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Identification of bitter pit protein markers in Malus domestica using differential in-gel electrophoresis (DIGE) and LC–MS/MS

AutorKrawitzky, Michael ; Orera Utrilla, Irene ; López-Millán, Ana Flor ; Oria, Rosa; Val Falcón, Jesús
Palabras claveBitter pit
Apple
Pathogenesis-related (PR) proteins
DIGE
LTQ-Orbitrap
Fecha de publicaciónene-2016
EditorElsevier
CitaciónKrawitzky M, Orera I, López-Millán AF, Oria R, Val J. Identification of bitter pit protein markers in Malus domestica using differential in-gel electrophoresis (DIGE) and LC–MS/MS. Postharvest Biology and Technology 111: 224–239 (2016)
ResumenBitter pit is a physiological disorder that occurs in apple, pear and quince and has long been associated with calcium uptake or lack thereof. In the present study, pooled biological Malus domestica proteins were collected from healthy and naturally occurring bitter pit fruit. Protein samples (bitter pit and healthy) were analyzed with differential in-gel electrophoresis (DIGE) and SameSpots software was used to compare gel spots by intensity. Identified spots (p < 0.05) were spot picked and trypsin digested. Peptides were separated by liquid chromatography (LC) and submitted to LTQ-Orbitrap mass spectrometer to infer protein identification. A total of 200 ± 5 protein spots were detected, 41 spots classified as having p < 0.05 and were successfully identified by their peptide sequence listed in an online M. domestica database. Thirteen spots were identified as having p < 0.05 and a minimum 2-fold change. Several pathogenesis-related (PR) proteins belonging to three PR families, PR-5 (10.3-fold), PR-8/chitinase (35.5-fold) and PR-10 (6.8-fold) were upregulated in bitter pit sampled tissues. Three proteins involved in several metabolic processes including ethylene biosynthesis (3.3-fold), glycosyltransferase reactions in metabolism (2.3-fold) and metal biding (11.1-fold) were found to be overly expressed in healthy sampled tissues. This research provides a significant advance in the knowledge of protein expression alterations occurring in bitter pit in comparison with sound tissues.
Descripción36 Pags.- 7 Figs.- 3 Tabls. The definitive version is available at: http://www.sciencedirect.com/science/journal/09255214
Versión del editorhttp://dx.doi.org/10.1016/j.postharvbio.2015.09.006
URIhttp://hdl.handle.net/10261/123001
DOI10.1016/j.postharvbio.2015.09.006
ISSN0925-5214
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