ISOLATION OF APOPLASMIC FLUID FROM SUNFLOWER LEAVES AND ITS USE FOR STUDIES ON INFLUENCE OF NITROGEN SUPPLY ON APOPLASMIC PH

被引：63

作者：

DANNEL, F

PFEFFER, H

MARSCHNER, H

机构：

[1] Institut für Pflanzenernahrung, Universität Hohenheim, Stuttgart

来源：

JOURNAL OF PLANT PHYSIOLOGY | 1995年 / 146卷 / 03期

关键词：

HELIANTHUS ANNUUS L CV FRANKASOL; APOPLASMIC PH; ISOLATION APOPLASMIC FLUID; MARKER ENZYMES; NITROGEN SUPPLY;

D O I：

10.1016/S0176-1617(11)82053-5

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

A centrifugation method for collection of apoplasmic fluid from isolated sunflower leaves has been developed. As indicated by the marker enzymes hexose phosphate isomerase EC 5.3.1.9, glyceraldehyde-3-phosphate dehydrogenase EC 1.2.1.13 and malate dehydrogenase EC 1.1.1.37, the symplasmic contamination of the obtained fluid is smaller than 1.6%. Experiments with the apoplasmic tracer Sulphorhodamine G showed that no dilution with membrane filtered cell sap occurred. The described method allows quick and easy collection of apoplasmic fluid that is representative of its composition in intact leaves. The amounts of fluid obtained enable most analyses to be carried out. The method has been used to study the influence of ammonium or nitrate supply to roots on the apoplasmic pH in sunflower leaves. In plants supplied with either 2 mol m(-3) nitrate or ammonium the pH of the xylem exudate at the stem base was 5.80 and 5.79, respectively, and apoplasmic pH was 6.77 and 6.87, respectively. In plants supplied with 4 mol m(-3) nitrate, pH of the xylem exudate was 5.87 and apoplasmic pH 7.42. The results suggest that high apoplasmic pH in leaves is caused by high proton consumption during proton-anion cotransport across the plasma membrane.

引用

页码：273 / 278

页数：6

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