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1.
Phys Rev Lett ; 130(5): 050801, 2023 Feb 03.
Article in English | MEDLINE | ID: mdl-36800447

ABSTRACT

We study the implementation of arbitrary excitation-conserving linear transformations between two sets of N stationary bosonic modes, which are connected through a photonic quantum channel. By controlling the individual couplings between the modes and the channel, an initial N-partite quantum state in register A can be released as a multiphoton wave packet and, successively, be reabsorbed in register B. Here we prove that there exists a set of control pulses that implement this transfer with arbitrarily high fidelity and, simultaneously, realize a prespecified N×N unitary transformation between the two sets of modes. Moreover, we provide a numerical algorithm for constructing these control pulses and discuss the scaling and robustness of this protocol in terms of several illustrative examples. By being purely control-based and not relying on any adaptations of the underlying hardware, the presented scheme is extremely flexible and can find widespread applications, for example, for boson-sampling experiments, multiqubit state transfer protocols, or in continuous-variable quantum computing architectures.

2.
Phys Rev Lett ; 130(5): 050601, 2023 Feb 03.
Article in English | MEDLINE | ID: mdl-36800450

ABSTRACT

In this Letter, we provide analytical and numerical evidence that the single-layer quantum approximate optimization algorithm on universal Ising spin models produces thermal-like states. We find that these pseudo-Boltzmann states can not be efficiently simulated on classical computers according to the general state-of-the-art condition that ensures rapid mixing for Ising models. Moreover, we observe that the temperature depends on a hidden universal correlation between the energy of a state and the covariance of other energy levels and the Hamming distances of the state to those energies.

3.
CRISPR J ; 4(3): 400-415, 2021 06.
Article in English | MEDLINE | ID: mdl-34152221

ABSTRACT

The discovery of CRISPR has revolutionized the field of genome engineering, but the potential of this technology is far from reaching its limits. In this review, we explore the broad range of applications of CRISPR technology to highlight the rapid expansion of the field beyond gene editing alone. It has been demonstrated that CRISPR technology can control gene expression, spatiotemporally image the genome in vivo, and detect specific nucleic acid sequences for diagnostics. In addition, new technologies are under development to improve CRISPR quality controls for gene editing, thereby improving the reliability of these technologies for therapeutics and beyond. These are just some of the many CRISPR tools that have been developed in recent years, and the toolbox continues to diversify.


Subject(s)
CRISPR-Cas Systems , Clustered Regularly Interspaced Short Palindromic Repeats , Gene Editing/methods , Gene Expression , Genetic Techniques , Genome , Pathology, Molecular/methods , RNA, Guide, Kinetoplastida/genetics , Reproducibility of Results
4.
Med. oral patol. oral cir. bucal (Internet) ; 26(3): e361-e367, May. 2021. ilus, tab, graf
Article in English | IBECS | ID: ibc-224561

ABSTRACT

Background: Primary stability is an important key determinant of implant osseointegration. We investigated ap-proaches to improve primary implant stability using a new drilling technique termed osseodensification (OD),which was compared with the conventional under-drilling (UD) method utilized for low-density bones.Material and Methods: We placed 55 conical internal connection implants in each group, in 30 low-density sec-tions of pig tibia. The implants were placed using twist drill bits in both groups; groups Under Drilling (UD)and Osseodensification (OD) included bone sections subjected to conventional UD and OD drilling, respectively.Before placing the implants, we randomized the bone sections that were to receive these implants to avoid samplebias. We evaluated various primary stability parameters, such as implant insertion torque and resonance fre-quency analysis (RFA) measurements.Results: The results showed that compared with implants placed using the UD technique, those placed using theOD technique were associated with significantly higher primary stability. The mean insertion torque of the im-plants was 8.87±6.17 Ncm in group 1 (UD) and 21.72±17.14 Ncm in group 2 (OD). The mean RFA was 65.16±7.45ISQ in group 1 (UD) and 69.75±6.79 ISQ in group 2 (OD).Conclusions: The implant insertion torque and RFA values were significantly higher in OD group than in UD.Therefore, compared with UD, OD improves primary stability in low-density bones (based on torque and RFAmeasurements).(AU)


Subject(s)
Animals , Dental Implants , Bone Density , Resonance Frequency Analysis , Swine , Dental Prosthesis Retention , Oral Health , Oral Medicine , Pathology, Oral , Surgery, Oral
5.
Nat Biomed Eng ; 5(7): 713-725, 2021 07.
Article in English | MEDLINE | ID: mdl-33820980

ABSTRACT

Simple and fast methods for the detection of target genes with single-nucleotide specificity could open up genetic research and diagnostics beyond laboratory settings. We recently reported a biosensor for the electronic detection of unamplified target genes using liquid-gated graphene field-effect transistors employing an RNA-guided catalytically deactivated CRISPR-associated protein 9 (Cas9) anchored to a graphene monolayer. Here, using unamplified genomic samples from patients and by measuring multiple types of electrical response, we show that the biosensors can discriminate within one hour between wild-type and homozygous mutant alleles differing by a single nucleotide. We also show that biosensors using a guide RNA-Cas9 orthologue complex targeting genes within the protospacer-adjacent motif discriminated between homozygous and heterozygous DNA samples from patients with sickle cell disease, and that the biosensors can also be used to rapidly screen for guide RNA-Cas9 complexes that maximize gene-targeting efficiency.


Subject(s)
Biosensing Techniques/methods , CRISPR-Associated Protein 9/metabolism , DNA/genetics , Polymorphism, Single Nucleotide , Anemia, Sickle Cell/genetics , Anemia, Sickle Cell/pathology , Biosensing Techniques/instrumentation , CRISPR-Associated Protein 9/chemistry , DNA/metabolism , Genome, Human , Graphite/chemistry , Heterozygote , Homozygote , Humans , Immobilized Proteins/chemistry , Immobilized Proteins/metabolism , RNA, Guide, Kinetoplastida/metabolism , Superoxide Dismutase-1/genetics , Transistors, Electronic
6.
Phys Rev Lett ; 126(2): 023603, 2021 Jan 15.
Article in English | MEDLINE | ID: mdl-33512234

ABSTRACT

Coherent photon-emitter interfaces offer a way to mediate efficient nonlinear photon-photon interactions, much needed for quantum information processing. Here we experimentally study the case of a two-level emitter, a quantum dot, coupled to a single optical mode in a nanophotonic waveguide. We carry out few-photon transport experiments and record the statistics of the light to reconstruct the scattering matrix elements of one- and two-photon components. This provides direct insight to the complex nonlinear photon interaction that contains rich many-body physics.

7.
New Phytol ; 227(4): 1222-1234, 2020 08.
Article in English | MEDLINE | ID: mdl-32259283

ABSTRACT

Ovules are essential for sexual plant reproduction and seed formation, and are fundamental for agriculture. However, our understanding of the molecular mechanisms governing ovule development is far from complete. In Arabidopsis, ovule identity is determined by homeotic MADS-domain proteins that define the floral C- (AG) and D- (SHP1/SHP2, STK) functions. Pre-mRNA processing of these genes is critical and mediated by HUA-PEP activity, composed of genes encoding RNA-binding proteins. In strong hua-pep mutants, functional transcripts for C- and D-function genes are reduced, resulting in homeotic transformation of ovules. Thus, hua-pep mutants provide an unique sensitized background to study ovule morphogenesis when C- and D-functions are simultaneously compromised. We found that hua-pep ovules are morphologically sepaloid and show ectopic expression of the homeotic class-A gene AP1. Inactivation of AP1 or AP2 (A-function genes) in hua-pep mutants reduced homeotic conversions, rescuing ovule identity while promoting carpelloid traits in transformed ovules. Interestingly, increased AG dosage led to similar results. Our findings strongly suggest that HUA-PEP activity is required for correct C and D floral functions, which in turn prevents ectopic expression of class-A genes in ovules for their proper morphogenesis, evoking the classic A-C antagonism of the ABC model for floral organ development.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Dissection , Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , Ovule/genetics , Ovule/metabolism , Plant Proteins/genetics
8.
Proc Natl Acad Sci U S A ; 116(50): 25333-25342, 2019 12 10.
Article in English | MEDLINE | ID: mdl-31757847

ABSTRACT

Fruit have evolved a sophisticated tissue and cellular architecture to secure plant reproductive success. Postfertilization growth is perhaps the most dramatic event during fruit morphogenesis. Several studies have proposed that fertilized ovules and developing seeds initiate signaling cascades to coordinate and promote the growth of the accompanying fruit tissues. This dynamic process allows the fruit to conspicuously increase its size and acquire its final shape and means for seed dispersal. All these features are key for plant survival and crop yield. Despite its importance, we lack a high-resolution spatiotemporal map of how postfertilization fruit growth proceeds at the cellular level. In this study, we have combined live imaging, mutant backgrounds in which fertilization can be controlled, and computational modeling to monitor and predict postfertilization fruit growth in Arabidopsis We have uncovered that, unlike leaves, sepals, or roots, fruit do not exhibit a spatial separation of cell division and expansion domains; instead, there is a separation into temporal stages with fertilization as the trigger for transitioning to cell expansion, which drives postfertilization fruit growth. We quantified the coordination between fertilization and fruit growth by imaging no transmitting tract (ntt) mutants, in which fertilization fails in the bottom half of the fruit. By combining our experimental data with computational modeling, we delineated the mobility properties of the seed-derived signaling cascades promoting growth in the fruit. Our study provides the basis for generating a comprehensive understanding of the molecular and cellular mechanisms governing fruit growth and shape.


Subject(s)
Arabidopsis/cytology , Fruit/growth & development , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cell Division , Fertilization , Fruit/cytology , Fruit/genetics , Fruit/metabolism , Gene Expression Regulation, Plant , Seeds/cytology , Seeds/growth & development , Seeds/metabolism
9.
Development ; 145(13)2018 07 09.
Article in English | MEDLINE | ID: mdl-29914969

ABSTRACT

Ovule formation is a complex developmental process in plants, with a strong impact on the production of seeds. Ovule primordia initiation is controlled by a gene network, including components of the signaling pathways of auxin, brassinosteroids and cytokinins. By contrast, gibberellins (GAs) and DELLA proteins, the negative regulators of GA signaling, have never been shown to be involved in ovule initiation. Here, we provide molecular and genetic evidence that points to DELLA proteins as novel players in the determination of ovule number in Arabidopsis and in species of agronomic interest, such as tomato and rapeseed, adding a new layer of complexity to this important developmental process. DELLA activity correlates positively with ovule number, acting as a positive factor for ovule initiation. In addition, ectopic expression of a dominant DELLA in the placenta is sufficient to increase ovule number. The role of DELLA proteins in ovule number does not appear to be related to auxin transport or signaling in the ovule primordia. Possible crosstalk between DELLA proteins and the molecular and hormonal network controlling ovule initiation is also discussed.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/embryology , Gibberellins/metabolism , Ovule/embryology , Arabidopsis/cytology , Ovule/cytology
10.
Phys Rev Lett ; 120(15): 153602, 2018 Apr 13.
Article in English | MEDLINE | ID: mdl-29756880

ABSTRACT

We study the scattering of individual photons by a two-level system ultrastrongly coupled to a waveguide. The scattering is elastic for a broad range of couplings and can be described with an effective U(1)-symmetric Hamiltonian. This simple model allows the prediction of scattering resonance line shapes, validated up to α=0.3, and close to the Toulouse point α=1/2, where inelastic scattering becomes relevant. Our predictions model experiments with superconducting circuits [P. Forn-Díaz et al., Nat. Phys. 13, 39 (2017)NPAHAX1745-247310.1038/nphys3905] and can be extended to study multiphoton scattering.

11.
PLoS Genet ; 14(1): e1007182, 2018 01.
Article in English | MEDLINE | ID: mdl-29329291

ABSTRACT

Ovules are fundamental for plant reproduction and crop yield as they are the precursors of seeds. Therefore, ovule specification is a critical developmental program. In Arabidopsis thaliana, ovule identity is redundantly conferred by the homeotic D-class genes SHATTERPROOF1 (SHP1), SHP2 and SEEDSTICK (STK), phylogenetically related to the MADS-domain regulatory gene AGAMOUS (AG), essential in floral organ specification. Previous studies have shown that the HUA-PEP activity, comprised of a suite of RNA-binding protein (RBP) encoding genes, regulates AG pre-mRNA processing and thus flower patterning and organ identity. Here, we report that the HUA-PEP activity additionally governs ovule morphogenesis. Accordingly, in severe hua-pep backgrounds ovules transform into flower organ-like structures. These homeotic transformations are most likely due to the dramatic reduction in SHP1, SHP2 and STK activity. Our molecular and genome-wide profiling strategies revealed the accumulation of prematurely terminated transcripts of D-class genes in hua-pep mutants and reduced amounts of their respective functional messengers, which points to pre-mRNA processing misregulation as the origin of the ovule developmental defects in such backgrounds. RNA processing and transcription are coordinated by the RNA polymerase II (RNAPII) carboxyl-terminal domain (CTD). Our results show that HUA-PEP activity members can interact with the CTD regulator C-TERMINAL DOMAIN PHOSPHATASE-LIKE1 (CPL1), supporting a co-transcriptional mode of action for the HUA-PEP activity. Our findings expand the portfolio of reproductive developmental programs in which HUA-PEP activity participates, and further substantiates the importance of RNA regulatory mechanisms (pre-mRNA co-transcriptional regulation) for correct gene expression during plant morphogenesis.


Subject(s)
Arabidopsis , Cell Differentiation/genetics , Ovule/physiology , RNA Precursors/metabolism , RNA Processing, Post-Transcriptional , Arabidopsis/embryology , Arabidopsis/genetics , Flowers/genetics , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Genes, Plant , Morphogenesis/genetics , Ovule/embryology , Plants, Genetically Modified , RNA-Binding Proteins/genetics , Transcription Factors/genetics
12.
New Phytol ; 217(2): 813-827, 2018 01.
Article in English | MEDLINE | ID: mdl-29105090

ABSTRACT

Root knot nematodes (RKNs) penetrate into the root vascular cylinder, triggering morphogenetic changes to induce galls, de novo formed 'pseudo-organs' containing several giant cells (GCs). Distinctive gene repression events observed in early gall/GCs development are thought to be mediated by post-transcriptional silencing via microRNAs (miRNAs), a process that is far from being fully characterized. Arabidopsis thaliana backgrounds with altered activities based on target 35S::MIMICRY172 (MIM172), 35S::TARGET OF EARLY ACTIVATION TAGGED 1 (TOE1)-miR172-resistant (35S::TOE1R ) and mutant (flowering locus T-10 (ft-10)) lines were used for functional analysis of nematode infective and reproductive parameters. The GUS-reporter lines, MIR172A-E::GUS, treated with auxin (IAA) and an auxin-inhibitor (a-(phenyl ethyl-2-one)-indole-3-acetic acid (PEO-IAA)), together with the MIR172C AuxRE::GUS line with two mutated auxin responsive elements (AuxREs), were assayed for nematode-dependent gene expression. Arabidopsis thaliana backgrounds with altered expression of miRNA172, TOE1 or FT showed lower susceptibility to the RKNs and smaller galls and GCs. MIR172C-D::GUS showed restricted promoter activity in galls/GCs that was regulated by auxins through auxin-responsive factors. IAA induced their activity in galls while PEO-IAA treatment and mutations in AuxRe motifs abolished it. The results showed that the regulatory module miRNA172/TOE1/FT plays an important role in correct GCs and gall development, where miRNA172 is modulated by auxins.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/parasitology , Feeding Behavior , Gene Regulatory Networks , MicroRNAs/metabolism , Tylenchoidea/physiology , Animals , Arabidopsis/drug effects , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Base Sequence , Crops, Agricultural/genetics , Crops, Agricultural/parasitology , Disease Progression , Feeding Behavior/drug effects , Gene Expression Regulation, Plant/drug effects , Gene Regulatory Networks/drug effects , Giant Cells/metabolism , Giant Cells/parasitology , Glucuronidase/metabolism , Indoleacetic Acids/pharmacology , MicroRNAs/genetics , Models, Biological , Plant Diseases/parasitology , Plant Tumors/parasitology , Promoter Regions, Genetic/genetics , Tylenchoidea/drug effects , Up-Regulation/drug effects
13.
Phys Rev Lett ; 119(15): 153601, 2017 Oct 13.
Article in English | MEDLINE | ID: mdl-29077426

ABSTRACT

In this work we develop an experimental procedure to interrogate the single- and multiphoton scattering matrices of an unknown quantum system interacting with propagating photons. Our proposal requires coherent state laser or microwave inputs and homodyne detection at the scatterer's output, and provides simultaneous information about multiple-elastic and inelastic-segments of the scattering matrix. The method is resilient to detector noise and its errors can be made arbitrarily small by combining experiments at various laser powers. Finally, we show that the tomography of scattering has to be performed using pulsed lasers to efficiently gather information about the nonlinear processes in the scatterer.

14.
Elife ; 62017 10 23.
Article in English | MEDLINE | ID: mdl-29058667

ABSTRACT

Plant meristems carry pools of continuously active stem cells, whose activity is controlled by developmental and environmental signals. After stem cell division, daughter cells that exit the stem cell domain acquire transit amplifying cell identity before they are incorporated into organs and differentiate. In this study, we used an integrated approach to elucidate the role of HECATE (HEC) genes in regulating developmental trajectories of shoot stem cells in Arabidopsis thaliana. Our work reveals that HEC function stabilizes cell fate in distinct zones of the shoot meristem thereby controlling the spatio-temporal dynamics of stem cell differentiation. Importantly, this activity is concomitant with the local modulation of cellular responses to cytokinin and auxin, two key phytohormones regulating cell behaviour. Mechanistically, we show that HEC factors transcriptionally control and physically interact with MONOPTEROS (MP), a key regulator of auxin signalling, and modulate the autocatalytic stabilization of auxin signalling output.


Subject(s)
Arabidopsis/physiology , Cell Differentiation/drug effects , Gene Expression Regulation, Plant/drug effects , Plant Cells/physiology , Plant Growth Regulators/metabolism , Stem Cells/physiology , Genes, Plant , Plant Cells/drug effects , Plant Shoots/physiology , Stem Cells/drug effects , Transcription, Genetic
15.
PLoS Genet ; 13(4): e1006726, 2017 04.
Article in English | MEDLINE | ID: mdl-28388635

ABSTRACT

Fruits and seeds are the major food source on earth. Both derive from the gynoecium and, therefore, it is crucial to understand the mechanisms that guide the development of this organ of angiosperm species. In Arabidopsis, the gynoecium is composed of two congenitally fused carpels, where two domains: medial and lateral, can be distinguished. The medial domain includes the carpel margin meristem (CMM) that is key for the production of the internal tissues involved in fertilization, such as septum, ovules, and transmitting tract. Interestingly, the medial domain shows a high cytokinin signaling output, in contrast to the lateral domain, where it is hardly detected. While it is known that cytokinin provides meristematic properties, understanding on the mechanisms that underlie the cytokinin signaling pattern in the young gynoecium is lacking. Moreover, in other tissues, the cytokinin pathway is often connected to the auxin pathway, but we also lack knowledge about these connections in the young gynoecium. Our results reveal that cytokinin signaling, that can provide meristematic properties required for CMM activity and growth, is enabled by the transcription factor SPATULA (SPT) in the medial domain. Meanwhile, cytokinin signaling is confined to the medial domain by the cytokinin response repressor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFERASE 6 (AHP6), and perhaps by ARR16 (a type-A ARR) as well, both present in the lateral domains (presumptive valves) of the developing gynoecia. Moreover, SPT and cytokinin, probably together, promote the expression of the auxin biosynthetic gene TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA1) and the gene encoding the auxin efflux transporter PIN-FORMED 3 (PIN3), likely creating auxin drainage important for gynoecium growth. This study provides novel insights in the spatiotemporal determination of the cytokinin signaling pattern and its connection to the auxin pathway in the young gynoecium.


Subject(s)
Arabidopsis Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Cytokinins/metabolism , Meristem/genetics , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/metabolism , Basic Helix-Loop-Helix Transcription Factors/metabolism , Flowers/genetics , Flowers/growth & development , Fruit/genetics , Fruit/growth & development , Gene Expression Regulation, Plant , Indoleacetic Acids/metabolism , Meristem/growth & development , Seeds/genetics , Seeds/growth & development , Signal Transduction , Tryptophan Transaminase/genetics
16.
Proc Natl Acad Sci U S A ; 114(9): 2419-2424, 2017 02 28.
Article in English | MEDLINE | ID: mdl-28202720

ABSTRACT

Plants have evolved adaptive strategies that involve transcriptional networks to cope with and survive environmental challenges. Key transcriptional regulators that mediate responses to environmental fluctuations in nitrate have been identified; however, little is known about how these regulators interact to orchestrate nitrogen (N) responses and cell-cycle regulation. Here we report that teosinte branched1/cycloidea/proliferating cell factor1-20 (TCP20) and NIN-like protein (NLP) transcription factors NLP6 and NLP7, which act as activators of nitrate assimilatory genes, bind to adjacent sites in the upstream promoter region of the nitrate reductase gene, NIA1, and physically interact under continuous nitrate and N-starvation conditions. Regions of these proteins necessary for these interactions were found to include the type I/II Phox and Bem1p (PB1) domains of NLP6&7, a protein-interaction module conserved in animals for nutrient signaling, and the histidine- and glutamine-rich domain of TCP20, which is conserved across plant species. Under N starvation, TCP20-NLP6&7 heterodimers accumulate in the nucleus, and this coincides with TCP20 and NLP6&7-dependent up-regulation of nitrate assimilation and signaling genes and down-regulation of the G2/M cell-cycle marker gene, CYCB1;1 TCP20 and NLP6&7 also support root meristem growth under N starvation. These findings provide insights into how plants coordinate responses to nitrate availability, linking nitrate assimilation and signaling with cell-cycle progression.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Gene Expression Regulation, Plant , Plant Roots/genetics , Transcription Factors/genetics , Amino Acid Sequence , Arabidopsis/drug effects , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Binding Sites , Cyclin B/genetics , Cyclin B/metabolism , G2 Phase Cell Cycle Checkpoints , Gene Expression Regulation, Developmental , Nitrate Reductase/genetics , Nitrate Reductase/metabolism , Nitrates/metabolism , Nitrates/pharmacology , Plant Roots/drug effects , Plant Roots/growth & development , Plant Roots/metabolism , Promoter Regions, Genetic , Protein Binding , Protein Isoforms/genetics , Protein Isoforms/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Signal Transduction , Transcription Factors/metabolism , Transcription, Genetic
17.
New Phytol ; 213(1): 250-263, 2017 01.
Article in English | MEDLINE | ID: mdl-27513887

ABSTRACT

Several physiological functions have been attributed to class III peroxidases (PRXs) in plants, but the in planta role of most members of this family still remains undetermined. Here, we report the first functional characterization of PRX17 (At2g22420), one of the 73 members of this family in Arabidopsis thaliana. Localization of PRX17 was examined by transient expression in Nicotiana benthamiana. Loss- and gain-of-function mutants in A. thaliana were studied. Regulation at the gene and protein levels was analyzed using ß-glucuronidase (GUS) activity, quantitative reverse transcriptase (qRT)-PCR, zymography, and chromatin immunoprecipitation. Phenotypes were characterized including lignin and xylan contents. PRX17 was expressed in various tissues, including vascular tissues, and PRX17 was localized to the cell wall. In prx17, the lignin content was reduced in the stem and siliques and bolting was delayed, while the opposite phenotype was observed in 35S:PRX17 plants, together with a significant increase of lignin and xylan immunofluorescence signal. Finally, we demonstrated that the transcription factor AGAMOUS-LIKE15 (AGL15) binds to the PRX17 promoter and regulates PRX17 expression level. This converging set of structural, transcriptomic and physiological data suggests that PRX17, under the control of AGL15, contributes to developmental programs by playing an essential role in regulating age-dependent lignified tissue formation, including changes in cell wall properties.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Cell Wall/metabolism , Lignin/metabolism , MADS Domain Proteins/metabolism , Peroxidase/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Base Sequence , DNA, Bacterial/genetics , Flowers/physiology , Gene Expression Regulation, Plant , Genes, Plant , MADS Domain Proteins/genetics , Mutation/genetics , Peroxidases , Phylogeny , Plants, Genetically Modified , Promoter Regions, Genetic , Protein Binding , RNA, Messenger/genetics , RNA, Messenger/metabolism , Subcellular Fractions/metabolism
18.
PLoS Genet ; 11(2): e1004983, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25658099

ABSTRACT

Post-transcriptional control is nowadays considered a main checking point for correct gene regulation during development, and RNA binding proteins actively participate in this process. Arabidopsis thaliana FLOWERING LOCUS WITH KH DOMAINS (FLK) and PEPPER (PEP) genes encode RNA-binding proteins that contain three K-homology (KH)-domain, the typical configuration of Poly(C)-binding ribonucleoproteins (PCBPs). We previously demonstrated that FLK and PEP interact to regulate FLOWERING LOCUS C (FLC), a central repressor of flowering time. Now we show that FLK and PEP also play an important role in the maintenance of the C-function during floral organ identity by post-transcriptionally regulating the MADS-box floral homeotic gene AGAMOUS (AG). Previous studies have indicated that the KH-domain containing protein HEN4, in concert with the CCCH-type RNA binding protein HUA1 and the RPR-type protein HUA2, facilitates maturation of the AG pre-mRNA. In this report we show that FLK and PEP genetically interact with HEN4, HUA1, and HUA2, and that the FLK and PEP proteins physically associate with HUA1 and HEN4. Taken together, these data suggest that HUA1, HEN4, PEP and FLK are components of the same post-transcriptional regulatory module that ensures normal processing of the AG pre-mRNA. Our data better delineates the roles of PEP in plant development and, for the first time, links FLK to a morphogenetic process.


Subject(s)
AGAMOUS Protein, Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis/genetics , Flowers/genetics , MADS Domain Proteins/genetics , RNA-Binding Proteins/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/biosynthesis , Flowers/growth & development , Gene Expression Regulation, Plant , MADS Domain Proteins/biosynthesis , Microscopy, Electron, Scanning , Morphogenesis , Phenotype , RNA-Binding Proteins/biosynthesis , Reproduction/genetics
19.
Bio Protoc ; 5(19): e1604, 2015 Oct 05.
Article in English | MEDLINE | ID: mdl-27034968

ABSTRACT

Production of functional eukaryotic RNA is a very elaborate process that involves a complex interplay between transcription and various RNA processing activities, including splicing, 5' capping, and 3' cleavage and polyadenylation (Bentley, 2014). Accurate mapping of RNA ends provides a valuable tool to assess transcriptional and post-transcriptional events giving rise to different gene transcripts. The abundance of such transcripts most likely depends on exogenous and developmental cues, or mutations. In the reference plant Arabidopsis, perturbation of the HUA-PEP post-transcriptional regulatory factors (Rodríguez-Cazorla et al., 2015) leads to the accumulation of aberrant transcripts of the key floral homeotic gene AGAMOUS (AG) (Yanofsky et al., 1990) that retain intronic sequence. It was determined by 3' RACE reactions that such erroneous transcripts correspond to premature processing and polyadenylation events taking place at the AG intron region. Here we describe a protocol that is suitable for analysis of relatively abundant transcripts and also for detecting aberrant RNA species that are likely prone to rapid turnover. Likewise, the method, here adapted to Arabidopsis reproductive tissues, can be applied to characterize RNA species from other organs (leaf, root) and/or other plant species. We provide a detailed protocol of our 3' RACE procedure comprising four major parts: Total RNA extraction, RNA amount determination and quality control, the RACE procedure itself, and isolation of the resulting RACE products for cloning and sequencing.

20.
Phys Rev Lett ; 113(19): 193601, 2014 Nov 07.
Article in English | MEDLINE | ID: mdl-25415906

ABSTRACT

We show how a pair of superconducting qubits coupled to a microwave cavity mode can be used to engineer a single-atom laser that emits light into a nonclassical state. Our scheme relies on the dressing of the qubit-field coupling by periodic modulations of the qubit energy. In the dressed basis, the radiative decay of the first qubit becomes an effective incoherent pumping mechanism that injects energy into the system, hence turning dissipation to our advantage. A second, auxiliary qubit is used to shape the decay within the cavity, in such a way that lasing occurs in a squeezed basis of the cavity mode. We characterize the system both by mean-field theory and exact calculations. Our work may find applications in the generation of squeezing and entanglement in circuit QED, as well as in the study of dissipative few- and many-body phase transitions.

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