查詢結果分析
相關文獻
- Improvement of Techniques for Purification of Leafroll Associated Closterovirus from Affected Grapevines and the Preparation of Antisera for Disease Indexing
- The Occurrence of Grapevine Leafroll Disease Among the Main Grapevine Cultivars and Breeding Stocks in Taiwan
- Purification and Characterization of a Tospovirus Systemically Infecting Cucurbits in Taiwan
- 百合Plantago asiatica mosaic virus檢測試劑製備與提升病毒檢出率之血清檢定法研發
頁籤選單縮合
題 名 | Improvement of Techniques for Purification of Leafroll Associated Closterovirus from Affected Grapevines and the Preparation of Antisera for Disease Indexing=葡萄捲葉毒素病病毒純化技術之改進及其抗血清之製備與應用 |
---|---|
作 者 | 陳慧璘; 陳脈紀; 曾德賜; | 書刊名 | Botanical Bulletin of Academia Sinica |
卷 期 | 40:4 1999.10[民88.10] |
頁 次 | 頁295-304 |
分類號 | 435.38 |
關鍵詞 | 血清製備; 中藥乾物粉碎機; 病害檢診; 葡萄捲葉病; 病毒純化; Antiserum preparation; Chinese herb grinder; Disease indexing; Grapevine leafroll disease; Virus purification; |
語 文 | 英文(English) |
中文摘要 | 本研究主要目的在發展一改良之技術, 利用其純化葡萄捲葉病毒, 並製備其對應多元及單元抗血清,以供病害檢測之用。此改良方法包括利用中藥乾物粉碎機, 將罹病葡萄樹皮材料於液態氮中磨碎,由於此一方法只需很有限的人力,且於短短一、兩分鐘內即可將材料研磨成細粉,應用於大量葡萄樹皮材料之研磨時,可避免因材料研磨困難且所需時間較長, 以致造成病毒分解、收量減低等現象,對於自葡萄組織中純化濃度極低之捲葉病毒,純化結果與再現性均可獲得改善。利用此一方法,並參照及修飾前人之病毒純化方法,本研究已成功由台灣的巨峰、黑后及金香等主要栽培品種葡萄材料純化出捲葉病毒,經電子顯微鏡檢視發現, 所純化病毒之收量與純度,確與前人研究所得病毒製備相當或更好。所純化之病毒經注射大白兔及 Bdlb/C 小白鼠, 已製備成功單元及多元抗體, 利用直接及間接酵素連接抗體之血清檢測法檢定並發現,危害本省葡萄之捲葉病毒,主要為第三與第四血清型。利用所建立的血清檢診技術,本研究於1987~1994年間,檢測菸試所所保存的葡萄品種,結果發現 204 個保存的葡萄品種中,大約有 50% 罹患捲葉病,其中 30% 感染第三型,另31% 感染第四型, 本省田間主要栽培品種如巨峰等複合感染第三及第四型捲葉病毒之現象相當普遍。此外調查結果並指出本病毒於田間有明顯蔓延之現象,顯示在重要種源保存上,嚴格的病原檢測與清潔苗木工作極為必要。 |
英文摘要 | An improved technique was developed for the purification of leafroll associated closterovirus (GLRaV) and successfully applied for the preparation of polyclonal and monoclonal antisera for the virus detection. The technique improvement mainly involves the use of a Chinese herb grinder for the homogenization of diseased sample. It requires very limited labor, is repeatable in regard to the virus yield, and can process considerable quantities within a short period. By this sample homogenization technique and a method modified from one in the available literature, GLRaV was successfully purified from diseased Kyoho, Black Queen, and Golden Mutscate grapevines cultivated in Taiwan. The electron microscopic examination of the obtained virus preparations revealed that the yield and purity were comparable or better than that reported by previous workers. With the obtained virus samples, polyclonal and monoclonal antisera were prepared. The results obtained from direct ELISA (enzyme linked immunosorbent assay) and indirect ELISA revealed that the prepared antisera detected mainly type III and type IV GLRaV. With the immunological method established, a disease survey was conducted between the years 1987 and 1994. A total of 204 grapevine cultivars or breeding lines were examined, and approximately 52% of them were found affected by GLRaV. Among the surveyed grapevines, about 30% were affected by Type III virus and another 31% by Type IV. A complexed infection of Types III and IV appeared to be common among Kyoho grapevines. The results also indicated the natural spread of GLRaV in tested field and suggested the need of a strict sanitation program for important breeding stock conservat |
本系統中英文摘要資訊取自各篇刊載內容。