Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/28862
標題: 以園藝性狀及逢機增殖多型性DNA標誌鑑定百慕達草栽培種之研究
Identification of Cynodon dactylon (L.) Pers. by Morphological Traits and Randomly Amplified Polymorphic DNA (RAPD) Markers.
作者: 劉怡君
Liu, Yi-Chin
關鍵字: Bermudagrass
百慕達草
Morphological traits
RAPD
性狀特徵
逢機增殖多型性DNA標誌
出版社: 園藝學系
摘要: 本試驗以百慕達草(Cynodon dactylon (L.) Pers..)為材料,探討應用性 狀及性狀遺傳連鎖群分析,與逢機增殖多型性DNA標誌(RAPD marker)二 種方法,對百慕達草品種鑑別的應用。RAPD為初次使用於百慕達草的品種 鑑別上,所以此一試驗的目的在:(1)建立百慕達草完全的性狀資料,並 以性狀遺傳連鎖群分析建立性狀親緣資料;(2)與性狀對照,測試RAPD在 百慕達草應用上的可信度;(3)建立快速以及正確的百慕達草品種鑑別的 技術。百慕達草品種的性狀調查,參考禾草類的分類標準,根據外部的葉 耳、葉舌、葉領邊緣纖毛、葉面及葉背茸毛以及莖的顏色、株型大小、莖 葉質地和高度等性狀,建立16種百慕達草品種外觀性狀的詳細資料。利用 性狀外觀資料,加上長期測量的莖葉長度寬度值,進行性狀的連鎖群分析 ;可將所有百慕達草品種區分為四個彼此有差異的性狀群。逢機增殖多型 性DNA標誌的實驗,自100種的逢機引子(random primer)中篩選出4種可 以產生多形性DNA片段的引子,藉著四個引子,可以辨別N1、0035及0036 三個品種;而對所有的品種能以遺傳連鎖群分析,將其區分為普通型、中 度矮生型及矮生型三個不同族群,在矮生型的群中,所有的品種完全沒有 歧異存在。根據實驗的結果,在所蒐集到被稱為Tifway(419)的品種上, 發現可能有名稱誤用的現象。對照性狀與RAPD的結果,RAPD在百慕達草上 所表現的多型性,與性狀的基本資料對照後,證明RAPD標誌與性狀間,確 實有相當大程度的相關性;除了Tifway品種外,二者都可區分百慕達草為 普通型、半矮生型及矮生型。但在性狀歸群上,與RAPD標誌的歸群結果因 Tifway種,有小部分不符合而無法解釋之處。這原因應在所測試的引子數 量不足,因此增加測試隨機引子的數目實有其必要。RAPD應用於百慕達草 的品種鑑別上,應是一可靠且簡便的方法,在建立充足的性狀與RAPD的相 關資訊以後,RAPD篩選的適當引子,將能夠預測一個草種未來形成的草皮 性狀。
Summary The clusis analysis with both the morphological traits and the RAPD marker data were used to identify 16 bermudagrass cultivars. It was the first time RAPD method been used on bermudagrass for cultivar identification. The goals for this experiment are: 1)To build up a database for bermudagrass morphological characters, and use the data for cluster analysis to construct relationship data between cultivars. 2)Testing the reliability of RAPD test compare to the morphological traits. 3)Develop an accurate and rapid techniquefor cultivar identification. The morphological traits were measured based on the categories for Gramineaegrasses. Both qualitative and quantitative characters including: ligual, hairs on the edges of collar, pubescence on leaf blade and on leaf back, stem color,plant height, texture of the leaf and the canopy of the grass were used to describe the characteristics of 16 bermudagrass collected. We quantitized those qualitative characters then added up with the quantitative charactersfor similarity cluster analysis. With the cluster analysis, 16 bermudagrass cultivars were separated into three groups. Selected from 100 primers tested, four were chosen to be useful forcultivar identification using RAPD marker. Four primers were able to produce polymorphic DNA fragments to clearly identify N1, 0035 and 0036 varieties.With cluster analysis, 16 bermudagrass cultivars were separated into threegroups. Menbers of the three groups are corresponding to three types of bermudagrass according to their plant heitht and genetic constituent. The use of RAPD marker data for bermudagrass identification were proved to be useful by comparing the results with morphological traits data. Except for Tifway,15 cultivars were easily categorized into three types (common, dwarf, or semi-dwarf) by either RAPD or morphological traits method. The primerbank may need to be expanded for the selection of right primer used to separate every bermudagrass cultivars. From our experiments, use of the RAPD marker for cultivar identification is a quick and reliable method. A large scale selection of primers suitable for RAPD on bermudagrass has to be held before the method can be efficiently used.
URI: http://hdl.handle.net/11455/28862
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