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頁籤選單縮合
題名 | 火鶴花細菌性葉枯病病原菌PCR引子之研發與應用=Development and Application of PCR Primer for Xanthomonas Axonopodis pv. Dieffenbachiae, the Causal Agent of Anthurium Blight |
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作者 | 林貝珊; 林長平; | 書刊名 | 植物病理學會刊 |
卷期 | 11:2 2002.06[民91.06] |
頁次 | 頁97-106 |
分類號 | 433.4 |
關鍵詞 | 火鶴花; 火鶴花細菌性葉枯病; 聚合酵素連鎖反應; Anthurium; Anthurium blight; Polymerase chain reaction; |
語文 | 中文(Chinese) |
中文摘要 | 本實驗利用聚合酵素連鎖反應(polymerase chain reaction,簡稱PCR)技術偵測火鶴花細菌性葉枯病原菌(Xanthomonas axonopodis pv. dieffenbachiae,原名X. campestris pv. dieffenbachiae,簡稱XCD)。實驗中使用之引子對是依據葉枯病菌16S-23S rDNA 基因區間(spacer region)之核啟酸序列而設計。藉由16S-23S rRNA 基因的保守特性,在16S rDNA 序列之3 '端和23S rDNA 序列之5 '端設計廣效性引子對Bf1/Br1,以增幅出包含有基因區間之585 bp PCR產物,並選殖入質體pCR II完成核啟酸解序,XCD之16S-23S rDNA基因區間核啟酸序列顯示包含了tRNAAla和tRNAIle的基因序列。利用XCD及其他Xanthomonas spp.細菌之16S-23S rDNA基因區間核啟酸序列比對後,依據其序列差異設計出P C R引子對 Dif1/Dir1,其可從XCD DNA模版中專一地擴增出一大小為409b p之PCR產物,而不會由其他測試之 Xanthomonas spp . 細菌、其他種類細菌及火鶴花葉表細菌之DNA模版中擴增出任何產物。PCR引子對Dif1/Dir1的偵測極限為1.5 pg之純化DNA或3-4個XCD細胞。 |
英文摘要 | One pair of primers for the polymerase chain reaction-based assay were devel oped for the detection of the causal agent of anthurium blight, Xanthomonas axonopodis pv. dieffenbachiae (XCD). Universal primers targeting conserved sequences flanking the 3' end of the 16S and the 5'end of the 23S rRNA genes were synthesized and used to amplify the 16S-23S rDNA internal transcribed spacer of XCD. A 585 bpPCR product containing spacer region thus amplified was cloned in pCRII for nucleotide sequence determination. Sequences for tRNAAla and tRNAIle genes were revealed in the 16S-23S rDNA spacer region. Primer pair Difl/Dirl, synthesized according to the sequence of the spacer region, could effectively amplify a 409 bp DNA fragment from the DNA template prepared from XCD, but not from any other tested Xanthomonas spp., other bacterial species or epiphytic bacteria isolated from anthurium. A minimum of 1.5 pg DNA or 3-4 XCD cells was sufficient to amplify the specific XCD PCR fragments using the primer pair Difl/Dirl. Rapid detection of XCD from anthurium samples by PCR was also evaluated. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。