Robinson S J, Deroo C S, Yocum C F
Department of Cell and Molecular Biology, The University of Michigan, Ann Arbor, Michigan 48109.
Plant Physiol. 1982 Jul;70(1):154-61. doi: 10.1104/pp.70.1.154.
Electron transfer activity in intact trichomes of Spirulina platensis (Nordst.) Geitl. can be observed with either CO(2) or methylviologen as the Hill acceptor. Ferricyanide cannot penetrate the intact trichomes, but photoreduction of this oxidant can be observed when mediated by lipophilic oxidants such as p-phenylenediamine or 2,5-dimethyl-p-benzoquinone. The insensitivity of these reactions to dibromothymoquinone indicates that they are due largely to the activity of photosystem II. Direct photoreduction of ferricyanide can be observed in spheroplasts of Spirulina, indicating that such preparations have altered permeability properties when compared with intact trichomes. Preparation of these spheroplasts, which are osmotically fragile, requires that intact trichomes be washed with KCl and EDTA to induce lysozyme sensitivity and thereby allow digestion of the cell wall. The KCl/EDTA washing procedure used for spheroplast preparation alters the permeability of Spirulina trichomes, as evidenced by the ability of these preparations to photoreduce ferricyanide. This photoreduction reaction is insensitive to dibromothymoquinone, and is stimulated by high concentrations of divalent cations. During assays, the reaction is inhibited by the inclusion of polyethyleneglycol as an osmotic protectant. Photoreduction of methylviologen and NADP(+) is also observed in the washed trichomes, along with an endogenously catalyzed photoreduction of O(2) to H(2)O(2). Photophosphorylation cannot be observed in the washed preparations, but cyclic photophosphorylation with phenazinemethosulfate is observed after mild sonication. These results indicate that KCl/EDTA-washed trichomes of S. platensis retain the full range of energy transducing capacities associated with thylakoid membranes of the intact trichomes; the washing procedure facilitates spheroplast formation and alters, but does not abolish, permeability barriers in these preparations.
钝顶螺旋藻(Nordst.)Geitl.完整藻丝体中的电子传递活性,可通过以CO₂或甲基紫精作为希尔受体来观察。铁氰化物无法穿透完整的藻丝体,但当由诸如对苯二胺或2,5 - 二甲基对苯醌等亲脂性氧化剂介导时,可观察到该氧化剂的光还原。这些反应对二溴百里香醌不敏感,表明它们主要归因于光系统II的活性。在螺旋藻原生质球中可观察到铁氰化物的直接光还原,这表明与完整藻丝体相比,此类制剂的通透性特性发生了改变。制备这些对渗透压敏感的原生质球,需要用KCl和EDTA洗涤完整的藻丝体以诱导溶菌酶敏感性,从而使细胞壁得以消化。用于原生质体制备的KCl/EDTA洗涤程序改变了螺旋藻藻丝体的通透性,这些制剂能够光还原铁氰化物就证明了这一点。这种光还原反应对二溴百里香醌不敏感,并受到高浓度二价阳离子的刺激。在测定过程中,该反应会因加入聚乙二醇作为渗透保护剂而受到抑制。在洗涤过的藻丝体中还观察到甲基紫精和NADP⁺的光还原,以及O₂内源性催化光还原为H₂O₂。在洗涤过的制剂中未观察到光合磷酸化,但在轻度超声处理后,可观察到与硫酸吩嗪甲酯的循环光合磷酸化。这些结果表明,钝顶螺旋藻经KCl/EDTA洗涤的藻丝体保留了与完整藻丝体类囊体膜相关的全套能量转换能力;洗涤程序促进了原生质体的形成,并改变但并未消除这些制剂中的通透性屏障。