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Proteomics Analysis of Round and Wrinkle Pea (Pisum sativum L.) Seeds during Different Development Periods

ABSTRACT: Seed development is a complex process controlled by many factors, including genetics, plant growth regulators, and the environment. Understanding the different compositions and abundances of proteins in various types of seeds at different developmental stages is fundamental for improving seed quality and enhancing nutritional and food security. In this study, we applied label-free quantitative proteomics to analyze round and wrinkled pea seeds at five different growth stages: 4, 7, 12, 15 days after anthesis (DAA), and at maturity. Wrinkled peas had lower starch content (29.5%) compared to round peas (~47-55%). Proteomic analysis identified 3,659 protein groups, of which approximately 21-24% proteins were shared across growth stages. Relatively more proteins were identified during early seed development compared to later stages, and the protein profiles were also drastically different between early and late stages. Statistical analysis identified 735 significantly different proteins between wrinkled and round seeds, regardless of the growth stage. These detected proteins were characterized into different functional classes including metabolic enzymes, proteins involved in protein biosynthesis and homeostasis, carbohydrate metabolism, vesicle trafficking, cell division, and cell wall organization. Cell division-related proteins were more abundant in the early stages of seed development, whereas storage proteins and proteins implicated in lipid metabolism were more abundant at the later stages. Wrinkled seeds had a significantly lower abundance of starch-branching enzyme than did round seeds. This enzyme is involved in the biosynthesis of the amylopectin component of starch. Seed storage proteins such as legumin and a form of albumin (PA2) accumulated more in round pea genotypes, whereas vicilin was more abundant in the wrinkled pea genotype. In summary, this study can enhance our understanding of pea seed development, highlighting key differences in protein profiles between round and wrinkled pea seeds.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Pisum Sativum (ncbitaxon:3888)

SUBMITTER: Rebecca J. McGee

PROVIDER: MSV000095262 | MassIVE | Fri Jul 05 13:58:00 BST 2024

REPOSITORIES: MassIVE

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Project description:Barley (Hordeum vulgare) is one of the major food sources for humans and forage source for animal livestock. Barley endosperm is structured into three distinct cell layers: the starchy endosperm, which acts essentially as storage tissue for starch, the subaleurone, which is characterized by a high accumulation of endoplasmic reticulum (ER)-derived seed storage proteins (SSP) and finally the aleurone beside the seed coat with a prominent role during seed germination. Prolamins account for more than 50% of the total protein amount in mature seeds. Together with other seed storage proteins (SSPs) they are important for both grain quality and flour quality. Prolamins are synthesized on the rough ER, translocated into the ER lumen and accumulate in distinct, ER-derived protein bodies (PBs) that are most abundant in the SE. PB formation is regulated by the protein disulfide isomerase (PDI) that is involved in the disulfide transfer pathway. Here, we used laser microdisection (LMD) to characterize spatio-temporal molecular and morphological differences of the ER during barley endosperm development. We revealed by nanoLC-MS/MS proteomic analyses performed on whole seeds and collected tissues at different seed development stages that the protein level of the protein disulfide isomerase HvPDIL1-1 is spatio-temporally regulated in developing barley endosperm. Our microscopic studies showed that HvPDIL1-1 preferentially accumulates in SE, especially at 12 days after pollination (dap). HvPDIL1-1 re-localized from PBs to the protein matrix at the periphery of starch granules along grain filling process. Detailed analysis of SE proteome dynamics identified clusters of proteins with similar expression pattern as HvPDIL1-1, which were analysed in a protein-protein network. It revealed a strong functional interconnection between transcription and translation, protein folding and amino acid synthesis with sucrose and starch metabolism. Our data indicate a role of HvPDIL1-1 in the coordination of protein synthesis and prolamins deposition during grain filling processes in developing barley endosperm. These results are discussed in relation to the putative role of HvPDIL1-1 for cereal food end-product quality and recombinant protein production in cereal seeds.

Project description:Jatropha curcas, a tropical shrub, is an attractive biofuel crop, which produces seeds with a high content of oil and protein. Most J. curcas accessions contain a range of toxins and anti-nutritional compounds, which are unfavorable for human consumption and animal feed. With the goal of better understanding the development of its seeds and to improve Jatropha`s agronomic performance, a two-step approach was performed: 1) generation of the entire transcriptome of six different developmental stages of J. curcas seeds using 454-Roche sequencing of a cDNA library, 2) comparison of transcriptional expression levels in six different developmental stages of seeds using a custom Agilent 8x60K oligonucleotide gene expression microarray. A total of 793,875 high quality reads with an average length of 358 bp were generated and assembled into 19,841 unique full-length contigs, of which 13,705 could be annotated with GO terms. Microarray data analysis identified a total of 9,111 contigs which were differentially expressed between the six developmental stages (based on P-value of < 1e-8). The microarray-based differential expression results were validated for 192 putative genes using high-throughput quantitative real-time PCR. Results from Gene Ontology (GO) and pathway enrichment analysis showed that the upstream biosynthesis pathways were relatively active among differentially expressed sequences (DESs) during seed development, such as starch and sucrose metabolism, glycolysis/gluconeogenesis, glycerophospholipid metabolism, amino sugar and nucleotide sugar metabolism. Further, DESs related to hormone and seed development, flavonoid biosynthesis-related pathways were over-represented. A pairwise comparison was used to discover particular seed maturation dependent patterns of gene expression. Genes related to fatty acid biosynthesis were over represented in early stage, while flavonoid biosynthesis in the late stage. The data provide a useful resources and references for the seed transcriptome of J. curcas and other related species with oil producing seeds. The identified transcripts related to seed development provide a starting point for further investigation into the molecular mechanisms underlying this process.

Dataset Information

Proteomics Analysis of Round and Wrinkle Pea (Pisum sativum L.) Seeds during Different Development Periods

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