Dataset Information

Oryza sativa egg, sperm, callus, pollen and seedling proteome

ABSTRACT: In angiosperms, female gamete differentiation, fertilization, and subsequent zygotic development occur in embryo sacs deeply embedded in the ovaries. Despite their importance in plant reproduction and development, how the egg cell is specialized, fuses with the sperm cell, and converts into an active zygote for early embryogenesis remains unclear. This lack of knowledge is partly attributable to the difficulty of direct analyses of gametes in angiosperms. In this study, proteins from egg and sperm cells obtained from Oryza sativa flowers were separated by one-dimensional polyacrylamide gel electrophoresis and globally identified by highly sensitive liquid chromatography coupled with tandem mass spectroscopy. Proteome analyses were also conducted for seedlings, callus, and pollen grains to compare their protein expression profiles to those of gametes. Database was searched using Mascot software (ver.2.2.1, Matrix Science, MA, USA) with the following parameters. The fixed modification was propionasmide (Cys) and variable modification parameters were pyro-Glu, acetylation (protein N-terminus), and oxidation (Met). The maximum missed cleavage was set at 3 with a peptide mass tolerance of +/– 15 ppm. Peptide charges from +2 to +4 states and MS/MS tolerances of +/– 0.8 Da were allowed.

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Oryza Sativa (rice)

TISSUE(S): Callus, Seedling, Egg, Semen, Sperm, Pollen

SUBMITTER: Masato Taoka

LAB HEAD: Masato Taoka

PROVIDER: PXD000265 | Pride | 2013-08-02

REPOSITORIES: Pride

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Publications

Identification of proteins enriched in rice egg or sperm cells by single-cell proteomics.

Abiko Mafumi M Furuta Kensyo K Yamauchi Yoshio Y Fujita Chiharu C Taoka Masato M Isobe Toshiaki T Okamoto Takashi T

PloS one 20130725 7

In angiosperms, female gamete differentiation, fertilization, and subsequent zygotic development occur in embryo sacs deeply embedded in the ovaries. Despite their importance in plant reproduction and development, how the egg cell is specialized, fuses with the sperm cell, and converts into an active zygote for early embryogenesis remains unclear. This lack of knowledge is partly attributable to the difficulty of direct analyses of gametes in angiosperms. In the present study, proteins from egg ...[more]

PMID: 23936051

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Project description:The formation of a zygote by the fusion of egg and sperm involves the two gametic transcriptomes. In flowering plants, the embryo sac embedded within the ovule contains the egg cell, while the pollen grain contains two sperm cells inside a supporting vegetative cell. The difficulties of collecting isolated gametes and consequent low recovery of RNA have restricted in-depth analysis of gametic transcriptomes in flowering plants. We isolated living egg cells, sperm cells, and pollen vegetative cells from rice, and identified transcripts for ~36,000 genes by deep sequencing. The three transcriptomes are highly divergent, with about three quarters of those genes differentially expressed in the different cell types. Distinctive expression profiles were observed for genes involved in chromatin conformation, including an unexpected expression in the sperm cell of genes associated with active chromatin. Furthermore, both the sperm cell and the pollen vegetative cell were deficient in expression of key RNAi components. Differences in gene expression were also observed for genes for hormonal signaling and cell cycle regulation. The egg cell and sperm cell transcriptomes reveal major differences in gene expression to be resolved in the zygote, including pathways affecting chromatin configuration, hormones and cell cycle. The sex-specific differences in expression of RNAi components suggest that epigenetic silencing in the zygote might act predominantly through female-dependent pathways. More generally, this study provides a detailed gene expression landscape for flowering plant gametes, enabling the identification of specific gametic functions and their contributions to zygote and seed development. The gene expression profiles of the three gametic cells, egg cell (EC), sperm cell (Sp) and vegetative cell (Ve) were compared via RNA-seq using three biological replicates for each. Differential expression (DE) analysis was performed using edgeR and the resulting DE genes were assessed for Gene Ontology term enrichment.

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