2014-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/682468摘要：&nbsp;&nbsp;&nbsp; 台灣重要柑橘系統性病原包括黃龍病細菌(HLBB)、萎縮病毒(CTV)、破葉病毒(CTLV)與鱗砧次病毒(CEVd)等，這些病原經由帶病種苗普遍傳播而危害嚴重，生產無病種苗搭配敏感且正確的檢疫技術乃是防治這些病害的首要之道。本計畫以原本初步研發成功的柑橘病原檢疫技術為基礎，進一步再研發與改善，目標在於建立一套方便且快速的整合性偵測套組。本年度具體工作目標包括：(1) 研發可同時萃取CTV、CTLV與CEVd等三種病毒RNA的通用性核酸萃取法；(2) 研發更經濟、快速且簡易的黃龍病細菌DNA的核酸萃取法；(3) 完成可同時偵測CTV、CTLV與CEVd等三種病毒的三合一Multiplex RT-PCR快速檢測技術之研發與改良；(4) 完成柑橘黃龍病菌與三種病毒的商品化整合性偵測套組之研製並實際應用於田間柑橘樣本之檢測。本計畫所得研究成果不但可供國內防檢疫之應用，更因這些病原皆為國際性重要病原，未來可申請專利而推展於國際。<br> Abstract: &nbsp; The systemic pathogens such as the bacteria (Candidatus Liberibacter asiaticus = Las) causing citrus huanglongbing, Citrus tristeza virus (CTV), Citrus tatter leaf virus (CTLV) and Citrus exocortis viroid (CEVd) are predominant and destructive to citrus industry in Taiwan. These pathogens are mainly transmitted by vegetative propagation. Establishment of pathogen-free nursery systems and development of rapid and sensitive diagnostic probes are considered to be the most important methods for the pest control. This research project is dedicated to develop new diagnostic probes and improve several devised diagnostic methods using immunological and molecular techniques. An integrated diagnostic kit will be produced through combination of all our devised detection methods for these citrus systemic pathogens. Several targets will be achieved this year: (1) development of&nbsp;a common RNA extraction method simultaneously adequate for the extraction of CTV, CTLV and CEVd; (2) development of the more economic, rapid and simple method for the extraction of Las DNA; (3) development and improvement of&nbsp;3 in 1 multiplex RT-PCR&nbsp;assays for the simultaneous detection of CTV, CTLV and CEVd; (4)&nbsp;development of the commercialized diagnostic kit and its application in the promotion of pathogen-free nursery system of citrus and the control of citrus diseases. These devised methods will be helpful to the inspection and management of the citrus pests, and it is worthy of international patent for world-wide extension.分子偵測黃龍病柑橘黃龍病細菌柑橘萎縮病毒柑橘破葉病毒柑橘鱗砧類病毒Molecular DetectionHuanglongbingCitrusCandidatus Liberibacter asiaticusCitrus Tristeza VirusCitrus Tatter Leaf VirusCitrus Exocortis Viroid