In situ analysis and structural elucidation of sainfoin (Onobrychis viciifolia) tannins for high-throughput germplasm screening

A rapid thiolytic degradation and cleanup procedure was developed for analyzing tannins directly in chlorophyll-containing sainfoin (Onobrychis viciifolia) plants. The technique proved suitable for complex tannin mixtures containing catechin, epicatechin, gallocatechin, and epigallocatechin flavan-3-ol units. The reaction time was standardized at 60 min to minimize the loss of structural information as a result of epimerization and degradation of terminal flavan-3-ol units. The results were evaluated by separate analysis of extractable and unextractable tannins, which accounted for 63.6−113.7% of the in situ plant tannins. It is of note that 70% aqueous acetone extracted tannins with a lower mean degree of polymerization (mDP) than was found for tannins analyzed in situ. Extractable tannins had between 4 and 29 lower mDP values. The method was validated by comparing results from individual and mixed sample sets. The tannin composition of different sainfoin accessions covered a range of mDP values from 16 to 83, procyanidin/prodelphinidin (PC/PD) ratios from 19.2/80.8 to 45.6/54.4, and cis/trans ratios from 74.1/25.9 to 88.0/12.0. This is the first high-throughput screening method that is suitable for analyzing condensed tannin contents and structural composition directly in green plant tissue.

Binding of pentagalloyl glucose to two globular proteins occurs via multiple surface sites

The interaction between pentagalloyl glucose (PGG) and two globular proteins, bovine serum albumin (BSA) and ribulose-1,5-bisphosphate carboxylase oxygenase (rubisco), was investigated by isothermal titration calorimetry (ITC). ITC data fit to a binding model consisting of two sets of multiple binding sites, which reveal similarities in the mode of binding of PGG to BSA and rubisco. In both cases, the interaction is characterized by a high number of binding sites, which suggests that binding occurs by a surface adsorption mechanism that leads to coating of the protein surface, which promotes aggregation and precipitation of the PGG-protein complex. This model was confirmed by turbidimetry analysis of the PGG-BSA interaction. Analysis of tryptophan fluorescence quenching during the interaction of PGG with BSA suggests that binding of PGG leads to some conformational changes that are energetically closer to the unfolded state of the BSA structure, because small red shifts in the resulting emission spectra were observed.

Structural diversity of sainfoin (Onobrychis viciifolia) tannins

Tannins are oligomeric and polymeric polyphenols that are produced by many plants. The study of their biological activities is of interest because they can generate valuable nutritional, veterinary and environmental effects in ruminant livestock production [1]. Isolated tannin fractions from sainfoin (Onobrychis viciifolia), which is potentially a very useful animal feed, were characterised by MALDI-TOF MS and thiolytic degradation with benzyl mercaptan. Condensed tannins were analysed and characterised in more than different 40 sainfoin varieties to provide guidelines for future plant breeding programmes. Several different techniques were used to study these complex tannin mixtures.

Simple solution for a complex problem: Proanthocyanidins, galloyl glucoses and ellagitannins fit on a single calibration curve in high performance-gel permeation chromatography

This study was undertaken to explore gel permeation chromatography (GPC) for estimating molecular weights of proanthocyanidin fractions isolated from sainfoin (Onobrychis viciifolia). The results were compared with data obtained by thiolytic degradation of the same fractions. Polystyrene, polyethylene glycol and polymethyl methacrylate standards were not suitable for estimating the molecular weights of underivatized proanthocyanidins. Therefore, a novel HPLC-GPC method was developed based on two serially connected PolarGel-L columns using DMF that contained 5% water, 1% acetic acid and 0.15 M LiBr at 0.7 ml/min and 50 degrees C. This yielded a single calibration curve for galloyl glucoses (trigalloyl glucose, pentagalloyl glucose), ellagitannins (pedunculagin, vescalagin, punicalagin, oenothein B, gemin A), proanthocyanidins (procyanidin B2, cinnamtannin B1), and several other polyphenols (catechin, epicatechin gallate, epicallocatechin gallate, amentoflavone). These GPC predicted molecular weights represented a considerable advance over previously reported HPLC-GPC methods for underivatized proanthocyanidins. (C) 2011 Elsevier B.V. All rights reserved.

Geoarchaeological investigations of midden-formation processes in the early to late Neolithic levels at Catalhoyuk, Turkey ca. 8550-8370 cal BP.

Anthropogenic midden deposits are remarkably well preserved at the Neolithic settlement of atalhöyük and provide significant archaeological information on the types and nature of activities occurring at the site. To decipher their complex stratigraphy and to investigate formation processes, a combination of geoarchaeological techniques was used. Deposits were investigated from the early ceramic to late Neolithic levels, targeting continuous sequences to examine high resolution and broader scale changes in deposition. Thin-section micromorphology combined with targeted phytolith and geochemical analyses indicates they are composed of a diverse range of ashes and other charred and siliceous plant materials, with inputs of decayed plants and organic matter, fecal waste, and sedimentary aggregates, each with diverse depositional pathways. Activities identified include in situ burning, with a range of different fuel types that may be associated with different activities. The complexity and heterogeneity of the midden deposits, and thus the necessity of employing an integrated microstratigraphic approach is demonstrated, as a prerequisite for cultural and palaeoenvironmental reconstructions.

The ecological impact of conquest and colonization on a medieval frontier landscape: combined palynological and geochemical analysis of lake sediments from Radzyń Chełminski, northern Poland

Slavic and German colonization of the southern Baltic between the 8th and 15th centuries A.D. is well-documented archaeologically and historically. Despite the large number of pollen profiles from Poland, few palaeoecological studies have examined the ecological impact of a process that was central to the expansion of European, Christian, societies. This study aims to redress this balance through multiproxy analysis of lake sediments from Radzyń Chełminski, Northern Poland, using pollen, element geochemistry (Inductively Coupled-Optical Emission Spectroscopy [ICP-OES]), organic content, and magnetic susceptibility. The close association between lake and medieval settlements presents the ideal opportunity to reconstruct past vegetation and land-use dynamics within a well-documented archaeological, historical, and cultural context. Three broad phases of increasing landscape impact are visible in the pollen and geochemical data dating from the 8th/9th, 10th/11th, and 13th centuries, reflecting successive phases of Slavic and German colonization. This involved the progressive clearance of oak-hornbeam dominated woodland and the development of an increasingly open agricultural landscape. Although the castles and towns of the Teutonic Order remain the most visible signs of medieval colonization, the palynological and geochemical data demonstrate that the major phase of woodland impact occurred during the preceding phase of Slavic expansion; Germans colonists were entering a landscape already significantly altered.

Sampling and analyzing metals in soils for archaeological prospection: a critique

This paper presents a critique of current methods of sampling and analyzing soils for metals in archaeological prospection. Commonly used methodologies in soil science are shown to be suitable for archaeological investigations, with a concomitant improvement in their resolution. Understanding the soil-fraction location, concentration range, and spatial distribution of autochthonous (native) soil metals is shown to be a vital precursor to archaeological-site investigations, as this is the background upon which anthropogenic deposition takes place. Nested sampling is suggested as the most cost-effective method of investigating the spatial variability in the autochthonous metal concentrations. The use of the appropriate soil horizon (or sampling depth) and point sampling are critical in the preparation of a sampling regime. Simultaneous extraction is proposed as the most efficient method of identifying the location and eventual fate of autochthonous and anthropogenic metals, respectively.