台灣原生杜鵑核醣體核酸內轉錄間隔區之選殖及分析

Cloning and Characterization of Internal Transcribed Spacer Region of rDNA in Rhododendron Species in Taiwan

蔡奇助(Chi-Chu Tsai) ； 蔡素蕙(Suh-Huey Tsai) ； 黃勝忠(Sheng-Chung Huang)

《臺中區農業改良場研究彙報》 64期 (1999/09) Pp. 13-26

https://doi.org/10.6956/BTDAIS.199909.0013

摘要

本研究設計一組核苷酸引子（primer），序列分別為IT1：5'TCGTAACAAGGTTTCCGTAGGT 3'和IT2：5' GTAAGTTTCTTCTCCTCCGCT 3'，利用聚合酵素連鎖反應（polymerase chain reaction，PCR），將14種台灣原生杜鵑（Rhododendron spp.）的5.8S核糖體核酸（rRNA）基因與內轉錄間隔區（internal transcribed spacer，ITS）選殖。將上述PCR產物定序，14種台灣原生杜鵑內轉錄間隔區（ITS）長在642～648 bp間。其中5.8S rRNA基因區各種長皆為164 bp，且序列上並無差異。在ITS1區域中，長度具253、254及255 bp三類，其中有29個核苷酸具多型性。在ITS2區域中，長度具225、226及229 bp三類，且序列中有33個核苷酸具多型性。本研究之核酸引子能將台灣各原生杜鵑之ITS加以選殖，藉由分析ITS序列，可獲取有用的分子標誌，供探討杜鵑花屬的微演化（microevolution）及親緣研究上。未來在杜鵑花育種上，亦可供為品種鑑別，及品種專利之應用。

關鍵字

杜鵑 ； 聚合酵素連鎖反應 ； 5.8S核糖體核酸基因 ； 核糖體核酸內轉錄間隔區

並列摘要

The entire nucleotide sequence of internal transcribed spacer (ITS) region between 18S and 26S ribosomal RNA (rRNA) genes among 14 Rhododendron species in Taiwan were amplified by polymerase chain reaction (PCR). The primers of PCR, IT1: 5' TCGTAACAAGGTTTCCGTAGGT 3' and IT2: 5' GTAAGTTTCTTCTCCTCCGCT 3' were designed for amplification. The PCR products of 14 Rhododendron species were sequenced. To compare with the sequence of ITS region of other higher plant species, the length of ITS in 14 Rhododendron species were ranged from 642 to 648 bp. These 5.8S rRNA gene, ITS1, and ITS2 were included within ITS region. In 5.8S rRNA gene, no variation was revealed between 14 Rhododendron species. In ITS1 and ITS2 regions, variable length and sequence, generated from deletion, insertion, and substitution, were introduced between 14 Rhododendron species. Therefore, the microevolution and phylogeny of Rhododendron species could be studied based on the sequence of ITS. For further application, ITS region is a good marker for cultivar identification and protection as well.

並列關鍵字

Rhododendron ； polymerase chain reaction (PCR) ； 5.8S rRNA gene ； internal transcribed spacer (ITS)