Abstract
BackgroundBreeding for strong red skin color is an important objective of the pear breeding program. There are few reports of proteome research in green skin pear and its red skin bud mutation. The manuscript at hand is one of the first studies dealing with 2D-PAGE-based analysis of pear fruits and leaves, establishing a suitable sample preparation and testing different 2D-PAGE protocols. Therefore, it may grant a basis for further studies on the pear proteome being the studies main goal. A proteomic analysis was conducted on leaves and fruits of ‘Zaosu’ pear (Pyrus bretschneideri Rehd.) and its red skin bud mutation in order to reveal their genetic differences in the protein level.ResultsIn the present study, the optimized two-dimensional (2-D) gel electrophoresis system of pear leaf and fruit was set up, and applied to analyze the leaves and fruit protein. The interesting peptide fragments were determined using 4800 Plus MALDI TOF/TOFTM Analyzer mass spectrometer, and the sequence obtained was blasted in NCBInr to identify the differentially-expressed protein. In the 1.5-fold differently-expressed proteins between ‘Zaosu’ pear and its mutant, 10 out of 35 proteins in fruit and 12 out of 24 ones in leaves were identified successfully. Among the 22 identified proteins, 7 protein spots were related to photosynthesis and energy metabolism; 4 were associated with environmental stress; 4 with disease defense; 2 with amino acid metabolism; 2 with cytoskeleton; 1 with antioxidant function; 1 with calcium metabolism; and 1 with unknown function. Moreover, related physiological index, such as chlorophyll content, Rubisco content and polyphone oxidase activity, were different between ‘Zaosu’ pear and its mutant.ConclusionA 2-D gel electrophoresis system of pear leaves and fruits was established, which was suitable for the analysis of proteome comparison. To the best of our knowledge, we have performed the first analysis of the proteomic changes in leaves and fruits of ‘Zaosu’ pear and its red skin bud mutation. Our study provides important information on the use of proteomic methods for studying protein regulation of ‘Zaosu’ pear and its red skin bud mutation.
Highlights
Breeding for strong red skin color is an important objective of the pear breeding program
The proteomic analysis with pH 3-10 showed that the pear protein spots distributed mainly within the range of pH 4-7 (Figure 2 A)
282 protein spots were detected on pH 3-10 IPG strip, while 377 spots were on pH 4-7 IPG strip (Figure 2 B)
Summary
Breeding for strong red skin color is an important objective of the pear breeding program. The manuscript at hand is one of the first studies dealing with 2D-PAGE-based analysis of pear fruits and leaves, establishing a suitable sample preparation and testing different 2D-PAGE protocols. It may grant a basis for further studies on the pear proteome being the studies main goal. A proteomic analysis was conducted on leaves and fruits of ‘Zaosu’ pear (Pyrus bretschneideri Rehd.) and its red skin bud mutation in order to reveal their genetic differences in the protein level. Gamay) cell suspension in response to elicitation with methylated cyclodextrins (MBCD) and methyl jasmonate, and identified 25 proteins by MALDI-TOF These studies provided valuable experience for the subsequent researchers. No studies have been reported to date of the differential expression of protein in green skin pear and its red skin bud mutation
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