Bacteria Associated with Acute Oak Decline: Where Did They Come From? We Know Where They Go.

Acute oak decline is a high-impact disease causing necrotic lesions on the trunk, crown thinning and the eventual death of oak. Four bacterial species are associated with the lesions-Brenneria goodwinii, Gibbsiella quercinecans, Rahnella victoriana and Lonsdalea Britannica-although an epi-/endophytic lifestyle has also been suggested for these bacteria. However, little is known about their environmental reservoirs or their pathway to endophytic colonisation. This work aimed to investigate the ability of the four AOD-associated bacterial species to survive for prolonged periods within rhizosphere soil, leaves and acorns in vitro, and to design an appropriate method for their recovery. This method was trialled on field samples related to healthy and symptomatic oaks. The in vitro study showed that the majority of these species could survive for at least six weeks within each sample type. Results from the field samples demonstrated that R. victoriana and G. quercinecans appear environmentally widespread, indicating multiple routes of endophytic colonisation might be plausible. B. goodwinii and L. britannica were only identified from acorns from healthy and symptomatic trees, indicating they may be inherited members of the endophytic seed microbiome and, despite their ability to survive outside of the host, their environmental occurrence is limited. Future research should focus on preventative measures targeting the abiotic factors of AOD, how endophytic bacteria shift to a pathogenic cycle and the identification of resilient seed stock that is less susceptible to AOD.

Influence of soil chemical composition on U, 226Ra and 210Pb uptake in leaves and fruits of Quercus ilex L.

To determine their transfer factors, activity concentrations of natural radionuclides were measured in the leaves and acorns of holm oak (Quercus ilex L.) trees collected from seven locations with different soil properties and radionuclide activity concentrations. The chemical and mineralogical compositions of the soils were also analysed to investigate the effect these had on radionuclide absorption by the trees. Soil chemistry showed significant effects on radionuclide incorporation into Quercus ilex L. tissues. A significant relationship was established between activity concentrations and soil content of Ca and P with 238U and 226Ra in the leaves and acorns of Quercus ilex L. Differentiated transfer was found for 40K, which showed greater transfer to the leaves than the other radionuclides. The activity concentration of U and 226Ra was higher in the fruits than in the leaves, with the opposite effect being observed for 40K. The risk of U and 226Ra transfer into the food chain through acorn consumption by livestock is predicted to increase in soils poor in Ca and rich in P.

Traits variation of acorns and cupules during maturation process in Quercus variabilis and Quercus aliena.

Quercus variabilis and Quercus aliena are two native tree species in China, which have similar habitats, and their regeneration mainly depends on acorn dispersal. This study analyzed the contents of water, soluble sugar, starch, soluble protein, and total phenolics in acorns and cupules during the whole development process to explore the difference between species. Thereinto, starch and total phenol occupied the dominant roles as their high contents. The acorn starch contents increased sharply during development in both species, but the contents in Q. variabilis were almost twice those of Q. aliena when mature. Similarly, high expression levels of starch synthase, soluble starch synthase 2 (SSS2) were also found in the acorns of Q. variabilis. The total phenol contents in Q. variabilis acorns were high at the early stages, and decreased sharply to similar contents in Q. aliena when mature. Additionally, the cupules in Q. variabilis had high contents of total phenols during the whole development period. Similar trends were also found in the expression patterns of UGT84A13 and SDH. The high total phenols in acorns and cupules of Q. variabilis probably protect the acorns from Mechoris ursulus, as only Q. aliena suffered a severe pest infestation in the early development stages. This study not only clarifies the interspecific difference between storage and defense substances during the development process in acorns and cupules, but also deepens understanding the specialized mechanisms of plant-pest/animal interactions in Quercus.

Assessing how well students understand the molecular basis of evolution by natural selection.

Researchers have called for undergraduate courses to update teaching frameworks based on the Modern Synthesis with insights from molecular biology, by stressing the molecular underpinnings of variation and adaptation. To support this goal, we developed a modified version of the widely used Assessing Conceptual Reasoning of Natural Selection (ACORNS) instrument. The expanded tool, called the E-ACORNS, is explicitly designed to test student understanding of the connections among genotypes, phenotypes, and fitness. E-ACORNS comprises a slight modification to the ACORNS open-response prompts and a new scoring rubric. The rubric is based on five core concepts in evolution by natural selection, with each concept broken into elements at the novice, intermediate, and expert-level understanding. Initial tests of the E-ACORNS showed that (1) upper-level undergraduates can score responses reliably and quickly, and (2) students who were just starting an introductory biology series for majors do not yet grasp the molecular basis of phenotypic variation and its connection to fitness.

Authentication of the Montanera Period on Carcasses of Iberian Pigs by Using Analytical Techniques and Chemometric Analyses.

The potential of two complementary analytical techniques (near infrared spectroscopy, NIRS and gas chromatography-ion mobility spectrometry, GC-IMS) was used to establish the time that Iberian pigs have been fed on acorns and pasture and to verify their genetic purity. For both techniques it was neither necessary to carry out any chemical treatment in advance nor to identify individual compounds. The results showed that both the NIR spectrum and the spectral fingerprint obtained by GC-IMS were affected by the time that the Iberian pig feeds on natural resources. High percentages of correct classification were achieved in the calibration for both techniques: >98% for the days of montanera and >96% for the breed by NIRS and >99% for the days of montanera and >98% for the breed by GC-IMS. The results obtained showed that NIR spectra taken from intact samples is a quick classification method according to the time of montanera and breed.

Simultaneous determination of selected catechins and pyrogallol in deer intoxications by HPLC-MS/MS.

A selective and sensitive analytical method for the determination of selected catechins (catechin, epicatechin, gallocatechin, epigallocatechin) and pyrogallol in biological matrices by HPLC-MS/MS was developed. The utilized sample preparation technique was a two-stage liquid-liquid extraction using ethyl acetate. The HPLC-system was equipped with a Phenomenex Luna Pentafluorophenyl Column (150 × 2 mm, 5 µm) and operated with an acetonitrile-water gradient as a mobile phase system. Detection was performed with a 3200 Q Trap mass spectrometer. For analysis the mass spectrometer was used in the MRM-mode with negative ionization. The method validation was performed with serum as matrix. The selectivity of the method as well as the linearity of calibration was successfully proven for all analytes. The limits of quantification were between 5.3 and 11.2 ng/mL and the recovery rates were above 50 % for all analytes. Results from the samples of three deer poisoning cases demonstrated that the developed HPLC-MS/MS method is applicable to real biological samples.

The Quality and Health-Promoting Value of Meat from Pigs of the Native Breed as the Effect of Extensive Feeding with Acorns.

The popularity of meat from animals of native breeds is growing all over the world, due to consumer belief regarding its higher quality compared to meat from industrial farm animals. In addition, the living conditions (welfare) are of great importance for consumers. We observed the effect of different ways of keeping and feeding pigs of the same conservative breed on the quality of meat and its health benefits. The aim of the study was to compare the meat quality from pigs of the native Zlotnicka Spotted breed, fattened intensively or extensively (with conventional farm-produced compound feed and acorns). The meat from free-range pigs extensively fed on silage and small amounts of acorns was characterized by a higher content of fat, which acts as a carrier for flavor and juiciness, as well as higher monounsaturated fatty acid (MUFA) content (p ≤ 0.05) and lower atherogenic, thrombogenic, and peroxidability indices (p ≤ 0.05). It may be stated that the meat quality of the native pig breed is significantly dependent on the housing and feeding method. A more beneficial effect on the quality of meat and its dietetic value, as well as its susceptibility to rancidity, can be obtained throughextensive pig feeding with roughage and the addition of acorns.

Phytochemical composition and variability in Quercus ilex acorn morphotypes as determined by NIRS and MS-based approaches.

Quercus acorns have been used as a staple food since archeological times; currently, there is a renewed interest in the direct use of the acorns as nuts or as a source of consumable flour. In this regard, the phytochemical composition and variability of various morphotypes of acorns from Quercus ilex have been determined by using two platforms, near infrared spectroscopy (NIRS) and mass spectrometry (MS)-based metabolomics. According to NIRS, starch was the most abundant compound (50-60%). UHPLC-QToF analysis was used for untargeted metabolomic analysis and 192 metabolites were annotated. Principal component analysis (PCA) discriminated the morphotypes; 50 compounds out of 192 identified had the highest load over the first two PCA components (explaining 67.2% variability) and can be potential markers of variability. The data presented here support the value of acorns as a source of nutritionally rich compounds thus justifying their use a good alternative to other nuts.

Dissecting the Seed Maturation and Germination Processes in the Non-Orthodox Quercus ilex Species Based on Protein Signatures as Revealed by 2-DE Coupled to MALDI-TOF/TOF Proteomics Strategy.

Unlike orthodox species, seed recalcitrance is poorly understood, especially at the molecular level. In this regard, seed maturation and germination were studied in the non-orthodox Quercus ilex by using a proteomics strategy based on two-dimensional gel electrophoresis coupled to matrix-assisted laser desorption ionization/time of flight (2-DE-MALDI-TOF).Cotyledons and embryo/radicle were sampled at different developmental stages, including early (M1-M3), middle (M4-M7), and late (M8-M9) seed maturation, and early (G1-G3) and late (G4-G5) germination. Samples corresponding to non-germinating, inviable, seeds were also included. Protein extracts were subjected to 2-dimensional gel electrophoresis (2-DE) and changes in the protein profiles were analyzed. Identified variable proteins were grouped according to their function, being the energy, carbohydrate, lipid, and amino acid metabolisms, together with protein fate, redox homeostasis, and response to stress are the most represented groups. Beyond the visual aspect, morphometry, weight, and water content, each stage had a specific protein signature. Clear tendencies for the different protein groups throughout the maturation and germination stages were observed for, respectively, cotyledon and the embryo axis. Proteins related to metabolism, translation, legumins, proteases, proteasome, and those stress related were less abundant in non-germinating seeds, it related to the loss of viability. Cotyledons were enriched with reserve proteins and protein-degrading enzymes, while the embryo axis was enriched with proteins of cell defense and rescue, including heat-shock proteins (HSPs) and antioxidants. The peaks of enzyme proteins occurred at the middle stages (M6-M7) in cotyledons and at late ones (M8-M9) in the embryo axis. Unlike orthodox seeds, proteins associated with glycolysis, tricarboxylic acid cycle, carbohydrate, amino acid and lipid metabolism are present at high levels in the mature seed and were maintained throughout the germination stages. The lack of desiccation tolerance in Q. ilex seeds may be associated with the repression of some genes, late embryogenesis abundant proteins being one of the candidates.

Acorn size and tolerance to seed predators: the multiple roles of acorns as food for seed predators, fruit for dispersal and fuel for growth.

Fitness of parents and offspring is affected by offspring size. In oaks (Quercus spp.), acorns vary considerably in size across, and within, species. Seed size influences dispersal and establishment of oaks, but it is not known whether size imparts tolerance to seed predators. Here, we examine the relative extent to which cotyledon size serves as both a means for sustaining partial consumption and energy reserves for developing seedlings during early stages of establishment. Acorns of 6 oak species were damaged to simulate acorn predation by vertebrate and invertebrate seed predators. Seedling germination/emergence and growth rates were used to assess seedling performance. We predicted that if cotyledons are important for dispersal, acorns should show tolerance to partial seed consumption. Alternatively, if the cotyledon functions primarily as an energy reserve, damage should significantly influence seedling performance. Acorns of each species germinated and produced seedlings even after removing >50% of the cotyledon. Seed mass explained only some of the variation in performance. Within species, larger acorns performed better than smaller acorns when damaged. Undamaged acorns performed as well or better than damaged acorns. There was no pattern among individual species with increasing amounts of damage. In some species, simulated invertebrate damage resulted in the poorest performance, suggesting alternative strategies of oaks to sustain damage. Large cotyledons in acorns may be important for attracting seed dispersers and sustaining partial damage, while also providing energy to young seedlings. Success of oak establishment may follow from the resilience of acorns to sustain damage at an early stage.

Anti-Inflammatory Oleanolic Triterpenes from Chinese Acorns.

Acorns play an important role in human history and are a source of food and recipes for many cultures around the world. In this study, eleven oleanolic triterpenes, one of which was novel, were isolated from Chinese acorns (Quercus serrata var. brevipetiolata). The chemical structure of the novel triterpene, which was identified as 2α,3ß,19α-trihydroxy-24-oxo-olean-12-en-28-oic acid (1), was established based on the interpretation of chemical and spectroscopic analyses, including IR, HR-ESI-MS, and NMR experiments (¹H, (13)C NMR, DEPT, ¹H-¹H COSY, HSQC, HMBC, and NOESY). All isolated compounds were tested for their inhibitory effects on LPS-induced nitric oxide (NO) production in RAW 264.7 macrophages. Compared with the positive control drug indomethacin (IC50 = 47.4 µM), compounds 1, 3, 6 and 8 exhibited remarkable anti-inflammatory activities with IC50 values of 5.4, 7.8, 4.0 and 8.9 µM, respectively. Besides, compounds 2, 4, 7 and 9 also showed moderate anti-inflammatory activities with IC50 values of 10.1, 13.0, 20.1 and 17.2 µM, respectively. Furthermore, Compound 1 could inhibit TNF-α-induced IL-6 and IL-8 production in MH7A cells.

Seventy-five years of masting and rodent population peaks in Norway: Why do wood mice not follow the rules?

Wood mouse (Apodemus sylvaticus) populations are expected to show a peak in autumn in the year after a mast year of sessile oak (Quercus petraea), because stored acorns increase winter survival. In Aust-Agder, South Norway, only 16 of 34 mast years from 1939-2014 were followed by a year with a peak in the wood mouse population. For many of the remaining instances, there rather was a minor peak 2 or 3 years after the mast. In multiple logistic regression models, the probability of a wood mouse population peak after a mast year of sessile oak was positively related to a snow-corrected temperature index of the previous winter and negatively to a small rodent population index of the previous autumn. The present study thus supports the hypothesis that longer periods with snow-free ground and subzero temperatures negatively affect wood mouse winter survival. Because it may be difficult for wood mice to survive on a diet consisting of acorns alone, the negative relationship with the rodent population index of the previous year is most likely caused by an over-exploitation of necessary alternative food resources, such as other plant seeds and arthropods. Stored acorns not utilized during one winter are assumed to benefit wood mice in a succeeding winter, giving a delayed population peak relative to the mast year.

A New Age for Quercus spp. Fruits: Review on Nutritional and Phytochemical Composition and Related Biological Activities of Acorns.

The current global food system must adapt to the expected growth of world population (about 9 billion individuals by 2050). This adaptation will probably include an increased consumption of edible wild foods, due to their richness in micronutrients and bioactive compounds, besides providing a cost-effective and sustainable way of improving caloric food security. A striking example of such natural matrices is the Quercus genus, which has the additional advantage of being widespread throughout the Northern Hemisphere. In a traditional sense, Quercus fruits (acorns) were mainly used in animal feeding, despite their potentially important role on the rural economy. But this preconception is changing. In fact, their nutritional value, high contents in phytochemical compounds, biological activity (such as antioxidant, anticarcinogenic, and cardioprotective properties) and use in the treatment of specific diseases (such as atherosclerosis, diabetes, or Alzheimer's disease) have raised the interest in integrating acorns into the human diet. Accordingly, this comprehensive overview was designed to provide an evidence-based review of the literature, with the objective to achieve useful conclusions regarding the nutritional properties, methodologies of extraction, identification, and characterization of a wide variety of bioactive compounds and scientifically validated bioactivities in Quercus species worldwide. The industrial by-products from acorn oil extraction or flour production are also included. Data regarding the analytical techniques, individual compounds, and their bioactivities, are organized in tables. The reported data are discussed and directions for further investigations are suggested, highlighting the use of acorns in food, nutraceutical, and pharmaceutical applications.

Adsorption of Disperse Orange 30 dye onto activated carbon derived from Holm Oak (Quercus Ilex) acorns: A 3(k) factorial design and analysis.

In this study, samples of activated carbon were prepared from Holm Oak acorns by chemical activation with H3PO4, ZnCl2 and KOH as activating agents. The samples were characterized by SEM, BET, FTIR and elemental analysis, and were then evaluated for the removal of Disperse Orange 30 (DO30) dyes from aqueous solutions. A 3(k) factorial design was used to determine the interaction effects of carbonization temperature, pH, dosage of adsorbent and type of activating agent on the amount of dye removal. Also, level of effectiveness factors were determined by conducting regression models for maximum adsorption efficiency. Of all the samples, the sample generated using ZnCl2 as an activating agent showed a maximum dye removal efficiency of 93.5% at a carbonization temperature of 750 °C, a pH of 2 and an adsorbent dosage of 0.15 g/25 ml. The analysis shows that the adsorption process depends significantly on the type of activating agent used in the preparation of activated carbon.