Data from: Monitoring of plant-environment interactions by high throughput FTIR spectroscopy of pollen

1. Fourier transform infrared (FTIR) spectroscopy enables chemical analysis of pollen samples for plant phenotyping to study plant-environment interactions, such as influence of climate change or pathogens. However, current approach, such as microspectroscopy and Attenuated Total Reflection spectroscopy, does not allow for high-throughput protocols. The paper at hand suggests a new spectroscopic method for high-throughput characterization of pollen.

2. Samples were measured as thin films of pollen fragments using a Bruker FTIR spectrometer with a high-throughput eXTension (HTS-XT) unit employing 384-well plates.

In total, 146 pollen samples, belonging to 31 different pollen species of Fagaceae and Betulaceae and collected during three consecutive years (2012-2014) at locations in Croatia, Germany and Norway, were analyzed. Critical steps in the sample preparation and measurement, such as variabilities between technical replicates, between microplates and between spectrometers, were studied.

3. Measurement variations due to sample preparation, microplate holders and instrumentation were low, and thus allowed differentiation of samples with respect to phylogeny and biogeography. The spectral variability for a range of Fagales species (Fagus, Quercus, Betula, Corylus, Alnus and Ostrya) showed high species-specific differences in pollen's chemical composition due to either location or year. Statistically significant inter-annual and locational differences in the pollen spectra indicate that pollen chemical composition has high phenotypic plasticity and is influenced by local climate conditions. The variations in composition are connected to lipids, proteins, carbohydrates and sporopollenins that play crucial roles in cold and desiccation tolerance, protection against UV radiation, and as material and energy reserves.

4. The results of the study demonstrate the value of high throughput FTIR approach for the systematic collection of data on ecosystems. The novel FTIR approach offers fast, reliable and economical screening of large number of samples by semi-automated methodology. The high-throughput approach could provide crucial understanding on plant-climate interactions with respect to biochemical variation within genera, species and populations.

1. 傅里叶变换红外（Fourier transform infrared, FTIR）光谱技术可用于花粉样品的化学分析，以开展植物表型组学研究，进而解析植物与环境的互作机制，例如气候变化或病原菌的影响。然而当前主流方法（如显微光谱法与衰减全反射光谱技术）均无法适配高通量实验流程。本研究提出了一种全新的光谱学方法，可实现花粉样品的高通量表征。
2. 实验以花粉碎片制成的薄膜为样品，使用搭载高通量扩展模块（HTS-XT）的布鲁克（Bruker）FTIR光谱仪进行检测，该设备采用384孔板承载样品。本研究共分析了146份花粉样品，这些样品隶属于壳斗科（Fagaceae）与桦木科（Betulaceae）的31个不同花粉物种，采集时间为2012至2014年连续三年，采样地点覆盖克罗地亚、德国与挪威。研究同时考察了样品制备与检测流程中的关键环节，包括技术重复间、微孔板间以及光谱仪间的检测变异情况。
3. 由样品制备、微孔板托架与仪器设备带来的检测变异均处于较低水平，因此该方法可依据系统发育与生物地理学特征对样品进行区分。针对壳斗目（Fagales）多个属的物种（包括山毛榉属、栎属、桦木属、榛属、桤木属与铁木属），其光谱变异分析显示，花粉化学组成存在显著的物种特异性差异，且差异与采样地点或采集年份相关。花粉光谱中存在显著的年际与空间差异，这表明花粉化学组成具有较高的表型可塑性，且受当地气候条件的调控。花粉组成的变异与脂类、蛋白质、碳水化合物以及孢粉素（sporopollenins）密切相关，这些物质在植物应对低温与干旱胁迫、抵御紫外线辐射以及作为物质与能量储备方面发挥关键作用。
4. 本研究结果证实了高通量FTIR光谱技术在生态系统数据系统化采集方面的应用价值。该新型FTIR光谱技术可通过半自动化流程，实现大批量样品的快速、可靠且经济的筛选。这种高通量检测方法可帮助我们深入理解属、物种与种群水平上的生化变异与植物-气候互作之间的关联。