Department of Bioresource Engineering, McGill University, Macdonald Campus, 21,111 Lakeshore Road, Ste-Anne-De-Bellevue, Quebec, Canada; Department of Electro-Chemistry Engineering, Dexcom, Inc., 6340 Sequence Dr., San Diego, CA, USA.
Broad Institute of Harvard and MIT, Stanley Center for Psychiatric Research, 75 Ames Street, Cambridge, MA, USA.
J Proteomics. 2022 Aug 15;265:104635. doi: 10.1016/j.jprot.2022.104635. Epub 2022 Jun 1.
Incident light is a central modulator of plant growth and development. However, there are still open questions surrounding wavelength-specific plant proteomic responses. Here we applied tandem mass tag based quantitative proteomics technology to acquire an in-depth view of proteome changes in Arabidopsis thaliana response to narrow wavelength blue (B; 450 nm), amber (A; 595 nm), and red (R; 650 nm) light treatments. A total of 16,707 proteins were identified with 9120 proteins quantified across all three light treatments in three biological replicates. This enabled examination of changes in the abundance for proteins with low abundance and important regulatory roles including transcription factors and hormone signaling. Importantly, 18% (1631 proteins) of the A. thaliana proteome is differentially abundant in response to narrow wavelength lights, and changes in proteome correlate well with different morphologies exhibited by plants. To showcase the usefulness of this resource, data were placed in the context of more than thirty published datasets, providing orthogonal validation and further insights into light-specific biological pathways, including Systemic Acquired Resistance and Shade Avoidance Syndrome. This high-resolution resource for A. thaliana provides baseline data and a tool for defining molecular mechanisms that control fundamental aspects of plant response to changing light conditions, with implications in plant development and adaptation. SIGNIFICANCE: Understanding of molecular mechanisms involved in wavelength-specific response of plant is question of widespread interest both to basic researchers and to those interested in applying such knowledge to the engineering of novel proteins, as well as targeted lighting systems. Here we sought to generate a high-resolution proteomic profile of plant leaves, based on exposure to specific narrow-wavelength lights. Although changes in plant physiology in response to light spectral composition is well documented, there is limited knowledge on the roles of specific light wavelengths and their impact. Most previous studies have utilized relatively broad wavebands in their experiments. Such multi-wavelengths lights trigger diverse and complex signaling networks that pose major challenges in inference of wavelength-specific molecular processes that underly the plant response. Moreover, most studies have compared the effect of blue and red wavelengths comparing with FL, as control. As FL light consists the mixed spectra composition of both red and blue as well as numerous other wavelengths, comparing undeniably results in inconsistent and overlapping responses that will hamper effects to elucidate the plant response to specific wavelengths [1, 2]. Monitoring plant proteome response to specific wavelengths and further contrasting the changes with one another, rather than comparing plants proteome to FL, is thus necessary to gain detailed insights on underlying biological pathways and their consequences in plant physiology. Here, we employed narrow wavelength LED lights in our design to eliminate a potential overlap in molecular responses by ensuring non-overlapping wavelengths in the light treatments. We further applied TMT-labeling technology to gain a high-resolution view on the proteome changes. Our proteomics data provides an in-depth coverage suitable for system-wide analyses, providing deep insights on plant molecular response particularly because of the tremendous increase in the coverage of identified proteins which outreach the other biological data.
入射光是植物生长和发育的主要调节剂。然而,对于特定于波长的植物蛋白质组学反应,仍存在一些悬而未决的问题。在这里,我们应用串联质量标签(Tandem Mass Tag)定量蛋白质组学技术,深入了解拟南芥对窄波长蓝光(B;450nm)、琥珀光(A;595nm)和红光(R;650nm)处理的蛋白质组变化。在三个生物学重复中,总共鉴定到 16707 种蛋白质,其中 9120 种蛋白质在三种光照处理中均有定量。这使得我们能够检查低丰度和具有重要调控作用的蛋白质(包括转录因子和激素信号)的丰度变化。重要的是,18%(1631 种蛋白质)的拟南芥蛋白质组对窄波长光有差异丰度,并且蛋白质组的变化与植物表现出的不同形态很好地相关。为了展示这个资源的有用性,数据被置于三十多个已发表数据集的背景下,提供了正交验证,并进一步深入了解了光特异性的生物学途径,包括系统获得性抗性和避阴综合征。这个拟南芥的高分辨率资源为控制植物对不断变化的光照条件的基本方面的分子机制提供了基线数据和工具,这对植物的发育和适应具有重要意义。
对植物的特定于波长的反应的分子机制的理解是一个广泛关注的问题,无论是对基础研究人员还是对那些有兴趣将这些知识应用于新型蛋白质工程以及有针对性的照明系统的人都是如此。在这里,我们试图根据特定的窄波长光暴露,生成植物叶片的高分辨率蛋白质组图谱。尽管植物对光光谱组成的生理反应已有广泛的记载,但对特定波长的作用及其影响的了解有限。大多数先前的研究在实验中都使用了相对较宽的波段。这种多波长光触发了不同的和复杂的信号网络,这在推断植物反应背后的波长特异性分子过程方面带来了重大挑战。此外,大多数研究比较了蓝色和红色波长与 FL(作为对照)的效果。由于 FL 光由红色和蓝色以及许多其他波长的混合光谱组成,因此不可避免地会导致不一致和重叠的反应,这将阻碍对特定波长下植物反应的阐明。因此,监测植物对特定波长的蛋白质组反应,并进一步相互对比这些变化,而不是将植物蛋白质组与 FL 进行比较,对于深入了解潜在的生物学途径及其对植物生理学的影响是必要的。在这里,我们在设计中使用了窄波长 LED 灯,以通过确保光处理中不存在重叠的波长来消除分子反应中的潜在重叠。我们进一步应用 TMT 标记技术来获得蛋白质组变化的高分辨率视图。我们的蛋白质组学数据提供了适合系统范围分析的深入覆盖范围,为植物分子反应提供了深刻的见解,特别是因为鉴定出的蛋白质的覆盖率大大增加,超出了其他生物学数据。