RESUMEN
Carrot yellows disease has been associated for many years with the Gram-positive, insect-vectored bacteria, 'Candidatus Phytoplasma' and Spiroplasma citri. However, reports in the last decade also link carrot yellows symptoms with a different, Gram-negative, insect-vectored bacterium, 'Ca. Liberibacter solanacearum'. Our study shows that to date 'Ca. L. solanacearum' is tightly associated with carrot yellows symptoms across Israel. The genetic variant found in Israel is most similar to haplotype D, found around the Mediterranean Basin. We further show that the psyllid vector of 'Ca. L. solanacearum', Bactericera trigonica, is highly abundant in Israel and is an efficient vector for this pathogen. A survey conducted comparing conventional and organic carrot fields showed a marked reduction in psyllid numbers and disease incidence in the field practicing chemical control. Fluorescent in situ hybridization and scanning electron microscopy analyses further support the association of 'Ca. L. solanacearum' with disease symptoms and show that the pathogen is located in phloem sieve elements. Seed transmission experiments revealed that while approximately 30% of the tested carrot seed lots are positive for 'Ca. L. solanacearum', disease transmission was not observed. Possible scenarios that may have led to the change in association of the disease etiological agent with carrot yellows are discussed. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .
Asunto(s)
Daucus carota/microbiología , Hemípteros/microbiología , Insectos Vectores/microbiología , Enfermedades de las Plantas/microbiología , Rhizobiaceae/fisiología , Animales , Daucus carota/ultraestructura , Haplotipos , Hibridación Fluorescente in Situ , Israel , Hojas de la Planta/microbiología , Hojas de la Planta/ultraestructura , Rhizobiaceae/genética , Rhizobiaceae/aislamiento & purificación , Rhizobiaceae/ultraestructura , Semillas/microbiología , Semillas/ultraestructuraRESUMEN
The vascular pathogen Clavibacter michiganensis subsp. michiganensis is responsible for bacterial wilt and canker of tomato. Pathogenicity of this bacterium is dependent on plasmid-borne virulence factors and serine proteases located on the chromosomal chp/tomA pathogenicity island (PAI). In this study, colonization patterns and movement of C. michiganensis subsp. michiganensis during tomato infection was examined using a green fluorescent protein (GFP)-labeled strain. A plasmid expressing GFP in C. michiganensis subsp. michiganensis was constructed and found to be stable in planta for at least 1 month. Confocal laser-scanning microscopy (CLSM) of inoculated stems showed that the pathogen extensively colonizes the lumen of xylem vessels and preferentially attaches to spiral secondary wall thickening of the protoxylem. Acropetal movement of the wild-type strain C. michiganensis subsp. michiganensis NCPPB382 (Cmm382) in tomato resulted in an extensive systemic colonization of the whole plant reaching the apical region after 15 days, whereas Cmm100 (lacking the plasmids pCM1 and pCM2) or Cmm27 (lacking the chp/tomA PAI) remained confined to the area surrounding of the inoculation site. Cmm382 formed biofilm-like structures composed of large bacterial aggregates on the interior of xylem walls as observed by CLSM and scanning electron microscopy. These findings suggest that virulence factors located on the chp/tomA PAI or the plasmids are required for effective movement of the pathogen in tomato and for the formation of cellular aggregates.
Asunto(s)
Actinomycetales/fisiología , Proteínas Bacterianas/metabolismo , Pared Celular/microbiología , Enfermedades de las Plantas/microbiología , Plásmidos/genética , Solanum lycopersicum/microbiología , Actinomycetales/genética , Actinomycetales/ultraestructura , Proteínas Bacterianas/genética , Biopelículas/crecimiento & desarrollo , Pared Celular/ultraestructura , Cromosomas Bacterianos/genética , Regulación Bacteriana de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Islas Genómicas , Proteínas Fluorescentes Verdes , Solanum lycopersicum/ultraestructura , Organismos Modificados Genéticamente , Factores de Tiempo , Virulencia , Factores de Virulencia , Xilema/microbiología , Xilema/ultraestructuraRESUMEN
The molecular interactions between Clavibacter michiganensis subsp. michiganensis and tomato plant were studied by following the expression of bacterial virulence and host-defense genes during early stages of infection. The C. michiganensis subsp. michiganensis genes included the plasmid-borne cellulase (celA) and the serine protease (pat-1), and the serine proteases chpC and ppaA, residing on the chp/tomA pathogenicity island (PAI). Gene expression was measured following tomato inoculation with Cmm382 (wild type), Cmm100 (lacking the plasmids pCM1 and pCM2), and Cmm27 (lacking the PAI). Transcriptional analysis revealed that celA and pat-1 were significantly induced in Cmm382 at initial 12 to 72 h, whereas chpC and ppaA were highly expressed only 96 h after inoculation. Interdependence between the expression of chromosomal and of plasmid-located genes was revealed: expression of celA and pat-1 was substantially reduced in the absence of the chp/tomA PAI, whereas chpC and ppaA expressions were reduced in the absence of the virulence plasmids. Transcription of chromosomal genes involved in cell wall degradation (i.e., pelA1, celB, xysA, and xysB), was also induced at early stages of infection. Expression of the host-defense genes, chitinase class II and pathogenesis-related protein-5 isoform was induced in the absence of the PAI at early stages of infection, suggesting that PAI-located genes are involved in suppression of tomato basal defenses.
Asunto(s)
Regulación Bacteriana de la Expresión Génica/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Bacterias Grampositivas/fisiología , Bacterias Grampositivas/patogenicidad , Solanum lycopersicum/microbiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Pared Celular/metabolismo , Cromosomas Bacterianos , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , VirulenciaRESUMEN
We present a novel method for multiple alignment of protein structures and detection of structural motifs. To date, only a few methods are available for addressing this task. Most of them are based on a series of pairwise comparisons. In contrast, MASS (Multiple Alignment by Secondary Structures) considers all the given structures at the same time. Exploiting the secondary structure representation aids in filtering out noisy results and in making the method highly efficient and robust. MASS disregards the sequence order of the secondary structure elements. Thus, it can find non-sequential and even non-topological structural motifs. An important novel feature of MASS is subset alignment detection: It does not require that all the input molecules be aligned. Rather, MASS is capable of detecting structural motifs shared only by a subset of the molecules. Given its high efficiency and capability of detecting subset alignments, MASS is suitable for a broad range of challenging applications: It can handle large-scale protein ensembles (on the order of tens) that may be heterogeneous, noisy, topologically unrelated and contain structures of low resolution.
Asunto(s)
Algoritmos , Modelos Moleculares , Proteínas/química , Alineación de Secuencia/métodos , Análisis de Secuencia de Proteína/métodos , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Conformación Proteica , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Homología de Secuencia de Aminoácido , Programas Informáticos , Interfaz Usuario-ComputadorRESUMEN
This essay studies the convergence of brain research with the physiology of emotions during the early twentieth century. It argues that the brain entered the laboratory of emotions not as an object of knowledge, but as a technique for producing emotions, in spite of the laboratory. The new brain-generated emotion signaled an epistemic break in the nature of studied emotion. It restructured the relationships between physiological and psychological forms of knowledge. It embodied the historical and political concerns of physiologists with pain. And it excluded the affectively experiencing subject from the study of "emotion." The essay also suggests that the brain-generated emotion was an object suspended in time and abstracted from history. Its unique a-temporal and de-contextualized characteristic transformed emotion into a product of a laboratory whose mode of production mimicked the modern factory. The constitutive elements that were assembled in creating the brain as emotion-generator were instrumental for the important studies of James Papez, Paul MacLean, and for the modern concept of Limbic System.
Asunto(s)
Encéfalo , Emociones , Neurociencias/historia , Historia del Siglo XIX , Historia del Siglo XXRESUMEN
The author argues that during the late nineteenth and early twentieth centuries Anglo-American physiologists discovered that the emotional experiences of their laboratory animals influenced their experiments. Asserting that previous generations had ignored the affective dimension of experimentation, these physiologists developed practices for recording, reporting, managing, and controlling the emotions of laboratory animals during physiological encounters. The author also argues that emotion became a powerful conceptual, rhetorical, political, and practical tool of the modern laboratory and that physiologists invoked emotion in order to contain and interpret numerous physiological facts and artifacts, promulgate alternative forms of knowledge-making, privilege new knowledge claims, and diffuse mounting political pressures. The study of the affect of experiment aims to contribute to the history of experimentation, knowledge, and emotion.