U.S.-Japan Cooperative Science: Characterization of Alkaloid Biosynthetic Genes in Nicotiana Tabacum

该奖项支持弗吉尼亚州布莱克斯堡弗吉尼亚理工学院的John Jelesko教授和日本埼玉日立高级研究实验室的Masaki Furuya博士之间为期两年的合作研究项目。研究人员将对烟草中生物碱生物合成基因的特征进行研究。Tropane生物碱是参与植物化学生态的重要次生代谢物。尽管植物生物碱具有重要的作用，但人们对生物碱合成基因的分子生物学及其调控知之甚少。本研究项目的目的是利用一种称为荧光差异显示（FDD）的先进转录分析方法，鉴定和分离烟草（Nicotiana Tabacum）中的tropane生物碱合成基因。研究人员将进行几次大规模的FDD筛选，以鉴定烟草中的生物碱生物合成基因。他们将试图确定野生型烟草和突变型烟草之间表达差异的基因，突变型烟草具有显著降低tropane生物碱水平的双突变和单突变。通过利用特定的根培养条件和在近等基因背景下定义的突变，他们将确定与生物碱生物合成有关的差异表达基因。这种方法克服了传统生物化学和遗传方法在烟草物种中固有的困难。该项目汇集了两个具有互补专业知识和研究能力的实验室的努力。该研究将为植物生物碱合成提供重要信息，从而在植物生物技术中得到潜在的应用。这项研究也将引起制药行业的兴趣。该项目通过一名博士后和一名研究生的参与来促进国际人力资源的发展。通过思想和技术的交流，该项目将扩大我们的基础知识基础，促进国际了解与合作。研究结果将发表在国际科学期刊上，并将在美国和国外的科学会议上发表。

9912099JeleskoThis award supports a two-year collaborative research project between Professor John Jelesko of Virginia Polytechnic Institute in Blacksburg, Virginia and Dr. Masaki Furuya of the Hitachi Advanced Research Laboratory in Saitama, Japan. The researchers will be undertaking a study of the characterization of alkaloid biosynthetic genes in Nicotiana Tabacum. Tropane alkaloids are important secondary plant metabolites involved in plant chemical ecology. Despite the importance of plant alkaloids little is known about the molecular biology of alkaloid biosynthetic genes or their regulation. The goal of this research project is to use an advanced transcription profiling method, called Fluorescent Differential Display (FDD), to identify and isolate tropane alkaloid biosynthetic genes in Nicotiana Tabacum (tobacco). The researchers will perform several large-scale FDD screens to identify alkaloid biosynthetic genes in tobacco. They will attempt to identify genes that are differentially expressed between a wild type tobacco accession and mutant tobacco accessions that have defined double and single mutations that greatly reduce tropane alkaloid levels. By utilizing specific root culture conditions and defined mutations in near isogenic backgrounds they will identify differentially expressed genes specifically involved in alkaloid biosynthesis. This approach overcomes the difficulties inherent with traditional biochemical and genetic methods in Nicotiana species. The project brings together the efforts of two laboratories that have complementary expertise and research capabilities. The research should provide important information on alkaloid biosynthesis in plants, which should then result in potential applications in plant biotechnology. The research will also be of interest to the pharmaceutical industries. The project advances international human resources through the participation of a postdoc and a graduate student. Through the exchange of ideas and technology, this project will broaden our base of basic knowledge and promote international understanding and cooperation. Results of the research will be published in international scientific journals and will also be presented at scientific meetings in the U.S. and abroad.