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Characterization of secreted proteins by symbiont technology

ABSTRACT: By adding Agrobacterium plant growth regulator genes to a binary plant transformation vector also encoding a gene of interest, we engineered plant tissues capable of autonomous cell division and vascularization while expressing biomolecules on the stems of host plants and in vitro plant cell tissue culture cured of Agrobacterium. We refer to these repurposed Agrobacterium galls as symbionts to reflect their intended beneficial interaction with the host plant. When grown in vitro, symbionts efficiently export secreted proteins into the culture media and may serve as an advantageous platform for heterologous protein expression from plant cells. In this experiment we monitored accumulation of a secreted marker portein (Flowering Locus T, tagged with mCherry) in the media of tomato cell symbionts grown in-vitro and characterized the native proteins that also accumulated in the culture medium.

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ORGANISM(S): Solanum Lycopersicum

SUBMITTER: Samuel Coradetti

LAB HEAD: Michelle Heck

PROVIDER: PXD071454 | Pride | 2026-03-06

REPOSITORIES: Pride

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Project description:Bathymodiolus mussels inhabiting deep-sea hydrothermal vents harbor bacterial symbionts in their gills, which support the animals’ diet. While the basic mechanisms of energy generation and CO2 fixation that drive these symbioses are largely established, details of molecular interactions between the symbiotic partners and adaptations to their respective habitats remain unknown. In this study, we therefore comparatively examined the genomes and proteomes of two Bathymodiolus hosts and their respective symbionts from different geographical locations. Two mussel species were proteomically compared: i) B. thermophilus mussel containing sulfur-oxidizing symbiont from the east pacific rise. thermophilus and ii) B. azoricus containing thiotrophic and methanotrophic symbionts from the mid-atlantic ridge. Symbionts (for both species) and host components (for B. azoricus) were selectively enriched using a multi-step centrifugation procedure. Enriched host and symbiont fractions along with unenriched gill foot tissue were subject to in-depth semi-quantitative proteomic analyses using the orbitrap and velos mass spectrometers. Proteins were quantified based on their spectral counts using the normalized spectral abundance factor (NSAF) method. We identified common strategies of metabolic interactions that provide mutual nutritional support between host and symbionts, such as the detoxification of ambient sulfide by the Bathymodiolus host, which provides a stable thiosulfate reservoir for the thiotrophic symbionts, and a putative amino acid cycling mechanism that could supply the host with symbiont-derived amino acids. A suite of genes and proteins putatively related to virulence or defense functions was particularly abundant in the B. thermophilus symbiont, compared to its symbiont relatives, and may pose a host species-specific adaptation. Our results reveal both, a high degree of integration between the symbiotic partners, and great potential to adapt to the prevailing environment, which facilitate the holobiont’s survival in its hydrothermal vent habitat.

			Action	DRS
	F428185.mztab	Mztab
	F428185.raw	Raw
	F428186.mztab	Mztab
	F428186.raw	Raw
	F428187.mztab	Mztab

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Characterization of secreted proteins by symbiont technology

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