A DIGE-based quantitative proteomic analysis of grape berry flesh development and ripening reveals key events in sugar and organic acid metabolism

Abstract

Grapevine (Vitis vinifera L.) is an economically important fruit crop. Quality-determining grape components, such as sugars, acids, flavours, anthocyanins, tannins, etc., are accumulated during the different grape berry development stages. Thus, correlating the proteomic profiles with the biochemical and physiological changes occurring in grape is of paramount importance to advance the understanding of the berry development and ripening processes. Here, the developmental analysis of V. vinifera cv. Muscat Hamburg berries is reported at protein level, from fruit set to full ripening. A top-down proteomic approach based on differential in-gel electrophoresis (DIGE) followed by tandem mass spectrometry led to identification and quantification of 156 and 61 differentially expressed proteins in green and ripening phases, respectively. Two key points in development, with respect to changes in protein level, were detected: end of green development and beginning of ripening. The profiles of carbohydrate metabolism enzymes were consistent with a net conversion of sucrose to malate during green development. Pyrophosphate-dependent phosphofructokinase is likely to play a key role to allow an unrestricted carbon flow. The well-known change of imported sucrose fate at the beginning of ripening from accumulation of organic acid (malate) to hexoses (glucose and fructose) was well correlated with a switch in abundance between sucrose synthase and soluble acid invertase. The role of the identified proteins is discussed in relation to their biological function, grape berry development, and to quality traits. Another DIGE experiment comparing fully ripe berries from two vintages showed very few spots changing, thus indicating that protein changes detected throughout development are specific.

Influence of Fruit Ripening Stage and Harvest Period on the Antioxidant Content of Sweet Pepper Cultivars

“…In peach fruits, different proteins related to sucrose and ethylene metabolism and organic acid biosynthesis were found to augment during maturation, increasing the nutritional and quality value [24]. In grapevine, two specific key points in development have been detected by proteomic analysis: end of green development and beginning of ripening [25]. In pepper fruits, specific proteins as polygalacturonase, cellulase and β-galactosidase have been shown to change between harvest dates and among maturity groups.…”

A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media

“…The protein extracts from Vitis callus/suspensions and Taxus callus were prepared as described by Martínez-Esteso et al [50]. Briefly, plant material was homogenized in extraction buffer (50 mM HEPES, 0.25 M sucrose, 1% [wt/vol] PVPP, 5% glycerol, 10 mM EDTA, 10 mM Na 2 O 5 S 2 , 10 mM acid ascorbic, 1 mM PMSF and Sigma Protease inhibitor) at a ratio of 2 ml per gram of plant material at 4°C.…”

iTRAQ-based protein profiling provides insights into the central metabolism changes driving grape berry development and ripening

“…DIGE has been applied to study grape berry development. It has led to a better coverage and accuracy of the protein changes reported from a gel-based approach [30,31]. Lücker et al [32] studied grape berry ripening initiation by the quantitative MS-based iTRAQ approach, and reported a significantly larger number of total protein identification and protein changes than single-stain 2-DE-based studies.…”

“…[41], used a large-scale transcriptomic analysis to study the differential expression of genes among main tissues, seed, flesh and skin, and reported that 60% of these genes exhibited a significant differential expression in at least one of the three major tissue types. Several proteomic analyses at the tissue-specific level have been carried out [24,26,31,32,36,42]. Nevertheless on the whole, a gap still remains either because one only tissue is analyzed or berry development is not comprehensively covered.…”