Flavescence doree phytoplasma strain FD92 gene for variable membrane protein B
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PROVIDER: PRJEB32304 | ENA |
REPOSITORIES: ENA
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Project description:The adhesion of flavescence dorée phytoplasma to the midgut epithelium cells of their insect vectors is partially mediated by the Variable Membrane Protein A (VmpA), an adhesin which shows lectin properties. In order to identify the insect receptor for VmpA, we lokked for Euscelidius variegatus cells proteins interacting with recombinant VmpA-His6 by mass spectrometry analysis of VmpA-E. variegatus protein complexes formed upon in vitro interaction assays.
2023-11-06 | PXD046447 | Pride
Project description:Flavescence dorée is the most serious grapevine yellows disease in Europe. It is caused by phytoplasmas which are transmitted from grapevine to grapevine by the leafhopper Scaphoideus titanus. Differences in susceptibility among grapevine varieties suggest the existence of specific genetic features associated with resistance to the phytoplasma and/or possibly with its vector. In this work, RNA-Seq was used to compare early transcriptional changes occurring during the three-trophic interaction between the phytoplasma, its vector and the grapevine, represented by two different cultivars, one very susceptible to the disease and the other scarcely susceptible. Background: Flavescence dorée is the most serious grapevine yellows disease in Europe. It is caused by phytoplasmas which are transmitted from grapevine to grapevine by the leafhopper Scaphoideus titanus. Differences in susceptibility among grapevine varieties suggest the existence of specific genetic features associated with resistance to the phytoplasma and/or possibly with its vector. In this work, RNA-Seq was used to compare early transcriptional changes occurring during the three-trophic interaction between the phytoplasma, its vector and the grapevine, represented by two different cultivars, one very susceptible to the disease and the other scarcely susceptible. The comparison of the transcriptomic responses highlighted both passive and active defense mechanisms against the vector and/or the pathogen in the scarcely-susceptible variety, as well as the capacity of the phytoplasmas to repress the defense reaction against the insect in the susceptible variety.
2019-06-13 | E-MTAB-7710 | biostudies-arrayexpress
Project description:Flavescence dorée phytoplasma has multiple ftsH genes differentially expressed in plants and insects
| PRJEB26323 | ENA
Project description:BackgroundFlavescence dorée (FD) of grapevine is a phloem bacterial disease that threatens European vineyards. The disease is associated with a non-cultivable mollicute, a phytoplasma that is transmitted by the grapevine leafhopper Scaphoideus titanus in a persistent, propagative manner. The specificity of insect transmission is presumably mediated through interactions between the host tissues and phytoplasma surface proteins comprising the so-called variable membrane proteins (Vmps). Plant spiroplasmas and phytoplasmas share the same ecological niches, the phloem sieve elements of host plants and the hemocoel of insect vectors. Unlike phytoplasmas, however, spiroplasmas, and Spiroplasma citri in particular, can be grown in cell-free media and genetically engineered. As a new approach for studying phytoplasmas-insect cell interactions, we sought to mimic phytoplasmas through the construction of recombinant spiroplasmas exhibiting FD phytoplasma Vmps at the cell surface.ResultsHere, we report the expression of the FD phytoplasma VmpA in S. citri. Transformation of S. citri with plasmid vectors in which the vmpA coding sequence was under the control of the S. citri tuf gene promoter resulted in higher accumulation of VmpA than with the native promoter. Expression of VmpA at the spiroplasma surface was achieved by fusing the vmpA coding sequence to the signal peptide sequence of the S. citri adhesin ScARP3d, as revealed by direct colony immunoblotting and immunogold labelling electron microscopy. Anchoring of VmpA to the spiroplasma membrane was further demonstrated by Triton X-114 protein partitioning and Western immunoblotting. Using the same strategy, the secretion of free, functionally active β-lactamase (used as a model protein) into the culture medium by recombinant spiroplasmas was achieved.ConclusionsConstruction of recombinant spiroplasmas harbouring the FD phytoplasma variable membrane protein VmpA at their surface was achieved, which provides a new biological approach for studying interactions of phytoplasma surface proteins with host cells. Likewise, the secretion of functional β-lactamase by recombinant spiroplasmas established the considerable promise of the S. citri expression system for delivering phytoplasma effector proteins into host cells.
| S-EPMC4392738 | biostudies-literature
Project description:The variable membrane protein VmpA of flavescence dorée phytoplasma acts as an adhesin binding cells of the insect vector Euscelidius variegatus
| PRJEB22700 | ENA
Project description:A comprehensive transcriptional landscape of Flavescence dorée phytoplasma infecting in field-grown Vitis vinifera leaves
| PRJEB6982 | ENA
Project description:Phytoplasmas are uncultivated plant pathogens and cell wall-less bacteria and are transmitted from plant to plant by hemipteran insects. The phytoplasma's circulative propagative cycle in insects requires the crossing of the midgut and salivary glands, and primary adhesion to cells is an initial step toward the invasion process. The flavescence dorée (FD) phytoplasma possesses a set of variable membrane proteins (Vmps) exposed on its surface, and this pathogen is suspected to interact with insect cells. The results showed that VmpA is expressed by the flavescence dorée phytoplasma present in the midgut and salivary glands. Phytoplasmas cannot be cultivated at present, and no mutant can be produced to investigate the putative role of Vmps in the adhesion of phytoplasma to insect cells. To overcome this difficulty, we engineered the Spiroplasma citri mutant G/6, which lacks the ScARP adhesins, for VmpA expression and used VmpA-coated fluorescent beads to determine if VmpA acts as an adhesin in ex vivo adhesion assays and in vivo ingestion assays. VmpA specifically interacted with Euscelidius variegatus insect cells in culture and promoted the retention of VmpA-coated beads to the midgut of E. variegatus In this latest case, VmpA-coated fluorescent beads were localized and embedded in the perimicrovillar membrane of the insect midgut. Thus, VmpA functions as an adhesin that could be essential in the colonization of the insect by the FD phytoplasmas.IMPORTANCE Phytoplasmas infect a wide variety of plants, ranging from wild plants to cultivated species, and are transmitted by different leafhoppers, planthoppers, and psyllids. The specificity of the phytoplasma-insect vector interaction has a major impact on the phytoplasma plant host range. As entry into insect cells is an obligate process for phytoplasma transmission, the bacterial adhesion to insect cells is a key step. Thus, studying surface-exposed proteins of phytoplasma will help to identify the adhesins implicated in the specific recognition of insect vectors. In this study, it is shown that the membrane protein VmpA of the flavescence dorée (FD) phytoplasma acts as an adhesin that is able to interact with cells of Euscelidius variegatus, the experimental vector of the FD phytoplasma.
| S-EPMC5881044 | biostudies-literature
Project description:Transcriptional profiling of phytoplasma grown in plant (Chrysanthemum coronarium) and grown in insect (Macrosteles striifrons). Two-condition experiment, phytoplasma-infected plant and phytoplasma-infected insect. Biological replicates: 6 phytoplasma-infected plants and 6 phytoplasma-infected insects, independently grown and harvested. One replicate per array.
2012-01-16 | E-GEOD-30302 | biostudies-arrayexpress
Project description:Transcriptional profiling of phytoplasma grown in plant (Chrysanthemum coronarium) and grown in insect (Macrosteles striifrons).
2012-01-16 | GSE30302 | GEO
Project description:Transcriptional profiling of phytoplasma grown in plant (Chrysanthemum coronarium) and grown in insect (Macrosteles striifrons). Two-condition experiment, phytoplasma-infected plant and phytoplasma-infected insect. Biological replicates: 4 phytoplasma-infected plants and 4 phytoplasma-infected insects, independently grown and harvested. One replicate per array.
2012-01-16 | E-GEOD-29274 | biostudies-arrayexpress