Abstract
<p>Bananas and plantains are a major staple food and export product in many countries.<strong> </strong>Banana fruit tissues contain large amounts of secondary compounds, polysaccharides and other plant polymers which will interfere with protein extraction for gel-based proteomic analysis. Due to presence of very small amount of proteins (approximately 1%) in a large matrix of fruit tissues, it classified as ‘recalcitrant’. In this connection, assessment of different protein extraction protocols and extraction of high quality proteins from banana fruit tissues is crucial for successful gel-based proteome analysis. In this study, two different protein extraction protocols were validated to isolate proteins from banana (cv.Grand Naine) fruit peel and pulp tissues, and proteins were resolved by using SDS-PAGE. A comparative study showed that phenol based method is effective in extracting proteins over TCA-acetone method. Isolated proteins were further subjected to two-dimensional gel electrophoresis separation and stained with colloidal coomassie blue to visualize protein spots. On average 380 protein spots could be detected on coomassie stained two-dimensional gel and our study clearly demonstrated the differential protein accumulation during pre-climacteric and climacteric stages of banana fruit. <strong></strong></p>
Highlights
Bananas and plantains are major fruit crops in many countries
Phenol extraction of the proteins from banana pulp and peel tissues at climacteric stage resulted in the preparation of high-quality proteins for SDS-PAGE separation
Banana (Musa spp.) is a good representative of a difficult plant species for protein extraction since it contains many interfering metabolites.To date, only classicalTCAacetone precipitation and phenol extraction methods proved useful as standard methods for protein extraction from different plant species and organs (Sebastiana et al, 2013; Wu et al, 2014; Singh et al, 2015; Vaganan et al, 2015)
Summary
Bananas and plantains are major fruit crops in many countries. India stands as the largest producer of banana with an annual production of 28.4 million tons on 796,500 ha, which contribute to 27% of the world production and about 38% of the total fruit crop production in the nation (FAOSTAT, 2011). Post-harvest losses of fruits and vegetables account for almost 50% of the produce. The world’s second largest producer of fruits and vegetables, loses 35-40% of the produce due to excessive softening. Excessive softening exacerbates the damage incurred during handling, decreases the shelf life of fruits, and increases the susceptibility toward postharvest pathogens. Fruit ripening is a highly coordinated, genetically programmed, irreversible phenomenon that involves a complex series of physiological and biochemical events. Proteomic investigations on banana fruits during development and ripening are still limited
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