PCR/RFLP-based method for molecular characterization of ‘Candidatus Phytoplasma prunorum’ strains using the aceF gene.

New molecular typing tools for phytoplasmas belonging to the 16SrX phytoplasma group have recently been developed based on the non-ribosomal genes aceF, pnp, imp, and SecY. In the present work we chose to perform a PCR-RFLP method based on the aceF gene. This genetic marker had previously shown high variability among strains of the 16SrX group, moreover, it had allowed for the differentiation of French hypovirulent ‘Candidatus Phytoplasma prunorum’ strains from virulent ones.Most of the stone fruit samples were collected in north-east Italy, although a few samples from Bosnia and Herzegovina, and Turkey were also included in the work to explore variability. French hypovirulent and virulent strains, one Azerbaijan strain and ‘Ca. P. prunorum’ strains maintained in periwinkles were used as reference strains. Some of the Italian samples were not collected in the field and they became infected by Cacopsylla pruni under controlled conditions.Sequencing of the aceF gene was performed on some of the samples tested and based on the alignment, a few restriction enzymes were selected for ‘Ca. P. prunorum’ strain differentiation. Nested PCR was performed using previously developed primers on all samples and RFLP analyses were carried out with BpiI, HaeIII and Tsp509I enzymes. BpiI and HaeIII enzymes generated two different profiles, one profile was undigested and the second one constituted by two different fragments. The Tsp509I enzyme enabled three different pattern types to be distinguished. Combining the results obtained with the three restriction enzymes, it was possible to distinguish between the ‘Ca. P. prunorum’ strains investigated in this study: 6 different RFLP subgroups AceF-A, -B, -C, -D, -E and –F. We confirmed that strains belonging to 4 subgroups, AceF-A, -B, -C and -E were present in north-east Italy, where a large number of the samples were processed. The strains of AceF-A and -E subgroups were the predominant ones (21.6% and 17.0%, respectively) and mixed infections of AceF-A+E subgroups (17.0%), and AceF-B+E (14.8%) subgroups were quite common. Keywords: phytoplasma, European stone fruit yellows, molecular differentiation, sequencin

Evaluation of the noise properties of a dual-frequency VECSEL for compact Cs atomic clocks (Poster)

International audienceWe evaluate a dual-frequency and dual-polarization optically-pumped semiconductor laser emitting at 852 nm as a new laser source for compact atomic clocks based on the coherent population trapping (CPT) technique. The frequency difference between the laser modes is tunable to 9.2 GHz corresponding to the ground state hyperfine-split of Cs. Impact of the laser noise has been investigated. Laser relative intensity noise is limited by the pump-RIN transfer to a level of-110 dB/Hz. Laser frequency noise shows excess mechanical and technical noise resulting in a laser linewidth of 1 MHz at 1 s in lock operation. The noise performance and spectral properties of the laser are already adequate to realize CPT experiments and should result in Allan standard-deviation of the clock below 1 × 10-12 at 1 second

Dual-frequency VECSEL for atomic clocks using coherent population trapping

Workshop on Laser Diodes for Space Applications, Nov 2015, Palaiseau, FranceAtomic frequency references provide high-precision stable signals, which are crucial in the most demanding applications as high bitrate communication networks, high-end inertial navigation, or satellite positioning. One way to obtain those laser fields with low intensity-and frequency-noise is to use the dual-frequency and dual-polarization emission of an optically-pumped vertical external-cavity semiconductor laser (OP-VECSEL)

Laser à semiconducteur à 852 nm bifrequence pompé optiquement pour les horloges atomiques CPT (poster)

National audienceNous présentons un laser à semiconducteur en cavité externe pompé optiquement, émettant sur deux fréquences optiques polarisées perpendiculairement, destiné au piégeage cohérent d'atomes (CPT) de Cs. L'émission est accordable autour de 852 nm. La différence de fréquence est ajustée grâce à une lame électro-optique autour de 9,2 GHz. La longueur d'onde du mode ordinaire est stabilisée sur la raie D2 du Cs et la différence de fréquence est asservie sur un signal de référence RF. En fonctionnement stabilisé, nous caractérisons les sources de bruits du laser afin d'évaluer les performances du laser en vue de son application dans une horloge atomique CPT

CANELC: constructing an e-language corpus

This paper reports on the construction of CANELC: the Cambridge and Nottingham e-language Corpus.3 CANELC is a one million word corpus of digital communication in English, taken from online discussion boards, blogs, tweets, emails and SMS messages. The paper outlines the approaches used when planning the corpus: obtaining consent; collecting the data and compiling the corpus database. This is followed by a detailed analysis of some of the patterns of language used in the corpus. The analysis includes a discussion of the key words and phrases used as well as the common themes and semantic associations connected with the data. These discussions form the basis of an investigation of how e-language operates in both similar and different ways to spoken and written records of communication (as evidenced by the BNC - British National Corpus). 3 CANELC stands for Cambridge and Nottingham e-language Corpus. This corpus has been built as part of a collaborative project between The University of Nottingham and Cambridge University Press with whom sole copyright of the annotated corpus resides. CANELC comprises one-million words of digital English taken from SMS messages, blogs, tweets, discussion board content and private/business emails. Plans to extend the corpus are under discussion. The legal dimension to corpus ‘ownership’ of some forms of unannotated data is a complex one and is under constant review. At the present time the annotated corpus is only available to authors and researchers working for CUP and is not more generally available

When a Palearctic bacterium meets a Nearctic insect vector: Genetic and ecological insights into the emergence of the grapevine Flavescence dorée epidemics in Europe

Flavescence dorée (FD) is a European quarantine grapevine disease transmitted by the Deltocephalinae leafhopper Scaphoideus titanus. Whereas this vector had been introduced from North America, the possible European origin of FD phytoplasma needed to be challenged and correlated with ecological and genetic drivers of FD emergence. For that purpose, a survey of genetic diversity of these phytoplasmas in grapevines, S. titanus, black alders, alder leafhoppers and clematis were conducted in five European countries. Out of 132 map genotypes, only 11 were associated to FD outbreaks, three were detected in clematis, whereas 127 were detected in alder trees, alder leafhoppers or in grapevines out of FD outbreaks. Most of the alder trees were found infected, including 8% with FD genotypes M6, M38 and M50, also present in alders neighboring FD-free vineyards and vineyard-free areas. The Macropsinae Oncopsis alni could transmit genotypes unable to achieve transmission by S. titanus, while the Deltocephalinae Allygus spp. and Orientus ishidae transmitted M38 and M50 that proved to be compatible with S. titanus. Variability of vmpA and vmpB adhesin-like genes clearly discriminated 3 genetic clusters. Cluster Vmp-I grouped genotypes only transmitted by O. alni, while clusters Vmp-II and -III grouped genotypes transmitted by Deltocephalinae leafhoppers. Interestingly, adhesin repeated domains evolved independently in cluster Vmp-I, whereas in clusters Vmp-II and-III showed recent duplications. Latex beads coated with various ratio of VmpA of clusters II and I, showed that cluster II VmpA promoted enhanced adhesion to the Deltocephalinae Euscelidius variegatus epithelial cells and were better retained in both E. variegatus and S. titanus midguts. Our data demonstrate that most FD phytoplasmas are endemic to European alders. Their emergence as grapevine epidemic pathogens appeared restricted to some genetic variants pre-existing in alders, whose compatibility to S. titanus correlates with different vmp gene sequences and VmpA binding properties

A corpus-assisted study of the discourse marker well as an indicator of judges' institutional roles in court cases with litigants in person

In this paper, I concentrate on court cases with litigants in person (lay people who act on their own behalf in legal proceedings without a counsel or solicitor) and discuss the challenges of building a corpus of courtroom discourse where it is crucial to distinguish between speakers due to their distinct institutional roles. The corpus incorporates seven sub-corpora of verbatim transcripts from different court cases with litigants in person and comprises over eleven-million tokens. The focus of this paper is on the interplay between the legal and lay discourse types and how judges project their institutional roles through well-initiated turns directed at litigants in person and counsels. As a versatile discourse marker, well provides a good opportunity to explore how judges have to adapt their roles to ensure lay litigants in person receive the necessary support and that their lack of competence does not impede on the fairness of the proceedings. Given the breadth and importance of the topic of litigation in person, I discuss how the tools and approaches of corpus linguistics can be helpful in this multi-disciplinary area where multiple functions and uses of individual linguistic features need to be explored in depth

Spectral and noise characterization of a 852 nm dual-frequency VECSEL (orale)

3rd Workshop VECSEL, Nov 2015, Montpellier, FranceCoherent population trapping (CPT) is a common technique used in compact atomic clocks which requires two-phase coherent laser modes with a frequency difference in the GHz range for alkali atoms. To improve the performance vs size trade-off of Cs atomic clocks, we develop a laser source generating two cross-polarized coherent laser fields at 852 nm. It relies on the dual-frequency and dual-polarization operation of an optically-pumped vertical external-cavity semiconductor laser