Proteomics of the ripening of blackberry (Rubus sp.) fruits grown in Mexico, a first approximation
DOI:
https://doi.org/10.35197/rx.08.03.e1.2012.15.acKeywords:
proteins, denaturing electrophoresis, two-dimensional electrophoresisAbstract
Blackberry production in Mexico has increased by 200% in the last decade. The main varieties used have been introduced and their establishment under the climatic conditions of Mexico has required considerable adaptations to the agronomic management observed in the regions of origin of these varieties. The essential challenge of managing this product is based on the intrinsic characteristics of its soft fruits and its short shelf life, so knowledge of its ripening process under the growing conditions in Mexico is essential to successfully manipulate and improve the productivity and quality of the fruits until their final destination. With the intention of initially addressing this problem, the objective of this work was to establish the conditions for protein analysis protocols that allow the evaluation of protein components in blackberry fruits during ripening. For this, six stages of fruit ripening of the commercial variety ‘Brazos’ were identified (considering from green and underdeveloped fruits to fruits ripe for consumption), a protein extraction method was selected and a protein profile was performed by electrophoresis under denaturing conditions. In addition, the requirements for two-dimensional electrophoresis (2-DE) of the extracts obtained were established, evaluating the isoelectric focusing conditions and staining methods. According to the results obtained, it was determined to use 400 µg of total protein in 7 cm IPG strips with a pH range of 3 to 10, at a maximum voltage of 50,000 V and staining with Coomassie blue. Using the KODAK MI software version 4.5, a preliminary analysis of the distribution and abundance of the peptides expressed in the six stages of ripening was performed, observing that stage 2 presented the highest number of peptide spots (158), that the highest percentage of peptide spots in all stages were presented in a pH range of 5.0 to 6.9 and molecular weight of 30 to 50 kDa. Four spots of similar intensity during ripening were identified; suggesting their constitutive expression, three of decreasing intensity; which could be involved in initial processes of fruit growth, four that are induced during ripening and six of oscillating intensity.
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Copyright (c) 2012 Ana Tztzqui Chávez Bárcenas, Cristina Alonso Ojeda, Pedro Antonio García Saucedo
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