Quantification of Mycorrhiza from Taiwania Using Real-Time PCR

自六龜鳳岡山苗圃附近的台灣杉(Taiwaniana cryptomerioides)造林地根域土壤，以濕篩及糖液離心法，分離出Scutellospora calospora和Glomus sp.兩種較具優勢的叢枝菌根菌孢子(AMF)。利用realtime PCR配合菌種18S rRNA專一性引子（產物約649 bp）螢光劑SYBR(上标 ®) Green I，可個別偵測並定量台灣杉共生的菌根菌，分別為S. calospora與Glomus sp.。定量所依據的標準曲線為個別帶有菌種部分SSU rRNA不同拷貝數之質體DNA的對數值(log)與定量PCR threshold循環數所繪成的；相關係數分別為r^2=0.998及r^2=0.999。已可順利定量出溫室台灣杉苗木菌根的族群量，顯示該方法的可行性。未來在菌根研究的領域裡，real-time PCR將是一個有效率且精確的定量菌種技術。

關鍵字

台灣杉；螢光劑SYBR(上标 ®) green I ； real-time PCR ； Glomus sp. ； Scutellospora calospora

並列摘要

Scutellospora calospora and Glomus sp. were 2 dominant arbuscular mycorrhizal fungi (AMF) species extracted from rhizosphere soil of Taiwania (Taiwania cryptomerioides) tree stands near Feng-Gang Mountain nursery at Liukuei in Taiwan by wet sieving and sucrose centrifugation. To efficiently detect and quantify the population of the AMF, S. calospora and Glomus sp., associated with the roots of Taiwania, a real-time polymerase chain reaction (PCR) technique was used to amplify the 18S rRNA gene (ca. 649 bp) with specific primers, and it was detected using fluorescent SYBR(superscript ®) green I dye. Standard curves showed a linear relation (r^2=0.998 and r^2=0.999 each) between log values of the starting copy numbers of target sequences and real-time PCR threshold cycles. The real-time PCR quantification assay can also be used to monitor individual AMF communities in the greenhouse. The results demonstrate that the real-time PCR technique is a powerful universal tool for quantifying individual AMF species in modern mycorrhizal research.

並列關鍵字

Taiwania cryptomerioides ； fluorescent SYBR(superscript ®) green I dye ； real-time PCR ； Glomus sp. ； Scutellospora calospora

國際替代計量

Quantification of Mycorrhiza from Taiwania Using Real-Time PCR

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