應用增殖限制酵素DNA片段多型性分析於蝴蝶蘭花色性狀與變異鑑別的研究

Studies of Using Amplified Fragment Length Polymorphism (AFLP) Technique on Identification of Flower Color and Somaclonal Variation in Phalaenopsis

將紅花、白花與白花紅心三種花色的蝴蝶蘭，利用16組不同引子對進行增殖限制酵素DNA片段多型性分析。其中E1M6和E3M4兩組引子對所產生之圖譜可將各品種之母本及｀W1-3＇和｀WR1-3＇的子代分生苗依花色區分為三個叢集。但所有試驗之引子對在相同花色性狀內，無法進一步鑑別品種。以白花紅心蝴蝶蘭｀WR3-2＇的母株及區分為外型正常與變異的子代組織培養苗為樣本，找到三組引子對E4M5、E6M8和E7M5產生的多型性條帶，有些只出現在變異株，卻不見於外型正常株和母株，這些條帶可能為分生苗變異株的分子標誌，且從條帶圖譜分析比較，變異株與母本之間有較低的相似性。 Sixteen primer pairs were applied to amplified fragment length polymorphism (AFLP) analyses of different Phalaenopsis cultivars grouped by flower colors as red, white and white with red-lips. AFLP patterns generated by E1M6 and E3M4 primer pairs could successfully separate all mother plants and regenerants of ‘W1-3’ and ‘WR1-3’ into 3 flower color groups by similarity analysis. However, none of tested primer pairs was able to distinguish cultivars within same flower color character. Mother plant of Phalaenopsis ‘WR3-21’ and its tissue-cultured derivatives classified as normal plants and somaclonal variants by their phenotypes were used for genetic fidelity test. Clear polymorphic banding patterns from E4M5, E6M8 and E7M5 primer pairs were observed on samples of somaclonal variant but not on normal regenerants. These polymorphic bands could become potential molecular markers on detection of somaclonal variations. In addition, banding patterns of somaclonal variants were more diverse from mother plant in comparing patterns of normal regenerants.