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Development and Application of Quarantine Technique for Seed-Borne Pathogens on the Imported and Exported Vegetable Seeds
隨著全球各地進出口種子或植物品種的增加及其由於地理分佈或生態特性的異動,容易造成輸入地沒有發生過之新病害出現.種子傳播性病害在田間引起的病害發展迅速,但其病徵在幼苗時卻不易被發現.傳統偵測種傳性病害的方法包括: (1) 直接觀察 (2) 浸潤種子鏡檢 (3) 沖洗法檢查 (4) 播種後檢查 (5) 幼苗期檢查.而最近十年的診斷技術進步很多,包括了:以DNA為主軸之診斷技術(DNA-based methods)、酵素連結免疫抗體法(ELISA)、脈衝電泳分析(PFGE)、以基因體核酸為主之基因庫或隨機引子核酸增幅多型性分析(RAPD)等技術.由於我國進出口種子的增加,本研究擬由國外進口或田間與國內各種苗公司所販售茄科、葫蘆科以及十字花科蔬菜種子檢測不同種媒真菌菌株,進行茄科、葫蘆科以及十字花科蔬菜重要種媒真菌專一性分子標記,並發展以聚合酵素連鎖反應為主的種媒病原檢測方法,配合種子濕濾紙檢查等方法,以供檢防疫人員參考使用.而十字花科種傳之XCC亦為重要檢疫病菌,十字花科蔬菜黑腐病菌(Xanthomonas campestris pv. campestris)簡稱XCC,在田間可危害多種十字花科作物.十字花科蔬菜黑腐病菌為種子傳播性病菌,污染或感染本病原菌的種子為病害最主要的初次感染源,為世界各國檢防疫單位所重視之重要植物病原細菌.我國為十字花科種子之進出口國,十字花科蔬菜種子出口時,進口國常要求業者加註種子未有XCC之感染,為協助業者生產十字花科蔬菜健康種子,本計畫擬發展準確、快速和靈敏之XCC核酸偵測技術,以供檢防疫工作快速檢測之用.
Imported and exported seeds and plant varieties from one geographical region or ecological to another is increasing, and this practice may introduce new diseases where they had not been encountered previously. Seed-borne inoculum may give rise to progressive disease development in the crop, and symptoms of disease may not be apparent immediately in the germination seed. Traditional methods in the recognition and detection of seed-borne organisms include: (i) direct observation; (ii) microscopic examination of imbibed seeds; (iii) examination of cultured removed by washing; (iv) examination of seed after incubation; (v) examination of growing plants. The last decade has been great advances in diagnostic technology including: DNA-based methods, ELISA (enzyme-linked immunosorbent assay), pulse-field gel electrophoresis (PFGE), PCR amplification of various genome and random amplified polymorphic DNA (RAPD).Because the the increasing of imported and exported seeds in our country, in this study, we want to isolate and identify the seed-borne fungi from imported and exported vegetable seeds (Solanaceae, Cucurbitaceae and Cruciferae). Meanwhile, we will establish the molecular markers of different isolates for the further study and design of specific primers to set up the quarantine techniques for the seed-borne pathogens as well as PCR-ELISA and blotter method.In general, the seed-borne bacterial pathogens Xcc is an important quarantine pathogen, black rot crucifers caused by Xanthomonas campestris pv. campestris (X. c. pv. campestris) is a seedborne bacterial disease that affects cruciferous crops worldwide. The primary inoculum source of X.c. pv. campestris is usually from infected or contaminated seeds. X.c. pv. campestris has been considered as an important quarantine plant pathogen for several countries, however, a rapid and sensitive detection technique for this pathogen has not been developed yet. The objective of this project is to develop and establish rapid, sensitive and specific detection techniques for. X.c. pv. campestris.
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