Nowicka Beatrycze, Strzalka Wojciech, Strzalka Kazimierz
Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Cracow, Poland.
J Plant Physiol. 2009 Jul 1;166(10):1045-56. doi: 10.1016/j.jplph.2008.12.010. Epub 2009 Mar 10.
Zeaxanthin epoxidase (ZE, E.C. 1.14.13.90), an enzyme belonging to the lipocalin superfamily, catalyses the conversion of zeaxanthin to antheraxanthin and violaxanthin. These reactions are part of the xanthophyll biosynthetic pathway and the xanthophyll cycle. The role of carotenoids in the dissipation of excessive light energy has been widely studied using mutants with a disabled carotenoid biosynthetic pathway. In this paper, the transgenic line MaZEP7 with partially disabled ZE activity is described and compared with wild-type plants and npq2 mutant lacking active ZE. We examined the presence and the abundance of aba1 transcripts, measured pigment composition, xanthophyll cycle functioning and chlorophyll fluorescence in all three lines. The MaZEP7 line contains additional copies of the aba1 gene introduced by agroinfiltration, but no enhanced aba1 transcript level was observed. In addition, ZE activity in MaZEP7 was impaired, resulting in an altered xanthophyll profile. In dark-adapted plants, violaxanthin and neoxanthin levels were lower than in wild-type plants, whereas antheraxanthin and zeaxanthin levels were considerably higher. The presence of lutein epoxide was also observed. Violaxanthin levels changed only minimally during light exposition, whereas antheraxanthin was converted to zeaxanthin and there was no epoxidation during the course of the experiment indicating disturbed xanthophyll cycle functioning. The amounts of carotenoids and chlorophylls on a dry weight basis and chl a/chl b ratio were similar in all lines. The presence of epoxidated pigments in MaZEP7 plants indicates that epoxidation occurs, but it is likely very slow. Chlorophyll fluorescence measurements showed that the dependence of electron transport rates on light intensity for the MaZEP7 line resembled the npq2 mutant. Kinetic measurements showed that the MaZEP7 line exhibited very rapid induction and a high steady-state value of non-photochemical quenching.
玉米黄质环氧化酶(ZE,E.C. 1.14.13.90)是一种属于脂质运载蛋白超家族的酶,催化玉米黄质转化为花药黄质和紫黄质。这些反应是叶黄素生物合成途径和叶黄素循环的一部分。利用类胡萝卜素生物合成途径有缺陷的突变体,人们对类胡萝卜素在耗散过量光能中的作用进行了广泛研究。本文描述了ZE活性部分丧失的转基因系MaZEP7,并将其与野生型植物以及缺乏活性ZE的npq2突变体进行比较。我们检测了所有三个株系中aba1转录本的存在情况和丰度,测量了色素组成、叶黄素循环功能以及叶绿素荧光。MaZEP7株系包含通过农杆菌浸润导入的额外aba1基因拷贝,但未观察到aba1转录本水平升高。此外,MaZEP7中的ZE活性受损,导致叶黄素谱发生改变。在暗适应的植物中,紫黄质和新黄质水平低于野生型植物,而花药黄质和玉米黄质水平则显著更高。还观察到了环氧叶黄素的存在。在光照过程中,紫黄质水平变化极小,而花药黄质转化为玉米黄质,并且在实验过程中没有环氧化现象,这表明叶黄素循环功能受到干扰。所有株系中基于干重的类胡萝卜素和叶绿素含量以及叶绿素a/叶绿素b比值相似。MaZEP7植物中存在环氧化色素表明环氧化反应会发生,但可能非常缓慢。叶绿素荧光测量结果表明,MaZEP7株系的电子传递速率对光强的依赖性类似于npq2突变体。动力学测量表明,MaZEP7株系表现出非常快速的诱导以及非光化学猝灭的高稳态值。