Proteome Map of Pea (Pisum sativum L.) Embryos Containing Different Amounts of Residual Chlorophylls

Abstract

Due to low culturing costs and high seed protein contents, legumes represent the main global source of food protein. Pea (Pisum sativum L.) is one of the major legume crops, impacting both animal feed and human nutrition. Therefore, the quality of pea seeds needs to be ensured in the context of sustainable crop production and nutritional efficiency. Apparently, changes in seed protein patterns might directly affect both of these aspects. Thus, here, we address the pea seed proteome in detail and provide, to the best of our knowledge, the most comprehensive annotation of the functions and intracellular localization of pea seed proteins. To address possible intercultivar differences, we compared seed proteomes of yellow- and green-seeded pea cultivars in a comprehensive case study. The analysis revealed totally 1938 and 1989 nonredundant proteins, respectively. Only 35 and 44 proteins, respectively, could be additionally identified after protamine sulfate precipitation (PSP), potentially indicating the high efficiency of our experimental workflow. Totally 981 protein groups were assigned to 34 functional classes, which were to a large extent differentially represented in yellow and green seeds. Closer analysis of these differences by processing of the data in KEGG and String databases revealed their possible relation to a higher metabolic status and reduced longevity of green seeds.

Profiling of Seed Proteome in Pea (Pisum sativum L.) Lines Characterized with High and Low Responsivity to Combined Inoculation with Nodule Bacteria and Arbuscular Mycorrhizal Fungi

“…According to the results of the Bradford assay, yields of the protein extraction were in the range of 63.6–175.0 mg/g fresh weight (Table S1-5). This was confirmed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, performed with the sample load, calculated based on the results of the Bradford assay [31]; the whole lane average intensities obtained with equal sample amounts (5 µg of protein) were 1.77 × 10 4 ± 2.25 × 10 3 (RSD = 12.7%). Thereby, the patterns of the signals, observed in the electropherograms were similar between lanes and treatment groups.…”

“…Thereby, the patterns of the signals, observed in the electropherograms were similar between lanes and treatment groups. The subsequent tryptic digestion (Figure 1) was considered to be complete, as the bands of major pea storage proteins, as legumin (α- and β- subunits, ∼40 and ∼20 kDa, correspondingly), vicilin (subunits of ∼29, ∼35, and ∼47 kDa) and convicilin (subunit of ∼71 kDa), were not detectable [31], assuming a staining sensitivity better than 30 ng [32] and a legumin content of at least 80% of the total seed protein [33].…”

“…Therefore, here we decided on a non-redundant combined database relying on several legume proteomes, closely related to pea— Medicago truncatula Gaertn, Lotus japonicas (Regel) K. Larsen, and Phaseolus vulgaris L. Earlier, to confirm the applicability of this database, we manually evaluated the MS/MS spectra of confidently identified peptides with the lowest values of the SEQUEST function XCorr [34]. As the spectra were acquired with the mass accuracy within 5 ppm, peptide sequences could be unambiguously assigned by characteristic patterns of N- and C-terminal ion series ( b and y ions, respectively) [31].…”

Glycation of Plant Proteins: Regulatory Roles and Interplay with Sugar Signalling?

“…The combination of these post-translational modifications (PTMs) is often referred to as “carbonylation” and recognized as a marker for plant senescence and a tag for proteolysis [99]. In mature seeds, glycation represents one of the major markers of seed ageing, in concert with the decrease of their quality and longevity during natural and accelerated storage [100,101].…”

In-Depth Investigation of Low-Abundance Proteins in Matured and Filling Stages Seeds of Glycine max Employing a Combination of Protamine Sulfate Precipitation and TMT-Based Quantitative Proteomic Analysis

“…These HAPs depletion methods seem to be effective and are broadly applied for the depletion of HAPs from different plants including broad bean, pea, wild soybean, and peanut, which contain their own specific HAPs [5]. Previously, the PS precipitation method allowed the identification of only a few specific low abundance proteins (LAPs) related to various functions in pea seeds such as transcription factors and regulators, kinases, transporter proteins, and various enzymes related to energy metabolism [7].…”