Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/28714
標題: 逢機增殖多型性DNA標誌在蕙蘭鑑別之研究
Identification of Cymbidium spp. by randomly amplified polymorphic DNA markers
作者: 邱金春
Chiou, Chin-Chun
關鍵字: Cymbidium
蕙蘭
identification
linkage cluster analysis
鑑別
遺傳連鎖群分析
出版社: 園藝學系
摘要: 壹、中文摘要 本研究以台灣所栽培的蕙蘭屬種原為材料,調查其外觀形態,並以RAPD標 誌分析其種原,目的為(1) 建立區別蕙蘭屬種原的分子標誌,(2)建立種 原間遺傳連鎖群之關係;以期作為蕙蘭品種鑑定及育種之參考。調查十種 蕙蘭屬不同種植物(四季蘭、 寒蘭、觀音素心蘭、鳳蘭、報歲蘭、東亞蘭 、金稜邊、春蘭、菅草蘭與綠花竹柏蘭)、二種素心蘭(觀音素心蘭與鐵骨 素心蘭)、十一種報歲蘭品系(報歲蘭)、`瑞寶'、 `陽明錦'、`雪白爪'、 `金山'、 `聖紀光'、`旭晃'、`石門'、`桃姬、`金華山'與 `十八學士') 及二個春蘭品系(春蘭與細葉春蘭)之外表園藝性狀及生長習性。結果顯 示十種蕙蘭屬不同種植物可依葉緣構造、葉片形態、葉片大小及生長習性 等順序將其完全分類;十五種蕙蘭品系,依葉藝或花藝的變化及葉緣形態 構造等順序可將其 部份分類。以 UBC primer set 共計 300 組逢機引子 進行蕙蘭種原DNA 片段的增殖 ,經由 PCR 產物的分析結果,篩選出 20 組引子能有效地區別十種蕙蘭屬不同種間 植株,其中有 46 個多型性 DNA 片段為單一種原所特有,33 個多型性 DNA 片段為 兩種原所共有, 可以作為十種蕙蘭屬不同種間植株辨識種原的標誌。平均每一引子 產生 12.5 條 DNA 條帶,產生3.9個多型性 DNA 標誌。經由群體分析所得的遺 傳連 鎖關係顯示,十種蕙蘭屬不同種植物大致可以分為四群,地生蘭類 的蕙蘭種原包括 四季蘭、觀音素心蘭、寒蘭、報歲蘭、菅草蘭為第一群 ,第二群為綠花竹柏蘭與春 蘭,第三群為半著生性的金稜邊與東亞蘭, 著生性的鳳蘭則與其他九種種原的相似 性距離最遠。同時自300組逢機引 子篩選出 11 組引子可有效地區別十五種蕙蘭屬 不同品種,其中 11 個 多型性 DNA 片段為單一種原所特有,10 個多型性DNA 片段 為二至四個 種原所共有,可以作為十五種蕙蘭屬不同品種植株辨識品系的標誌。以 報歲蘭而言,平均每一引子產生 11 條 DNA 條帶,產生 1.9 個多型性 DNA 標誌。 經由群體分析所得的遺傳連鎖關係顯示,供試的十一種報歲 蘭品系大致可以分為四 群,第一群包括報歲蘭與`桃姬',第二群包括有` 瑞寶'、`陽明錦'、`金山'、`雪 白爪'、`旭晃'與`聖紀光'等藝蘭,第三 群亦為觀葉的`石門',第四群包括有觀花 的`金華山'與`十八學士'。
Morphological traits and randomly amplified polymorphic DNA (RAPD) markerswere used to study the identification and discrimination of Cymbidium species.The purposes of this study were to establish the molecular markers for discrimination of the Cymbidium spp. and cultivars and to explore the relationships of genetic linkage among the Cymbidium spp. and cultivars. The data of phenotype characters and growth habit showed that the tenCymbidium spp. can be discriminated followed the order of serrulated marginof leaf, leaf shape, leaf size, and growth habit. However, fifteen cultivars ofCymbidium can only be parly discriminated with leaf stripes and leaf shape. Twenty DNA primers, which generated clear and discriminative polymorphicDNA fragments, were selected from 300 random primers to discriminate ten Cymbidium species. Of 79 polymorphic DNA fragments, 46 fragment were species-specific and 33 fragments were presented between two Cymbidium spp. Therewere averaged 12.5 amplified DNA fragments a-nd 3.9 DNA polymorphic markersgenerated by each primer. The results of linkagof cluster analysis revealedthat the ten Cymbidium spp. can be classified into four groups, the first group includes terrestrial plants of C. rubrigemmum, C. ensifolium var. misericors, C. karan, C. sinense and C.tortisepalum, the second group includes C. lancifolium form aspidistrifolium and C. formosanum, the third group includes epiphytic plants of C. Sensation and C. pumilumm, and the fourth group includes C. dayanum var. austro-japonicum with the lowest similar-ity with other Cymbidium spp. Eleven DNA primers were selected from 300 random primers to discriminate fifteen cultivars of Cymbidium. Of 21 polymorphic DNA fragments which discriminated eleven cultivars of Cymbidiu sinense, 11 fragments were cultivar-specific and 10 were shared among two to four cultivars. There were averaged 11 amplified DNA fragments and 1.9 DNA polymorphic markers gener-ated by each primer. The results of linkage cluster analysis revealed that the eleven cultivars of Cymbidium sinense can be classified into four groups, the first group includes C. sinense and `taur-ji', the second group includes `rueih-baau', `yarng-ming-jiin', `jin- shan', `shyueh-bair-jaau' and`shyuh-huarng', thethird group includes ` shyr-mern', and the fourth groupincludes `jin-huah- shan' and `shih-ba-hsiieh-shih'.
URI: http://hdl.handle.net/11455/28714
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