Comparative genomic and transcriptome analyses of pathotypes of Xanthomonas citri subsp. citri provide insights into mechanisms of bacterial virulence and host range

Abstract Background Citrus bacterial canker is a disease that has severe economic impact on citrus industries worldwide and is caused by a few species and pathotypes of Xanthomonas. X. citri subsp. citri strain 306 (XccA306) is a type A (Asiatic) strain with a wide host range, whereas its variant X. citri subsp. citri strain Aw12879 (Xcaw12879, Wellington strain) is restricted to Mexican lime. Results To characterize the mechanism for the differences in host range of XccA and Xcaw, the genome of Xcaw12879 that was completed recently was compared with XccA306 genome. Effectors xopAF and avrGf1 are present in Xcaw12879, but were absent in XccA306. AvrGf1 was shown previously for Xcaw to cause hypersensitive response in Duncan grapefruit. Mutation analysis of xopAF indicates that the gene contributes to Xcaw growth in Mexican lime but does not contribute to the limited host range of Xcaw. RNA-Seq analysis was conducted to compare the expression profiles of Xcaw12879 and XccA306 in Nutrient Broth (NB) medium and XVM2 medium, which induces hrp gene expression. Two hundred ninety two and 281 genes showed differential expression in XVM2 compared to in NB for XccA306 and Xcaw12879, respectively. Twenty-five type 3 secretion system genes were up-regulated in XVM2 for both XccA and Xcaw. Among the 4,370 common genes of Xcaw12879 compared to XccA306, 603 genes in NB and 450 genes in XVM2 conditions were differentially regulated. Xcaw12879 showed higher protease activity than XccA306 whereas Xcaw12879 showed lower pectate lyase activity in comparison to XccA306. Conclusions Comparative genomic analysis of XccA306 and Xcaw12879 identified strain specific genes. Our study indicated that AvrGf1 contributes to the host range limitation of Xcaw12879 whereas XopAF contributes to virulence. Transcriptome analyses of XccA306 and Xcaw12879 presented insights into the expression of the two closely related strains of X. citri subsp. citri. Virulence genes including genes encoding T3SS components and effectors are induced in XVM2 medium. Numerous genes with differential expression in Xcaw12879 and XccA306 were identified. This study provided the foundation to further characterize the mechanisms for virulence and host range of pathotypes of X. citri subsp. citri.

Genome sequence of Xanthomonas fuscans subsp. fuscans strain 4834-R reveals that flagellar motility is not a general feature of xanthomonads

Abstract Background Xanthomonads are plant-associated bacteria responsible for diseases on economically important crops. Xanthomonas fuscans subsp. fuscans (Xff) is one of the causal agents of common bacterial blight of bean. In this study, the complete genome sequence of strain Xff 4834-R was determined and compared to other Xanthomonas genome sequences. Results Comparative genomics analyses revealed core characteristics shared between Xff 4834-R and other xanthomonads including chemotaxis elements, two-component systems, TonB-dependent transporters, secretion systems (from T1SS to T6SS) and multiple effectors. For instance a repertoire of 29 Type 3 Effectors (T3Es) with two Transcription Activator-Like Effectors was predicted. Mobile elements were associated with major modifications in the genome structure and gene content in comparison to other Xanthomonas genomes. Notably, a deletion of 33 kbp affects flagellum biosynthesis in Xff 4834-R. The presence of a complete flagellar cluster was assessed in a collection of more than 300 strains representing different species and pathovars of Xanthomonas. Five percent of the tested strains presented a deletion in the flagellar cluster and were non-motile. Moreover, half of the Xff strains isolated from the same epidemic than 4834-R was non-motile and this ratio was conserved in the strains colonizing the next bean seed generations. Conclusions This work describes the first genome of a Xanthomonas strain pathogenic on bean and reports the existence of non-motile xanthomonads belonging to different species and pathovars. Isolation of such Xff variants from a natural epidemic may suggest that flagellar motility is not a key function for in planta fitness.

A multilevel converter structure for grid-connected PV plants

A new conversion structure for three-phase grid-connected photovoltaic (PV) generation plants is presented and discussed in this Thesis. The conversion scheme is based on two insulated PV arrays, each one feeding the dc bus of a standard 2-level three-phase voltage source inverter (VSI). Inverters are connected to the grid by a traditional three-phase transformer having open-end windings at inverters side and either star or delta connection at the grid side. The resulting conversion structure is able to perform as a multilevel VSI, equivalent to a 3-level inverter, doubling the power capability of a single VSI with given voltage and current ratings. Different modulation schemes able to generate proper multilevel voltage waveforms have been discussed and compared. They include known algorithms, some their developments, and new original approaches. The goal was to share the grid power with a given ratio between the two VSI within each cycle period of the PWM, being the PWM pattern suitable for the implementation in industrial DSPs. It has been shown that an extension of the modulation methods for standard two-level inverter can provide a elegant solution for dual two-level inverter. An original control method has been introduced to regulate the dc-link voltages of each VSI, according to the voltage reference given by a single MPPT controller. A particular MPPT algorithm has been successfully tested, based on the comparison of the operating points of the two PV arrays. The small deliberately introduced difference between two operating dc voltages leads towards the MPP in a fast and accurate manner. Either simulation or experimental tests, or even both, always accompanied theoretical developments. For the simulation, the Simulink tool of Matlab has been adopted, whereas the experiments have been carried out by a full-scale low-voltage prototype of the whole PV generation system. All the research work was done at the Lab of the Department of Electrical Engineering, University of Bologna.

Studi epidemiologici su Pseudomonas syringae pv.actinidiae agente causale del cancro batterico in actinidia

Il cancro batterico dell’actinidia causato da Pseudomonas syringae pv.actinidiae (Psa) suscita grande interesse a livello globale a partire dal 2008. La malattia è comparsa in Giappone e in due anni ha avuto una diffusione epidemica in tutte le aree di coltivazione mondiale di actinidia. Gravi perdite economiche hanno attirato l’attenzione internazionale su questa problematica e grandi sforzi sono stati rivolti allo studio di questo patosistema ancora poco conosciuto. E’ emerso infatti che il patogeno può rimanere in fase latente per lunghi periodi senza causare sintomi caratteristici nelle piante infette, e che dalla comparsa dei sintomi la pianta muore nell’arco di un paio d’anni. Il monitoraggio ed il controllo della situazione è perciò di fondamentale importanza ed è ancora più importante prevenire la comparsa di nuovi focolai di infezione. A questo proposito sarebbe opportuno l’impiego di materiale vegetale di propagazione non infetto, ma in molti casi questo diventa difficile, dal momento che il materiale impiegato è generalmente quello asintomatico, non analizzato precedentemente per la presenza del patogeno. Negli ultimi anni sono state perciò messe a punto molte tecniche molecolari per l’identificazione di Psa direttamente da materiale vegetale. L’obiettivo di questo lavoro è stato quello di studiare l’epidemiologia di Psa in piante adulte infette e di verificare l’efficacia di metodi di diagnosi precoce per prevenire la malattia. A tale scopo il lavoro sperimentale è stato suddiviso in diverse fasi: i) studio della localizzazione, traslocazione e sopravvivenza di Psa nelle piante, a seguito di inoculazione in piante adulte di actinidia di ceppi marcati Psa::gfp; ii) studio della capacità di Psa di essere mantenuto in germogli di actinidia attraverso sette generazioni di micropropagazione dopo l’inoculazione delle piante madri con lo stesso ceppo marcato Psa::gfp; iii) studio ed applicazioni di un nuovo metodo di diagnosi precoce di Psa basato sull’analisi molecolare del “pianto”.

On the Co-Occurrence of Warm Conveyor Belt Outflows and PV Streamers*

The co-occurrence of warm conveyor belts (WCBs), strongly ascending moist airstreams in extratropical cyclones, and stratospheric potential vorticity (PV) streamers, indicators for breaking Rossby waves on the tropopause, is investigated for a 21-yr period in the Northern Hemisphere using Interim European Centre for Medium-Range Weather Forecasts (ECMWF) Re-Analysis (ERA-Interim) data. WCB outflows and PV streamers are respectively identified as two- and three-dimensional objects and tracked during their life cycle. PV streamers are more frequent than WCB outflows and nearly 15% of all PV streamers co-occur with WCBs during their life cycle, whereas about 60% of all WCB outflows co-occur with PV streamers. Co-occurrences are most frequent over the North Atlantic and North Pacific in spring and winter. WCB outflows are often located upstream of the PV streamers and form earlier, indicating the importance of diabatic processes for downstream Rossby wave breaking. Less frequently, PV streamers occur first, leading to the formation of new WCBs.

Detección de Xanthomonas cucurbitae (Bryan) Vauterin et al. afectando frutos de zapallo coreano en Mendoza y San Juan, Argentina

Durante las campañas 2007/08 y 2008/09, en explotaciones comerciales de zapallo coreano (Cucurbita moschata Duch) de las zonas hortícolas de Mendoza y San Juan, se observaron sobre los frutos numerosas lesiones circulares, de 3 a 6 mm de diámetro, de aspecto húmedo, algo deprimidas hacia el centro, con un exudado gomoso color ámbar. Cuando las lesiones superficiales se unían, se desarrollaba una podredumbre gelatinosa hacia el interior de los tejidos, que podía profundizar hasta la cavidad seminal. El resultado final de la afección era una podredumbre seca y costrosa, que abarcaba gran parte del fruto, inutilizándolo para ser comercializado. En follaje también se observaron síntomas de la enfermedad, como manchas cloróticas angulares de 2 a 3 mm de lado, que en ocasiones se fusionaban. El objetivo del presente trabajo fue determinar la etiología de la enfermedad. Para ello, se tomaron y analizaron muestras de frutos y hojas afectadas, durante ambas temporadas. Se concluyó que la sintomatología es causada por una bacteria, que de acuerdo con los estudios morfobioquímicos y patogénicos es Xanthomonas cucurbitae (Bryan) Vauterin et al. Esta investigación constituye la primera cita de este patógeno afectando frutos en Argentina.

Approche empirique des difficultés de compréhension en traduction, mises en évidence par la méthode d'observation du raisonnement á voix haute (Protocole Verbal - PV)

Fil: Daule, Gabriela. Universidad Nacional de La Plata. Facultad de Humanidades y Ciencias de la Educación; Argentina.

Approche empirique des difficultés de compréhension en traduction, mises en évidence par la méthode d'observation du raisonnement á voix haute (Protocole Verbal - PV)

Fil: Daule, Gabriela. Universidad Nacional de La Plata. Facultad de Humanidades y Ciencias de la Educación; Argentina.