Morphological evidence for phages in Xylella fastidiosa

Presumptive phage particles associated with Xylella fastidiosa strain Temecula-1 grown in PW broth were observed by transmission electron microscopy (TEM) in ultrathin sections of bacterial cell-containing low speed centrifugation pellets and in partially purified preparations from CsCl equilibrium centrifugation density gradients. Ultrathin-sectioned cell pellets contained icosahedral particles of about 45 nm in diameter. Samples collected from CsCl density gradients revealed mostly non-tailed icosahedral but also tailed particles. The icosahedral particles could be divided into two types: a large type (about 45 nm) and a small type (about 30 nm). Filamentous phage-like particles (17 × 120 to 6,300 nm) were also observed. The presence of different types of phage-like particles resembling to those in several bacteriophage families provides new physical evidence, in addition to X. fastidiosa genomic information, that X. fastidiosa possesses active phages. This is the first report of phage particles released in X. fastidiosa cultures

Molecular characterization of a mosaic locus in the genome of 'Candidatus Liberibacter asiaticus'

<p>Abstract</p> <p>Background</p> <p>Huanglongbing (HLB) is a highly destructive disease of citrus production worldwide. '<it>Candidatus </it>Liberibacter asiaticus', an unculturable alpha proteobacterium, is a putative pathogen of HLB. Information about the biology and strain diversity of '<it>Ca</it>. L. asiaticus' is currently limited, inhibiting the scope of HLB research and control.</p> <p>Results</p> <p>A genomic region (CLIBASIA_05640 to CLIBASIA_05650) of '<it>Ca</it>. L. asiaticus' showing hyper-sequence variation or locus mosaicism was identified and investigated using 262 bacterial strains (188 from China and 74 from Florida). Based on the characteristic electrophoretic profiles of PCR amplicons generated by a specific primer set, eight electrophoretic types (E-types) were identified, six E-types (A, B, C, D, E, and F) in China and four E-types (A, C, G, and H) in Florida. The '<it>Ca</it>. L. asiaticus' strains from China consisted predominately of E-type A (71.3%) and E-type B (19.7%). In contrast, the '<it>Ca</it>. L. asiaticus' strains from Florida was predominated by E-type G (82.4%). Diversity of '<it>Ca</it>. L. asiaticus' in China was also evidenced. Strains from the high altitude Yunnan Province consisted of five E-types with E-type B being the majority (62.8%), whereas strains from the low altitude coastal Guangdong Province consisted of only two E-types with E-type A as the majority (97.0%). Sequence analyses revealed that variation of DNA amplicons was due to insertion/deletion events at CLIBASIA_05650 and the downstream intergenic region.</p> <p>Conclusions</p> <p>This study demonstrated the genomic mosaicism of '<it>Ca</it>. L. asiaticus' resulted from active DNA insertion/deletion activities. Analyses of strain variation depicted the significant inter- and intra-continent diversity of '<it>Ca</it>. L. asiaticus'.</p

An Evolutionary Perspective of Pierce’s Disease of Grapevine, Citrus Variegated Chlorosis, and Mulberry Leaf Scorch Diseases

Xylella fastidiosa causes diseases on a growing list of economically important plants. An understanding of how xylellae diseases originated and evolved is important for disease prevention and management. In this study, we evaluated the phylogenetic relationships of X. fastidiosa strains from citrus, grapevine, and mulberry through the analyses of random amplified polymorphic DNAs (RAPDs) and conserved 16S rDNA genes. RAPD analysis emphasized the vigorous genome-wide divergence of X. fastidiosa and detected three clonal groups of strains that cause Pierce’s disease (PD) of grapevine, citrus variegated chlorosis (CVC), and mulberry leaf scorch (MLS). Analysis of 16S rDNA sequences also identified the PD and CVC groups, but with a less stable evolutionary tree. MLS strains were included in the PD group by the 16S rDNA analysis. The Asiatic origins of the major commercial grape and citrus cultivars suggest the recent evolution of both PD and CVC disease in North and South America, respectively, since X. fastidiosa is a New World organism. In order to prevent the development of new diseases caused by X. fastidiosa, it is important to understand the diversity of X. fastidiosa strains, how strains of X. fastidiosa select their hosts, and their ecological roles in the native vegetation

Movable Antenna-Empowered AirComp

A novel over-the-air computation (AirComp) framework, empowered by the incorporation of movable antennas (MAs), is proposed to significantly enhance computation accuracy. Within this framework, the joint optimization of transmit power control, antenna positioning, and receive combining is investigated. An efficient method is proposed to tackle the problem of computation mean-squared error (MSE) minimization, capitalizing on the approach of alternating optimization. Numerical results are provided to substantiate the superior MSE performance of the proposed framework, which establish its clear advantage over benchmark systems employing conventional fixed-position antennas (FPAs)

Origins of the Xylella fastidiosa Prophage-Like Regions and Their Impact in Genome Differentiation

Xylella fastidiosa is a Gram negative plant pathogen causing many economically important diseases, and analyses of completely sequenced X. fastidiosa genome strains allowed the identification of many prophage-like elements and possibly phage remnants, accounting for up to 15% of the genome composition. To better evaluate the recent evolution of the X. fastidiosa chromosome backbone among distinct pathovars, the number and location of prophage-like regions on two finished genomes (9a5c and Temecula1), and in two candidate molecules (Ann1 and Dixon) were assessed. Based on comparative best bidirectional hit analyses, the majority (51%) of the predicted genes in the X. fastidiosa prophage-like regions are related to structural phage genes belonging to the Siphoviridae family. Electron micrograph reveals the existence of putative viral particles with similar morphology to lambda phages in the bacterial cell in planta. Moreover, analysis of microarray data indicates that 9a5c strain cultivated under stress conditions presents enhanced expression of phage anti-repressor genes, suggesting switches from lysogenic to lytic cycle of phages under stress-induced situations. Furthermore, virulence-associated proteins and toxins are found within these prophage-like elements, thus suggesting an important role in host adaptation. Finally, clustering analyses of phage integrase genes based on multiple alignment patterns reveal they group in five lineages, all possessing a tyrosine recombinase catalytic domain, and phylogenetically close to other integrases found in phages that are genetic mosaics and able to perform generalized and specialized transduction. Integration sites and tRNA association is also evidenced. In summary, we present comparative and experimental evidence supporting the association and contribution of phage activity on the differentiation of Xylella genomes

Winter curing of Prunus dulcis cv ‘Butte,’ P. webbii and their interspecific hybrid in response to Xylella fastidiosa infections

Clonal replicates of Prunus dulcis cv ‘Butte,’ P. webbii and their interspecific hybrid P 63-61 were inoculated with Xylella fastidiosa strain M23 and evaluated for almond leaf scorch disease and subsequent winter curing of infections during three growing seasons. Initial inoculations established greater than 90% infection in each of the accessions, based on PCR diagnoses from petiole tissues sampled near the inoculation site. Classic leaf scorch symptoms were evident in each population during the first growing season in a controlled greenhouse environment. Trees were removed from the greenhouse during the winters to accumulate chill hours and to provide the possibility of winter curing X. fastidiosa infections. Both PCR diagnostics and in vitro cultivation were used during the second and third growing seasons to determine the persistence of X. fastidiosa in clones among the three populations. Tree survival and the degree of winter cured infections differed among the three populations, with P. webbii and P 63-61 demonstrating enhanced levels of survivorship over ‘Butte.’ After two cycles of ambient winter temperatures and subsequent growth, ‘Butte’ averaged 21.2% winter cured trees with 73.1% mean survival. Tree survival and winter cured infections were nearly 100% for both P. webbii and P 63-61, demonstrating the utility of P. webbii in almond breeding efforts aimed at reducing tree vulnerability to X. fastidiosa infections

A zinc, copper and citric acid biocomplex shows promise for control of Xylella fastidiosa subsp. pauca in olive trees in Apulia region (southern Italy)

The bacterium Xylella fastidiosa subsp. pauca is associated with the “olive quick decline syndrome” in the Apulia region of southern Italy. To investigate control of this phytopathogen, a compound containing zinc and copper complexed with citric-acid hydracids (Dentamet®) was evaluated for in vitro and in planta bactericidal activity. Confocal laser scanning microscopy, fluorescent quantification and atomic emission spectroscopy were then used to determine if the compound reached the xylem networks of leaves, twigs and branches of olive, to release zinc and copper within the xylem. A 3-year field trial in an olive orchard containing mature Cellina di Nardò and Ogliarola salentina olive trees, and officially declared infected by X. fastidiosa subsp. pauca,was also carried out o to determine if the compound affected severity of the disease. Each year, from early April to October (excluding July and August), six spray treatments of 0.5% (v:v) Dentamet® were applied on the olive tree crowns. The compound reduced severity of symptoms in both cultivars. Most untreated trees died by the end of the trial, whereas all treated trees survived with good vegetative status as assessed by a normalized difference vegetation index. Quantitative real-time PCR was performed from June 2016 to September 2017, following the official procedures established by the European and Mediterranean Plant Protection Organization. The analysis revealed a statistically significant reduction of X. fastidiosa cell densities within the leaves of treated trees. These promising results suggest that integrated management to reduce severity of X. fastidiosa that includes regular pruning and soil harrowing with spring and summer spray treatments with Dentamet®, is likely to effectively control the disease.

Robust linear parameter varying control of an unmanned aerial vehicle

The dynamic response characteristics of modern unmanned aerial vehicles (UAVs) are highly nonlinear and vary substantially with flight conditions due to their reduced dimensions compared to normal aircraft. In this thesis, design frameworks that are based on parameter dependent Lyapunov functions (PDLFs) are developed for UAV flight control systems. These design frameworks or procedures can systematically deal with aircraft systems with nonlinear and parameter dependent dynamics, and uncertainty in the mathematical models. To this end, we analyse robust stability and performance of LPV systems and present two LPV controller design methods using the PDLF approach: Two-Degree-of-Freedom (2DoF) and loop shaping with coprime factorisation. We formulate and solve the control problem for an LPV plant with measurable parameters and an output feedback structure. The solvability conditions are reduced to LMIs and can be solved approximately using finite-dimensional convex programming. A parameter dependent performance approach is used in a 2DoF/PDLF design and constitutes a flexible generalisation for calibrations of local performance. In loop shaping/PDLF design, a left coprime factorisation is derived by H2 filtering, and then a loop shaping design is implemented in the PDLF framework. We also incorporate pole placement constraints into the LMI synthesis to improve controller performance. To be able to use the robust gain-scheduling synthesis results, an LPV model of the UAV is developed and validated. The gain scheduling controller design of longitudinal/lateral-directional dynamics of the UAV is illustrated in the design example. It is shown that a flight control system can be built with satisfactory robust stability and performance