Characterization of the Conserved Pseudomonas Syringae Pv. Tomato DC3000 Effector Protein, HopAA1-1 PDF Download
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Author: Kathy Roberts Munkvold
Publisher:
ISBN:
Category :
Languages : en
Pages : 286
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Author: Kathy Roberts Munkvold
Publisher:
ISBN:
Category :
Languages : en
Pages : 286
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Author: Misty D. Janes Wehling
Publisher:
ISBN:
Category :
Languages : en
Pages : 328
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Author: Alexandre Robert-Seilaniantz
Publisher:
ISBN:
Category :
Languages : en
Pages : 234
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Author:
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Category :
Languages : en
Pages :
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Author: Jay N. Worley
Publisher:
ISBN:
Category :
Languages : en
Pages : 238
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Pseudomonas syringae pv. tomato DC3000 (DC3000) is a model plant pathogenic bacterium that infects tomato and Arabidopsis thaliana. It requires the phytotoxin coronatine and the delivery of type III effector proteins (T3Es) into the host cell cytoplasm for defense suppression and virulence. CmaL is a small protein found to be necessary for coronatine production. Coronatine is a potent molecular mimic of jasmonoyl-isoleucine, a plant hormone conjugate involved in regulating plant defenses. Coronatine is constructed of two amide bond-linked moieties, coronafacic acid and coronamic acid. CmaL was shown to be required for the production of L-allo-isoleucine, a precursor for coronamic acid biosynthesis. DC3000 mutants lacking both cmaL and the T3E gene hopAA1-1 are reduced in speck formation in tomato. hopAA1-1 is member of the conserved effector locus, a group of effector genes located adjacent to the genes encoding the type three secretion apparatus that are widespread among P. syringae strains. HopAA1-1 is toxic to both plants and yeast upon expression within them. To gain insight into the basis for its toxicity in eukaryotic cells, the subcellular localization of HopAA1-1 was investigated. HopAA1-1 was found to colocalize with plant peroxisomes. Truncated derivatives of HopAA1-1 that are not cytotoxic and cannot promote symptom formation do not localize with peroxisomes. Additionally, other truncated derivatives of HopAA1-1 colocalize with the endoplasmic reticulum in addition to peroxisomes, suggesting that HopAA1 -1 interacts with the endomembrane system. A DC3000 mutant with 28 T3E genes deleted (DC3000D28E) is a recently developed tool for investigating effector functions. DC3000D28E derivatives with small sets of effector genes progressively restored show increasing virulence wh en inoculated by infiltration with a blunt syringe into the model plant Nicotiana benthamiana. Because of its location in a cluster of effector genes, cmaL was inadvertently deleted in the construction of DC3000D28E. The importance of coronatine and its partial redundancy with HopAA1-1 in promoting an early stage of pathogenesis was revealed by restoring cmaL and hopAA1-1 to selected DC3000D28E derivatives and assaying the strains by dip inoculation of N. benthamiana leaves, which requires bacteria to follow a natural infection route through stomata.
Author: Libo Shan
Publisher:
ISBN:
Category :
Languages : en
Pages : 246
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Author: Diana Carolina Mazo Molina
Publisher:
ISBN:
Category :
Languages : en
Pages : 132
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Pseudomonas syringae pv. tomato (Pst) is a persistent pathogen of tomato that causes bacterial speck disease. On tomato, resistance conferred by the gene Pto is effective against race 0 Pst strains which express the effector proteins AvrPto and/or AvrPtoB; however, race 1 strains of Pst, which do not express AvrPto/AvrPtoB but rather a different repertoire of effectors, evade Pto-mediated resistance. Race 1 strains of Pst are becoming increasingly common, and no simply-inherited genetic resistance to such strains is known. It was discovered that a locus in Solanum lycopersicoides, termed Pseudomonas tomato race 1 (Ptr1), confers resistance to race 1 Pst strains by recognizing the type III effector AvrRpt2. In Arabidopsis and apple, strains of Pst and Erwinia amylovora expressing AvrRpt2 degrade the RIN4 protein, thereby activating RPS2 or Mr5-mediated immunity, respectively. Ptr1 also recognized homologs of AvrRpt2 from diverse bacteria including one in Ralstonia pseudosolanacearum and this correlated with the ability of AvrRpt2 to degrade RIN4. Using site-directed mutagenesis of AvrRpt2, we found that, like RPS2, activation of Ptr1 requires AvrRpt2 proteolytic activity. Ptr1 detection of AvrRpt2 activity suggests it likely encodes an NLR protein or possibly a guardee such as RIN4. Ptr1 was identified by cloning of candidate NLR-encoding genes located in the Ptr1 region and testing using Agrobacterium-mediated transient expression in Nicotiana glutinosa identified one gene for the ability to activate the plant immune system in response to AvrRpt2 in the presence of tomato Rin4. Interestingly, while overexpression of Ptr1 in N. glutinosa leaves caused localized cell death, co-expression of Ptr1 with tomato Rin4 prevented this cell death. The protein encoded by Ptr1 has little similarity to RPS2 or Mr5, which suggests that Ptr1 is a third example of convergent evolution in different plant species for recognition of AvrRpt2. In summary, the Ptr1 gene has the potential to become an important component (along with Pto) in controlling bacterial speck disease. Further research focused on studying the mechanism of action between Ptr1 and Rin4 may contribute to a better understanding of the recognition of the type III effector AvrRpt2 in tomato.
Author: Vanessa Revindran
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 262
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We have created a yeast madel system to study the action of the plant pathogen effector HopM1 in Saccharomyces cerevisiae. Pseudomonas syringae, causative agent of bacterial speck in tomatoes, utilizes the type III secretion system to shuttle the effector proteins into the host cell. When expressed in yeast, HopM1 is lethal on solid media at 21°C, but not at 30°C and 37°C. The same temperature sensitive ability of HopM1 to cause death on solid media is also observed in liquid. As demonstrated by SDS PAGE-Western blot analysis, HopM1 protein is present at 21°C, 30°C and 37°C. At 21°C, a full-length protein of 78kDA is observed. At 30°C and 37°C, the majority of HopM1 protein exists as degraded fragments. HopM1 containing strains were visualized using the V5 epitope and immunofluorescent microscopy. HopM1 localizes to mitochondria and secretory organelles. This result was confirmed using cellular fractionation and sucrose gradient density centrifugation. When plated on media containg glycerol, we observed no change in expression of HopM1, thus indicating that is it unlikely that binding to mitochondria results in the lethal phenotype. We have isolated 19 spontaneous suppressor strains that are capable of surviving the HopM1 imposed lethality at 21°C. All strains have been examined for HopM1 protein expression, of which 13 express full-length HopM1 at 21°C, and 5 do not. SupM1-16, showed a significant increse in growth rates as compared to the wild type strain expressing HopM1. None of the suppressor strains show a change in localization of HopM1 as compared to wild type. One of the suppressor stains, SupM1-16 was sequenced to identify the gene(s) responsible for the suppression phenotype. Six genes that may be the suppressor gene were identified. The most likely candidate is RSP5; an E3 Ubiquitin Ligase. RSP5 contains a single mutation that changes a Glycine to Valine in the HECT domain. Overall our findings suggest that HopM1 kills the yeast cell by perturbing a secrectory pathway regulator and that mutation of RSP5 alters HopM1 effects on this pathway to allow survival.
Author:
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ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 1044
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Author: K. Lindsey
Publisher: Springer Science & Business Media
ISBN: 9780792311157
Category : Plant cell culture
Languages : en
Pages : 120
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Book Description
Basic techniques - cells tissue culture of model species. Tissue culture & transformation of crop species. Propagation & conservation of germplasm. Direct gene transfer & protoplast fusion. Reproductive tissues. Mutant selection.
