以ISSR與RAPD分子標誌分析火鶴花栽培種間遺傳相似性

Comparative Analysis of Genetic Similarity among Anthurium Cultivars by Using ISSR and RAPD Markers

王昭月(Jau-Yueh Wang) ； 莊耿彰(Keng-Chang Chuang) ； 范明仁(Ming-Jen Fan)

《中華農業研究》 50卷1期 (2001/03) Pp. 54-67

https://doi.org/10.29951/JARC.200103.0006

摘要

本研究以農業試驗所『火鶴花育種』計畫中，主要雜交親本30個栽培種為材料，探討利用ISSR(inter simple sequence repeat)及RAPD(random amplified polymorphic DNA)兩種分子標誌技術進行親本間遺傳相似性之分析與比較其鑑別能力。試驗結果顯示，此兩種分析在火鶴花栽培種間，均表現高度多型性。ISSR分析結果，發現dinucleotides motif:(TC)(下標 n)明顯大量存在於火鶴花基因內，以其作為引子表現之多型性最高；自篩選得7條引子進行之PCR產物，共可獲153個多型性DNA片段；RAPD分析結果，則自篩選得7條引子反應產生131個多型性片段。此外，ISSR分析產物中發現32個特定的DNA多型性片段，可用以鑑別18個栽培種；而RAPD分析中，則有34個特定的DNA多型性片段，可用以鑑別19個栽培種。進一步將ISSR總產物159個片段，合併進行UPGMA(unweighted pair-group mean arithmetic)群集分析結果：30個栽培種分成四個主要群組，其中'Southern Blush'、'Fla-range'及'Lady Anne'為個別獨立的三群，明顯別於第四群組的27個栽培種；連鎖四個群組的30個栽培種，其遺傳相似性係數為053。在RAPD分析方面，其總產物為134個片段，經合併進行UPGMA分析結果也分成四個主要群組：第一群'Southern Blush'、第二群'Champion'及'Marian-Seefurthh'及第三群'Midori'等四品種，亦明顯相異於第四群組的26個栽培種，連鎖四個群組之遺傳相似性係數則為057。綜合以上兩分析結果顯不，ISSR或RAPD分子標誌，在火鶴花栽培種間遺傳相似性分析之結果相近，兩種分子標誌亦均可利用於鑑別部分栽培種。分子層次之分析結果，可以助益於育種親本遺傳背景之瞭解，對加速優良雜交組合之獲得及提升育種效率，應具實用之價值。

關鍵字

火鶴花 ； 育種 ； ISSR ； RAPD ； 鑑定 ； 遺傳相似性

並列摘要

Thirty Anthurium cultivars in the breeding program of Taiwan Agriculture Research Institute were evaluated and compared by using ISSR (inter simple sequence repeat) and RAPD (random amplified polymorphic DNA) molecular markers. Objectives were to assess genetic similarity within the cultivars and survey the identification markers. The results indicated both methods revealed high degree of genetic polymorphism. The result of ISSR suggested that among the dinucleotides motifs, the poly(TC)(subscript n) motif was more abundant and gave the largest number of polymorphism. There were 153 polymorphic bands produced by seven selected primers using ISSR analysis while 131 polymorphic bands were produced by RAPD. Additionally, 32 specific DNA fragments derived from ISSR analysis that could be used to distinguish 18 cultivars while 34 DNA fragments derived from RAPD analysis were used to distinguish 19 cultivars. Furthermore, analysis of genetic similarity by UPGMA (Un-weighted Pair-group Mean Arithmetic) using the markers of ISSR and RAPD revealed both were closely related. In the ISSR analysis, a dendrogram based on the UPGMA cluster analysis was constructed using the similarity matrix that derived from 159 polymorphic ISSR fragments generated by seven primers. These thirty cultivars could be divided into four major groups: The cultivars of 'Southern Blush' 'Fla-range' and 'Lady Anne' performed as three independent groups that appeared to be distinct from other cultivars of group four. The similarity coefficient calculated from thirty cultivars analysis by ISSR was 0.529. In the RAPD analysis, a dendrogram also constructed using UPGMA from 134 fragments that were amplified by seven selected primers, divided the thirty cultivars into four main groups: 1) 'Southern Blush' 2) 'Champion' and 'Marian-Seefurth' 3) 'Midori' that seems to be distinct from other cultivars of group forth too. The similarity coefficient calculated from all thirty cultivars analysis by RAPD was 0.57. It is suggested both 1SSR and RAPD markers are useful in current breeding program of Anthurium, and allowed to estimate the genetic similarity among genotypes as well as to identify cultivars. These quick and reliable molecular tools are useful in selecting the best parents and obtaining new combinations.

並列關鍵字

Anthurium ； Breeding ； ISSR ； RAPD ； Identification ； Genetic similarity