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      • How insect herbivores hijack plant chemical defenses for their own defense against natural enemies

        F. Beran,Z.-L. Yang,T. Sporer 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10

        Many herbivorous insects sequester plant defense compounds from their host plants to protect themselves from natural enemies. In plants, these defense compounds are often stored as protoxins separated from their activating enzymes. A well-known example is the glucosinolate-myrosinase defense system in plants of the order Brassicales. When plant tissue is ingested by herbivores, glucosinolates are hydrolyzed by the enzyme myrosinase to form highly reactive isothiocyanates. We previously reported that flea beetles of the genus Phyllotreta selectively sequester high amounts of glucosinolates from their crucifer host plants, and convergently evolved their own myrosinase which enables them to utilize sequestered glucosinolates for their own purposes (Beran et al., 2014). The presence of intact glucosinolates in these beetles suggests that despite tissue damage, ingested glucosinolates are not activated by the plant myrosinase. Rapid and efficient glucosinolate uptake from the gut lumen into gut epithelial cells can prevent hydrolysis and thus might be crucial to overcome this activated plant defense. We use the horseradish flea beetle Phyllotreta armoraciae as a model to study the molecular basis of sequestration in insects. In short-term feeding experiments, we showed that ingested glucosinolates are rapidly sequestered in the foregut. To identify the transporters that mediate glucosinolate import from the foregut lumen into gut epithelial cells, we focused on the MFS transporter family, which is known to transport a wide range of substrates. A phylogenetic analysis of putative MFS transporter sequences identified in P. armoraciae and other beetles revealed several specifically expanded clades in P. armoraciae. Out of 21 candidate genes that were heterologously expressed in Sf9 cells, nine showed glucosinolate transport activity in vitro. Interestingly, most candidate genes were exclusively expressed in the malpighian tubules, and two genes were additionally expressed in the foregut. We currently elucidate the function of these transporters in glucosinolate sequestration in vivo using RNAi. To better understand the function of sequestered glucosinolates, we performed bioassays with P. armoraciae larvae and the generalist predatory ladybird Harmonia axyridis. Upon predator attack, P. armoraciae larvae emitted high amounts of isothiocyanates and ladybird larvae stopped feeding within a few seconds and were highly irritated. However, silencing myrosinase gene expression in P. armoraciae larvae led to increased mortality compared to control larvae in survival assays with ladybird larvae. Our results demonstrate how Phyllotreta use plant defense metabolites to defend themselves against predators.

      • KCI등재

        Antimicrobial polyhydroxybutyrate submicron fiber mat loaded with extract of Hypericum perforatum

        Milos Beran,Ales Horna,Viktor Vorisek,Eliska Berkova,Radka Korinkova,Vojtech Trousil,Marketa Hrubanova 한국식물생명공학회 2022 JOURNAL OF PLANT BIOTECHNOLOGY Vol.49 No.3

        The aim of this work was to prepare a new biodegradable polyhydroxybutyrate (PHB) submicron fiber mat loaded with hypericin-rich Hypericum perforatum raw extract by centrifugal spinning technology, an alternative approach to the traditional method of electrospinning to fabricate nanofibers or microfibers from solutions at high speed and low cost. Hypericins in methanol/acetone extract of H. perforatum were determined by UHPLC-MS/MS and HPLC/PDA. Submicron fiber mats composed of pure PHB or PHB enriched with H. perforatum extract were prepared using a pilot plant demonstrator for the centrifugal spinning technology and characterized by SEM. Singlet oxygen production was quantified by the 1,3-diphenylisobenzofuran (DPIBF) method in hexane. The results proved a significant production of singlet oxygen by the prepared submicron fiber mat. We also found a significant antibacterial activity against the bacterial strain Escherichia coli CCM 5417 by a method in accordance with JIS Z 2801/ISO 22196 standards. The H. perforatum extract-enriched PHB submicron fiber mats showed potential for the development of self-cleaning and antimicrobial air filters.

      • KCI등재
      • The cabbage stem flea beetle uses several detoxification strategies to overcome the glucosinolate-myrosinase defense system in crucifers

        F. BERAN,T. Sporer,S.-J. Ahn,F. Betzin,C. Paetz,G. Kunert,M. Reichelt 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10

        Many insects are able to feed on crucifers despite the presence of a potent activated defense system known as the mustard oil bomb. In damaged tissue, mustard oil glucosides (glucosinolates) are hydrolyzed by the enzyme myrosinase to form toxic mustard oils (isothiocyanates). Here, we analyzed how the the cabbage stem flea beetle Psylliodes chrysocephala, a key pest of oilseed rape, copes with this chemical defense. First, we found that P. chrysocephala prevents the activation of ingested glucosinolates by two different strategies, a) by sequestering glucosinolates and b) by converting glucosinolates to desulfo-glucosinolates. Our next aim was to identify the sulfatase enzyme(s) responsible for the detoxification of glucosinolates in P. chrysocephala. Nine arylsulfatase-like genes were identified in the transcriptome of P. chrysocephala, and five of them showed glucosinolate sulfatase activity upon heterologous expression in Sf9 cells. By using RNAi, we confirmed that PcGSS1 and PcGSS2 are active towards benzenic and indolic glucosinolates in P. chrysocephala adults in vivo. However, in feeding experiments, the proportion of sequestered and desulfated glucosinolates ranged from 26 to 35% which suggests that these strategies alone are likely not sufficient to overcome the chemical plant defense. Indeed, P. chrysocephala additionally conjugates isothiocyanates to glutathione and metabolizes them via the conserved mercapturic acid pathway. In summary, the cabbage stem flea beetle avoids isothiocyanate formation by specialized strategies (sequestration and desulfation), but also relies on a conserved detoxification pathway to prevent toxicity of isothiocyanates.

      • KCI등재

        Effect of Composition on the Matrix Transformation of the Co-Re-Cr-Ta-C Alloys

        Přemysl Beran,Debashis Mukherji,Pavel Strunz,Ralph Gilles,Michael Hofmann,Lukas Karge,Oleksandr Dolotko,Joachim Rösler 대한금속·재료학회 2016 METALS AND MATERIALS International Vol.22 No.4

        Neutron diffraction measurement was performed in-situ at high temperatures on Co-Re-Ta-C alloys with and without Cr addition. This included alloys containing different C content with the C/Ta ratio varying between 0.5 and 1.0. The Co-Re-solid solution matrix of the experimental alloys is polymorphic (like in pure cobalt) and transformed from low temperature hexagonal ε phase to high temperature cubic γ phase on heating. This transformation is reversible and show hysteresis. The main alloying addition, Re, stabilizes the ε Co-phase and increases the transformation temperature to above 1273 K. The onset of the ε γ transformation during heating and cooling was found to differ depending on the alloy composition. In alloys without Cr addition the transformation was not completed on cooling and the high temperature γ phase was partly retained at room temperature in metastable state with the amount depending on the cooling rate from high temperature. The diffraction and microstructural results showed that Cr is ε stabilizer (similar as Re) but the role of Ta is not clear. The C/Ta ratio has no direct effect on the matrix phase transformation. Nevertheless, it influences indirectly by determining the amount of Ta which is freely available in the matrix.

      • KCI등재

        Identification of tumor necrosis factor-α levels around miniscrews during canine distalization

        Filiz Acun Kaya,Nihal Hamamcı,Ersin Uysal,Beran Yoku? 대한치과교정학회 2011 대한치과교정학회지 Vol.41 No.1

        Objective: The aim of this study was to measure tumor necrosis factor-α (TNF-α) levels around miniscrews used for anchorage during a 3-month period of canine distalization. Methods: Sixteen patients (8 boys, 8 girls; mean age, 16.6 ± 2.4 years) whose upper first premolars were extracted for orthodontic treatment were included in this study. Miniscrews were used as an anchorage unit in canine distalization. Thirty-two (32) miniscrew implants were placed bilaterally in the alveolar bone between the maxillary second premolars and first molars. The treatment, miniscrew, and control groups comprised upper canines, miniscrew implants, and upper first premolars, respectively. Peri-miniscrew implant crevicular fluid and gingival crevicular fluid were obtained before applying force and at 1, 24, and 48 hours, and at 7 and 21 days, and 3 months after applying force. Results: During the 3-month period, the TNF-α levels increased significantly at 24 hours only in the treatment group (p < 0.01). In the miniscrew and control groups, there were no statistically significant changes. No significant differences were observed between groups. Conclusions: Miniscrews can be conveniently used for anchorage in orthodontics. (Korean J Orthod 2011;41(1):36-41)

      • KCI등재

        Matrix Transformation in Boron Containing High-Temperature Co–Re–Cr Alloys

        Pavel Strunz,Debashis Mukherji,Přemysl Beran,Ralph Gilles,Lukas Karge,Michael Hofmann,Markus Hoelzel,Joachim Rösler,Gergely Farkas 대한금속·재료학회 2018 METALS AND MATERIALS International Vol.24 No.5

        An addition of boron largely increases the ductility in polycrystalline high-temperature Co–Re alloys. Therefore, the effect ofboron on the alloy structural characteristics is of high importance for the stability of the matrix at operational temperatures. Volume fractions of ε (hexagonal close-packed—hcp), γ (face-centered cubic—fcc) and σ (Cr 2 Re 3 type) phases were measuredat ambient and high temperatures (up to 1500 °C) for a boron-containing Co–17Re–23Cr alloy using neutron diff raction. The matrix phase undergoes an allotropic transformation from ε to γ structure at high temperatures, similar to pure cobaltand to the previously investigated, more complex Co–17Re–23Cr–1.2Ta–2.6C alloy. It was determined in this study that thetransformation temperature depends on the boron content (0–1000 wt. ppm). Nevertheless, the transformation temperaturedid not change monotonically with the increase in the boron content but reached a minimum at approximately 200 ppm ofboron. A probable reason is the interplay between the amount of boron in the matrix and the amount of σ phase, which bindshcp-stabilizing elements (Cr and Re). Moreover, borides were identifi ed in alloys with high boron content.

      • SCOPUSSCIEKCI등재

        Identification of tumor necrosis factor-${\alpha}$ levels around miniscrews during canine distalization

        Kaya, Filiz Acun,Hamamcl, Nihal,Uysal, Ersin,Yokus, Beran The Korean Association Of Orthodontists 2011 대한치과교정학회지 Vol.41 No.1

        Objective: The aim of this study was to measure tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) levels around miniscrews used for anchorage during a 3-month period of canine distalization. Methods: Sixteen patients (8 boys, 8 girls; mean age, $16.6{\pm}2.4$ years) whose upper first premolars were extracted for orthodontic treatment were included in this study. Miniscrews were used as an anchorage unit in canine distalization. Thirty-two (32) miniscrew implants were placed bilaterally in the alveolar bone between the maxillary second premolars and first molars. The treatment, miniscrew, and control groups comprised upper canines, miniscrew implants, and upper first premolars, respectively. Peri-miniscrew implant crevicular fluid and gingival crevicular fluid were obtained before applying force and at 1, 24, and 48 hours, and at 7 and 21 days, and 3 months after applying force. Results: During the 3-month period, the (TNF-${\alpha}$) levels increased significantly at 24 hours only in the treatment group (p < 0.01). In the miniscrew and control groups, there were no statistically Significant changes. No significant differences were observed between groups. Conclusions: Miniscrews can be conveniently used for anchorage in orthodontics.

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