TRANSGENIC PLANT AEQUORIN REPORTS THE EFFECTS OF TOUCH AND COLD-SHOCK AND ELICITORS ON CYTOPLASMIC CALCIUM
TRANSGENIC PLANT AEQUORIN REPORTS THE EFFECTS OF TOUCH AND COLD-SHOCK AND ELICITORS ON CYTOPLASMIC CALCIUM
复制标题
DOI：
10.1038/352524a0
复制
发表时间：
1991-08-08
期刊：
影响因子：
64.8
通讯作者：
TREWAVAS, AJ
中科院分区：
文献类型：
--
作者：
KNIGHT, MR;CAMPBELL, AK;TREWAVAS, AJ
使用微电极或微量注射荧光染料测量植物细胞质钙的方法存在大量技术问题，因此测量仅限于组织条或原生质体中的单个细胞或小群细胞[1,2]。水母发光蛋白是一种来自腔肠动物维多利亚多管水母（A. forskalea）的钙敏感发光蛋白[3]，它由相对分子质量约为22,000的脱辅基水母发光蛋白和疏水性发光团腔肠素组成[4]。微量注射的水母发光蛋白已广泛用于动物细胞的细胞内钙测量[4]，但它在植物中的应用仅限于特别大的细胞[5]。我们在此表明，水母发光蛋白可以在转基因植物中重组，并且它能报告由触摸、冷激和真菌激发子诱导的钙变化。重组的水母发光蛋白位于细胞质中且无干扰；可以对整株植物进行测量，并且每个活细胞中都可以有一个钙指示剂。既然脱辅基水母发光蛋白可以靶向特定的细胞器、细胞和组织，并且有具有不同钙敏感性和特性的腔肠素可供使用[6]，这种新方法对于确定钙在植物细胞内信号传导过程中的作用可能是有价值的。
METHODS for measuring plant cytoplasmic calcium using microelectrodes or microinjected fluorescent dyes are associated with extensive technical problems, so measurements have been limited to single or small groups of cells in tissue strips or protoplasts 1,2. Aequorin is a calcium-sensitive luminescent protein 3 from the coelenterate Aequorea victoria (A. forskalea) which is formed from apoaequorin, a polypeptide of relative molecular mass approximately 22,000, and coelenterazine, a hydrophobic luminophore 4. Microinjected aequorin has been widely used for intracellular calcium measurement in animal cells 4, but its use in plants has been limited to exceptionally large cells 5. We show here that aequorin can be reconstituted in transformed plants and that it reports calcium changes induced by touch, cold-shock and fungal elicitors. Reconstituted aequorin is cytoplasmic and nonperturbing; measurements can be made on whole plants and a calcium indicator can be constituted in every viable cell. Now that apoaequorin can be targeted to specific organelles, cells and tissues, with the range of coelenterazines with differing calcium sensitivities and properties available 6, this new method could be valuable for determining the role of calcium in intracellular signalling processes in plants.