摘要：黃龍病是柑橘極重要的病害，由無法培養的靭皮部侷限細菌所引起，主要經由帶病種苗來傳播，而在種植後又可經柑桔木蝨媒介傳播，屬於世界性的植物流行病，本研究室對此病已累積數十年之研究經驗。2005年柑橘黃龍病入侵美國佛羅里達州，引起美國政府重視，包括美國農業部(Animal and Plant Health Inspection Service=APHIS)與佛羅里達州政府(Florida Department of Agriculture & Consumer Services=FDACS)皆提出與本研究室合作研究的要求。本計畫之研提旨在建構台美學術合作關係，應用多項先進的分子生物學相關技術，研發出更高效能的柑橘黃龍病菌分子檢測技術，一方面用於協助美國的黃龍病防疫與檢疫工作，達到學術外交的目的；另一方面也期望透過國際專利申請與學術論文發表，建立一套具有國際認證性的黃龍病菌標準檢測方法，或生產出可行銷於世界的檢測試劑，以彰顯我們在柑橘黃龍病研究上的優勢地位，也有助於我國內防檢疫之應用。本計畫預計執行兩年，期間除了促成雙方研究人員互訪與技術交流外，將致力改良或研發更先進的核酸分子檢測技術，包括PCR快速檢測、Real-Time PCR定量偵測、迴路媒介恆溫增殖法(LAMP)與DNA Microarray等；另外也要進行蛋白分子檢測技術研發，以病原菌相關的蛋白分子為抗原，進行單株抗體製備，進而製成膠金抗體試劑，以期更能增加檢測的方便性。
Abstract: Huanglongbing (HLB), also known as greening, is a destructive disease of citrus. It is caused by a phloem-limited fastidious bacterium, and transmitted by vegetative propagation (grafting) as well as the insect vector, Asian citrus psyllid (Diaphorina citri Kuwayama). HLB bacterium has not yet been successfully cultured in vitro, and a tentative name of Candidatus Liberibacter asiaticus has been proposed. HLB has been discovered in Asia, Africa and America. It has become a very important epidemic in the world. We have been studying HLB for so many years and accumulated abundant experiences. This pathogen invaded Florida in 2005, which immediately caused official concern of the United States. Several months ago, both FDACS (Florida Department of Agriculture & Consumer Services) and APHIS (Animal and Plant Health Inspection Service, USDA) communicated with us to search for a cooperative opportunity in the HLB study. This research project was therefore proposed for the construction of collaboration between Taiwan and USA. The project is dedicated to develop more efficiently diagnostic methods for HLB using advanced molecular techniques. The newly devised methods will directly assist the inspection and quarantine for HLB in USA as well as Taiwan. They will be further published in international journals and then assigned to apply the patent. We are looking forward to establishing an excellent and standard protocol for the detection of C. Liberibacter asiaticus that can be widely adopted for international quarantine. This research project will be conducted for two years to achieve three major goals：first, creation of the exchange channel of scholar to share the experiences in HLB research between Taiwan and USA； Second, development of advanced molecular diagnostic methods including the simplified PCR-based assay, Real-Time PCR quantitative detection, Loop-mediated isothermal amplification(LAMP), and DNA microarray; third, preparation of the monoclonal antibodies against the specific protein molecules associated with C. Liberibacter asiaticus. The monoclonal antibodies showing high sensitivity and specificity will be selected to produce colloidal-gold labeled antibody strips for more convenient and economical detection.
citrus huanglongbing (greening)
cooperation between Taiwan and USA
molecular diagnostic methods