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頁籤選單縮合
題名 | 以Nested PCR偵測臺灣杉根部菌根菌種=Detection of Taiwania Mycorrhizal Fungi with Nested PCR |
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作者姓名(中文) | 顏江河; 吳羽婷; 王也珍; | 書刊名 | 臺灣林業科學 |
卷期 | 21:4 民95.12 |
頁次 | 頁409-420 |
分類號 | 436.193 |
關鍵詞 | 臺灣杉; Acaulospora mellea; Scutellospora calospora; Nested PCR; 18S rRNA; Taiwaniana cryptomerioides; |
語文 | 中文(Chinese) |
中文摘要 | 採自六龜鳳岡山臺灣杉 (Taiwania cryptomerioides) 造林地根域土壤,分離出Acaulospora mellea和Scutellospora calospora兩種叢枝菌根菌 (AMF)。應用nested PCR及菌根菌18S rRNA專一性引子(分別為GeoA1/ART4; GeoA2/Geo11),自上述兩種孢子萃取DNA,可增幅反應A. mellea和S. calospora的18S rRNA;另外也增幅得到臺灣杉的18S rRNA片段。將臺灣杉與此兩種叢枝菌根菌18S rRNA排序後,依據不保守區域設計個別菌種的專一性引子 (ASG1062-18S與Acau2/Scute2),其產物大小約649 bp。將溫室中臺灣杉幼苗接種單一菌種後,取根段萃取DNA,經專一性引子進行nested PCR及選殖,並應用限制性酵素片段多型性 (HinfI和HaeIII) 篩選至適當的載體進行定序。序列比對結果與原始接種的菌種一致,確定自菌根偵測菌根菌種的方法是可行的。進一步應用於偵測野外臺灣杉,證實野外共生菌種有S. calospora。臺灣杉幼苗接種成功的A. mellea菌種,在野外的臺灣杉偵測不到,但是與林下的其他植物共生;推測在本試驗地A. mellea與臺灣杉之間屬於較弱的共生關係。 |
英文摘要 | Acaulospora mellea and Scutellospora calospora are 2 dominant arbuscular mycorrhizal fungal (AMF) species isolated from rhizosphere soil of Taiwania (Taiwania cryptomerioides) stands near Feng-Gang Mountain nursery at the Liuguei. DNA was extracted from AMF spores and Taiwania roots. Nearly complete 18S rRNA genes of A. mellea and S. calospora were amplified by nested PCR with two 18S rRNA-specific primer pairs (GeoA1/ART4 and GeoA2/Geo11). The 18S rRNA gene of Taiwania was also amplified. Sequences of the 18S rRNA gene of Taiwania and both AMFs were aligned. Based on the conserved sequences of 18S rRNA genes, fungal-specific primers were designed. About 649 bp of a DNA fragment was PCR-amplified from each fungal species. DNA products were cloned and sequenced. The resulting sequences matched that of the inoculated AMF. The methodology was then applied to monitor AMF species within Taiwania roots from the field, and the association of the symbiotic fungus, S. calospora, and Taiwania was confirmed. However, A. mellea was only detected in the roots of other plants instead of Taiwania roots. This phenomenon suggests that the symbiotic relation between A. mellea and Taiwania is possibly weak in the field. |
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