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用电场脉冲融合原生质体的膜完整性的水力传导率作为标准。

The hydraulic conductivity as a criterion for the membrane integrity of protoplasts fused by an electric field pulse.

机构信息

Arbeitsgruppe Membranforschung am Institut für Medizin, Kernforschungsanlage Jülich GmbH, Postfach 1913, D-5170, Jülich.

出版信息

Planta. 1982 Jul;155(2):140-5. doi: 10.1007/BF00392544.

Abstract

The hydraulic conductivity of the membrane, Lp, of fused plant protoplasts was measured and compared to that for unfused cells, in order to identify possible changes in membrane properties resulting from the fusion process. Fusion was achieved by an electric field pulse which induced breakdown in the membranes of protoplasts in close contact. Close membrane contact was established by dielectrophoresis. In some experiments pronase was added during field application; pronase stabilizes protoplasts against high field pulses and long exposure times to the field. The Lp-values were obtained from the shrinking and swelling kinetics in response to osmotic stress. The Lp-values of fused mesophyll cell protoplasts of Avena sativa L. and of mesophyll and guard cell protoplasts of Vicia faba L. were found to be 1.9±0.9·10(-6), 3.2±2.2·10(-6), and 0.8±0.7·10(-6) cm·bar(-1)·s(-1), respectively. Within the limits of error, no changes in the Lp-values of fused protoplasts could be detected in comparison to unfused protoplasts. The Lp-values are in the range of those reported for walled cells of higher plants, as revealed by the pressure probe.

摘要

为了鉴定融合过程是否导致膜性质发生变化,我们测量并比较了融合和未融合植物原生质体的膜透性(Lp)。融合通过电脉冲诱导紧密接触的原生质体的膜破裂来实现。通过介电泳使原生质体紧密接触。在某些实验中,在施加电场时加入了 Pronase;Pronase 可稳定原生质体,使其能够耐受高强度的电场脉冲和长时间的电场暴露。通过原生质体对渗透压胁迫的收缩和膨胀动力学来获得 Lp 值。融合的燕麦(Avena sativa L.)叶肉细胞原生质体和融合的蚕豆(Vicia faba L.)叶肉细胞和保卫细胞原生质体的 Lp 值分别为 1.9±0.9·10(-6)、3.2±2.2·10(-6)和 0.8±0.7·10(-6) cm·bar(-1)·s(-1)。在误差范围内,与未融合的原生质体相比,融合原生质体的 Lp 值没有变化。通过压力探针揭示,Lp 值与已报道的高等植物有壁细胞的 Lp 值范围一致。

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