Quantitative Proteomic Analysis of Wheat Seeds during Artificial Ageing and Priming Using the Isobaric Tandem Mass Tag Labeling

Abstract

Wheat (Triticum aestivum L.) is an important crop worldwide. The physiological deterioration of seeds during storage and seed priming is closely associated with germination, and thus contributes to plant growth and subsequent grain yields. In this study, wheat seeds during different stages of artificial ageing (45°C; 50% relative humidity; 98%, 50%, 20%, and 1% Germination rates) and priming (hydro-priming treatment) were subjected to proteomics analysis through a proteomic approach based on the isobaric tandem mass tag labeling. A total of 162 differentially expressed proteins (DEPs) mainly involved in metabolism, energy supply, and defense/stress responses, were identified during artificial ageing and thus validated previous physiological and biochemical studies. These DEPs indicated that the inability to protect against ageing leads to the incremental decomposition of the stored substance, impairment of metabolism and energy supply, and ultimately resulted in seed deterioration. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the up-regulated proteins involved in seed ageing were mainly enriched in ribosome, whereas the down-regulated proteins were mainly accumulated in energy supply (starch and sucrose metabolism) and stress defense (ascorbate and aldarate metabolism). Proteins, including hemoglobin 1, oleosin, agglutinin, and non-specific lipid-transfer proteins, were first identified in aged seeds and might be regarded as new markers of seed deterioration. Of the identified proteins, 531 DEPs were recognized during seed priming compared with unprimed seeds. In contrast to the up-regulated DEPs in seed ageing, several up-regulated DEPs in priming were involved in energy supply (tricarboxylic acid cycle, glycolysis, and fatty acid oxidation), anabolism (amino acids, and fatty acid synthesis), and cell growth/division. KEGG and protein-protein interaction analysis indicated that the up-regulated proteins in seed priming were mainly enriched in amino acid synthesis, stress defense (plant-pathogen interactions, and ascorbate and aldarate metabolism), and energy supply (oxidative phosphorylation and carbon metabolism). Therefore, DEPs associated with seed ageing and priming can be used to characterize seed vigor and optimize germination enhancement treatments. This work reveals new proteomic insights into protein changes that occur during seed deterioration and priming.

Improvement effect of nitrogen fertilizer and plant density on wheat (Triticum aestivum L.) seed deterioration and yield

“…One of the most important factors in the production of wheat is the quality of the seeds that have been planted. The quality of wheat seeds is affected by many factors, including cultivar, genetic purity, physical purity, vitality, germination percentage, and seed viability (Lv et al, 2016).…”

Source-Sink Modifications Affect Leaf Senescence and Grain Mass in Wheat

“…Protein disulfide isomerases (PDIs) are enzymes found primarily in the endoplasmic reticulum (ER) in eukaryotes play a vital role in protein folding. The up-regulated levels of PDIs (A0A024FRN3, A0A1D5XQP7, A0A1D5XQP9, B9A8E3, F8THZ6, Q6JAB5, Q93XQ8) in the present study indicated the higher PDI activity (MF) and confirmed the active status of protein synthesis (Hayano et al ., 1995; Lv et al ., 2016; Kayum et al ., 2017) and higher resistance to abiotic stresses (Kayum et al ., 2017). This conclusion was strongly confirmed by the upregulation of 60S ribosomal protein L29 (A0A1D5RRD3), 40S ribosomal protein S12 (A0A1D5UFM0, A0A1D5UVS7) and Eukaryotic translation initiation factor 4B1 (Q9AUJ7) (Table S1).…”

“…In plants, acetyl-CoA carboxylase activity is found in the plastids and catalyzes the first committed step of de novo fatty acid biosynthesis by the carboxylation of acetyl-CoA to malonyl-CoA (Fazli et al ., 2005; Liu et al ., 2007; Lv et al ., 2016). In the present study, acetyl-CoA carboxylase (Q8L4W1), plastid acetyl-CoA carboxylase (fragment) (A0A068B0K3, A0A068B0L8, A0A068B0M7, A0A068B226, A0A068B2H1, A0A068B2I1, A0A068B3I5, A0A068B675, B1A0F0, D0V7D7, D0V7D9, D0V7E1, D0V7E3, D0V7F1) and acetyl-coenzyme A carboxylase (Fragment) (O48960), were down-regulated, indicating that CoA carboxylase activity (GO:0016421), acetyl-CoA carboxylase activity (GO:0003989) and biotin carboxylase activity (GO:0004075) were affected.…”

Peer Review #3 of "Dynamic proteomic changes in soft wheat seeds during accelerated ageing (v0.1)"

“…The original germination rate (Gr) was 97.0% (designated as YM97), and the average seed moisture content was 12.19%. A 400 g sample of seeds with a similar size and weight was stored in a 5 L plastic bottle at 45 ± 1°C and 50% relative humidity in a constant temperature-and humidity-controlled cabinet (Binder KMF720, Tuttingen, Germany) according to our previous research (Lv et al, 2016). Measurement of CO 2 from wheat respiration and calculation of Gr were performed as previously described (Dong et al, 2015;Zhang et al, 2014).…”

“…2.3. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis and database searches LC-MS/MS and database searches were performed as described previously (Lv et al, 2016).…”