分子標記在仙履蘭品種鑑定上之應用

Use of Molecular Markers for Species Identification in Paphiopedilum.

孫永偉(Yung-Wei Sun) ； 廖文毅(Wen-Yi Liao) ； 沈翰祖(Han-Tsu Shen) ； 劉明宗(Ming-Chung Liu) ； 廖玉珠(Yu-Ju Liao) ； 蔡瑜卿(Yu-Ching Tsai) ； 蕭吉雄(Chi-Hsiung Hsiao) ； 陳駿季(Junne-Jih Chen)

《植物種苗》 7卷3期 (2005/09) Pp. 1-9

https://doi.org/10.30077/SN.200509.0001

摘要

近年來以DNA為研究基礎的分子標誌技術已成功應用於可作為區分植物品種差異之指紋分析方式。這些DNA 技術包括逢機增幅多形性DNA(RAPD; random amplified polymorphism DNA)、增殖片段長度多型性(AFLP; amplified fragment length polymorphism)、SRAP(sequence-related amplified polymorphism)、內轉錄間隔區(ITS; internal transcribed spacer)、簡單重複序列(SSR; simple sequence repeats)等。仙履蘭(Paphiopedilum)由於花型奇特深受國內外賞花人士喜愛，育種者培育新品種不斷推陳出新，為台灣重要新興花卉作物。仙履蘭品種鑑定多以植株葉片形態色澤、花瓣及唇瓣形狀等外表形態上差異加以區分，此鑑定方式需累積豐富經驗且不同學者常有不同鑑定結果。因此，利用分子標記技術鑑定仙履蘭品種DNA層次差異性，配合UPGMA群叢分析方式，將可較精確區分各品種分類屬性。

關鍵字

仙履蘭 ； 逢機增幅多形性DNA ； 增殖片段長度多型性 ； 內轉錄間隔區

並列摘要

The polymerase chain reaction (PCR) is widely used in genomic DNA analysis. More recently, DNA-based techniques have been used successfully in DNA fingerprinting of plant genomes. These include random amplified polymorphism DNA (RAPD), amplified fragment length polymorphism (AFLP), internal transcribed spacer (ITS), sequence-related amplified polymorphism (SRAP), simple sequence repeat polymorphism (SSR) and a few others. RAPD procedures were first developed in 1990 using PCR to randomly amplify anonymous segments of nuclear DNA with an identical pair of primers 8-10 bp in length. Because the primers are short and relatively low annealing temperatures are used, the likelihood of amplifying multiple products is great, with each product a different locus. The nuclear ribosomal DNA (rDNA) gene family is multigene family. In most eukaryotes, the 5' to 3' organization of the gene family is an external transcribed spacer (ETS), the gene 18S, an ITS1, 5.8S, ITS2, 28S, and intergenic spacer (IGS). ITS, located between the repeating array of nuclear 18S and 28S ribosomal DNA genes, is a versatile genetic marker. The ITS region does not encode any product, permitting it to evolve at a faster rate than the ribosomal coding regions. The level of variation in this region makes it suitable for detecting genetic variation among genus, species, and within species. This results in variability in what they amplify, from genotype to genotype, but also from one DNA sample to another. SRAP is a simple marker technique aimed for the amplification of open reading frames (ORFs). AFLP is a PCR-based multi-locus fingerprinting technique, which efficiently identifies DNA polymorphisms without prior information on the DNA sequence of plants. The AFLP technique is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA. The technique involves three steps: (a) restriction of the DNA and ligation of adapters, (b) selective amplification of restriction fragments, (c) gel analysis of the amplified fragments. For Paphiopedilum, the molecular markers should provided a high number polymorphisms per run when compared to morphological characters.

並列關鍵字

Paphiopedilum ； random amplified polymorphism DNA RAPD ； amplified fragment length polymorphism AFLP ； internal transcribed spacer ITS