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Biosynthesis and Regulation of Wheat Amylose and Amylopectin from Proteomic and Phosphoproteomic Characterization of Granule-binding Proteins

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ABSTRACT

Waxy starch has an important influence on the qualities of breads. Generally, grain weight and yield in waxy wheat (Triticum aestivum L.) are significantly lower than in bread wheat. In this study, we performed the first proteomic and phosphoproteomic analyses of starch granule-binding proteins by comparing the waxy wheat cultivar Shannong 119 and the bread wheat cultivar Nongda 5181. These results indicate that reduced amylose content does not affect amylopectin synthesis, but it causes significant reduction of total starch biosynthesis, grain size, weight and grain yield. Two-dimensional differential in-gel electrophoresis identified 40 differentially expressed protein (DEP) spots in waxy and non-waxy wheats, which belonged mainly to starch synthase (SS) I, SS IIa and granule-bound SS I. Most DEPs involved in amylopectin synthesis showed a similar expression pattern during grain development, suggesting relatively independent amylose and amylopectin synthesis pathways. Phosphoproteome analysis of starch granule-binding proteins, using TiO2 microcolumns and LC-MS/MS, showed that the total number of phosphoproteins and their phosphorylation levels in ND5181 were significantly higher than in SN119, but proteins controlling amylopectin synthesis had similar phosphorylation levels. Our results revealed the lack of amylose did not affect the expression and phosphorylation of the starch granule-binding proteins involved in amylopectin biosynthesis.

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Comparison of the expression patterns of three starch granule-binding proteins in SN119 and ND5181.(A) Close-up views of expression levels for GBSS I, SS I and SS IIa protein spots on 2-DE gels. (B) Expression patterns and proportion of GBSS I, SS I and SS IIa proteins in five stages of seed development in SN119 and ND5181.
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f1: Comparison of the expression patterns of three starch granule-binding proteins in SN119 and ND5181.(A) Close-up views of expression levels for GBSS I, SS I and SS IIa protein spots on 2-DE gels. (B) Expression patterns and proportion of GBSS I, SS I and SS IIa proteins in five stages of seed development in SN119 and ND5181.

Mentions: The dynamic expression patterns of SS I, SS IIa and GBSS I at five grain developmental stages in SN119 and ND5181, detected by 2-DE, are shown in Fig. 1 and Supplemental Fig. 6. Multiple isoforms were present in SS I, SS IIa and GBSS I (Fig. 1A). GBSS I had at least 10 isoforms in ND5181, and SS IIa and SS I had at least 12 and 5 isoforms in both SN119 and ND5181, respectively (Fig. 1A). The waxy protein content increased from 10 DPA to 15 DPA in ND5181, and then reduced slightly in later periods. SS I and SS IIa exhibited small changes during five stages in both varieties and showed a down-up-down pattern in ND5181, with the greatest quantity at 25 DPA. SS IIa displayed an up-down pattern, while SS I showed a down-up-down pattern in SN119, with the highest quantity at about 20 DPA (Fig. 1B).


Biosynthesis and Regulation of Wheat Amylose and Amylopectin from Proteomic and Phosphoproteomic Characterization of Granule-binding Proteins
Comparison of the expression patterns of three starch granule-binding proteins in SN119 and ND5181.(A) Close-up views of expression levels for GBSS I, SS I and SS IIa protein spots on 2-DE gels. (B) Expression patterns and proportion of GBSS I, SS I and SS IIa proteins in five stages of seed development in SN119 and ND5181.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015113&req=5

f1: Comparison of the expression patterns of three starch granule-binding proteins in SN119 and ND5181.(A) Close-up views of expression levels for GBSS I, SS I and SS IIa protein spots on 2-DE gels. (B) Expression patterns and proportion of GBSS I, SS I and SS IIa proteins in five stages of seed development in SN119 and ND5181.
Mentions: The dynamic expression patterns of SS I, SS IIa and GBSS I at five grain developmental stages in SN119 and ND5181, detected by 2-DE, are shown in Fig. 1 and Supplemental Fig. 6. Multiple isoforms were present in SS I, SS IIa and GBSS I (Fig. 1A). GBSS I had at least 10 isoforms in ND5181, and SS IIa and SS I had at least 12 and 5 isoforms in both SN119 and ND5181, respectively (Fig. 1A). The waxy protein content increased from 10 DPA to 15 DPA in ND5181, and then reduced slightly in later periods. SS I and SS IIa exhibited small changes during five stages in both varieties and showed a down-up-down pattern in ND5181, with the greatest quantity at 25 DPA. SS IIa displayed an up-down pattern, while SS I showed a down-up-down pattern in SN119, with the highest quantity at about 20 DPA (Fig. 1B).

View Article: PubMed Central - PubMed

ABSTRACT

Waxy starch has an important influence on the qualities of breads. Generally, grain weight and yield in waxy wheat (Triticum aestivum L.) are significantly lower than in bread wheat. In this study, we performed the first proteomic and phosphoproteomic analyses of starch granule-binding proteins by comparing the waxy wheat cultivar Shannong 119 and the bread wheat cultivar Nongda 5181. These results indicate that reduced amylose content does not affect amylopectin synthesis, but it causes significant reduction of total starch biosynthesis, grain size, weight and grain yield. Two-dimensional differential in-gel electrophoresis identified 40 differentially expressed protein (DEP) spots in waxy and non-waxy wheats, which belonged mainly to starch synthase (SS) I, SS IIa and granule-bound SS I. Most DEPs involved in amylopectin synthesis showed a similar expression pattern during grain development, suggesting relatively independent amylose and amylopectin synthesis pathways. Phosphoproteome analysis of starch granule-binding proteins, using TiO2 microcolumns and LC-MS/MS, showed that the total number of phosphoproteins and their phosphorylation levels in ND5181 were significantly higher than in SN119, but proteins controlling amylopectin synthesis had similar phosphorylation levels. Our results revealed the lack of amylose did not affect the expression and phosphorylation of the starch granule-binding proteins involved in amylopectin biosynthesis.

No MeSH data available.


Related in: MedlinePlus