RAPID HIGH-YIELD ISOLATION OF METABOLICALLY INTACT PROTOPLASTS OF LEAVES FROM BETULA-PENDULA ROTH

被引：2

作者：

BIELZER, M ^{[1
]}

BENDL, HB ^{[1
]}

NADER, WF ^{[1
]}

BRINKMANN, K ^{[1
]}

机构：

[1] UNIV BONN,INST BOT,KIRSCHALLEE 1,W-5300 BONN,GERMANY

来源：

PLANT SCIENCE | 1991年 / 75卷 / 01期

关键词：

BETULA-PENDULA; PROTOPLAST ISOLATION; PHOTOSYNTHESIS; HOMOGENEITY; DIURNAL CYCLE;

D O I：

10.1016/0168-9452(91)90011-V

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A short duration isolation method has been developed achieving high yields of protoplasts from leaves of Betula pendula (2-4 x 10(7) protoplasts/g fresh wt.). After optimizing growth conditions of donor plants and the composition of digestion medium, intact protoplasts were isolated within 3 h. The rate of photosynthesis of purified protoplasts is comparable to those of the donor tissue. The maintenance of physiological intactness of that fraction is demonstrated by a diurnal cycle of photosynthetic capacity immediately following the preparation. The homogeneity of the protoplast fractions has been investigated by green and red autofluorescence using a flow fluorescence cytometer. Highest degrees of homogeneity (cytometer record), as highest intactness of plasmalemma (enzyme assay) and highest capacity of photosynthetic activity (oxygen polarography), occurred in the 20-30% Percoll fraction.

引用

页码：73 / 82

页数：10