摘要：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。具體工作項目包括：1. 黃龍病細菌(Candidatus Liberibacter asiaticus)之Real-time PCR定量偵測技術之改良與應用；2. 鱗砧類病毒(CEVd)偵測技術之改良；3. 柑橘萎縮病毒(CTV)膠金抗體試劑之製備與應用；4. 以人工合成的多肽鏈製備柑橘破葉病毒(CTLV)之診斷用抗體試劑；5.診斷技術實際應用於台灣柑橘健康種苗制度之推展；6.診斷技術實際應用於台灣柑橘新興品種與新育成品種的罹病性評估；7. 柑橘黃龍病菌與萎縮病毒的偵測技術應用在病媒昆蟲的診斷鑑定與傳病特性之研究。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。
Abstract: The systemic pathogens such as the bacteria 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 items will be conducted this year: (1) improvement and application of the Real-time PCR quantitative detection for huanglongbing bacteria (HLBB)；(2) improvement of the detection method for Citrus exocortis viroid (CEVd); (3) development and application of the diagnostic strips with colloidal-gold antibodies against Citrus tristeza virus (CTV); (4) preparation of the diagnostic antibodies for Citrus tatter leaf virus (CTLV) using the artificially synthetic oligo-peptides against HLBB; (5) application of the integrated diagnostic kit in the promotion of pathogen-free nursery system of citrus; (6) developed diagnostic techniques applied in the evaluation of susceptibility to HLBB, CTV, CTLV and CEVd for new citrus cultivars; (7) developed diagnostic techniques of HLBB and CTV applied in the study of transmission and detection of pathogens in insect vectors. 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.
Candidatus Liberibacter asiaticus
Citrus tristeza virus
Citrus tatter leaf virus
Citrus exocortis viroid