長度多型性（restriction fragment length polymorphism）及 DNA 定
序 法，探討南方靈芝 Ganoderma australe (Fr.) Pat. 種內兩個雜
交不孕性群(intersterility group）的遺傳變異，並以同亞屬的樹舌靈
芝 G. applanatum (Pers. ex Gray) 純培養菌株 CBS 187.31 及不同亞
屬的新日本靈芝 G. neo-japonicum Imaz. 純培養菌株 CCRC 36049 做為
比較研究。以互補於酵母菌 17S rRNA 基因及 25S rRNA 基因的引子與熱
穩定性的 DNA 聚合酵素，將 2.1 Kb 的 17S rRNA 基因片段及 1.85 Kb
的 25S rRNA 基因 5 端片段選殖出來。再以限制酵 素 HhaI 切割 2.1
Kb 片段或 AvaII、HinfI 切割 1.85 Kb 片段後,多聚丙烯醯膠體電泳分
析切割片段條紋型，可將南方靈芝的兩群及同亞屬的樹舌靈芝、不同亞屬
的新日本靈芝明顯區別出來。以這些限制酵素做不完全切割（partial
digest）後，利用南方墨點法(Southern blot）及 Dig-ddUTP 標記的引
子分別定出 2.1 Kb 及 1.85 Kb 片段的限制酵素圖譜。最後將 rDNA 重
覆單位上部分變異集中的內部轉錄間隔片段（internal transcribed
spacer）選殖出來，置入 pGEM-T 載體中，以進行 DNA 序列分析。各菌
株序列經排序（alignment）後，再以 PAUP、MEGA 等軟體，分析其 ITS
序列的 GC 含量及核甘酸置換（nucleotide substitution）等，並繪出
演化樹狀圖。本研究結果顯示南方靈芝種內之兩個「雜交不孕性群」﹞w
呈現顯著的遺傳分化。

australe (Fr. ) Pat. was investigated by polymerase chain
reaction,restriction fragment length polymorphism, and DNA
sequencing.Two cultures of G. applanatum (Pers. ex Grau) Pat.
CBS 187.31 and G. neo-japonicum Imaz. CCRC 36049 were selected
for comparative studies. By using conserved primers
complementary to rRNA genes of Saccharomyces cerevisiae, a 2.1
Kb fragment containing 17S rRNA gene and a 1.85 Kb fragment
containing 5` half of 25S rRNA gene were efficiently amplified.
Electrophoresis of HhaI, AvaII or HinfI digested PCR products
produced restriction phenotypes. The two intersterility groups
of G. arstrale can be differentiated by AvaII and HinfI
restriction phenotypes. Detail mapping of restriction
sites within rDNA locus by partial digestion and Southern
hybridization reveals conserved and variable restriction sites
for each species or group. The regions containing internal
transcribed spacers were further amplified and subcloned into
pGEM-T for DNA sequencing. Phylogenetic structures were
constructed from analysis of ITS regions by PAUP and MEGA
programs. Our data suggest the two groups of G. australe are
genetically differentiated.
australe (Fr. ) Pat. was investigated by polymerase chain