Sweet pepper ferredoxin-like protein (pflp) gene as a novel selection marker for orchid transformation

• Published in: Planta 2003-05, Vol.217 (1), p.60-65
• Main Authors: You, Su-Juan, Liau, Chia-Hui, Huang, Hsiang-En, Feng, Teng-Yung, Prasad, Venkatesh, Hsiao, Hsin-hao, Lu, Jian-Cheng, Chan, Ming-Tsair
• Format: Article
• Language: English
• Subjects: Agrobacterium tumefaciens - genetics; Antibiotics; Biological and medical sciences; Biotechnology; Cinnamates; Complementary DNA; DNA; Exchange rates; Ferredoxins - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant - drug effects; Genetic engineering; Genetic Markers - genetics; Genetic technics; Hygromycin B - analogs & derivatives; Hygromycin B - pharmacology; Immunity, Innate - genetics; Leaves; Methods. Procedures. Technologies; Mutation; Orchidaceae - genetics; Orchidaceae - metabolism; Pectobacterium carotovorum - growth & development; Peppers; Plant cells; Plant Diseases - microbiology; Plant Proteins - genetics; Plants; Plants, Genetically Modified; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transformation, Genetic - genetics; Transgenes; Transgenic animals and transgenic plants; Transgenic plants
• ISSN: 0032-0935; 1432-2048
• DOI: 10.1007/s00425-002-0970-7

A novel method for selection of transgenic plants utilizing the sweet pepper (Capsicum annuum L.) ferredoxin-like protein (pflp) gene as selection marker and Erwinia carotovora as the selection agent has been developed. An expression vector containing a pflp cDNA driven by a cauliflower mosaic virus 35S promoter was successfully transformed into protocorm-like bodies of Oncidium orchid by Agrobacterium tumefaciens and particle bombardment, respectively. Erwinia carotovora was used as a selection agent to screen transformants, thereby obtaining transgenic plants without the use of an antibiotic selection agent. A total of 32 independent transgenic orchid lines were obtained, out of which 9 transgenic lines (β-glucuronidase positive) were randomly selected and confirmed by Southern and northern blot analyses. The transgenic orchid plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest the novel use of the pflp gene as a resistance selection marker in plant genetic engineering strategies. In the future, the use of the pflp gene as a selection marker may facilitate the use of smaller gene constructs due to removal of bulky antibiotic selection and reporter genes. These constructs can then be used to incorporate additional genes of choice.