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

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. This study at hand suggests a new spectroscopic method for high-throughput characterization of pollen. 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 analysed. Critical steps in the sample preparation and measurement, such as variabilities between technical replicates, between microplates and between spectrometers, were studied. 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. The results of this 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.傅里叶变换红外（Fourier transform infrared, FTIR）光谱技术可用于花粉样本的化学分析，以开展植物表型组学研究，进而探究植物与环境的相互作用，例如气候变化或病原体带来的影响。然而当前的研究方法，如显微光谱法和衰减全反射光谱法，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，可实现花粉的高通量表征。 样本以花粉碎片薄膜的形式进行检测，实验采用布鲁克（Bruker）FTIR光谱仪，并搭配高通量延伸（high-throughput eXTension, HTS-XT）单元及384孔板。本研究共分析了146份花粉样本，这些样本隶属于壳斗科（Fagaceae）和桦木科（Betulaceae）的31个不同花粉物种，采集自克罗地亚、德国和挪威的多个采样点，时间跨度为连续三年（2012年至2014年）。研究还考察了样本制备与检测过程中的关键环节，包括技术重复间、微孔板间以及光谱仪间的差异。 样本制备、微孔板托架与仪器设备带来的测量变异均处于较低水平，因此可依据系统发育和生物地理学特征对样本进行区分。对壳斗目（Fagales）多个物种（水青冈属、栎属、桦木属、榛属、桤木属和铁木属）的光谱变异分析显示，花粉的化学组成存在显著的物种特异性差异，这种差异与采样地点或采样年份相关。花粉光谱中存在统计学意义上显著的年际和地域差异，这表明花粉的化学组成具有较高的表型可塑性，且受当地气候条件的影响。花粉组成的差异与脂类、蛋白质、碳水化合物以及孢粉素密切相关，这些物质在植物抵御低温和干旱胁迫、抵御紫外线辐射以及作为物质和能量储备方面发挥着至关重要的作用。 本研究结果证实了高通量FTIR方法在系统性收集生态系统数据方面的应用价值。这种新颖的FTIR方法可通过半自动化实验流程，实现对大量样本的快速、可靠且经济的筛选。该高通量方法有望为揭示植物与气候之间的相互作用提供关键认知，助力学界深入理解属、物种及种群层面的生化变异。