以RAPD標誌鑑別原生石斛蘭之研究

Abstract

Native Dendrobium species, including originated in Taiwan such as Dendrobium chameleon, D. crumenatum, D. falconeri, D. leptocladum, D. linawianum, D. miyakei, D. moniliforme, D. somai, D. tosaense, D. sanseiense, Flickingeria comata and F. tairukounia and originated in China such as D. candidum, D. falconeri and F. fimbriata and originated in Philippines such as D. crumenatum, D. miyakei, were used as experimental materials at present studies.The Phalaenopsis amabilis seedlings derived from tissue culture were used as control. The morphology and horticulture characteristics of the plants were investigated. The relationship of the Dendrobium spp. were distinguished and identified by means of Randomly Polymorphism DNA analysis techniques. The similarity of DNA bands was observed few in the native Dendrobium by RAPDs. There were six groups of native Dendrobium species could be grouped in accordance with the results dendrogram of RAPD analysis at present materials. This result indicated that D. crumenatum could be an independent genus due to the similarity was lower than control. The Ⅱ and Ⅴ groups were also possible another independent genus.以原生於台灣地區，包括屏龍山之長爪石斛(Dendrobium chameleon)等14種，菲律賓地區原生之鴿石斛(D. crumenatum)等2種及大陸地區原生之駐皮石斛(D. candidum)等3種，共計19種之原生石斛蘭為試驗材料，並以產自台東大武之白花蝴蝶蘭(Phalaenopsis amabilis)組織分生苗為遠端對照，利用分子層次之逢機增殖多形性DNA片段(RAPD)標誌鑑別原生石斛蘭種原。 以OPERON之隨機引子組OPE-01~20進行RAPD分析，結果顯示：原生石斛蘭種間的條帶圖譜相同的地方少，種間變異性相當大且無法得到種間一致性或是石斛蘭屬間特有的條帶，然而在不同來源之相同種間條帶圖譜的一致性就顯的相當高。繪製出之親緣關係樹狀圖顯示：本試驗中的原生石斛蘭可以分為六群，分別是第Ⅰ群─鴿石斛(菲律賓及綠島兩種)，第Ⅱ群─紅花石斛(菲律賓及蘭嶼兩種)、長爪石斛、三星石斛，第Ⅲ群─白花石斛、大陸鐵皮石斛、黃花石斛、紅鸝石斛(谷關、杉林溪、大陸雲南三種)，第Ⅳ群─金草石斛、櫻石斛，第Ⅴ群─ 小雙花石斛、細莖石斛，第Ⅵ群─鬚唇暫花蘭、尖葉暫花蘭、流蘇金石斛。就中第Ⅰ群之鴿石斛較遠端對照組之白花蝴蝶蘭相似度更低，理當獨自成一屬；第Ⅱ群之紅花石斛、長爪石斛、三星石斛以及第Ⅴ群之雙花石斛、細莖石斛等依相關資料顯示亦有獨立成屬的條件。同種但不同地域生長者其遺傳相似度雖較他種接近，但仍存在有差異性，外觀形態也有些不同；以紅鸝石斛為例：產於台灣杉林溪、谷關以及來自大陸雲南三者，以台灣自產者親緣相似度高於產自大陸者，故地域的遠近也會影響同種間之親緣相似度