Kaldenhoff R, Kölling A, Meyers J, Karmann U, Ruppel G, Richter G
Institut für Botanik, Universität Hannover, Germany.
Plant J. 1995 Jan;7(1):87-95. doi: 10.1046/j.1365-313x.1995.07010087.x.
According to our previous studies the Arabidopsis gene AthH2 which is inducible by blue light and phytohormones codes for an intrinsic membrane protein. It bears a resemblance to several distinct channel proteins of plant and animal species classified as the MIP/NOD-26/GlpF family. In the present study biochemical analyses and electron microscopic immunochemistry were used to elucidate the subcellular location of the AthH2 protein. The results clearly demonstrate that it is an exclusive constituent of the plasmalemma. Furthermore, the expression of the AthH2 gene in transgenic Arabidopsis plants containing the promoter region of AthH2 fused to the beta-glucuronidase (gus) reporter gene was studied. The in situ localization of gus activity revealed that the specific promoter is temporally activated by light in expanding and/or differentiating cells comprising newly formed tissues and organs: root elongation zone, guard cells of stomata, vascular bundle sheaths, filaments of stamen and young siliques. Several sites of gus expression coincide spatially with those of in situ hybridization and the immunocytochemical reaction, respectively, suggesting that the AthH2 promoter had correctly responded to light as an important exogenous factor with relevance to the complex pattern of differentiation. Studies with protoplasts from plants transformed with an antisense construct revealed a water transport capacity of the AthH2 protein.
根据我们之前的研究,拟南芥基因AthH2可被蓝光和植物激素诱导,编码一种内在膜蛋白。它与植物和动物物种中几种不同的通道蛋白相似,这些通道蛋白被归类为MIP/NOD - 26/GlpF家族。在本研究中,采用生化分析和电子显微镜免疫化学方法来阐明AthH2蛋白的亚细胞定位。结果清楚地表明,它是质膜的唯一成分。此外,还研究了AthH2基因在含有与β - 葡萄糖醛酸酶(gus)报告基因融合的AthH2启动子区域的转基因拟南芥植物中的表达。gus活性的原位定位显示,特定启动子在包括新形成的组织和器官的正在扩展和/或分化的细胞中被光暂时激活:根伸长区、气孔保卫细胞、维管束鞘、雄蕊花丝和幼嫩角果。gus表达的几个位点分别在空间上与原位杂交和免疫细胞化学反应的位点一致,这表明AthH2启动子作为与复杂分化模式相关的重要外源因子已正确响应光。对用反义构建体转化的植物原生质体的研究揭示了AthH2蛋白的水运输能力。