华南8号木薯及其四倍体块根淀粉及蛋白表达谱的差异分析

doi:10.3864/j.issn.0578-1752.2015.13.017

Abstract

Abstract: 【Objective】The aim of the study was to better understand the change of starch content, starch structure and its proteins regulatory mechanisms of cassava tuberous roots of diploid and its tetraploid.【Method】Tuberous roots of 10 months after planting were used as the materials, starch content was measured by weight in air and water, the proportion of amylose and amylopectin was determined by spectrophotometry, Excel 2013 and DPS v7.05 statistical software were used to analyze the data, the significant difference was determined by using Duncan method, the size, shape and number of amyloplast were observed by scanning electron microscopy, Western blot was used to verify some enzymes involved in starch synthesis and degradation, roots proteins of cassava tetraploid and diploid genotypes were excised and purified from 2D gels, Delta 2D software were used to analyze the different proteins with more than 2.0 fold changes and assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-TOF-MS/MS) were used to identify them, and the proteins were classified according to their functions combined with KEGG database.【Result】After chromosome doubling in tetraploid cassava, dry matter content, starch content and fresh roots weight were significantly decreased by 12.18%, 11.41%, and 35.34%, respectively. No significant difference was observed in amylase and amylopectin proportion. No significant difference in amyloplast morphology, mainly spherical, irregular sphere and ellipsoid also exist, the size was uniform, while the loose degree of amyloplast and space gap were increased, the number was decreased. The level of sucrose phosphate synthase (SPS) significantly decreased, β-amylase expression was increased, while the level of granule bound starch synthase I (GBSSI) was not changed. Twenty differential expressed protein spots were showed and analyzed by software, including 2 up-regulated spots and 18 down-regulated spots, 19 were successfully matched in database, while spot 16, which was down-regulated, was no match. The functions of the 17 down regulated proteins were associated with carbohydrate and energy metabolism (4), structure (3), DNA and RNA metabolism (2), chaperones (2), HCN metabolism (2), antioxidant (1), protein synthesis (1) and unknown function (2), 2 up-regulated proteins were stem-specific proteins (TSJT1).【Conclusion】Protein related with carbohydrate and energy metabolism, nucleic acid metabolism and chaperones were down-regulated, the amyloplast loosely arranged and number decreased, the expression level of SPS decreased, β-amylase expression increased, indicating the low starch synthesis ability and high starch decomposition ability in tetraploid cassava, then decreased starch content, while no significant changes in amylose and amylopection proportion was found.

Key words: cassava, tetraploid, tuberous roots, starch, proteins

AN Fei-fei, CHEN Song-bi, LI Geng-hu, ZHOU Kai, LI Kai-mian. Comparison Analysis of Starch and Protein Expression Profiles on Cassava Tuberous Roots cv. SC8 and Its Tetraploid[J].Scientia Agricultura Sinica, 2015, 48(13): 2656-2665.

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