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頁籤選單縮合
題 名 | 植物菌質體廣效型PCR引子之評估=Evaluation of Universal PCR Primers for the Detection of Phytoplasmas |
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作 者 | 林翠淳; 林長平; | 書刊名 | 植物病理學會刊 |
卷 期 | 7:1 1998.03[民87.03] |
頁 次 | 頁33-42 |
分類號 | 433.4 |
關鍵詞 | 植物菌質體; 聚合酶連鎖反應; 廣效型引子對; Phytoplasma; Polymerase chain reaction; Universal PCR primer; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究乃利用植物病原菌質體之核糖體 RNA 基因序列,設計出多組之聚合�t連鎖 反應引子序列,並加以評估此類廣效型 (universal) 聚合�t連鎖反應引子 (primer pairs) 偵測十一種植物病原菌質體病害與本省所發生之疑似梨衰弱病病株中植物菌質體之效果。所 評估之 f �� /r �窗BR �� F �� /R �� R �砥BR �� F �� / R �� rP �窗BR �� F �� /L �� 、R �� F �� /rH �陛BfD �� /L �紫奶輔梩s效型聚合�t連鎖反應引子對, 各有不同廣效程 度, 而聚合�t連鎖反應產物大小分別為 0.63 kb、1.25 kb、1.35 kb、1.6 kb、1.7 kb 及 1.7 kb。另以十三重細菌菌株全 DNA 為模板 DNA 進行 PCR 反應,結果引子對 f �� /r �� 及 R �� F �� /R �� R �祠蒬怢峇妤M一性。在疑似梨衰弱病病株植物菌質體之檢測時,此 六對引子對均曾在一些梨樹病組織採樣中增幅出與以植物菌質體 DAN 為模板時之 PCR 產物 大小相同之 DNA 片段, 但以國外報告之梨樹衰弱病菌質體專一性引子對 fPD/rPDS 進行測 試時,卻無任何產物被增幅出。 在以 R �� F �� /R �� R �窄i行偵測敏感度試驗時,對疑 似梨衰弱病之病株全 DNA, 其可偵測到 100 ng,對絲瓜簇葉病、甘藷簇葉病大陸品系、甘 藷簇葉病、翠菊黃化病西岸品系、翠菊黃化病紐澤西品系、花生簇葉病及榆樹黃化病之罹病 日日春全 DNA,依序分別可偵測到 100ng、100ng、3.1ng、50 ng、6.25 ng 及 12.5 ng; 而在 f �� / r �答滌輕�敏感度方面,對次疑似梨衰弱病之罹病梨樹全 DNA 可測到 6.25 ng,對絲瓜簇葉病、甘藷族葉病大陸品系、甘藷簇葉病、翠菊黃化病西岸品系、花生簇葉病 及榆樹黃化病之罹病日日春全 DNA 則分別可測到 4 ng、 1.5 ng、 12.5 ng、 25 ng、 0.16 ng、 6.25ng 及 0.8ng,而對甘蔗白葉病罹病甘蔗與水稻黃萎病罹病水稻之全 DNA 之 偵測敏感度則分別為 0.8 ng 及 32 pg。 |
英文摘要 | Six universal PCR primer pairs, f �� /r �窗BR �� F �� /R �� R �砥BR �� F �� / R �� rP �窗B R �� F �� /L ��, R �� F �� /rH �軒nd fD �� /L �� for the detection of phytoplasmas were evaluated in this study. All the primer pairs tested can amplify the phytoplasma-specific PCR products of 0.63 kb, 1.25 kb, 1.35 kb, 1.6 kb, 1.7 kb, and 1.7 kb in length from phytoplasma infected plants, respectively. Besides, total DNA of 13 bacteria species were also used to evaluated the specificity of these six universial primer pairs applied in the PCR amplification. Among these, primer pairs f �� /r �� and R �� F �� /R �� R �� have the best specificity. PCR products of the size similar to that of phytoplasmaspecific fragment had ever been amplified with all of the six universal PCR primer pairs using DNA templates prepared from pear-decline-like pear tree in Taiwan. On the other hand, the PCR primers specific for pear decline phytolasma, fPD/rPDS, had never amplified any fragment from diseased pear tissue collected. The DNA templates prepared from diseased pear trees that gave PCR products were collected in October and December in 1994 and March, Auguest and October in 1995. The sensitivity for primer pair R �� F �� /R �� R �� to amplify the specific fragment from the total DNA of the pear-decline-like plants is 100 ng, and that for primer pair f �� /r �� is 6.25 ng. |
本系統中英文摘要資訊取自各篇刊載內容。