行政院農業委員會台南區農業改良場  研究彙報第46號
 

十字花科蔬菜黑腐病菌血清偵測技術

吳雅芳、陳紹崇、鄭安秀

摘  要

吳雅芳、陳紹崇、鄭安秀。 2005。十字花科蔬菜黑腐病病原細菌血清偵測技術之研發。臺南區農業改良場研究彙報 46:10-19。

十字花科蔬菜黑腐病病原細菌 ( Xanthomonas campestris pv. campestris ) 供試菌株分別分離自台灣各主要栽培區,並以甘藍分離菌株XCC214細菌全細胞製備抗血清。利用SDS免疫擴散反應進行抗血清力價測定,結果顯示 未稀釋和稀釋1/2倍後的抗血清均能與十字花科蔬菜黑腐病菌供試菌株形成一條明顯弧形反應帶,但均不與檬果黑斑病等15種病原細菌菌株形成任何反應帶。在間接酵素連結抗體檢測法(ELISA)試驗中,亦可偵測到所有黑腐病菌的供試菌株,而檬果黑斑病等15種病原細菌菌株均無反應。 在間接ELISA檢測法中,以IgG濃度0.16μg/ml為最適宜,可偵測到10 4 -10 5 c fu/ml濃度之黑腐病病原細菌。由西方墨點反應分析結果證實, X. campestris pv. campestris 菌株XCC214細菌全細胞製備之抗血清是由一種抗原蛋白免疫反應而製成,分子量大小約63.6kDa。

關鍵詞:甘藍、黑腐病病原細菌、酵素連結抗體免疫檢測法

接受日期:2005年9月2日


Development of a Serum Methodology For of the Detection Bacteria Causing Black Rot in Crucifers

Wu, Y. F., S. C. Chen and A. S. Cheng

Summary

   Cabbage black rot disease was infected by Xanthomonas campestris pv. c ampestris. To collect the cabbage black rot bacteria strains from the different area in Taiwan and detect the pathogenicity in green house. The high pathogenicity strain XCC214 was used for producing antiserum. The titer of antiserum was 1/2 dilution of crude antiserum in SDS-double diffusion test with one visible reaction band. The optimum concentrations of immunoglobulin G ( IgG ) was 0.16 μg/ml in indirect enzyme-linked immunosorbent assay ( indirict ELISA ) . The concentrations of sensitivity assay were 10 5 and 10 4 cfu/ml of strain XCC 214 in indirect ELISA. All isolates of X. campestris pv. c ampestris from different brassica crops were also reacted to antiserum with one precipitation band in SDS-double diffusion test and indirect ELISA. However the antiserum did not react with other kinds of bacteria including X. campestris pv. mangiferaeindicae, X. axonopodis pv. vesicatoria , X. oryzae pv. oryzae , X. axonopodis pv. diffenbachiae , Pseudomonas syringae pv averrhoi , Erwinia cypripedii , E. carotovora subsp. carotovora , E. chrysanthemi , E. sinocalami , Acidovorax avenae subsp. citrulli , A. avenae subsp. cattleyae , Ralstonia solanacearum , Burkholderia glumae , B. caryophylli in SDS- double diffusion test and indirect ELISA . The result of SDS-polyacrylamide gel electrophoresis and western bolting showed that one reacting band was observed with protein molecular weight about 63.6 kDa. The result was also consistent with that of in SDS-double diffusion test.

Keywords: Cabbage, Black rot disease, ELISA


 

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