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Proteomic characterization of plasma membrane-enriched fraction from tobacco (Nicotiana tabacum L. cv Samsun) pollen tubes.

ABSTRACT: Pollen germination and subsequent pollen tube elongation are essential for successful land plant reproduction. These processes are achieved through well-documented activation of membrane trafficking and cell metabolism. Despite this, our knowledge of the dynamics of cellular phospholipids remains scarce. In this project, we analyzed the turnover of the glycerolipid composition during the establishment of cell polarity and elongation processes in tobacco pollen and described the proteo-lipid composition of pollen plasma membrane-enriched. To achieve this, we have combined several techniques, such as lipidomics, live-cell microscopy, and plasma membrane isolation coupled with mass spectrometry-based proteomic characterization.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Nicotiana Tabacum (common Tobacco)

TISSUE(S): Plant Cell, Pollen

DISEASE(S): Disease Free

SUBMITTER: Martin Hubalek

LAB HEAD: Martin Hubálek

PROVIDER: PXD037046 | Pride | 2022-10-27

REPOSITORIES: Pride

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Project description:As in animals, cell-cell communication plays pivotal role in male-female recognition during plant sexual reproduction. Prelaid peptides secreted from the female reproductive tissues guide pollen tubes towards ovules for fertilization. However, the elaborate mechanisms for this dialogue have remained elusive, particularly from the male perspective. We perform genome-wide quantitative liquid chromatography coupled tandem mass spectrometry of a pistil-stimulated pollen tube secretome and identify 801 pollen tube-secreted proteins. Interestingly, in silico analysis reveals that the pollen tube-secretome is dominated by unconventional-type secreted proteins representing 57% of the total secretome. In support, we show that unconventional-type protein, translationally controlled tumor protein, is secreted to the apoplast. Remarkably, we discover that this protein could be secreted by infiltrating through the initial phases of the conventional secretory pathway and could reach the apoplast via exosomes as demonstrated by co-localization with Oleisin1 exosome marker. We demonstrate that Arabidopsis thaliana translationally controlled tumor protein-knockdown plants have pollen tubes that poorly navigate to the target ovule, and theÂ knocked down allele is poorly transmitted through the male. We show that regulators of the endoplasmic reticulum-trans-golgi network protein secretory pathway control secretion of pollen tube-secreted cysteine-rich proteins, including pollen tube attractants, and are essential for pollen tube growth and guidance, as well as ovule-targeting competence. This work, the first pollen tube secretome study, identifies novel genome-wide pollen tube-secreted proteins with potential function in pollen tube-ovule guidance for sexual reproduction. Functional analysis highlights a potential mechanism for pollen tube unconventional protein secretion and reveals likely regulators of pollen tube protein secretion. The association of pollen tube-secreted proteins with marker proteins shown to be secreted via exosomes in other species suggest secretion via exosomes as a possible mechanism for cell-cell communication between the pollen tube and female reproductive cells. For processed dataset with quantitative information, see Hafidh S, Potesil D, Fila J et al. Genome Bilogy 2016.

			Action	DRS
	210720_PMR3.mzML	Mzml
	210720_PMR3.mzid.gz	Mzid
	210720_PMR3.raw	Raw
	220614_PM_3_4.mzML	Mzml
	220614_PM_3_4.mzid.gz	Mzid

Dataset Information

Proteomic characterization of plasma membrane-enriched fraction from tobacco (Nicotiana tabacum L. cv Samsun) pollen tubes.

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