Isolation and transformation of genes from woody plants that have high ability to clean up air polllutants

该项目旨在从木本植物中分离出对二氧化氮（一种典型的空气污染物）具有高清除能力的基因，如杜鹃花（Rhododendron mucronatum）。利用蛋白质的二维凝胶电泳，我们发现p1 5.6和MW 25-26kD的多肽点在二氧化氮熏蒸后强度增加了4到5倍。用气相法测定了这些多肽氨基端约20个氨基酸序列。同源性研究表明，这些多肽与拟南芥中所谓的gemin有57-68%的同源性。研究了编码该多肽的cDNA全长的克隆。利用差异显示法，我们发现在二氧化氮熏蒸处理下，长叶杜鹃（Rhododendron mucronatum）几种mRNA的表达强度有所增加或减少。测定了其中几个条带的DNA序列，同源性搜索表明，这些DNA与报道的蛋白质或cdna没有明显的同源性，提示其中一些是新的基因，在二氧化氮熏蒸后表达被诱导或降低。利用粒子轰击法对拟南芥和无花果进行了转化研究。

This project is aimed at the isolation of genes from woody plants such as Rhododendron mucronatum that have high ability to clean up nitrogen dioxide, a typical air pollutant. Using 2-D gel electrophoresis of proteins, we have found that polypeptide spots of p1 5.6 and MW 25-26kD increased their intensities by four to five times in response to fumigation with nitrogen dioxide. About 20 amino acid sequences at the amino terminals of these polypeptides were determined by vapor phase method. Homology search indicated that these polypeptide had 57-68% identity with so-called gemin from Arabidopsis thaliana. Cloning of the full length of cDNA that encodes this polypeptide was investigated. Using differential display method, we have found that several mRNA from the leaves Rhododendron mucronatum increased or decreased their intensities in response to fumigation with nitrogen dioxide. The DNA sequences of several bands were determined, and homology search indicated these DNAs did not have any distinct homology with reported proteins or cDNAs, suggesting that some of them are novel genes whose expression is induced or decreased upon fumigation with nitrogen dioxide. Transformation of Arabidopsis thaliana and Ficus thunbergii using particle bombardment method has also been investigated.