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I Barash - One of the best experts on this subject based on the ideXlab platform.
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revealing the inventory of type iii effectors in pantoea agglomerans gall forming pathovars using draft genome sequences and a machine learning approach
2018Co-Authors: Gal Nissan, Laura Chalupowicz, Guido Sessa, I Barash, Shulamit Manulissasson, Michael Gershovits, Michael Morozov, Tal PupkoAbstract:Pantoea agglomerans, a widespread epiphytic bacterium, has evolved into a hrp-dependent and host-specific gall-forming pathogen by acquiring a pathogenicity plasmid containing a type III secretion system (T3SS) and its effectors (T3Es). P. agglomerans pv. betae (Pab) elicits galls on beet (Beta vulgaris) and Gypsophila (Gypsophila paniculata), whereas P. agglomerans pv. Gypsophilae (Pag) incites galls on Gypsophila and hypersensitive response (HR) on beet. Draft genome sequences were generated and employed in combination with a machine learning approach and a translocation assay into beet roots to identify the pools of T3Es in the two pathovars. The genomes of the sequenced Pab4188 and Pag824-1 strains have a similar size (∼5 MB) and GC content (∼55%). Mutational analysis revealed that in Pab4188 eight T3Es (HsvB, HsvG, PseB, DspA/E, HopAY1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on beet and Gypsophila. In Pag824-1, nine T3Es (HsvG, HsvB, PthG, DspA/E, HopAY1, HopD1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on Gypsophila, whereas the PthG effector triggers HR on beet. HsvB, HsvG, PthG and PseB appear to endow pathovar specificities to Pab and Pag and no homologous T3Es were identified for these proteins in other phytopathogenic bacteria. Conversely, the remaining T3Es contributed to virulence of both pathovars and homologous T3Es were found in other phytopathogenic bacteria. Remarkably, HsvG and HsvB, which act as host-specific transcription factors, displayed the largest contribution to disease development. This article is protected by copyright. All rights reserved.
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characterization of nuclear localization signals in the type iii effectors hsvg and hsvb of the gall forming bacterium pantoea agglomerans
2011Co-Authors: Dan M. Weinthal, Victor Gaba, Guido Sessa, I Barash, Tzvi Tzfira, Doron Teper, Shulamit ManulissassonAbstract:HsvG and HsvB, two paralogous type III effectors of the gall-forming bacteria Pantoea agglomerans pv. Gypsophilae and P. agglomerans pv. betae, determine host specificity on Gypsophila and beet, respectively. They were previously shown to be DNA-binding proteins imported into host and non-host nuclei and might act as transcriptional activators. Sequence analysis of these effectors did not detect canonical nuclear localization signals (NLSs), but two basic amino acid clusters designated putative NLS1 and NLS2 were detected in their N-terminal and C-terminal regions, respectively. pNIA assay for nuclear import in yeast and bombardment of melon leaves with each of the NLSs fused to a 2xYFP reporter indicated that putative NLS1 and NLS2 were functional in transport of HsvG into the nucleus. A yeast two-hybrid assay showed that HsvB, HsvG, putative NLS1, putative NLS2, HsvG converted into HsvB, or HsvB converted into HsvG by exchanging the repeat domain, all interacted with AtKAP-α and importin-α3 of Arabidopsis thaliana. Deletion analysis of the NLS domains in HsvG suggested that putative NLS1 or NLS2 were required for pathogenicity on Gypsophila cuttings and presumably for import of HsvG into the nucleus. This study demonstrates the presence of two functional NLSs in the type III effectors HsvG and HsvB.
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the type iii effectors hsvg and hsvb of gall forming pantoea agglomerans determine host specificity and function as transcriptional activators
2006Co-Authors: Gal Nissan, Dan M. Weinthal, Guido Sessa, Shulamit Manulissasson, Henia Mor, I BarashAbstract:Pantoea agglomerans pv. Gypsophilae (Pag) elicits galls on Gypsophila and a hypersensitive response on beet, whereas P. agglomerans pv. betae (Pab) induces galls on both beet and Gypsophila. The pathogenicity of both pathovars is dependent on the presence of a plasmid harbouring type III secretion system (TTSS) components and effectors. The HsvG TTSS effectors of Pag (HsvG-Pag) and Pab (HsvG-Pab) determine the host specificity of both pathovars on Gypsophila. Here we describe a novel HsvG homologue, HsvB, which determines the host specificity of Pag and Pab on beet. HsvG requires two direct amino acid repeats for pathogenicity on Gypsophila, whereas one repeat in HsvB is sufficient for pathogenicity on beet. Exchanging repeats between HsvG-Pag and HsvB-Pab resulted in a switch of host specificities. Transient expression of GFP-HsvG or GFP-HsvB fusions in Gypsophila, beet or melon leaves showed that HsvG and HsvB were localized to the nuclei of host and non-host plants. A yeast one-hybrid assay revealed that a single repeat of HsvG or HsvB was sufficient to activate transcription. By employing random binding-site selection and gel-shift assay HsvG was demonstrated to be a double-stranded DNA-binding protein with an ACACC/aAA consensus binding site. These results suggest that HsvG and HsvB are host-specificity determinants and bear the potential to affect the host transcriptional machinery.
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comparative anatomy of gall development on Gypsophila paniculata induced by bacteria with different mechanisms of pathogenicity
2006Co-Authors: Laura Chalupowicz, I Barash, M Schwartz, Roni Aloni, S ManulisAbstract:Galls induced on Gypsophila paniculata by Pantoea agglomerans pv. Gypsophilae (Pag) and Agrobacterium tumefaciens (At), bacteria with different mechanisms of pathogenicity, were compared morphologically and anatomically. The pathogenicity of Pag is dependent on the presence of an indigenous plasmid that harbors hrp gene cluster, genes encoding Hop virulence proteins and biosynthetic genes for auxin (IAA) and cytokinins (CKs), whereas that of At involves host transformation. The Pag-induced gall was rough, brittle and exhibited limited growth, in contrast to the smooth, firm appearance and continuous growth of the At-induced gall. Anatomical analysis revealed the presence of cells with enlarged nuclei and multiple nucleoli, giant cells and suberin deposition in Pag that were absent from At-induced galls. Although circular vessels were observed in both gall types, they were more numerous and the vascular system was more organized in At. An aerenchymal tissue was observed in the upper part of the galls. Ethylene emission from Pag galls, recorded 6 days after inoculation, was eight times as great as that from non-infected controls. In contrast, a significant decrease in ethylene production was observed in Gypsophila cuttings infected with Pag mutants deficient in IAA and CK production. The results presented are best accounted for by the two pathogens having distinct pathogenicity mechanisms that lead to their differential recognition by the host as non-self (Pag) and self (At).
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cloning and characterization of iaam and iaah from erwinia herbicola pathovar Gypsophilae
1993Co-Authors: Ellen M Clark, Shulamit Manulis, Yakir Ophir, I Barash, Yedidya GafniAbstract:Erwinia herbicola pv. Gypsophilae induces galls on its host, Gypsophila paniculata. A 16-kb DNA fragment derived from a 78-Md native plasmid with homology to the iaa operon of Pseudomonas syringae pv. savastanoi was isolated from an EMBL3 library of E. h. Gypsophilae, strain PD713, DNA. A 7.5-kb EcoRI fragment was subcloned into pUC118 to generate pEG101. Escherichia coli DH5a cells transformed with pEG101 produced indole-3-acetic acid (IAA) when cultured in medium supplemented with L-tryptophan (TRP) []
Inoñan Zeña, Henry Omar - One of the best experts on this subject based on the ideXlab platform.
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Exportación de Gypsophila al mercado estadounidense en el periodo 2008 – 2015
2016Co-Authors: Inoñan Zeña, Henry OmarAbstract:En los últimos años, las exportaciones de Gypsophila en el mercado internacional no han tenido una buena acogida debido a diferentes factores externos e internos del país. Pero en la actualidad existen diferentes tratados o convenios internacionales firmados por el estado peruano, que favorecen a las exportaciones de la gyspophila como una opción dentro de los productos de exportación no tradicionales, aumentando el valor y volumen en las exportaciones anuales. El objetivo de la presente investigación ha sido determinar la tendencia positiva de las exportaciones de Gypsophila al mercado de los Estados Unidos mediante el periodo 2008 – 2015. Para analizar estos datos hemos utilizado el método estadístico que consiste en organizar la información mediante cuadros, luego presentar la información mediante gráficos de línea y luego describirlos mediante los estadísticos correspondientes
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Exportación de Gypsophila al mercado estadounidense en el periodo 2008 – 2015
2016Co-Authors: Inoñan Zeña, Henry OmarAbstract:TesisLima NorteEscuela Académico Profesional de Negocios InternacionalesComercio InternacionalEn los últimos años, las exportaciones de Gypsophila en el mercado internacional no han tenido una buena acogida debido a diferentes factores externos e internos del país. Pero en la actualidad existen diferentes tratados o convenios internacionales firmados por el estado peruano, que favorecen a las exportaciones de la gyspophila como una opción dentro de los productos de exportación no tradicionales, aumentando el valor y volumen en las exportaciones anuales. El objetivo de la presente investigación ha sido determinar la tendencia positiva de las exportaciones de Gypsophila al mercado de los Estados Unidos mediante el periodo 2008 – 2015. Para analizar estos datos hemos utilizado el método estadístico que consiste en organizar la información mediante cuadros, luego presentar la información mediante gráficos de línea y luego describirlos mediante los estadísticos correspondientes
Pérez Padilla, Katherine Milagros - One of the best experts on this subject based on the ideXlab platform.
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Comercio Internacional y Competitividad de la Gypsophila peruana 2008-2015
2016Co-Authors: Pérez Padilla, Katherine MilagrosAbstract:El presente trabajo fue realizado con el objetivo de determinar el comercio internacional y la competitividad de la Gypsophila peruana en el comercio internacional durante el periodo 2008-2015.La metodología empleada es de tipo descriptivo, y de diseño no experimental, puesto que no existe manipulación de las variables, además se observan antecedentes históricos y se consultaran fuentes. Una vez recopilados los datos, utilizamos e método estadístico que consiste en organizarlos a través de cuadros, gráficos de líneas y describirlo a través de la estimación de la línea de tendencia y el coeficiente e determinación. Se obtuvo como principal conclusión que la Gypsophila tuvo una tendencia de exportación negativa dado que el volumen tuvo tendencia negativa y el valor tuvo tendencia de exportación negativa dentro de los años 2008-2015
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Comercio Internacional y Competitividad de la Gypsophila peruana 2008-2015
2016Co-Authors: Pérez Padilla, Katherine MilagrosAbstract:TesisLima NorteEscuela Académico Profesional de Negocios InternacionalesComercio InternacionalEl presente trabajo fue realizado con el objetivo de determinar el comercio internacional y la competitividad de la Gypsophila peruana en el comercio internacional durante el periodo 2008-2015.La metodología empleada es de tipo descriptivo, y de diseño no experimental, puesto que no existe manipulación de las variables, además se observan antecedentes históricos y se consultaran fuentes. Una vez recopilados los datos, utilizamos e método estadístico que consiste en organizarlos a través de cuadros, gráficos de líneas y describirlo a través de la estimación de la línea de tendencia y el coeficiente e determinación. Se obtuvo como principal conclusión que la Gypsophila tuvo una tendencia de exportación negativa dado que el volumen tuvo tendencia negativa y el valor tuvo tendencia de exportación negativa dentro de los años 2008-2015
Max Henry - One of the best experts on this subject based on the ideXlab platform.
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Triterpenoid Saponins From the Roots of Gypsophila Trichotoma Wender
2013Co-Authors: Laurence Voutquenne-nazabadioko, Alexander Weng, Reneta Gevrenova, Nicolas Bories, Dominique Harakat, Charlotte Sayagh, Mayank Thakur, Maya Zaharieva, Max HenryAbstract:Eleven triterpenoid saponins were isolated from the roots of Gypsophila trichotoma Wender. (G. trichotoma Wender. var. trichotoma) (Caryophyllaceae), together with one known compound. The structures were established on the basis of extensive NMR analysis ((1)H, (13)C NMR, COSY, TOCSY, ROESY, HSQC, and HMBC), completed by analysis of HR-ESI-MS and ESI-MS(n). The saponins have the commonly found gypsogenin as the aglycone substituted at C-3 with trisaccharide and at C-28 with oligosaccharide through a fucose residue, as saponins isolated from Gypsophila perfoliata L. originated from China. The oligosaccharide attached to C-28 is substituted with acetyl and (or) sulfate groups. Тhe cytotoxicity of the saponin extract from G. trichotoma was evaluated against a rat alveolar macrophage-like cell line NR8383 and human leukemia cell lines U937 and BV-173. The synergistic effect of the aminoacyl saponins, previously isolated from G. trichotoma, was tested for its ability to enhance the cytotoxicity of the targeted toxin in HER14 cells.
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root in vitro cultures of six Gypsophila species and their saponin contents
2010Co-Authors: Reneta Gevrenova, Tanya Stancheva, Yulian Voynikov, Dominique Laurainmattar, Max HenryAbstract:A simple and rapid method of excised root cultures from six Gypsophila species was performed allowing continuous growth without phytohormones. Established on MH3 medium from solid-grown seedlings these roots were subcultured for 1 year on a solid medium before being transferred in a liquid medium to obtain substantial biomass for saponin content analysis. Morphologically, the different root lines presented different growth behaviors and different physical aspects: some have linear growth by the tip of the main axial root from the original seedling; others have additional lateral root initiations producing a hairy root-system more or less dense. A marked increase of biomass was observed in the light by comparison with dark conditions. Significant growth for Gypsophila glomerata was achieved within 3 weeks on liquid medium; biomass grew up to 50-fold in batch cultures reaching 10 g DW. The fingerprints of the saponin HPLC profiles of the six Gypsophila species were drastically different with at least up to 30 different saponins detected for some of them. The roots of Gypsophila elegans accumulated saponins up to 65 mg/g DW. These amounts were higher than in Gypsophila paniculata roots classically found as the best producing ones. On the contrary the root lines of G. glomerata showed a smaller quantitative amount of saponins (between 1.3 and 7.10 mg/g DW) than those of G. elegans but nearly the same HPLC profiles as for root extracts of G. paniculata plants grown directly in the fields.
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complete 1h and 13c nmr assignments of saponins from roots of Gypsophila trichotoma wend
2006Co-Authors: Reneta Gevrenova, Dominique Harakat, Laurence Voutquennenazabadioko, Elise Prost, Max HenryAbstract:The assignments of 1H and 13C NMR spectra of two new aminoacyl triterpene saponins from roots of Gypsophila trichotoma Wend. are reported. In addition to 1D NMR methods, 2D NMR techniques (COSY, TOCSY, ROESY, HSQC, HMBC, and HSQC-TOCSY) were used for the assignments. The structures were completed by analysis of HR-ESI-MS and ESI-MS(n).
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effects of Gypsophila saponins on bacterial growth kinetics and on selection of subterranean clover rhizosphere bacteria
2003Co-Authors: Francoise Fons, N Amellal, Corinne Leyval, N Saintmartin, Max HenryAbstract:Plant secondary metabolites, such as saponins, have a considerable impact in agriculture because of their allelopathic effects. They also affect the growth of soil microorganisms, especially fungi. We investigated the influence of saponins on rhizosphere bacteria in vitro and in soil conditions. The effects of Gypsophila saponins on the growth kinetics of rhizosphere bacteria were studied by monitoring the absorbance of the cultures in microtiter plates. Gypsophila saponins (1%) increased the lag phase of bacterial growth. The impact of Gypsophila saponins on subterranean clover rhizosphere was also investigated in a pot experiment. The addition of Gypsophila saponins did not modify clover biomass but significantly increased (twofold with 1% saponins) the weight of adhering soil. The number of culturable heterotrophic bacteria of the clover rhizosphere was not affected by the addition of Gypsophila saponins. Nevertheless, the phenotypical characterization of the dominant Gram-negative strains of the clover rhizosphere, using the Biolog system, showed qualitative and quantitative differences induced by 1% saponins. With the addition of saponins, the populations of Chryseomonas spp. and Acinetobacter spp., the two dominant culturable genera of control clover, were no longer detectable or were significantly decreased, while that of Aquaspirillum dispar increased and Aquaspirillum spp. became the major genus. Aquaspirillum dispar and Aquaspirillum spp. were also the dominant rhizosphere bacteria of Gypsophila paniculata, which greatly accumulates these saponins in its roots. These results suggest that saponins may control rhizosphere bacteria in soil through rhizodeposition mechanisms.
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effect of gypsogenin 3 o glucuronide pretreatment of Gypsophila paniculata and saponaria officinalis cell suspension cultures on the activities of microsomal 2 3 oxidosqualene cycloartenol and amyrin cyclases
1992Co-Authors: Max Henry, Alain Rahier, Maryse TatonAbstract:Abstract The regulation of triterpenoid saponin biosynthesis in Gypsophila paniculata and Saponaria officinalis suspension cells has been investigated by studying the effects of adding gypsogenin 3,O-glucuronide, a ubiquitous saponin precursor in plants of the genus Gypsophila which contain gypsogenin saponins, to the cultures. Treatment of suspended G. paniculata cells with gypsogenin 3,O-glucuronide 24 hr before administration of [ 14 C]acetate resulted in a marked reduction in the incorporation of radioactivity into saponins and their precursors but not into sterols and steryl glucosides. 2,3-Oxidosqualene-cycloartenol and -amyrin cyclase activities were measured in microsomal fractions obtained from suspension cultures of the two species. Pretreatment of cultures with gypsogenin 3,O-glucuronide was shown to lead to inhibition (in G. paniculata cells) or stimulation (in S. officinalis cells) of the 2,3-oxidosqualene-amyrin cyclase activity and was without any marked effect on 2,3-oxidosqualene-cycloartenol cyclase activity. These results strongly suggest that in these species 2,3-oxidosqualene-cyclases are regulating steps in the isoprenoid pathway orienting the biosynthetic flux towards either tetracyclic or pentacyclic triterpenes.
Guido Sessa - One of the best experts on this subject based on the ideXlab platform.
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revealing the inventory of type iii effectors in pantoea agglomerans gall forming pathovars using draft genome sequences and a machine learning approach
2018Co-Authors: Gal Nissan, Laura Chalupowicz, Guido Sessa, I Barash, Shulamit Manulissasson, Michael Gershovits, Michael Morozov, Tal PupkoAbstract:Pantoea agglomerans, a widespread epiphytic bacterium, has evolved into a hrp-dependent and host-specific gall-forming pathogen by acquiring a pathogenicity plasmid containing a type III secretion system (T3SS) and its effectors (T3Es). P. agglomerans pv. betae (Pab) elicits galls on beet (Beta vulgaris) and Gypsophila (Gypsophila paniculata), whereas P. agglomerans pv. Gypsophilae (Pag) incites galls on Gypsophila and hypersensitive response (HR) on beet. Draft genome sequences were generated and employed in combination with a machine learning approach and a translocation assay into beet roots to identify the pools of T3Es in the two pathovars. The genomes of the sequenced Pab4188 and Pag824-1 strains have a similar size (∼5 MB) and GC content (∼55%). Mutational analysis revealed that in Pab4188 eight T3Es (HsvB, HsvG, PseB, DspA/E, HopAY1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on beet and Gypsophila. In Pag824-1, nine T3Es (HsvG, HsvB, PthG, DspA/E, HopAY1, HopD1, HopX2, HopAF1 and HrpK) contribute to pathogenicity on Gypsophila, whereas the PthG effector triggers HR on beet. HsvB, HsvG, PthG and PseB appear to endow pathovar specificities to Pab and Pag and no homologous T3Es were identified for these proteins in other phytopathogenic bacteria. Conversely, the remaining T3Es contributed to virulence of both pathovars and homologous T3Es were found in other phytopathogenic bacteria. Remarkably, HsvG and HsvB, which act as host-specific transcription factors, displayed the largest contribution to disease development. This article is protected by copyright. All rights reserved.
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Polar auxin transport is essential for gall formation by Pantoea agglomerans on Gypsophila.
2012Co-Authors: Laura Chalupowicz, Dan M. Weinthal, Victor Gaba, Guido Sessa, Isaac Barash, Shulamit Manulis-sassonAbstract:Summary The virulence of the bacterium Pantoea agglomerans pv. Gypsophilae (Pag) on Gypsophila paniculata depends on a type III secretion system (T3SS) and its effectors. The hypothesis that plant-derived indole-3-acetic acid (IAA) plays a major role in gall formation was examined by disrupting basipetal polar auxin transport with the specific inhibitors 2,3,5-triiodobenzoic acid (TIBA) and N-1-naphthylphthalamic acid (NPA). On inoculation with Pag, galls developed in Gypsophila stems above but not below lanolin rings containing TIBA or NPA, whereas, in controls, galls developed above and below the rings. In contrast, TIBA and NPA could not inhibit tumour formation in tomato caused by Agrobacterium tumefaciens. The colonization of Gypsophila stems by Pag was reduced below, but not above, the lanolin–TIBA ring. Following Pag inoculation and TIBA treatment, the expression of hrpL (a T3SS regulator) and pagR (a quorum-sensing transcriptional regulator) decreased four-fold and that of pthG (a T3SS effector) two-fold after 24 h. Expression of PIN2 (a putative auxin efflux carrier) increased 35-fold, 24 h after Pag inoculation. However, inoculation with a mutant in the T3SS effector pthG reduced the expression of PIN2 by two-fold compared with wild-type infection. The results suggest that pthG might govern the elevation of PIN2 expression during infection, and that polar auxin transport-derived IAA is essential for gall initiation.
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characterization of nuclear localization signals in the type iii effectors hsvg and hsvb of the gall forming bacterium pantoea agglomerans
2011Co-Authors: Dan M. Weinthal, Victor Gaba, Guido Sessa, I Barash, Tzvi Tzfira, Doron Teper, Shulamit ManulissassonAbstract:HsvG and HsvB, two paralogous type III effectors of the gall-forming bacteria Pantoea agglomerans pv. Gypsophilae and P. agglomerans pv. betae, determine host specificity on Gypsophila and beet, respectively. They were previously shown to be DNA-binding proteins imported into host and non-host nuclei and might act as transcriptional activators. Sequence analysis of these effectors did not detect canonical nuclear localization signals (NLSs), but two basic amino acid clusters designated putative NLS1 and NLS2 were detected in their N-terminal and C-terminal regions, respectively. pNIA assay for nuclear import in yeast and bombardment of melon leaves with each of the NLSs fused to a 2xYFP reporter indicated that putative NLS1 and NLS2 were functional in transport of HsvG into the nucleus. A yeast two-hybrid assay showed that HsvB, HsvG, putative NLS1, putative NLS2, HsvG converted into HsvB, or HsvB converted into HsvG by exchanging the repeat domain, all interacted with AtKAP-α and importin-α3 of Arabidopsis thaliana. Deletion analysis of the NLS domains in HsvG suggested that putative NLS1 or NLS2 were required for pathogenicity on Gypsophila cuttings and presumably for import of HsvG into the nucleus. This study demonstrates the presence of two functional NLSs in the type III effectors HsvG and HsvB.
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the type iii effectors hsvg and hsvb of gall forming pantoea agglomerans determine host specificity and function as transcriptional activators
2006Co-Authors: Gal Nissan, Dan M. Weinthal, Guido Sessa, Shulamit Manulissasson, Henia Mor, I BarashAbstract:Pantoea agglomerans pv. Gypsophilae (Pag) elicits galls on Gypsophila and a hypersensitive response on beet, whereas P. agglomerans pv. betae (Pab) induces galls on both beet and Gypsophila. The pathogenicity of both pathovars is dependent on the presence of a plasmid harbouring type III secretion system (TTSS) components and effectors. The HsvG TTSS effectors of Pag (HsvG-Pag) and Pab (HsvG-Pab) determine the host specificity of both pathovars on Gypsophila. Here we describe a novel HsvG homologue, HsvB, which determines the host specificity of Pag and Pab on beet. HsvG requires two direct amino acid repeats for pathogenicity on Gypsophila, whereas one repeat in HsvB is sufficient for pathogenicity on beet. Exchanging repeats between HsvG-Pag and HsvB-Pab resulted in a switch of host specificities. Transient expression of GFP-HsvG or GFP-HsvB fusions in Gypsophila, beet or melon leaves showed that HsvG and HsvB were localized to the nuclei of host and non-host plants. A yeast one-hybrid assay revealed that a single repeat of HsvG or HsvB was sufficient to activate transcription. By employing random binding-site selection and gel-shift assay HsvG was demonstrated to be a double-stranded DNA-binding protein with an ACACC/aAA consensus binding site. These results suggest that HsvG and HsvB are host-specificity determinants and bear the potential to affect the host transcriptional machinery.