本研究以台灣所分離之殺手酵母菌Barnettozyma californica及Cyberlindnera saturnus complex為對象，其中Barnettozyma californica利用三段基因序列(LSU D1/D2 domain；Internal transcribed spacer of rDNA, ITS；Cytochrome C oxidase subunit II, CoxII)組合，而Cyberlindnera saturnus complex則以細胞骨架(actin as cytoskeleton)基因做為分子標誌，進行種內之親緣性關係及遺傳結構分析。
B. californica及Cyber. saturnus complex皆分離自台灣山區土壤、植物葉面及菇類子實體，所分離而來的菌株，經由大單元核糖體DNA(LSU ribosomal DNA)D1/D2區域序列比對分析，分別鑑定出B. californica及其他未描述菌種sp2, sp3, sp13, sp27及親緣關係性相關之菌株-Candida sp50.及Candida sp53.共45株；另有56株鑑定為Cyber. saturnus complex之5種分類項及相關無性世代Candida菌種，並進行親緣關係探討籍資料分析與殺手毒性分析。
將分子變異做階層體制分析(hierarchical analysis)後，發現兩菌種具有明顯的族群結構。B. californica可劃分為8群、39個個體、22個基因單型、369個基因多型位、基因單型歧異度(haplotype diversity, h)為0.907±0.001%，而核苷酸歧異度(nucleotide diversity, π)則為2.73%。另Cyber. saturnus complex可劃分為4群、48個個體、31個基因單型、166個基因多型位、基因單型歧異度(h)為0.916±0.001%、核苷酸歧異度(π)則為2.91%，兩菌屬均具有高歧異度則表示該兩菌屬，屬內種間具有有效族群量。另外根據遺傳分化值(Fst)及基因交流值(Nm)也顯示遺傳分化明顯的結果(B. californica, Fst = 0.3~0.7, Nm = 0~1；Cyber. saturnus complex, Fst = 0.8, Nm = 0~0.07)，兩菌在種內顯示高度遺傳分化及低的基因交流表示同種之間其種內存在一定的差別，意指種內各群可以明顯區分。進一步以中性試驗檢測探討導致兩菌種種內分群之因素，利用Tajima`s D和Fu and Lis D* and F*值評估，結果顯示B. californica 部分族群和Cyber. saturnus complex以Cyberlindnera saturnus及Cyberlindnera mrakii偏離中性理論及根據高基因單型歧異度和低核苷酸歧異度，表示這兩菌種有族群擴張現象，而且可能曾經經歷過開拓者效應或者是瓶頸效應且是在近期內族群建立後並快速擴散。本實驗結果，根據高基因單型歧異度、低核苷酸歧異度及低基因交流值與高程度的遺傳分化，顯示出Barnettozyma californica與Cyberlindnera saturnus complex具有達到變種及種化的程度。

The aim of this study is to identify the yeasts of Barnettozyma californica and Cyberlindnera saturnus complex and to analyze intraspecific phylogenetic relationship and population structure of the kill yeasts, isolated from Taiwan. The Phylogenetic trees of B. californic were constructed based on multiple sequences of large subunit (LSU) D1/D2 domain, internal transcribed spacer (ITS) of rDNA ITS and cytochrome C oxidase subunit II (CoxII), and actin gene for Cyber. saturnus complex. All the strains of B. californica and Cyber. saturnus complex isolated from soil, leaves, and fruiting body of murshrooms in the mountain area of Taiwan, were identified based on sequence of the LSU D1/D2 domain of (LSU) rRNA gene, morphology and physiology. Among the strains isolated, 38 strains of B. californica, 8 of Barnettozyma sp2, sp3, sp13, sp27 and sp50 were authenti cated as Barnettozyma species, and 56strains were identified as one of five species in Cyberlindnera saturnus complex or related Candida species. A hierarchical analysis of molecular variance revealed that differentiation of genetic structure are significant among populations of the species.
B. californica is classidfied into 8 groups, 39 unity, 22 haplotypes and 369 polymorphism, and haplotype diversity(h) and nucleotide diversity(π) are (h)= 0.907±0.001%, (π)= 2.73%. Strains of C. saturnus complex are classidfied into 4 groups, 48 unity, 31 haplotypes and 166 polymorphism, and haplotype diversity(h) and nucleotide diversity(π) are (h)= 0.916±0.001%, (π)= 2.91%. According to the data, they are large effective population sizes, respectively. Moreover, the two species reveal significant in gene differentiation (Fst) and gene flow (Nm). (B. californica：Fst = 0.3~1, Nm = 0~1；Cyber. saturnus complex：Fst = 0.8, Nm = 0~0.07), because the two species demonstrate low gene flow and high gene differentiation, indicating that they has intraspecific differentiation. The differentiation among genetic population were furtherly evaluated by Tajima`s D and Fu and Lis D* and F* values with neutral tests. The results show that the group3 of B. californica, Cyber. saturnus and Cyber. mrakii showed significant deviation neutral model, because of the high haplotype diversity and the low nucleotide diversity.The results show the significant deviation neutral tests proposed that the two species happened resulting rapid population expansion from founder events or bottleneck events.
According to high haplotype diversity(h), low nucleotide diversity(π), high gene differentiation (Fst) and low gene flow (Nm). The results show that B. californica and Cyber. saturnus complex with achieve variety and specisity.

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