- 學位論文
以RAPD與ISSR分子標誌鑑別台灣區採集台灣白及之族群變異
Using RAPD and ISSR molecular markers analyze genetic variation of Bletilla formosana (Hayata) Schltr. population in Taiwan
摘要
台灣白及(Bletilla formosana (Hayata) Schltr.) 為台灣特有之蘭科植物，與常用之中藥白及(B. striata (Thunb.) Reichb. f.) 為同屬植物，兼具有藥用及觀賞之價值。本研究以RAPD (Random Amplified Polymorphic DNA)與ISSR(Inter Simple Sequence Repeat)分子標誌對白及屬不同種間以及採集自台灣不同地區之台灣白及個體樣品共25個樣品進行分析，之後並據此結果對其遺傳相似性進行評估。試驗之白及屬植物包括有分別採自於台灣不同地域生長之20處台灣白及(B. formosana (Hayata) Schltr. f.)族群、白及(B. striata)以及黃花白及(B. ochracea )。經由80個RAPD引子與100個ISSR引子之初步篩選，共選出18個具有高再現性的引子，其中由12個RAPD引子可獲得165個片段，多型性條帶占161個；由6個ISSR引子中可獲得90個片段，多型性條帶占83個。結果顯示所用之RAPD引子具有區分不同種白及之能力，但對於台灣白及同種內不同階級的蘭嶼白及則無法加以區別；ISSR標誌對白及屬不同種間及台灣白及種內之物種皆能區分。以所得之條帶及多型性條帶進行UPGMA (Unwighted Pair-Group Mean Arithmetical) 之遺傳相似性分析，依據樣品個體之遺傳分歧度結果來看，推測台灣白及的族群差異可能來自於地域間的區隔。本研究結果除可供白及屬植物品種之鑑別外，亦可作為輔助台灣白及育種之參考。
並列摘要
Bletilla formosana (Hayata) Schltr., with medicinal and ornamental value, is a unique Orchidaceae plant in Taiwan. It is in the same genus with Bletilla striata (Thunb.) Reichb. f., a commonly known Chinese medicine. Genetic makers of RAPD and ISSR were used to analyze different species in Bletilla genus and plants of B. formosana collected from different regions in Taiwan. Species of B. striata, B.ochracea and 20 plant samples of B. fomosana (Hayata) Schltr. collected form different regions of Taiwan were used in this study. A total 18 primers was selected by its high reproduction from 80 pairs of RAPD primers and 100 pairs of ISSR primers for the consecutive experimens. A total of 165 molecular markers was detected, of them, 161 markers were polymorphic by using 12 pairs of RAPD primers. A total of 90 molecular markers, of them, 83 markers were polymorphic by using the 6 pairs of ISSR primers. All RAPD msrkers were found unique to distinguish among Bletilla. genus species used in this study. However, only ISSR markers were able to reveal differences of class under B. formosana species. A unwighted pair-group mean arithmetical (UPGMA) analysis based on RAPD and ISSR bands from 25 tested samples for genetic similarity among detected samples was conducted The phylogenetic tree showed that genetic differences among ethnic groups of B. formosans (Hayata) Schltr. f. kotoensis (Hayata) T. P. Lin may related with the geographical segregation. The results from this study may provide not only unique markers for identification species among Bletilla genus but also provide useful genetic information for breeding new varieties of B. frommosana.