TY - JOUR A1 - Yan, Wenhao A1 - Chen, Dijun A1 - Kaufmann, Kerstin T1 - Efficient multiplex mutagenesis by RNA-guided Cas9 and its use in the characterization of regulatory elements in the AGAMOUS gene JF - Plant Methods N2 - Results: Here, an RNA-guided Cas9 system was optimized to enable efficient multiplex editing in Arabidopsis thaliana. We demonstrate the flexibility of our system for knockout of multiple genes, and to generate heritable large-fragment deletions in the genome. As a proof of concept, the function of part of the second intron of the flower development gene AGAMOUS in Arabidopsis was studied by generating a Cas9-free mutant plant line in which part of this intron was removed from the genome. Further analysis revealed that deletion of this intron fragment results 40 % decrease of AGAMOUS gene expression without changing the splicing of the gene which indicates that this regulatory region functions as an activator of AGAMOUS gene expression. Conclusions: Our modified RNA-guided Cas9 system offers a versatile tool for the functional dissection of coding and non-coding DNA sequences in plants. KW - RNA-guided Cas9 KW - Multiplex mutagenesis KW - Large fragment deletion KW - Germline transmission Y1 - 2016 U6 - https://doi.org/10.1186/s13007-016-0125-7 SN - 1746-4811 VL - 12 SP - 2381 EP - 2389 PB - BioMed Central CY - London ER - TY - JOUR A1 - Yan, Wenhao A1 - Chen, Dijun A1 - Schumacher, Julia A1 - Durantini, Diego A1 - Engelhorn, Julia A1 - Chen, Ming A1 - Carles, Cristel C. A1 - Kaufmann, Kerstin T1 - Dynamic control of enhancer activity drives stage-specific gene expression during flower morphogenesis JF - Nature Communications N2 - Enhancers are critical for developmental stage-specific gene expression, but their dynamic regulation in plants remains poorly understood. Here we compare genome-wide localization of H3K27ac, chromatin accessibility and transcriptomic changes during flower development in Arabidopsis. H3K27ac prevalently marks promoter-proximal regions, suggesting that H3K27ac is not a hallmark for enhancers in Arabidopsis. We provide computational and experimental evidence to confirm that distal DNase. hypersensitive sites are predictive of enhancers. The predicted enhancers are highly stage-specific across flower development, significantly associated with SNPs for flowering-related phenotypes, and conserved across crucifer species. Through the integration of genome-wide transcription factor (TF) binding datasets, we find that floral master regulators and stage-specific TFs are largely enriched at developmentally dynamic enhancers. Finally, we show that enhancer clusters and intronic enhancers significantly associate with stage-specific gene regulation by floral master TFs. Our study provides insights into the functional flexibility of enhancers during plant development, as well as hints to annotate plant enhancers. Y1 - 2019 U6 - https://doi.org/10.1038/s41467-019-09513-2 SN - 2041-1723 VL - 10 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Yang, Lei A1 - Perrera, Valentina A1 - Saplaoura, Eleftheria A1 - Apelt, Federico A1 - Bahin, Mathieu A1 - Kramdi, Amira A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Sokolowska, Ewelina A1 - Zhang, Wenna A1 - Li, Runsheng A1 - Pitzalis, Nicolas A1 - Heinlein, Manfred A1 - Zhang, Shoudong A1 - Genovesio, Auguste A1 - Colot, Vincent A1 - Kragler, Friedrich T1 - m(5)C Methylation Guides Systemic Transport of Messenger RNA over Graft Junctions in Plants JF - Current biology N2 - In plants, transcripts move to distant body parts to potentially act as systemic signals regulating development and growth. Thousands of messenger RNAs (mRNAs) are transported across graft junctions via the phloem to distinct plant parts. Little is known regarding features, structural motifs, and potential base modifications of transported transcripts and how these may affect their mobility. We identified Arabidopsis thalianam RNAs harboring the modified base 5-methylcytosine (m(5)C) and found that these are significantly enriched in mRNAs previously described as mobile, moving over graft junctions to distinct plant parts. We confirm this finding with graft-mobile methylated mRNAs TRANSLATIONALLY CONTROLLED TUMOR PROTEIN 1 (TCTP1) and HEAT SHOCK COGNATE PROTEIN 70.1 (HSC70.1), whose mRNA transport is diminished in mutants deficient in m(5)C mRNA methylation. Together, our results point toward an essential role of cytosine methylation in systemic mRNA mobility in plants and that TCTP1 mRNA mobility is required for its signaling function. Y1 - 2019 U6 - https://doi.org/10.1016/j.cub.2019.06.042 SN - 0960-9822 SN - 1879-0445 VL - 29 IS - 15 SP - 2465 EP - 2476.e5 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Yannelli, Florencia A. A1 - Karrer, Gerhard A1 - Hall, Rea A1 - Kollmann, Johannes A1 - Heger, Tina T1 - Seed density is more effective than multi-trait limiting similarity in controlling grassland resistance against plant invasions in mesocosms JF - Applied vegetation science : official organ of the International Association for Vegetation Science N2 - QuestionDisturbed areas offer great opportunities for restoring native biodiversity, but they are also prone to invasion by alien plants. Following the limiting similarity hypothesis, we address the question of whether or not similarity of plant functional traits helps developing seed mixtures of native communities with high resistance to invasive species at an early stage of restoration. LocationCentre of Greenhouses and Laboratories Durnast, Technische Universitat Munchen, Freising, Germany. MethodsUsing a system of linear equations, we designed native communities maximizing the similarity between the native and two invasive species according to ten functional traits. We used native grassland plants, two invasive alien species that are often problematic in disturbed areas (i.e., Ambrosia artemisiifolia and Solidago gigantea) and trait information obtained from databases. The two communities were then tested for resistance against establishment of the two invaders separately in a greenhouse experiment. We measured height of the invasive species and above-ground biomass, along with leaf area index, 4 and 8months after sowing respectively. ResultsBoth invasive species were successfully reduced by the native community designed to suppress S. gigantea dominated by small-seeded species. These results could be considered as partial support for the limiting similarity hypothesis. However, given the success of this mixture against both invasive species, suppression was better explained by a seed density effect resulting from the smaller seed mass of the native species included in this mixture. Further, the dominance of a fast-developing competitive species could also contribute to its success. ConclusionsThere was no unequivocal support for the limiting similarity hypothesis in terms of the traits selected. Instead we found that increasing seeding density of native species and selecting species with a fast vegetative development is an effective way to suppress invasive plants during early stages of restoration. If limiting similarity is used to design communities for restoration, early life-history traits should be taken into account. KW - Achillea millefolium KW - Ambrosia artemisiifolia KW - biotic resistance KW - competition KW - density-driven suppression KW - disturbed areas KW - restoration KW - seed mixtures KW - Solidago gigantea Y1 - 2018 U6 - https://doi.org/10.1111/avsc.12373 SN - 1402-2001 SN - 1654-109X VL - 21 IS - 3 SP - 411 EP - 418 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Yarman, Aysu T1 - Electrosynthesized Molecularly Imprinted Polymer for Laccase Using the Inactivated Enzyme as the Target JF - Bulletin of the Korean chemical society N2 - The first molecularly imprinted polymer (MIP) for the recognition of the copper-enzyme laccase was successfully prepared by electropolymerizing scopoletin in the presence of alkaline-inactivated enzyme. Laccase-MIP and the control polymer without laccase (nonimprinted polymer, NIP) were characterized by voltammetry using the redox marker ferricyanide. After electropolymerization, the signals for ferricyanide for both the MIP and the NIP were almost completely suppressed and increased after removal of the target from the polymer layer. Rebinding of both inactivated and active laccase decreased the ferricyanide peak currents to almost equal extent. The relative decrease of signal suppression approached saturation above 10 nM. Furthermore, the surface activity of rebound laccase toward the oxidation of catechol was investigated. The surface activity approached saturation above 10 nM, a value close to the value of the measurements with ferricyanide. Interaction of NIP with laccase brought about a six times smaller signal of catechol oxidation. KW - Molecularly imprinted polymers KW - Biomimetic sensors KW - Laccase KW - Electropolymerization KW - Scopoletin Y1 - 2018 U6 - https://doi.org/10.1002/bkcs.11413 SN - 1229-5949 VL - 39 IS - 4 SP - 483 EP - 488 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Yarman, Aysu T1 - Development of a molecularly imprinted polymer-based electrochemical sensor for tyrosinase JF - Turkish journal of chemistry N2 - For the first time a molecularly imprinted polymer (MIP)-based sensor for tyrosinase is described. This sensor is based on the electropolymerization of scopoletin or o-phenylenediamine in the presence of tyrosinase from mushrooms, which has a high homology to the human enzyme. The template was removed either by treatment with proteinase Kor by alkaline treatment. The measuring signal was generated either by measuring the formation of a product by the target enzyme or by evaluation of the permeability of the redox marker ferricyanide. The o-phenylenediamine-based MIP sensor has a linear measuring range up to 50 nM of tyrosinase with a limit of detection of 3.97 nM (R 2 = 0.994) and shows good discrimination towards other proteins, e.g., bovine serum albumin and cytochrome c. KW - Molecularly imprinted polymers KW - biomimetic sensors KW - tyrosinase KW - electropolymerization KW - scopoletin KW - ophenylenediamine Y1 - 2017 U6 - https://doi.org/10.3906/kim-1708-68 SN - 1300-0527 VL - 42 IS - 2 SP - 346 EP - 354 PB - Türkiye Bilimsel ve Teknik Araştırma Kurumu CY - Ankara ER - TY - JOUR A1 - Yarman, Aysu A1 - Badalyan, Artavazd A1 - Gajovic-Eichelmann, Nenad A1 - Wollenberger, Ursula A1 - Scheller, Frieder W. T1 - Enzyme electrode for aromatic compounds exploiting the catalytic activities of microperoxidase-11 JF - Biosensors and bioelectronics : the principal international journal devoted to research, design development and application of biosensors and bioelectronics N2 - Microperoxidase-11 (MR-11) which has been immobilised in a matrix of chitosan-embedded gold nanoparticles on the surface of a glassy carbon electrode catalyzes the conversion of aromatic substances. This peroxide-dependent catalysis of microperoxidase has been applied in an enzyme electrode for the first time to indicate aromatic compounds such as aniline. 4-fluoroaniline, catechol and p-aminophenol. The electrode signal is generated by the cathodic reduction of the quinone or quinoneimine which is formed in the presence of both MP-II and peroxide from the substrate. The same sensor principle will be extended to aromatic drugs. KW - Microperoxidase-11 KW - Nanoparticles KW - p-Aminophenol KW - Aniline KW - Catechol KW - 4-Fluoroaniline KW - Biosensors Y1 - 2011 U6 - https://doi.org/10.1016/j.bios.2011.09.004 SN - 0956-5663 VL - 30 IS - 1 SP - 320 EP - 323 PB - Elsevier CY - Oxford ER - TY - JOUR A1 - Yarman, Aysu A1 - Gröbe, Glenn A1 - Neumann, Bettina A1 - Kinne, Mathias A1 - Gajovic-Eichelmann, Nenad A1 - Wollenberger, Ursula A1 - Hofrichter, Martin A1 - Ullrich, Rene A1 - Scheibner, Katrin A1 - Scheller, Frieder W. T1 - The aromatic peroxygenase from Marasmius rutola-a new enzyme for biosensor applications JF - Analytical & bioanalytical chemistry N2 - The aromatic peroxygenase (APO; EC 1.11.2.1) from the agraric basidomycete Marasmius rotula (MroAPO) immobilized at the chitosan-capped gold-nanoparticle-modified glassy carbon electrode displayed a pair of redox peaks with a midpoint potential of -278.5 mV vs. AgCl/AgCl (1 M KCl) for the Fe(2+)/Fe(3+) redox couple of the heme-thiolate-containing protein. MroAPO oxidizes aromatic substrates such as aniline, p-aminophenol, hydroquinone, resorcinol, catechol, and paracetamol by means of hydrogen peroxide. The substrate spectrum overlaps with those of cytochrome P450s and plant peroxidases which are relevant in environmental analysis and drug monitoring. In M. rotula peroxygenase-based enzyme electrodes, the signal is generated by the reduction of electrode-active reaction products (e.g., p-benzoquinone and p-quinoneimine) with electro-enzymatic recycling of the analyte. In these enzyme electrodes, the signal reflects the conversion of all substrates thus representing an overall parameter in complex media. The performance of these sensors and their further development are discussed. KW - Unspecific peroxygenase KW - Cytochrome P450 KW - Biosensors KW - Phenolic substances Y1 - 2012 U6 - https://doi.org/10.1007/s00216-011-5497-y SN - 1618-2642 VL - 402 IS - 1 SP - 405 EP - 412 PB - Springer CY - Heidelberg ER - TY - JOUR A1 - Yarman, Aysu A1 - Kurbanoğlu, Sevinç A1 - Zebger, Ingo A1 - Scheller, Frieder W. T1 - Simple and robust BT - the claims of protein sensing by molecularly imprinted polymers JF - Sensors and actuators : B, Chemical : an international journal devoted to research and development of chemical transducers N2 - A spectrum of 7562 publications on Molecularly Imprinted Polymers (MIPs) has been presented in literature within the last ten years (Scopus, September 7, 2020). Around 10 % of the papers published on MIPs describe the recognition of proteins. The straightforward synthesis of MIPs is a significant advantage as compared with the preparation of enzymes or antibodies. MIPs have been synthesized from only one up to six functional monomers while proteins are made up of 20 natural amino acids. Furthermore, they can be synthesized against structures of low immunogenicity and allow multi-analyte measurements via multi-target synthesis. Electrochemical methods allow simple polymer synthesis, removal of the template and readout. Among the different sensor configurations electrochemical MIP-sensors provide the broadest spectrum of protein analytes. The sensitivity of MIP-sensors is sufficiently high for biomarkers in the sub-nanomolar region, nevertheless the cross-reactivity of highly abundant proteins in human serum is still a challenge. MIPs for proteins offer innovative tools not only for clinical and environmental analysis, but also for bioimaging, therapy and protein engineering. KW - Molecularly imprinted polymer KW - Plastibodies KW - Functional scaffolds KW - Biomimetic sensors KW - Proteins Y1 - 2021 U6 - https://doi.org/10.1016/j.snb.2020.129369 SN - 0925-4005 SN - 1873-3077 VL - 330 PB - Elsevier Science CY - Amsterdam [u.a.] ER - TY - JOUR A1 - Yarman, Aysu A1 - Scheller, Frieder W. T1 - Coupling biocatalysis with molecular imprinting in a biomimetic sensor JF - Angewandte Chemie : a journal of the Gesellschaft Deutscher Chemiker ; International edition KW - biomimetic sensors KW - electropolymers KW - enzymes KW - hierarchical structures KW - molecularly imprinted polymers Y1 - 2013 U6 - https://doi.org/10.1002/anie.201305368 SN - 1433-7851 SN - 1521-3773 VL - 52 IS - 44 SP - 11521 EP - 11525 PB - Wiley-VCH CY - Weinheim ER -