水稻叶片淀粉的分离与理化性质分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0628

摘要/Abstract

摘要：

目的分离高质量的叶片淀粉有助于研究叶片淀粉的理化特性及发育调控机制。利用已报道的叶片淀粉提取方法分离水稻叶片淀粉，步骤繁琐，操作要求高，且分离的水稻叶片淀粉纯度不高。因此，需要建立一种适合水稻叶片淀粉提取的方法。方法优化获得一种操作简单的水稻叶片淀粉分离和纯化方法，主要包括叶片磨粉、细胞裂解、淀粉释放、过筛、杂质去除、静置沉降、水洗淀粉、乙醇脱水和烘干保存9个步骤。该方法改良的核心在于利用静置沉降替代拟南芥叶片淀粉提取方法中的离心收集淀粉，通过3次悬浮沉降即可有效分离水稻叶片淀粉。结果利用该方法分离纯化3种水稻叶片的淀粉，经测定沉淀中淀粉纯度可以达到87%。与多角形的水稻胚乳淀粉粒形态相比，水稻叶片淀粉粒呈现扁平状和长的椭球形，且淀粉粒显著小于胚乳淀粉粒。热力学特性分析显示水稻叶片淀粉的糊化起始温度、峰值温度和终止温度显著高于胚乳淀粉，而热焓值低于胚乳淀粉。结论优化的叶片淀粉分离和提纯方法操作过程简单并且环保，获得的高纯度水稻叶片淀粉可以用于淀粉形态和理化特性的研究。

关键词: 水稻, 叶片淀粉, 淀粉粒, 淀粉提纯, 淀粉形态

Abstract:

Objective Separating high-quality leaf starch is conducive to studying the physicochemical properties and developmental regulation mechanisms of leaf starch. The separation of rice leaf starch using reported leaf starch extraction methods is complicated, requires high operational requirements, and the purity of the separated rice leaf starch is not high. Therefore, it is necessary to establish a method suitable for extracting starch from rice leaves. Method We optimized and obtained a simple method for separating and purifying rice leaf starch, which mainly includes nine steps: leaf grinding, cell splitting, starch release, filtration, impurity removal, standing sedimentation, starch washing, ethanol dehydration and drying preservation. The improvement of this method lies in using standing sedimentation to replace the centrifugal collection of starch in Arabidopsis leaf starch extraction method, which can effectively separate rice leaf starch through 3 times of suspension settling. Result Using this method to separate and purify starch from three types of rice leaves, the purity of starch in the precipitation reached 87%. Compared with the polygonal shape of endosperm starch granules, the leaf starch granules were flat and long ellipsoidal, whose size were significantly smaller than that of endosperm starch granules in rice. The thermodynamic property assay showed that the gelatinization onset temperature, gelatinization peak temperature and gelatinization completion temperature of leaf starch were significantly higher than those of endosperm starch, while the gelatinization enthalpy was lower than that of endosperm starch in rice. Conclusion The optimized method for separating and purifying rice leaf starch is simple and environment-friendly, by which the high-purity leaf starch is obtained for the study of starch morphology and physicochemical properties.

Key words: rice, leaf starch, starch granule, starch purification, starch morphology

方慧敏, 顾艺枢, 张晶, 张龙. 水稻叶片淀粉的分离与理化性质分析[J]. 生物技术通报, 2025, 41(2): 51-57.

FANG Hui-min, GU Yi-shu, ZHANG Jing, ZHANG Long. Isolation and Physicochemical Properties Analysis of Starch from Rice Leaves[J]. Biotechnology Bulletin, 2025, 41(2): 51-57.

图/表 5

图1 淀粉纯度测定Kit表示粳稻品种Kitaake，Dular表示籼稻品种Dular，GLXN表示糯稻品种广陵香糯（Guanglingxiangnuo），数值为平均值±标准差（n=3），相同字母表示数据之间没有显著性差异（P>0.05）

Fig. 1 Determination of starch purityKit indicates the japonicarice variety Kitaake, Dular indicates the indica rice variety Dular, and GLXN indicates the glutinous rice variety Guanglingxiangnuo, the data are of means ± standard deviations (n=3), and the same letter indicates no significant difference (P>0.05) among the data

图2 三个水稻品种淀粉的扫描电镜照片A-C： Kit， Dular和GLXN叶片淀粉； D-F： Kit， Dular和GLXN胚乳淀粉；红色箭头指的是空腔结构

Fig. 2 Scanning electron microscope photographs of starches from three rice varietiesA-C: The leaf starch of Kit, Dular and GLXN. D-F: The endosperm starch of Kit, Dular and GLXN. The red arrows refer to the cavity structure

图3 三个水稻品种淀粉的粒径分布A-C：叶片淀粉粒径分布图； D-F：胚乳淀粉粒径分布图

Fig. 3 Granule size distribution of starches in three rice varietiesA-C： Granule size distribution of leaf starch. D-F： Granule size distribution of endosperm starch

图4 Kit叶片淀粉和胚乳淀粉的DSC热谱图

Fig. 4 DSC thermograms of leaf starch and endosperm starch in Kit

表1 Kit叶片淀粉和胚乳淀粉的热力学参数

Table 1 Thermal properties of leaf starch and endosperm starch in Kit

淀粉组织

Starch tissue

起始温度

T_onset/℃

峰值温度

T_peak/℃

终止温度

T_completion/℃

糊化温度范围

T_range/℃

糊化焓值

Enthealpy/（J·g^-1）

叶片淀粉

Leaf starch

胚乳淀粉

Endosperm starch

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