Dataset Information

Functional analysis of phytoplasma effectors in Vitis vinifera pathogenesis

ABSTRACT: The aim of the experiment was to identify important protein targets of phytoplasma infection, by isolation of plant proteins, interacting with phytoplasma effectors. Nicotiana benthamiana plants were transformed with YFP (Yellow Fluorescent Protein) tagged phytoplasma effector PoStoSP28, the leaves of transformed plants were homogenized and the effector was extracted together with its interactors through the YFP tag with a pull-down assay. As a control, the same procedure was performed with mock transformed plants and the samples analyzed with mass spectrometry.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Nicotiana Tabacum (common Tobacco)

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Stefane Wienkoop

LAB HEAD: Stefanie Wienkoop

PROVIDER: PXD015374 | Pride | 2023-10-24

REPOSITORIES: Pride

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Publications

Candidate pathogenicity factor/effector proteins of 'Candidatus Phytoplasma solani' modulate plant carbohydrate metabolism, accelerate the ascorbate-glutathione cycle, and induce autophagosomes.

Dermastia Marina M Tomaž Špela Š Strah Rebeka R Lukan Tjaša T Coll Anna A Dušak Barbara B Anžič Barbara B Čepin Timotej T Wienkoop Stefanie S Kladnik Aleš A Zagorščak Maja M Riedle-Bauer Monika M Schönhuber Christina C Weckwerth Wolfram W Gruden Kristina K Roitsch Thomas T Pompe Novak Maruša M Brader Günter G

Frontiers in plant science 20230818

The pathogenicity of intracellular plant pathogenic bacteria is associated with the action of pathogenicity factors/effectors, but their physiological roles for most phytoplasma species, including 'Candidiatus Phytoplasma solani' are unknown. Six putative pathogenicity factors/effectors from six different strains of 'Ca. P. solani' were selected by bioinformatic analysis. The way in which they manipulate the host cellular machinery was elucidated by analyzing Nicotiana benthamia ...[more]

PMID: 37662165

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Project description:Stomata are pores in the epidermis of plants that can open and close and allow for gas exchange vital for photosynthesis and regulate transpiration. Stomatal development is driven by a set of conserved bHLH transcription factors (SPCH, MUTE, FAMA, and their heterodimerization partners ICE1, SCRM2), that initiate and promote progression of cell fates in the stomatal lineage. Due to the shared ancestry of SPCH, MUTE and FAMA (subgroup Ia) and ICE1 and SCRM2 (subgroup IIIb) their DNA binding specificity is similar and there is some functional redundancy. However, individual bHLHs also have unique functions. For example, SPCH is required for initiation of the stomatal lineage, while FAMA is responsible for terminal differentiation of the guard cell pair. In grasses, the stomatal complex comprises the guard cell pair, and two flanking subsidiary cells. Recruitment of the latter from neighboring cell filed during development requires expression of MUTE. Remarkably, while MUTE is absolutely required for the promotion of guard mother cell fate in maize and rice, this is not the case in Brachypodium. This suggests that another TF can at least partially substitute for MUTE in this function. While different expression profiles of the bHLH dimers within the stomatal lineage may partially responsible for distinct functions of each pair, it is likely that each pair forms different transcriptional complexes and that interaction with other transcriptional regulators affects the dimer’s binding DNA binding and gene regulation properties. Given the differences in bHLH function between dicots and grasses and even within the grass family, we were interested in elucidating the protein interaction networks of the stomatal lineage regulators in Brachypodium. To this end, we performed co-immunoprecipitation coupled to LC-MS/MS of BdSPCH2-YFP, YFP-BdMUTE, YFP-BdFAMA, YFP-ICE1 and YFP-SCRM2 from the developmental zone of young B. distachyon leaves using GFP-Trap beads. All YFP-fusion proteins were expressed under the endogenous promoter in the Bd21-3 background. The only exception was BdICE1, which was expressed under the ZmUBI promoter, but was nevertheless mostly restricted to the stomatal lineage. As control lines we use the Bd21-3 wild type and a line expressing 3x YFPnls (nuclear YFP) under the MUTE promoter. Comparison of proteins enriched with the bHLH fusion proteins vs. the controls revealed overlapping and distinct putative interactors, which is in agreement with the assumption that these transcription factors have both shared and unique functions. Notably, in addition to the presumed hetero-dimerization partners, we found a number of other bHLH transcription factors that were identified with one or more of the bait proteins. This suggests the presence of a larger bHLH network acting in the stomatal lineage.

Dataset Information

Functional analysis of phytoplasma effectors in Vitis vinifera pathogenesis

Publications

Candidate pathogenicity factor/effector proteins of '<i>Candidatus</i> Phytoplasma solani' modulate plant carbohydrate metabolism, accelerate the ascorbate-glutathione cycle, and induce autophagosomes.

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