開發鑑別台灣咖啡品種之分子標誌與外表性狀之調查

Development of Molecular Markers for Variety Identification of Taiwan Coffee and Morphological Traits

陳怡伶(Yi-Ling Chen)

指導教授 ： 王裕文

國立臺灣大學/生物資源暨農學院/農藝學研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.00281

摘要

本試驗採集台灣現有栽培的咖啡莊園共160個種源，調查其10個外觀性狀進行分析，配合從夏威夷種源中心所獲得的6個已知C. arabica品種及C. canephora，利用ISSR及SSR分子標誌探討台灣咖啡品種鑑定之可行性。調查的10個外觀性狀，葉片性狀包含葉長、寬、長寬比、及葉面積；植株性狀包含節間長度及主幹與一次分支之夾角；種子性狀包含帶殼種子長、寬、帶殼種子長寬比及帶殼種子厚的調查結果，變異係數在葉面積、節間長度及帶殼種子厚，分別為33.57%、37.62%及34.12%，其餘的性狀的變異係數則小於30%。分析結果顯示葉面積大小可做為判斷為C. arabica的重要指標。以6個外觀性性狀進行主成份分析，結果在採集區域、及老欉新苗間分群雖略有群聚狀況，但種源間仍呈現彼此混雜而致分群不明顯；但高海拔種源在葉寬、葉面積顯著大於低海拔種源；而以10個外觀性狀進行主成分分析，結果在採集區域間及海拔分布分群間均勻分布、分群情況不顯著。
在ISSR與SSR兩種分子標誌分析下，ISSR分析由100個ISSR引子中篩選出在C. arabica與C. canephora 物種間具多型性的32個分子標誌，就採集的樣品所顯示的遺傳相似度介於0.19至1之間，可區別C. arabica與其他物種；而SSR分析則由53個SSR引子中篩選出在C. arabica品種間具多型性的2個引子，共獲得4個標誌，並依其所得的遺傳相似性可將台灣咖啡種源分成4群，包含Bourbon群、Typica群、Caturra群及4個台灣樣品，並顯示台灣現有栽培的咖啡種源其變異有限，或許仍有引種的需求。由於所取得的對照品種為數較少，與篩選所獲取的具多型性標誌不足，因此品種鑑別力仍有不足，僅能區分主要的分群。

關鍵字

咖啡 ； ISSR ； SSR ； 品種鑑別 ； 主成分分析 ； 群聚分析

並列摘要

The purpose of this study was to establish the variety identification system of coffee in Taiwan by utilizing 10 morphological traits and ISSR and SSR molecular markers . The six C. arabica and C. canephora of HARC germplasm were used as reference, the DNA profile was established for the 160 coffee germplasm in Taiwan. According to the result of 10 morphological traits investigated, the frequency distribution of leaf length, leaf width, leaf shape, leaf area, length of plagiotropic internodes, the angle of the secondary branch at the insertion with the main one, seed length, seed width, seed shape, and seed thickness, except for leaf area, length of plagiotropic internodes, and seed thickness, the coefficient of variation of the others were estimated to be under 30%. Meanwhile, the trait of leaf area was a good indicator for discriminating between C. arabica and other species.
Thirty-two markers which showed polymorphism between species were identified from the 100 ISSR primers. The genetic similarity estimated by ISSR markers ranged from 0.19 to 1 between C. canephora and C. arabica accessions from HARC and Taiwan. The results showed ISSR markers could efficiently differentiate the Coffea species. In order to make up for the insufficient polymorphism between C. arabica cultivars, four polymorphic markers were identified after screened set of 53 SSR primers, which were then scored and used to estimate the genetic similarity and then clustering the C. arabica of HARC and of Taiwan. The result indicated the C. arabica germplasm could be divided into 4 groups: including Typica group, Bourbon group, Caturra group, and the unknown accessions, with the name after the known variety in the same cluster. Because of the narrow genetic base of C. arabica cultivars, the limited availability of the reference cultivars, the identified polymorphic markers, these markers were only able to discriminate the collected Taiwan C. arabica accessions into the same group as the reference Typica, Bourbon and Caturra variety.

並列關鍵字

coffee ； ISSR ； SSR ； variety identification ； principal component analysis ； cluster analysis

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