Dataset Information

Proteomic analysis of acorn cotyledon tissues from Quercus ilex with GeLC-orbitrap/MS (Non-Osthodox seed)

ABSTRACT: The protein content in the cotyledon was analyzed in 2 stages of maturity (CM6, CM9) and one of germination (CG5) using the GeLC-Orbitrap/MS technique in Quercus ilex Non-Orthodox seeds.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Quercus Ilex Subsp. Ilex

TISSUE(S): Cotyledon

SUBMITTER: Monica Escandon

LAB HEAD: Jesus V. Jorrin-Novo

PROVIDER: PXD020603 | Pride | 2021-01-07

REPOSITORIES: Pride

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Publications

GeLC-Orbitrap/MS and 2-DE-MALDI-TOF/TOF comparative proteomics analysis of seed cotyledons from the non-orthodox Quercus ilex tree species.

Sghaier-Hammami Besma B Castillejo María Ángeles MÁ Baazaoui Narjes N Jorrín-Novo Jesús V JV Escandón Mónica M

Journal of proteomics 20201225

Gel electrophoresis-based and shotgun approaches are the most employed proteomic platforms in plant biology research, with the latter replacing the former in the last years. We have compared 2-DE-MALDI-TOF/TOF and GeLC-Orbitrap/MS analyses using the same protein extracts from Quercus ilex cotyledons at different development stages. The results obtained (ProteomeXchange available data, PXD020603) showed that both platforms were complementary, showing common and specific proteins identified in eac ...[more]

PMID: 33359940

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Project description:Proteases and protease inhibitors have been analyzed in the non-orthodox Quercus ilex forest tree by using in silico and wet approaches. The in silico analysis showed that the species-specific transcriptome database included 2240 and 97 transcripts annotated as, respectively, proteases and protease inhibitors, with members of the different families according to the MEROPS protease database (http://www.merops.ac.uk) classification, mostly of serine- and metallo-type. Data for Q. ilex are compared with those previously published for other Quercus species, namely, Q. suber, Q. lobata, and Q. robur. A second approach was performed to assess the evolution of proteases and protease inhibitors along with seed germination in cotyledon and embryo tissues by determining changes at proteomics and activity levels, the last by mean of in vitro and in gel assays. Shotgun (LC-MSMS) analysis of acorn embryos and cotyledons of three developmental stages (NV, non-viable; T1, mature; and T3 germinated) allowed the identification of a total of 177 proteases and 12 protease inhibitors. In vitro protease activity was determined by using azocasein as substrate; the total activity was higher in cotyledons than embryos, showing the latter significant differences throughout germination. Gel protease activity assays showed common and differential bands between tissues and developmental stages. Shotgun analysis of 10 differential activity bands supported the results obtained from the whole proteome analysis, with some proteins highly represented in the same tissues and stages by both strategies. In overall, the combination of shotgun proteomics and protease activity approaches allowed the identification of tissue-specific (such as cysteine protease inhibitors), and stage-specific proteins (such as those related to storage proteins mobilization), with others being present in all the analyzed organs. Some of the proteases and protease inhibitors differentially identified between tissues and stages in this study could be used as indicators of seed quality and viability.

			Action	DRS
	EG5-1-1.RAW	Raw
	EG5-1-2.RAW	Raw
	EG5-1-3.RAW	Raw
	EG5-2-1.RAW	Raw
	EG5-2-2.RAW	Raw