應用核糖體DNA片段鑑別木棲性大型真菌

Application of Ribosomal DNA Sequence to Identify Wood-inhabiting Macrofungi

游婉婷(Wan-Ting You) ； 林亞立(Ya-Lih Lin)

《宜蘭大學生物資源學刊》 8卷1期 (2012/07) Pp. 45-58

https://doi.org/10.6175/job.2012.81.15

摘要

本研究以採集並經形態鑑定所得之木棲性大型真菌子實體為材料，萃取基因組DNA，進行PCR反應，增幅rDNA序列部分，包含ITS、18S rDNA、28S rDNA（D1～D3 domain）之片段，並利用NCBI網站進行序列比對。以43支菌株之rDNA序列比對與形態、鑑定結果相比，序列比對至種名者28株，至屬名者12株，形態鑑定至種名及屬名者分別為22及6株。由形態鑑定至屬以上分類階層的15支菌株，經由序列比對得知其中7株的種名及5株的屬名。同時由形態鑑定及序列比對分別得知種名者有17支菌株，其中16株種名相同。由此可知，rDNA序列部分只段可應用於菌種鑑定分類，雖然鑑定結果不一定與子實體形態鑑定結果相同，但確實可有效輔助形態鑑定上的不足。以增幅之rDNA序列部分為標記（marker），子囊菌群為外群（outgroup），採用PAUP* version 4.0 beta 10軟體進行序列演算及建構樹型圖。分析結果顯示，本實驗所使用的菌株大致分為多孔菌群（Polyporales group）及傘菌群（Agaricales group）兩類群；外群部分，子囊菌群（Ascomycota group）皆形成一穩定共群。三段序列所建構的樹圖中，以栓菌群（Trametes group）分群最為穩定，且支持Trametes屬及Pycnoporus屬歸為一群。

關鍵字

宜蘭大學實驗林場 ； 木棲性大型真菌

並列摘要

The genome extracted from the colonies obtained from collected and morphologically identified macrofungal fruiting bodies were amplified at the rDNA segments of ITS, 18S rDNA and 28S rDNA (D1~D3 domain) and the nucleotide sequences of these segments were matched with the DNA sequence data base of NCBI to reveal the names of these macrofungi. Among the 43 kinds of macrofungi specimens used for comparison in this study, 28 species and 12 genus names were revealed by rDNA sequences matching, and 22 species and 6 genus names were identified by morphological characteristics recognition. Seven species names and 5 genus names were suggested by rDNA sequences matching of the 15 specimens identified morphologically to classification hierarchy of family and above. Only 1 of the 17 kinds of macrofungi identified to their species individually by morphological characteristics and rDNA sequences matching carries different species names. It appears that nucleotide sequences of rDNA segments used in this study are relevant to fungal identification, though the results might not be consistent with that from morphological identification, it still provides great help on fungi identification.Applying ITS region, partial region of 18S rDNA, and 28S rDNA (D1~D3 domain) sequences as markers, using ascomycota as outgroup, with PAUP * version 4.0 beta 10 software, tree diagrams were calculated and constructed. The specimens used in this study were divided into polyporales and agaricus groups; outgroup for ascomycota group indicates a stable group. The tree diagrams constructed with the three rDNA segments used for sequence matching denote Trametes group the most stable cluster, and genera Trametes and Pycnoporus could be classified into a group.

並列關鍵字

Experimental Forest of National Ilan University ； wood-inhabiting macrofungi ； ITS region ； 18S rDNA ； 28S rDNA ； D1~D3 domain