- 期刊
櫻絨花(Emilia fosbergii Nicolson)與紫背草(E. sonchifolia L. var. javanica)之分子鑑定
Identification of Florida Tasselflower (Emilia fosbergii Nicoison) and Sowthistle Tasselflower (E. sonchifolia L. var. javanica) through Molecular Markers
摘要
櫻絨花(Emilia fosbergii Nicolson)為菊科紫背草屬一年或二年生草本植物，原生於非洲地區，現已成為臺灣農地的入侵雜草。由於櫻絨花與同屬植物紫背草(E. sonchifolia L. var. javanica)的植株形態極為相似，未開花之前不易區別，造成鑑定上的困擾。分子標誌己普遍運用於植物種類之鑑別，本研究針對櫻絨花與紫背草5.8S rRNA-ITS核酸序列進行選殖及解序，並建立PCR-RFLP、multiplex PCR及ISSR的分子標誌。櫻絨花與紫背草的ITS1-5.8S rRNA-ITS2序列長度不同，分別為643 bp 及641 bp，一致性(identity)為90.7%。經由ITS 序列比對，2者之間有Nsi I、Sph I 及Xho I 3種限制酵素切位的差異，5.8S rRNA-ITS核酸片段經3種限制酵素反應，故發展PCR-RFLP鑑定方法；另於ITS 序列差異處設計專一性引子，建立multiplex-PCR鑑定方法，可分別於櫻絨花與紫背草增幅678 bp 及340 bp片段；此外利用ISSR 587引子增幅1.0-2.1 kb之間的多型性條帶，可明顯區別櫻絨花與紫背草。multiplex-PCR 及ISSR兩種分子標誌具有簡易、快速及明確等特點，可應用於入侵植物的監測及管理，進而維護臺灣原生物種之生態多樣性與平衡。
並列摘要
Florida tasselflower (Emilia fosbergii Nicolson) is an annual or biennial Compositae plant and native to Africa, became an invasive plant among cropland in Taiwan in recent decades. As the morphological characteristics are similar, Florida tasselflower has been easily misidentified with sowthistle tasselflower (E. sonchifolia L. var. javanica) before blossom. DNA-based molecular markers have been used to detect the genetic diversity of invaded alien species. Novel methods for the identification of the invasive plant Florida tasselflower and sowthistle tasselflower at the early stage of plant development have established in this study, based on direct sequencing of the internal transcribes spacer (ITS) region of 18S-26S ribosomal DNA (rDNA), PCR-restriction fragment length polymorphism (RFLP), multiplex PCR and inter-simple sequence repeat (ISSR) assay. The ITS1-5.8S rRNA-ITS2 regions of Florida tasselflower and sowthistle tasselflower was 643 and 641 bp, respectively, and showed 90.7% identity. the amplified 5.8S rRNA-ITS fragments were compared using RFLP analysis with Nsi I、Sph I and Xho I endonucleases, allowing the detection of characteristic patterns of these two Emilia species. Fifty ISSR primers were screened in this study, 12 primers amplified 1-8 polymorphic markers. Amplified 1.0-2.1 kb bands from ISSR 857 primer and using multiplex PCR produced 678 or 340 bp distinct bands, both were clearly and easily distinguished Florida tasselflower and sowthistle tasselflower. These two markers may assist the effective management in invasion plant and maintain the balance of biodiversity in agricultural ecosystems.
被引用紀錄
延伸閱讀
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