Two-Dimensional Tannin Fingerprints by Liquid Chromatography Tandem Mass Spectrometry Offer a New Dimension to Plant Tannin Analyses and Help To Visualize the Tannin Diversity in Plants

Tannins are large-molecular-weight plant polyphenols that are produced in fruits, berries, leaves, flowers, seeds, stems, and roots of woody and non-woody plants. Hundreds and thousands of individual tannin structures are consequently found in many kinds of natural food and feed products. The huge structural variability in tannins is reflected as vast bioactivity differences between them but not in the accuracy of their typical analysis methods. Here, I show how the modern liquid chromatography mass spectrometry methods can be used to obtain new types of two-dimensional tannin fingerprints to better visualize both the tannin content and diversity in plants with just one 10 min analysis per sample

Changes in oak (Quercus robur) photosynthesis after winter moth (Operophtera brumata) herbivory are not explained by changes in chemical or structural leaf traits

Insect herbivores have the potential to change both physical and chemical traits of their host plant. Although the impacts of herbivores on their hosts have been widely studied, experiments assessing changes in multiple leaf traits or functions simultaneously are still rare. We experimentally tested whether herbivory by winter moth (Operophtera brumata) caterpillars and mechanical leaf wounding changed leaf mass per area, leaf area, leaf carbon and nitrogen content, and the concentrations of 27 polyphenol compounds on oak (Quercus robur) leaves. To investigate how potential changes in the studied traits affect leaf functioning, we related the traits to the rates of leaf photosynthesis and respiration. Overall, we did not detect any clear effects of herbivory or mechanical leaf damage on the chemical or physical leaf traits, despite clear effect of herbivory on photosynthesis. Rather, the trait variation was primarily driven by variation between individual trees. Only leaf nitrogen content and a subset of the studied polyphenol compounds correlated with photosynthesis and leaf respiration. Our results suggest that in our study system, abiotic conditions related to the growth location, variation between tree individuals, and seasonal trends in plant physiology are more important than herbivory in determining the distribution and composition of leaf chemical and structural traits

Relevance of the Concentrations and Sizes of Oligomeric Red Wine Pigments to the Color Intensity of Commercial Red Wines

Color is a major sensorial characteristic of red wines. Numerous monomeric and some small oligomeric pigments have been characterized from red wines but the contribution of larger oligomeric pigments to the color intensity has not been established by direct measurements. We measured the color intensity of 317 commercial red wines and semiquantified the malvidin glycosides and eight different adduct groups derived from the malvidin glycosides by ultra-performance liquid chromatography-tandem mass spectrometry. Two of these groups were oligomeric pigments consisting of proanthocyanidins and malvidin glycosides with either direct or methylmethine linkages. The carboxypyranomalvidins and the oligomeric pigments were found to be major contributors to the color intensity. Besides the concentrations, the sizes of the oligomeric pigments had a positive and significant connection to the color intensity. The 1-year-old wines were studied separately and, even in the youngest of wines, the adducts of the malvidin glycosides were the major contributors to the color intensity

Chemotaxonomic Markers for the Leaf Buds of Common Finnish Trees and Shrubs: A Rapid UHPLC MS Fingerprinting Tool for Species Identification

In this study, a chemotaxonomic tool was created on the basis of ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) for the identification of 13 common Finnish deciduous trees and shrubs from their leaf bud metabolites. The bud extracts were screened with UHPLC-ESI-QqQ-MS and UHPLC-ESI-Q-Orbitrap-MS to discover suitable markers for each species. Two approaches were tested in the marker selection: (1) unique species-specific markers to obtain selective fingerprints per species and (2) major markers to maximise the sensitivity of the fingerprints. The markers were used to create two selected ion-recording-based fingerprinting tools with UHPLC-ESI-QqQ-MS. The methods were evaluated for their selectivity, repeatability, and robustness in plant species identification by analysing leaf buds from several replicates of each species. The created chemotaxonomic tools were shown to provide unique chromatographic profiles for the studied species in less than 6 min. A variety of plant metabolites, such as flavonoids, triterpenoids, and hydroxycinnamic acid derivatives, were found to serve as good chemotaxonomic markers for the studied species. In 10 out of 13 cases, species-specific markers were superior in creating selective and repeatable fingerprints

Influence of the Hydrolyzable Tannin Structure on the Characteristics of Insoluble Hydrolyzable Tannin-Protein Complexes

Precipitation of bovine serum albumin (BSA) by 21 hydrolyzable tannins (HTs) and the characteristics of the insoluble complexes were studied stoichiometrically by ultra-performance liquid chromatography. With regard to HT monomers, the protein precipitation and the characteristic of the formed precipitates were unique for each studied HT and depended upon the functional groups present in the structures. The monomeric units comprising the oligomers formed the functional units important for the protein precipitation capacity, and small structural differences among the monomer units were less important than the overall oligomer size and flexibility. In addition, the greater tendency of certain HTs to form insoluble complexes when mixed with BSA was partially linked to the higher self-association and consequent stronger cooperative binding of these HTs with BSA

Binding of an oligomeric ellagitannin series to bovine serum albumin (BSA): analysis by isothermal titration calorimetry (ITC)

A unique series of oligomeric ellagitannins was used to study their interactions with bovine serum albumin (BSA) by isothermal titration calorimetry. Oligomeric ellagitannins, ranging from monomer to heptamer and a mixture of octamer–undecamers, were isolated as individual pure compounds. This series allowed studying the effects of oligomer size and other structural features. The monomeric to trimeric ellagitannins deviated most from the overall trends. The interactions of ellagitannin oligomers from tetramers to octa–undecamers with BSA revealed strong similarities. In contrast to the equilibrium binding constant, enthalpy showed an increasing trend from the dimer to larger oligomers. It is likely that first the macrocyclic part of the ellagitannin binds to the defined binding sites on the protein surface and then the “flexible tail” of the ellagitannin coats the protein surface. The results highlight the importance of molecular flexibility to maximize binding between the ellagitannin and protein surfaces

Insect community structure covaries with host plant chemistry but is not affected by prior herbivory

By feeding on plant tissue, insect herbivores can change several characteristics of their hosts. These changes have the potential to alter the quality of the plant for other herbivore species, potentially altering the structure of the community of species attacking the plant at a later point in time. We tested whether herbivory early in the season changes host plant performance, polyphenol chemistry, and the community structure of sessile herbivores later in the season. We experimentally manipulated densities of early‐season moth caterpillars on a set of young oak trees and measured tree growth, reproduction, leaf chemistry, and the abundance and community composition of leafmining and galling species later in the season. The experimental manipulations of early‐season herbivores did not affect late‐season leaf chemistry or tree performance. Early‐season herbivores had a weak negative effect on the abundance of gallers and a positive, tree‐dependent effect on the overall diversity of late‐season sessile herbivores. The chemical composition of leaves covaried with the species composition of the late‐season leafmining and galling community. Both the chemical composition of the host tree and the late‐season insect community structure were strongly affected by the growth location of the tree. Our results suggest that plant‐mediated indirect effects between herbivores might play a limited role in this system, whereas the underlying variation in plant chemistry is an important factor structuring the associated insect community. Our results emphasize that factors other than prior herbivory can be important determinants of plant chemistry and the community composition of herbivores

Plant Chemistry and Local Adaptation of a Specialized Folivore

Local adaptation is central for creating and maintaining spatial variation in plant-herbivore interactions. Short-lived insect herbivores feeding on long-lived plants are likely to adapt to their local host plants, because of their short generation time, poor dispersal, and geographically varying selection due to variation in plant defences. In a reciprocal feeding trial, we investigated the impact of geographic variation in plant secondary chemistry of a long-lived plant, Vincetoxicum hirundinaria, on among-population variation in local adaptation of a specialist leaf-feeding herbivore, Abrostola asclepiadis. The occurrence and degree of local adaptation varied among populations. This variation correlated with qualitative and quantitative differences in plant chemistry among the plant populations. These findings provide insights into the mechanisms driving variation in local adaptation in this specialized plant-herbivore interaction