Morphological, Molecular Structural and Physicochemical Characterization of Starch Granules Formed in Endosperm of Rice with Ectopic Overexpression of α-Amylase.

IF 1.2 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Journal of applied glycoscience Pub Date : 2024-04-20 eCollection Date: 2024-01-01 DOI:10.5458/jag.jag.JAG-2023_0016
Kuo Zhang, Sumiko Nakamura, Ken-Ichi Ohtsubo, Toshiaki Mitsui
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引用次数: 0

Abstract

The objective of this study was to characterize the endosperm starch in rice that ectopically overexpressed the α-amylase. Transgenic rice plants, transformed with cauliflower mosaic virus 35S promoter driven AmyI-1 (35S::AmyI-1) and AmyII-4 (35S::AmyII-4), and 10 kDa prolamin promoter driven AmyI-1 (P10::AmyI-1), were cultivated under standard conditions (23 °C, 12 h in the dark/ 26 °C, 12 h in the light), and brown grains were subsequently harvested. Each grain displayed characteristic chalkiness, while electron microanalyzer (EPMA)-SEM images disclosed numerous small pits on the surface of the starch granules, attributable to α-amylase activity. Fluorescence labeling and capillary electrophoresis analysis of starch chain length distribution revealed no significant alterations in the starches of 35S::AmyI-1 and 35S::AmyII-4 transgenic rice compared to the wild-type. Conversely, the extremely short α-glucan chains (DP 2-8) exhibited a dramatic increase in the P10::AmyI-1 starch. Rapid visco-analyzer analysis also identified variations in the chain length distribution of P10::AmyI-1 starch, manifesting as changes in viscosity. Moreover, 1H-NMR analysis uncovered dynamic modifications in the molecular structure of starch in rice grain transformed with P10::AmyI-1, which was found to possess unprecedented structural characteristics.

异位过表达α-淀粉酶的水稻胚乳中形成的淀粉颗粒的形态、分子结构和物理化学特征
本研究旨在描述异位过表达α-淀粉酶的水稻胚乳淀粉的特征。用花椰菜花叶病毒 35S 启动子驱动的 AmyI-1(35S::AmyI-1)和 AmyII-4(35S::AmyII-4)以及 10 kDa 脯氨酶启动子驱动的 AmyI-1(P10::AmyI-1)转化的转基因水稻植株在标准条件下(23 °C、12 小时黑暗/26 °C、12 小时光照)栽培,随后收获棕色谷粒。每粒淀粉都显示出特有的粉化现象,而电子显微分析仪(EPMA)-SEM 图像显示淀粉颗粒表面有许多小凹坑,这些凹坑可归因于α-淀粉酶的活性。淀粉链长度分布的荧光标记和毛细管电泳分析显示,与野生型相比,35S::AmyI-1 和 35S::AmyII-4 转基因水稻的淀粉没有明显变化。相反,在 P10::AmyI-1 淀粉中,极短的 α-葡聚糖链(DP 2-8)急剧增加。快速粘度分析仪分析也发现了 P10::AmyI-1 淀粉链长分布的变化,表现为粘度的变化。此外,1H-NMR 分析揭示了用 P10::AmyI-1 转化的稻谷中淀粉分子结构的动态变化,发现它具有前所未有的结构特征。
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来源期刊
Journal of applied glycoscience
Journal of applied glycoscience BIOCHEMISTRY & MOLECULAR BIOLOGY-
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9.10%
发文量
13
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