Compartmentalization of specialized metabolites across vegetative and reproductive tissues in two sympatric Psychotria species.

PREMISE: The specialized metabolites of plants are recognized as key chemical traits in mediating the ecology and evolution of sundry plant-biotic interactions, from pollination to seed predation. Intra- and interspecific patterns of specialized metabolite diversity have been studied extensively in leaves, but the diverse biotic interactions that contribute to specialized metabolite diversity encompass all plant organs. Focusing on two species of Psychotria shrubs, we investigated and compared patterns of specialized metabolite diversity in leaves and fruit with respect to each organ's diversity of biotic interactions. METHODS: To evaluate associations between biotic interaction diversity and specialized metabolite diversity, we combined UPLC-MS metabolomic analysis of foliar and fruit specialized metabolites with existing surveys of leaf- and fruit-centered biotic interactions. We compared patterns of specialized metabolite richness and variance among vegetative and reproductive tissues, among plants, and between species. RESULTS: In our study system, leaves interact with a far larger number of consumer species than do fruit, while fruit-centric interactions are more ecologically diverse in that they involve antagonistic and mutualistic consumers. This aspect of fruit-centric interactions was reflected in specialized metabolite richness-leaves contained more than fruit, while each organ contained over 200 organ-specific specialized metabolites. Within each species, leaf- and fruit-specialized metabolite composition varied independently of one another across individual plants. Contrasts in specialized metabolite composition were stronger between organs than between species. CONCLUSIONS: As ecologically disparate plant organs with organ-specific specialized metabolite traits, leaves and fruit can each contribute to the tremendous overall diversity of plant specialized metabolites.

Short-term exposure of dairy cows to pyrrolizidine alkaloids from tansy ragwort (Jacobaea vulgaris Gaertn.): effects on health and performance.

The increasing spread of ragworts is observed with concern. Ragworts like tansy ragwort (Jacobaea vulgaris Gaertn.) or marsh ragwort (J. aquatica) contain pyrrolizidine alkaloids (PA) which may induce hepatotoxic effects. Grazing animals usually avoid ragworts if their pasture management is appropriate. Preserved feed prepared from ragworts contaminated meadows may, however, lead to a significant exposure to PA. Previous studies on toxicity of PA for dairy cows revealed inconsistent results due to feeding ragwort plant material which was associated with heterogeneous PA exposure and thus failed to conclusively deduce critical PA doses. Therefore, the aim of the present study was to expose dairy cows (n = 4 per group) in a short-term scenario for 28 days with increasing PA doses (PA1: 0.47 mg PA/kg body weight (BW)/day (d); PA2: 0.95 mg PA/kg BW/d; PA3: 1.91 mg PA/kg BW/d) via oral administration by gavage of a defined PA-extract. While group PA3 was dosed with the PA-extract alone, groups PA2 and PA1 received PA-extracts blended in similar volumes with molasses to provide comparable amounts of sugar. Additionally, two control groups were treated either with water (CONWater) or with molasses (CONMolasses) to assess the effects of sugar without PA interference. While clinical traits including dry matter intake, milking performance, rectal body temperature, ruminal activity and body condition score (BCS) were not influenced by PA exposure, activities of enzymes indicative for liver damages, such as gamma-glutamyltransferase (GGT), aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH), increased significantly over time at an exposure of 1.91 mg total PA/kg BW/d.

Characterizing natural variability of lignin abundance and composition in fine roots across temperate trees: a comparison of analytical methods.

Lignin is an important root chemical component that is widely used in biogeochemical models to predict root decomposition. Across ecological studies, lignin abundance has been characterized using both proximate and lignin-specific methods, without much understanding of their comparability. This uncertainty in estimating lignin limits our ability to comprehend the mechanisms regulating root decomposition and to integrate lignin data for large-scale syntheses. We compared five methods of estimating lignin abundance and composition in fine roots across 34 phylogenetically diverse tree species. We also assessed the feasibility of high-throughput techniques for fast-screening of root lignin. Although acid-insoluble fraction (AIF) has been used to infer root lignin and decomposition, AIF-defined lignin content was disconnected from the lignin abundance estimated by techniques that specifically measure lignin-derived monomers. While lignin-specific techniques indicated lignin contents of 2-10% (w/w) in roots, AIF-defined lignin contents were c. 5-10-fold higher, and their interspecific variation was found to be largely unrelated to that determined using lignin-specific techniques. High-throughput pyrolysis-gas chromatography-mass spectrometry, when combined with quantitative modeling, accurately predicted lignin abundance and composition, highlighting its feasibility for quicker assessment of lignin in roots. We demonstrate that AIF should be interpreted separately from lignin in fine roots as its abundance is unrelated to that of lignin polymers. This study provides the basis for informed decision-making with respect to lignin methodology in ecology.

Untargeted In Silico Compound Classification-A Novel Metabolomics Method to Assess the Chemodiversity in Bryophytes.

In plant ecology, biochemical analyses of bryophytes and vascular plants are often conducted on dried herbarium specimen as species typically grow in distant and inaccessible locations. Here, we present an automated in silico compound classification framework to annotate metabolites using an untargeted data independent acquisition (DIA)-LC/MS-QToF-sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH) ecometabolomics analytical method. We perform a comparative investigation of the chemical diversity at the global level and the composition of metabolite families in ten different species of bryophytes using fresh samples collected on-site and dried specimen stored in a herbarium for half a year. Shannon and Pielou's diversity indices, hierarchical clustering analysis (HCA), sparse partial least squares discriminant analysis (sPLS-DA), distance-based redundancy analysis (dbRDA), ANOVA with post-hoc Tukey honestly significant difference (HSD) test, and the Fisher's exact test were used to determine differences in the richness and composition of metabolite families, with regard to herbarium conditions, ecological characteristics, and species. We functionally annotated metabolite families to biochemical processes related to the structural integrity of membranes and cell walls (proto-lignin, glycerophospholipids, carbohydrates), chemical defense (polyphenols, steroids), reactive oxygen species (ROS) protection (alkaloids, amino acids, flavonoids), nutrition (nitrogen- and phosphate-containing glycerophospholipids), and photosynthesis. Changes in the composition of metabolite families also explained variance related to ecological functioning like physiological adaptations of bryophytes to dry environments (proteins, peptides, flavonoids, terpenes), light availability (flavonoids, terpenes, carbohydrates), temperature (flavonoids), and biotic interactions (steroids, terpenes). The results from this study allow to construct chemical traits that can be attributed to biogeochemistry, habitat conditions, environmental changes and biotic interactions. Our classification framework accelerates the complex annotation process in metabolomics and can be used to simplify biochemical patterns. We show that compound classification is a powerful tool that allows to explore relationships in both molecular biology by "zooming in" and in ecology by "zooming out". The insights revealed by our framework allow to construct new research hypotheses and to enable detailed follow-up studies.

Evaluation of millets for physio-chemical and root morphological traits suitable for resilient farming and nutritional security in Eastern Himalayas.

Introduction: Millets are nutritionally superior and climate-resilient short-duration crops and hold a prominent place in cropping sequences around the world. They have immense potential to grow in a marginal environment due to diverse adaptive mechanisms. Methods: An experiment was conducted in an organic production system in the North Eastern Himalayan foothills of India for 3 consecutive years by evaluating high-yielding varieties (HYVs) of different millets, viz., finger millet, foxtail millet, little millet, barnyard millet, proso millet, and browntop millet, along with local landraces of finger millets (Sikkim-1 and Sikkim-2; Nagaland-1 and Nagaland-2) to identify stable, high-yielding, and nutritionally superior genotypes suited for the region. Results: Among the various millets, finger millet, followed by little millet and foxtail millet, proved their superiority in terms of productivity (ranging between 1.16 and 1.43 Mg ha-1) compared to other millets. Among different varieties of finger millets, cv. VL Mandua 352 recorded the highest average grain yield (1.43 Mg ha-1) followed by local landraces, Nagaland-2 (1.31 Mg ha-1) and Sikkim-1 (1.25 Mg ha-1). Root traits such as total root length, root volume, average diameter of roots, and root surface area were significantly higher in finger millet landraces Nagaland-1, Nagaland-2, and Sikkim-1 compared to the rest of the millet genotypes. The different millets were found to be rich sources of protein as recorded in foxtail millet cv. SiA 3088 (12.3%), proso millet cv. TNAU 145 (11.5%), and finger millet landraces, Sikkim-1 and Nagaland-2 (8.7% each). Finger millet landrace Sikkim-2 recorded the highest omega-6 content (1.16%), followed by barnyard millet cv. VL 207 (1.09%). Barnyard millet cv. VL 207 recorded the highest polyunsaturated fatty acid (PUFA) content (1.23%), followed by foxtail millet cv. SiA 3088 (1.09%). The local finger millet landraces Sikkim-1 and Sikkim-2 recorded the highest levels of histidine (0.41%) and tryptophan (0.12%), respectively. Sikkim-1 and Nagaland-2 recorded the highest level of thiamine (0.32%) compared to the HYVs. Conclusion: These findings indicate that finger millet has great potential in the organic production system of the North Eastern Himalayan Region (NEHR) of India, and apart from HYVs like VL Mandua 352, local landraces, viz., Nagaland-2 and Sikkim-1, should also be promoted for ensuring food and nutritional security in this fragile ecosystem.

Temperature sensitivity and decomposition rate of 101 leaf litter types from Mediterranean ecosystems.

Litter decomposition is a fundamental process, and the number of published studies has steadily increased in recent decades. However, few experiments have systematically compared a large number of litter types and evaluated their temperature sensitivity. We conducted a two-year experiment on the decomposition of litter bags containing 101 leaf litter sampled in Mediterranean ecosystems and incubated under laboratory conditions at 4 °C, 14 °C, and 24 °C. Litter was chemically characterized and analysed for carbon (C), nitrogen (N), cellulose and lignin concentration, C/N, and lignin/N ratios, which serve as predictors of decomposition rate. The sensitivity of litter decay rate to temperature was evaluated using Q10. Leaf litter chemistry varied widely in nitrogen content (range 0.52-6.80 %), lignin content (range 1.53-49.31 %), C/N ratio (range 5.21-77.78), and lignin/N ratio (range 0.34-34.90). Litter decomposition rate was negatively related to initial lignin concentration, lignin/N ratio, and C/N ratio, but only in the early stage. In the late stages of decomposition, litter decay rate was negatively correlated with initial N concentration but positively correlated with C/N and lignin/N ratios. Temperature sensitivity was negatively correlated with N concentration and positively correlated with lignin and lignin/N ratio. It is noteworthy that, contrary to expectations, temperature sensitivity exhibited a hump-shaped relationship with decay rate. N, C/N, and lignin/N ratios should be used with caution because their predictive power is reversed with respect to decomposition rate during the decomposition process. In addition, the new finding that temperature sensitivity has a hump-shaped relationship with decomposition rate deserves further confirmation and could be considered in ecosystem-level organic C modeling.

Metagenomic, microbiological, chemical and sensory profiling of Caciocavallo Podolico Lucano cheese.

In this study, Caciocavallo Podolico Lucano (CPL) cheese was deeply characterized for its bacterial community, chemical composition and sensory aspects. The entire cheese making process (from milk collection to ripened cheese) was performed by strictly applying the traditional protocol for CPL production in four dairy factories (A-D) representative of the production area. The vat made of wood represents the main transformation tool for CPL cheese production and the biofilms hosted onto the internal surfaces of all vats analyzed in this study were dominated by lactic acid bacteria. Total mesophilic microorganisms present in bulk milk (4.7-5.0 log CFU/ml) increased consistently after contact with the wooden vat surfaces (5.4-6.4 log CFU/ml). The application of Illumina sequencing technology identified barely 18 taxonomic groups among processed samples; streptococci and lactobacilli constituted the major groups of the wooden vat biofilms [94.74-99.70 % of relative abundance (RA)], while lactobacilli dominated almost entirely (94.19-100 % of total RA) the bacterial community of ripened cheeses. Except coagulase positive staphylococci, undesirable bacteria were undetectable. Among chemical parameters, significant variations were registered for unsaturated, monounsaturated, polyunsaturated fatty acids and antioxidant properties (significantly lower for CPL cheeses produced in factory B). The cheeses from factories A, C and D were characterized by a higher lactic acid and persistence smell attributes than factory B. This work indicated that the strict application of CPL cheese making protocol harmonized the main microbiological, physicochemical and sensory parameters of the final cheeses produced in the four factories investigated.

Changes in root chemical diversity along an elevation gradient of Changbai Mountain, China.

Root chemical traits play a critical role in plant resource use strategies and ecosystem nutrient cycling; however, the chemical diversity of multiple elements of fine root and community chemical assembly belowground are poorly understood. Here, we measured 13 elements (C, N, K, Ca, Mg, S, P, Al, Fe, Na, Mn, Zn, and Cu) in the fine roots of 204 plant species along elevational transect from 540 to 2357 m of Changbai Mountain, China to explore the variation, diversity, and community assembly of root chemical traits. At the species level, the concentrations of macronutrients (N, K, Ca, Mg, S, and P) decreased, whereas the trace metals (Fe, Mn, and Zn) increased with elevation. Root chemical traits at the community level systematically shifted along elevational gradients showing a pattern similar to that at the species level, which were mainly influenced by climate and soil rather than species diversity. In general, the interactions of climate and soil were the main drivers of root chemical assembly for woody layers, whereas soil factors played significant role for root chemical assembly for herb layer. The chemical assembly of rock-derived element P was mainly driven by soil factors. Meanwhile, root chemical diversities were mainly regulated by species diversity, the interactions of climate and soil, and soil factors in the tree, shrub, and herb layers, respectively. A better understanding of plant root chemical diversity and community chemical assembly will help to reveal the role of chemical traits in ecosystem functioning.

Different Combinations of Butchery and Vegetable Wastes on Growth Performance, Chemical-Nutritional Characteristics and Oxidative Status of Black Soldier Fly Growing Larvae.

Hermetia illucens larvae (five days old) were farmed on broiler feed (control diet), a vegetable diet (V100), a 50% of vegetable diet + 50% of butchery wastes (V50 + B50), and a 75% of vegetable diet + 25% of butchery wastes (V75 + B25) to evaluate their suitability. Ten kilograms of substrate and 6000 larvae composed each replicate (nine per group). Larvae were weighed and measured every two days until the 25% developed into prepupae. Larval mortality and growing indexes were calculated. Substrates, larvae, and frass chemical composition were analyzed. Larvae oxidative status and stability were measured in hemolymph and body. The V100 larvae showed the lowest live weight, length, thickness, and growth rate but had low mortality rate and high substrate reduction index and protein conversion ratio. The V100 larvae had similar protein to and lower lipids than the control ones, while the V50 + B50 and V75 + B25 larvae contained higher lipids and lower protein than the others. Despite the vegetable wastes, at different levels, the reactive oxygen species content decreased in hemolymph, and the V100 diet depressed growth performance and should be avoided. The use of butchery wastes combined with vegetable ingredients can be a suitable alternative to balance the high level of lipid and the low content of protein.

The Response of Critical Microbial Taxa to Litter Micro-Nutrients and Macro-Chemistry Determined the Agricultural Soil Priming Intensity After Afforestation.

Afforestation with trees and shrubs around cropland can effectively decrease soil degradation and avoid sand storms, but subsequent modification of litter quality accelerates the degradation of native organic matter via the soil priming effect (PE). Although carbon accumulation in agricultural soils after afforestation was widely studied, little is known about the extent to which soil organic carbon (SOC) mineralization is induced by complex residue input in agro-forest-grass composite ecosystems. Here, we mixed corn field soil and litter of afforestation tree and shrub species together in a micro-environment to quantify the effects of litter-mixture input on farmland soil priming associated with afforestation. Additionally, we studied the responses of bacterial and fungal species to litter chemistry, with the aim to identify the litter and microbial driver of soil priming. The results showed that soil priming was accelerated by different litter addition which varied from 24 to 74% of SOC mineralization, suggesting that priming intensity was relatively flexible and highly affected by litter quality. We also find that the macro-chemistry (including litter carbon, nitrogen, lignin, and cellulose) directly affects priming intensity, while micro-chemistry (including litter soluble sugar, water-soluble phenol, methanol-soluble phenol, and condensed tannin) indirectly influences priming via alteration to dominant bacterial taxa. The stepwise regression analysis suggested that litter nitrogen and cellulose were the critical litter drivers to soil priming (r 2 = 0.279), and the combination of bacterial phylum Proteobacteria, Firmicutes, Bacteroidetes, Acidobacteria, and fungal taxa Eurotiomycetes was a great model to explain the priming intensity (r 2 = 0.407).

Influence of Cold Storage on Pear Physico-Chemical Traits and Antioxidant Systems in Relation to Superficial Scald Development.

Superficial scald is the most common physiological disorder in apples and pears and causes huge economic losses worldwide. The aim of this study is to determine the different scald susceptibilities of seven pear cultivars/selections during five months of cold storage (CS). Four advanced pear selections and three commercial cultivars were harvested from an orchard located in Emilia-Romagna region, and cold stored at -1 °C and 85% relative humidity (RH).After 90, 120, and 150 days, fruits of each cultivar and selection were removed for ripening for 4 d, after which scald damage, physico-chemical and nutraceutical traits, and enzymatic antioxidant systems were evaluated on peel and pulp. 'Abbé Fétel', 'Falstaff', and 'CREA 171' did not showed superficial scald symptoms after 90 days, while 'Doyenne du Comice' and 'CREA 264' showed the highest susceptibility. After 90 days, CS 'Falstaff' and 'CREA 179' showed the highest total polyphenol content (TPH) in peel, followed by 'Doyenne du Comice' and 'AbbéFétel'; lowest TPH was detected in 'CREA 264'. After 120 and 150 days of CS, 'Abbé Fétel' and 'CREA 171' showed the highest peel TPH. 'CREA 264' and 'CREA 125' reached the lowest values of TPH during the three CS time periods. Superoxide dismutase and catalase activities were higher in the peel of scald-resistant than that in scald-susceptible pear cultivars/advanced selections. Superficial scald induced an increase in polyphenoloxidase, and guaiacol peroxidase activities involved in fruit-browning during CS. Furthermore, we observed an increase in lipoxygenase activity and consequent membrane damage in both the peel and flesh of the fruit. This study indicates that pear cultivars/advanced selections have different superficial scald susceptibilities that enable them to induce the activity of several antioxidant enzymes, following CS.

Disentangling dimensions of phytochemical diversity: alpha and beta have contrasting effects on an insect herbivore.

Phytochemical diversity is comprised of two main dimensions-the average (alpha) within-plant neighbors or the difference (beta) in the composition of chemicals between plant neighbors. Research, however, has primarily examined the consequences of phytochemical diversity on herbivore performance through a single dimension, even though diversity is multidimensional. Furthermore, the ecological role of phytochemical diversity is not well understood because each of these dimensions exhibits unique biological effects on herbivore performance. Therefore, it has been difficult to tease apart the relative importance of alpha and beta chemical diversities on plant-herbivore interactions. We experimentally manipulated alpha and beta diversities along a chemical gradient to disentangle the relative effects of these dimensions on the performance of a mobile generalist herbivore, Trichoplusia ni (Hübner), using 16 genotypes from the Solanum pennellii introgression lines. First, we found contrasting effects of alpha and beta diversities on herbivore performance. Second, when comparing diversity across and within chemical classes, herbivore performance was reduced when plant neighbors had greater diversity within chemical classes that are biologically inhibiting at higher quantities (i.e., quantitative defenses such as phenolics and acyl sugars). However, herbivore performance was enhanced when plant neighbors had higher levels of chemical classes that are biologically toxic (i.e., qualitative defenses such as alkaloids). Finally, herbivores performed better on plant dicultures compared to monocultures, and performance was positively associated with plant dicultures only when there were high levels of average alpha diversity within plant neighbors. Our results suggest T. ni generalist caterpillars do better when plant neighbors are chemically different because differences provide options for them to choose or to switch between plants to balance chemical uptake. Overall, herbivores interact with a large diversity of plant chemicals at multiple scales, and our results indicate that not all chemical diversity is equal: specific dimensions of phytochemical diversity have unique effects on the dynamics of herbivore performance.

Assessment of physico-chemical traits related to eating quality of young dairy bull beef at different ageing times using Raman spectroscopy and chemometrics.

Raman spectroscopy and chemometrics were investigated for the prediction of eating quality related physico-chemical traits of Holstein-Friesian bull beef. Raman spectra were collected on the 3rd, 7th and 14th days post-mortem. A frequency range of 1300-2800cm-1 was used for partial least squares (PLS) modelling. PLS regression (PLSR) models for the prediction of WBSF and cook loss achieved an R2CV of 0.75 with RMSECV of 6.82 N and an R2CV of 0.77 with RMSECV of 0.97%w/w respectively. For the prediction of intramuscular fat, moisture and crude protein content, R2CV values were 0.85, 0.91 and 0.70 with RMSECV of 0.52%w/w, 0.39%w/w and 0.38%w/w respectively. An R2CV of 0.79 was achieved for the prediction of both total collagen and hydroxyproline content, while for collagen solubility the R2CV was 0.88. All samples (100%) from 15- and 19-month old bulls were correctly classified using PLS discriminant analysis (PLS-DA), while 86.7% of samples from different muscles (longissimus thoracis, semitendinosus and gluteus medius) were correctly classified. In general, PLSR models using Raman spectra on the 3rd day post-mortem had better prediction performance than those on the 7th and 14th days. Raman spectroscopy and chemometrics have potential to assess several beef physical and chemical quality traits.