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

该项目旨在从杜鹃等木本植物中分离出对典型空气污染物二氧化氮具有很强净化能力的基因。利用蛋白质双向凝胶电泳法，我们发现p1 5.6和mw 25-26kD的多肽斑点对二氧化氮熏蒸的反应强度增加了4-5倍。用气相法测定了这些多肽的氨基末端约20个氨基酸序列。同源性搜索表明，这些多肽与拟南芥中的Gmin有57-68%的同源性。对编码该多肽的全长cDNA进行了克隆。采用差异显示的方法，我们发现在二氧化氮的熏蒸下，多个短毛杜鹃叶片中的几个基因的表达强度增加或降低。测定了几条带的DNA序列，同源性搜索表明这些DNA与已报道的蛋白质或cDNA没有明显的同源性，表明其中一些是新基因，其表达在二氧化氮熏蒸后被诱导或下调。还研究了用基因枪法转化拟南芥和无花果的情况。

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.