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題 名 | 萵苣嵌紋病毒之多元抗血清製作與應用=Preparation and Application of Polyclonal Antibody on Lettuce mosaic virus |
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作 者 | 曹麗玉; 林照能; | 書刊名 | 臺灣農業研究 |
卷 期 | 64:2 2015.06[民104.06] |
頁 次 | 頁159-166 |
分類號 | 435.252 |
關鍵詞 | 萵苣; 萵苣嵌紋病毒; 血清; 檢測; Lactuca sativa L.; Lettuce mosaic virus; Antiserum; Detection; |
語 文 | 中文(Chinese) |
中文摘要 | 引起嵌紋、皺葉病徵的萵苣樣品接種至紅藜、白藜等病毒指示植物,經過生物分析、寄主範圍測試、病斑特性觀察及血清學等證據顯示,病因為萵苣嵌紋病毒(Lettuce mosaic virus; LMV)。以人工方式接種單斑系病毒於7科27種植物,其中萵苣、菠菜、豌豆及圓葉菸草均產生系統性嵌紋病徵,奎藜表現系統性黃化斑點病徵,紅藜及千日紅則出現局部壞疽斑點病徵,其餘供試植株均未出現病徵。利用聚丙烯醯胺膠體電泳分離出病毒鞘蛋白(35 KDa)作為免疫源,注射白兔製作LMV抗血清。以間接酵素連結免疫抗體吸附分析法(indirect -ELISA)進行血清最適稀釋濃度評估,抗血清稀釋濃度可達5,000倍,以此濃度偵測罹病植株粗汁液,病葉汁液稀釋倍數靈敏度可達1/640倍。檢測市售10批商業品種萵苣種子及7批種苗,LMV檢出率分別介於0-28%及0-13.3%。調查雲林縣麥寮鄉的結球萵苣產區,隨機檢測疑似罹病萵苣113株,LMV病毒檢出率達37%。 |
英文摘要 | Lettuce mosaic virus was isolated and identified from infected lettuce plants (Lactuca sativa var. crisp. L.) by using host reactions, electron microscopy, biological and serological assays in Taiwan. Upon artificial inoculation to 27 plant species belonging to 7 families, the virus infected lettuces (Lactuca sativa var. asparagina, L. sativa var. capitata, L. sativa var. crispa, and L. scariola var. sativa), spinach (Spinacea oleracea), pea (Pisum sativum) and tobacco (Nicotiana benthamiana) showed systemically mosaic symptoms. It developed systemically yellow lesions on Chenopodium quinoa, but only local necrotic lesions displayed on C. amaranticolor and G. glodosa. No other tested plants with symptoms were observed. The virus coat protein was purified by SDS-PAGE, from which the antiserum against LMV was produced. The titer of the antiserum against homogenous antigen in the crude sap was 5,000, tested by the indirect ELISA. The dilution end point of the crude sap detected by the antiserum was 1/640. Ten commercial lettuce seed lots were tested with the antiserum and the incidence of LMV was determined in range of 0-28%. Seven lots of lettuce seedlings collected from markets. Among them, 0-13.3% of the seedlings were positive to the LMV when detected by the antiserum. By testing 113 lettuce plants randomly collected from Mailiao Township of Yunlin County, the incidence percentage of LMV was 37%. |
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