Proteomics

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Proteomics of isolated sieve tubes from N.tabacum


ABSTRACT: Symplasmicly connected cells called sieve elements form a network of tubes in the phloem of vascular plants. Sieve elements have essential functions as they provide routes for photoassimilate distribution, the exchange of developmental signals, and the coordination of defense responses. Nonetheless, they are the least understood main type of plant cells. They are extremely sensitive, possess a reduced endomembrane system without Golgi apparatus, and lack nuclei and translation machineries, so that transcriptomics and similar techniques cannot be applied. Moreover, the analysis of phloem exudates as a proxy for sieve element composition is marred by methodological problems. We developed a simple protocol for the isolation of sieve elements from leaves and stems of Nicotiana tabacum at sufficient amounts for large-scale proteome analysis. By quantifying the enrichment of individual proteins in purified sieve element relative to bulk phloem preparations, proteins of increased likelyhood to function specifically in sieve elements were identified. To evaluate the validity of this approach, YFP-constructs of genes encoding three of the candidate proteins were expressed in plants. Tagged proteins occurred exclusively in sieve elements. Two of them, a putative cytochrome b561/ferric reductase and a reticulon-like protein, appeared restricted to ER segments that were inaccessible to the luminal ER marker, HDEL-GFP, suggesting a previously unknown differentiation of the endomembrane system in sieve elements. Evidently, our list of promising candidate proteins provides a valuable exploratory tool for sieve element biology.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Nicotiana Tabacum (common Tobacco)

TISSUE(S): Plant Cell, Leaf, Stem

SUBMITTER: Joerg Hofmann  

LAB HEAD: Michael Knoblauch

PROVIDER: PXD030457 | Pride | 2022-02-17

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
10_TMT_stem_2ug.mzxml Mzxml
10_TMT_stem_2ug.raw Raw
10_TMT_vein_2ug.mzxml Mzxml
10_TMT_vein_2ug.raw Raw
11_TMT_stem_2ug.mzxml Mzxml
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Publications

Proteomics of isolated sieve tubes from <i>Nicotiana tabacum</i>: sieve element-specific proteins reveal differentiation of the endomembrane system.

Liu Yan Y   Vasina Viktoriya V VV   Kraner Max E ME   Peters Winfried S WS   Sonnewald Uwe U   Knoblauch Michael M  

Proceedings of the National Academy of Sciences of the United States of America 20220101 1


Symplasmicly connected cells called sieve elements form a network of tubes in the phloem of vascular plants. Sieve elements have essential functions as they provide routes for photoassimilate distribution, the exchange of developmental signals, and the coordination of defense responses. Nonetheless, they are the least understood main type of plant cells. They are extremely sensitive, possess a reduced endomembrane system without Golgi apparatus, and lack nuclei and translation machineries, so th  ...[more]

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