利用ISSR及細胞質DNA標誌探討薰衣草品種之遺傳歧異性

Using ISSR and Cytoplasmic DNA Markers to Study the Genetic Diversity of Lavender Varieties

簡銘錦(Ming-Chin Chien)

指導教授 ： 林順福

國立臺灣大學/生物資源暨農學院/農藝學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.01973

摘要

摘要
薰衣草之種類甚多且用途差異甚大，為避免薰衣草品種之錯誤命名與不當使用，本研究主要目的在建立薰衣草品種的鑑別系統及評估種原之遺傳變異。在分屬於五個不同sections的薰衣草品種中，本研究共收集三十個品種並進行葉片與香氣調查。在五個不同sections的葉片長寬比調查中，Lavandula (狹葉薰衣草)與 Pterostoechas (羽葉薰衣草) 這兩個sections分別具有最高（7.23）及最低的數值(2.01); 而Lavandula (狹葉薰衣草) (3.59)與Stoechas (法國薰衣草) (3.00) 兩個sections的葉長平均值相近; 且在葉寬的調查中，Lavandula (狹葉薰衣草) (0.49)與Stoechas (法國薰衣草) (0.47) 兩sections的平均值仍然非常相近，表示僅由葉片外觀性狀無法精確的判別薰衣草品種。而薰衣草品種香氣則被區分為六個等級﹝1代表香氣等級最優良，6代表香氣等級最低劣﹞，其中Lavandula (狹葉薰衣草) 與Pterostoechas (羽葉薰衣草) 這兩個sections的等級分別為2 (fine) 和6 (poor)。Dentata (齒葉薰衣草) 及Pterostoechas (羽葉薰衣草) 兩sections具有樟腦味，屬於Lavandula angustifolia (狹葉薰衣草) 的品種則具有高度香氣。
此外，本研究由100條引子中篩選出13個穩定且具有多型性的ISSR核心引子，可產生42個分子標誌做為薰衣草品種之DNA指紋資料。利用這些ISSR分子標誌計算品種得到品種之遺傳相似性介於4%∼95.5%，經主成分向量分析與分群分析可將全部品種分為兩群：其中一群屬於Lavandula (狹葉薰衣草) 與Dentata (齒葉薰衣草) 兩sections，另一群則屬於Stoechas (法國薰衣草) 與Pterostoechas (羽葉薰衣草) 兩sections，此分群與葉形分類及親緣分類相符合。因此，根據ISSR分子標誌的結果，不但可以鑑定出不同商品名稱之相同品種，且可以辨別出被誤認為狹業品種之狹業雜交品種。
另外，由13條mtDNA引子及16條cpDNA引子篩選出7條mtDNA與7條cpDNA引子，共可產生22個分子標誌。根據mtDNA與cpDNA分子標誌所得到的分群分析與主成分分析的結果，不但可以鑑別出胞器DNA相同的品種，且可利用於追蹤雜交種的母本。與ISSR分子標誌相比較， ISSR分子標誌較符合可雜交分類(Hybridization categories)與葉部特徵，較適合應用於薰衣草品種鑑定。mtDNA與cpDNA分子標誌因為可追蹤品種之母本來源，故可輔助ISSR分子鑑定技術。
在相同葉型的薰衣草品種中包含具有香氣佳或香氣差（含黃樟素）之變異，而以ISSR分子標誌進行品種分群結果，亦與香氣品質相關。本研究的結果顯示，葉型或葉片長寬比可供作品種初步鑑定的依據。另外，本研究所建立之品種分子指紋資料及鑑定流程，以及種原變異評估結果，可供薰衣草育種、栽培及產品選擇或使用之參考。

關鍵字

細胞質DNA標誌 ； 遺傳歧異性 ； ISSR ； 薰衣草

並列摘要

Abstract
Lavender is one of the most versatile species of all herbs and can be used for many purposes, but not all sections of lavenders are suitable for all applications. To solve the problems of the misnaming of varieties and inappropriate utilizations of lavender, the lavender variety identification system was established and the genetic diversity of the germplasm was evaluated. Thirty lavender varieties in five sections were investigated in leaf characters and fragrance. Sections Lavandula and Pterostoechas possessed the highest (7.23) and the lowest (2.01) value of the length/width, respectively. Because sections Lavandula (3.59cm) and Stoechas (3.00cm) possessed the approximate value of the leaf length and leaf width, lavender varieties couldn’t be identified only by the leaf character. The fragrance of lavender varieties was divided into six levels (score 1 represented fantasy fragrance, and score 6 represented poor fragrance), and sections Lavandula and Pterostoechas were evaluated with score 2 (fine) and score 6 (poor), respectively. Besides, both sections Dentata and Pterostoechas contained camphor, and the averaged fragrance scores of species Lavandula were ranged from 2 (fine) to 3 (superior).
Thirteen stable and polymorphic ISSR primers were selected from 100 primers. Forty-two ISSR markers amplified from primers those were applied in fingerprinting lavender varieties. The similarities among varieties were ranged from 4% to 95.5% based on ISSR markers. All varieties were grouped into two groups both by the principal component coordinate analysis and cluster analysis. One group belonged sections Lavandula and Dentata, and the other belonged to sections Stoechas and Pterostoechas. The result of the cluster analysis was corresponded to the leaf character and the hybridization categories. However, according to similarity analysis of ISSR markers, not only the same varieties with different names, but also the hybrid variety could be identified.
There were 7 mtDNA and 7 cpDNA makers selected from the 13 mtDNA and 16 cpDNA primer pairs, respectively. Twenty-two molecular markers were generated by those primer pairs. According to the results of the cluster analysis and the principle coordinate components analysis, it indicated that the cytoplasmic markers were able to identify the varieties with the same organelle DNA and to trace the female parent of the hybrid varieties. Therefore, the cytoplasmic markers could supplement ISSR markers in lavender varieties identification.
Variations in aromatic score among varieties of same leaf type were found, and the cluster analysis results of ISSR markers were also correlated to the fragrance analysis. According to the results of this study, the leaf characters could only be applied in the preliminary variety identification. Besides, the results of the DNA fingerprinting, variety identification, and genetic diversity analysis could provide information for lavender breeding, cultivation, and utilization.

並列關鍵字

Cytoplasmic DNA Markers ； ISSR ； Lavender ； Genetic Diversity

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