DNA Amplification Fingerprinting and Molecular Diagnosis for Detecting Bacterium-like Organisms Causative of Citrus Greening Disease, US-Nepal Cooperative Research

项目描述：本项目支持尼泊尔加德满都提尔布万大学植物系Bhaju K. Tamot博士在诺克斯维尔田纳西大学植物分子遗传学系Peter H. Gresshoff博士实验室的研究项目。采用DNA扩增指纹技术（DAF）对柑桔黄萎病病原菌进行DNA检测。DNA将分别从尼泊尔感染和健康柑橘的叶子、花芽和茎组织中分离出来。将尝试从任何细菌样生物（BLO）中分离DNA，并生成DAF模式。用特异性引物对感染组织和非感染组织的DNA进行扩增，产生DNA扩增产物的差异，即一个或多个多态性条带，将作为检测CGD的标记。研究范围：这项研究带来了一位来自尼泊尔的年轻科学家，刚获得博士学位，在DAF技术及其在植物育种中的应用方面有一些经验，以及一位在该领域发表过大量文章的美国资深科学家。Tamot博士在尼泊尔的实地工作，加上田纳西大学与Gresshoff博士合作开展的分析工作，应能产生两方面的好处：扩大DAF作为柑橘病早期检测手段有效性的研究数据和信息，并为尼泊尔科学家提供更好的培训，使他能够将该技术应用于本国的其他生物目标。这代表了一个与发展中国家的强有力的合作项目，以及对最近的外国博士后的支持。＊＊＊

9311802 Gresshoff Description: This project supports the participation of Dr. Bhaju K. Tamot, of the Department of Botany, Tirbhuvan University, Kathmandu, Nepal, in a research project at the laboratory of Dr. Peter H. Gresshoff of the Department of Plant Molecular Genetics at the University of Tennessee, Knoxville. The research is to use DNA amplification fingerprinting (DAF), a newly developed DNA- electrophoretic assay, to detect the DNA of the causative agent of citrus greening disease (CGD). The DNA will be isolated separately from leaf, flower bud and stem tissue of infected and healthy citrus in Nepal. Attempts will be made to isolate DNA from any Bacterium- Like Organism (BLO), and to generate DAF patterns. The difference in DNA amplification products, one or more polymorphic bands, generated from DNA of infected and non-infected tissues with specific primer will be used as markers for detecting CGD. Scope: This research brings a young scientist from Nepal, a recent Ph.D., who has had some experience in DAF technology and its application in plant breeding, with a senior U.S. scientist who has published extensively in the field. The field work by Dr. Tamot in Nepal, combined with the analytical work at the University of Tennessee in cooperation with Dr. Gresshoff should produce two benefits, an expanded research data and information about the validity of DAF as an early detector of the citrus disease, and a better training for the Nepalese scientist that should enable him to apply the technique to other biological objectives in his country. This represents a strong cooperative project with a developing country as well as support for a recent foreign Postdoctorate. ***