲醇、甲基叔丁基醚（MTBE）和水梯度的流动相，我们的方法在28分钟内分离上述色素和类异戊二烯。通过使用来自各种植物物种和组织类型的样本，如拟南芥和鳄梨植物的叶子、几种落叶树和针叶树物种、各种作物、寄生菟丝子的茎、番茄果实、胡萝卜根和小球藻，证明了我们方法的广泛适用性。与先前的方法相比，我们的方法非常经济实惠、快速且通用，可用于分析所有主要的光合色素，这些色素会导致叶片光学特性的变化，并且是大多数生态生理学研究感兴趣的对象。
