Legge R L, Cheng K H, Lepock J R, Thompson J E
Department of Biology, University of Waterloo, Waterloo, Ontario, Canada, N2L 3G1.
Plant Physiol. 1986 Aug;81(4):954-9. doi: 10.1104/pp.81.4.954.
Changes in the molecular organization of membranes in pericarp cells of ripening tomato fruit were examined by fluorescence depolarization after labeling with fluorescent lipid-soluble probes. The fluorescent labels were partitioned into isolated protoplasts and purified plastids from fruit at various stages of senescence. Values for steady-state anisotropy (r(ss)) of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labeled protoplasts rose progressively during the early stages of ripening over a time frame that overlapped the climacteric rise in ethylene production. This can be interpreted as reflecting a decrease in the lipid fluidity of primarily plasma membrane. By contrast, there was no significant change during ripening in r(ss) for plastid membranes labeled with DPH, 1-[4-trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH), and cis- or trans-parinaric acid. Nor was there any change during ripening in the limiting fluorescence anisotropy (r(oo)) and order parameter (S) for plastids labeled with DPH or TMA-DPH, parameters that are corrected for any differences in lifetime. Some degree of lifetime heterogeneity, possibly reflecting structurally distinct domains, was discerned in both young and senescent plastids that had been labeled with DPH or TMA-DPH, but this also did not change as ripening progressed. Thus membranes of the pericarp cells sustain different fates as the tomato fruit ripens, implying that there are distinguishable mechanisms of membrane deterioration in senescing tissues.
在用荧光脂溶性探针标记后,通过荧光去极化研究了成熟番茄果实果皮细胞中膜的分子组织变化。荧光标记被分配到处于不同衰老阶段果实的分离原生质体和纯化质体中。在成熟早期,1,6 - 二苯基 - 1,3,5 - 己三烯(DPH)标记的原生质体的稳态各向异性(r(ss))值在与乙烯产量的跃变上升重叠的时间范围内逐渐升高。这可以解释为主要反映了质膜脂质流动性的降低。相比之下,用DPH、1 - [4 - (三甲氨基)苯基] - 6 - 苯基 - 1,3,5 - 己三烯(TMA - DPH)以及顺式或反式十八碳四烯酸标记的质体膜在成熟过程中r(ss)没有显著变化。用DPH或TMA - DPH标记的质体的极限荧光各向异性(r(oo))和序参数(S)在成熟过程中也没有变化,这些参数是针对寿命的任何差异进行校正的。在用DPH或TMA - DPH标记的幼嫩和衰老质体中都发现了一定程度的寿命异质性,这可能反映了结构上不同的区域,但随着成熟的进行这也没有变化。因此,随着番茄果实成熟,果皮细胞的膜经历不同的命运,这意味着在衰老组织中存在可区分的膜降解机制。