Analysis of sugar accumulation in a single parenchyma cell of the fruit

果实单个薄壁细胞糖积累分析

基本信息

批准号：
11660027
负责人：
YONEMORI Keizo
金额：
$ 2.37万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

The analysis methods for an intact single parenchyma cell of the fruit was examined in this research. The method developed is firstly due to collect vacuolar sap from a single cell of the fruit, and then to analyze at picolitre levels for studying sugar accumulation in a single parenchyma cell of the fruit.1. The maicromanipulator system were used for collecting vacuolar sap. The glass micropipette was inserted into a targeted cell, and vacuolar sap was withdrawn from the cell by lowering the pressure. Then, the collected vacuolar sap was expelled into 5 μL of an oil drop and stored at -20℃ for sugar analysis. The content of glucose, fructose, sucrose, or sorbitol were determined by the intensity of NADPH, which was produced with enzymatic reaction of 15 pl vacuolar sap, using a fluorescence microscope equipped with a photometer produced. The invertase activities were also determined by the same method.2. The method for determining the osmotic pressure and potassium content of the vacuolar sap from a single cell were tested. Osmotic pressure was determined by measuring freezing point depression of the sap using a picolitre osmometer, and potassium content was determined by X-ray microanalysis using a scanning electron microscopy (SEM) with energy dispersive spectrometers.3. Using above methods mentioned, analysis of sugar accumulation was conducted to the fruit of persimmon, grape, peach, Japanese pear, and apple on a single parenchyma cell basis. It was revealed that sugar accumulation pattern of a single cell is coincident with that of bulk tissue of the fruit in every kind of fruit, and that increase in osmotic pressure is due to accumulation of the sugars in the cells. Furthermore, individual cells showed different sugar compositions in some fruit, but it had the same compositions in other fruit.

在这项研究中检查了完整的果实单实质细胞的分析方法。开发的方法首先是由于从水果的单个细胞中收集真空汁液，然后在Picolitre水平上分析以研究水果的单个实质细胞中的糖积累1。微型操纵器系统用于收集真空汁液。将玻璃微孔插入靶细胞中，并通过降低压力从细胞中撤出真空汁液。然后，将收集的真空汁液探索到5μl的油滴中，并存储在-20℃进行糖分析。葡萄糖，果糖，蔗糖或山梨糖醇的含量是由NADPH的强度确定的，NADPH的强度是用荧光显微镜等效的荧光显微镜与产生的光度计产生的。转化酶的活性也通过相同的方法确定。2。测试了从单个细胞中确定真空汁的渗透压和钾含量的方法。渗透压是通过使用PICOLITRE渗透压计测量SAP的冻结点抑制的渗透压，并使用X射线微分析使用扫描电子显微镜（SEM）确定钾含量。3。使用上述方法，以单个实质的基础对柿子，葡萄，桃子，日本梨和苹果的果实进行了糖积累的分析。据表明，单个细胞的糖积累模式与各种水果中果实的块状组织的糖积聚模式相吻合，并且渗透压的增加是由于细胞中糖的积累而引起的。此外，单个细胞在某些果实中显示出不同的糖成分，但在其他果实中具有相同的成分。