Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/29102
標題: 同功異構酵素與逢機增殖多型性DNA標誌在番椒品種鑑別之研究
Identifications of capsicum spp. by isozymes and randomly amplified polymorphic DNA (RAPD) markers
作者: 王昭月
Wang, Zhao-Yue
關鍵字: 番椒
CAPSICUM SPP
同功異構酵素
逢機增殖多型性DN
品種鑑別
園藝
農業
逢機增殖多型性DNA
ISOZYME
RANDOMLY AMPLIFIED POLYMORPHIC
IDENTIFICATION
HORTICULTURE
AGRICULTURE
Capsicum spp
Isozyme
Randomly amplified polymorphic DNA (RAPD)
Identification
Capsicum spp.
出版社: 園藝學研究所
摘要: 本試驗以番椒 (Capsicum spp) 為材料,探討利用植物性狀分類、同功異 構酵素及逢機增殖多型性 DNA 標誌 (RAPD marker) 三種方法,分析不同 自交系、雜交種及栽培種間的種原鑑別。目的為:(1)建立區別番椒品系 之分子標誌,(2) 建立品系間遺傳連鎖群之關係,(3) 建立快速及正確的 甜椒種子純度的鑑定技術。番椒品系之植物性狀鑑別,採用 IBPGR (International Board for Plant Genetic Resources) 番椒性狀分類標 準,調查結果顯示由花冠、花萼及每節位花朵數可區別番椒 (Capsicum) 屬 C. annuum L. 及 C. chinense Jacquin 二個種,其中供試番椒品系 中除 P1697 為 C. chinense Jacquin 外其餘均為 C. annuum L。調查果 實性狀則又可區分 C. annuum 中的 7品系。唯甜椒種內的F1雜交種無法 依外表性狀區別。同功異構酵素分析的結果顯示C. annuum L.與C. chinense Jacquin二個種間之 EST、GOT、PGI、PGM、PRX、SkDH 及 SOD 等 7種酵素圖譜明顯不同。其中在 C. annuum L.種內之 GOT、PRX、PGI 、SOD 則具有不同的多型性酵素圖譜,唯僅能清楚識別 1 ~ 3 個品系, 尚無法區分所有 C. annuum L. 中之品系。此外萃取緩衝液與取樣部位都 會影響同功異構酵素圖譜的解析度;以萃取過程不易褐化的緩衝液所得之 酵素解析力較高。不同取樣部位則視不同酵素而有不同表現,其中 PRX、 SOD、PGI 以下位葉之多型性表現較清晰, GOT、MDH、PGM、SkDH 則為中 位葉解析較好,而 EST則以上位葉的活性及解析度較高。自300種逢機引 子 (random primer) 中篩選具有多型性 DNA 標誌,獲得10種可產生多型 性的引子。其中有 6個引子 UBC313,UBC327,UBC346,UBC457 UBC483及 UBC484 產生的 RAPD 圖譜可以清晰的將供試的番椒 8品系加以區別,以 此引子的特定DNA片段的標誌可有效的用來鑑別甜椒品系之F1雜交種, 及12個甜椒栽培品種。連鎖群分析的結果顯示12個甜椒栽培品種的遺傳背 景很相似,可能近源於甜椒P852品系,5個甜椒自交系中以P1717及P852較 相似,P1657及P1740次之,而P859較遠;3個辣椒及5個甜椒自交系屬於不 同族群。本試驗結果顯示RAPD的分子標誌可有效的區別番椒種原及F1雜交 種,估算親緣關係及鑑定種子純度;尤其是親源接近的品系,更可增加鑑 定的準確性及敏感度。另外利用這些特定的多型性 DNA 標誌片段,可進 行種子純度檢測,以區別出混雜的異品種個體。 Morphological traits,isozymes and randomly amplified polymor- phic DNA (RAPD) markers were used to study the identification of Capsicum species.The purposes of this study were to establish molecular markers for classification of the Capsicum spp., to establish the genetic linkage relationships and a quick, technique for the identification of seed purity of Capsicum spp. The results of isozyme analysis indicated that there were significantly different in the zymograms of EST, GOT, PGI, PGM, PRX, SkDH and SOD between C. annuum L. and C. chinense Jacquin. Distinctly polymorphic patterns were exhibited among the C. annuum L., but only three lines can be distinguished. Besides, the isozyme patterns might vary with different extracting buffers and sampling positions. For example, the expressions of PRX, SOD and PGI were clearer in lower leaf, GOT, PGM and SkDH in middle leaf and EST in upper leaf. A total of 10 primers was selected from 300 random primers that showed the polymorphic patterns in Capsicum spp.. Among , the RAPD patterns of six primers,i.e."UBC 313","UBC 327" , "UBC 346", "UBC 457","UBC 483" and "UBC 484", could clearly eight tested Capsicum lines and might be used as an effective for the identification of F1 hybrids and cultivated varieties of Capsicum spp.. The results of linkage cluster analysis revealed that the genetic background of 12 cultivars was very similar, and might originate from P852. Among the five inbred lines of sweet pepper, P1717 had a closer relation with P852, next by P1657 and P1740, but P 859 was farthest.The three hot pepper lines belonged to different population, when compared with these five inbred lines of sweet pepper. The results suggested that the RAPD markers is preciser and more sensitive than morphological and isozyme identifications and can be used as an effective tool to distinguish plant germplasms.
URI: http://hdl.handle.net/11455/29102
Appears in Collections:園藝學系

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