Title: Screening promoters for Anthurium transformation using transient expression Authors
Submitted to: Plant Cell Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 12, 2012
Publication Date: January 5, 2013
Citation: Matsumoto Brower, T.K., Keith, L.M., Cabos, R.Y.M., Suzuki, J.Y., Gonsalves, D., Thilmony, R.L. 2013. Screening promoters for Anthurium transformation using transient expression. Plant Cell Reports. 32:443-451. Interpretive Summary: Anthurium plants are grown as cut flowers and potted plants for their attractive flowers and foliage. Genetic engineering is one method to improve traits of anthurium plants such as disease resistance. The desired trait introduced into the anthurium plant consists of the promoter, gene of interest and terminator. In general, the promoter controls the amount and specificity of the desired gene of interest. Here we describe a method in which we place DNA containing different promoters and a trait that encodes blue color onto small pieces of gold. These DNA coated gold particle are propelled into different anthurium tissues such as leaves, roots and embryos by high pressure. By measuring the intensity and amount of blue dots in each tissue we are able to determine which promoters would work best in each tissue. Our results show that promoters from rice and maize ubiquitin genes work best in all anthurium tissues tested. This research will help scientist design anthurium plants with single or multiple new traits.
Technical Abstract: Different promoters and tissue types were evaluated for transient '-glucoronidase (GUS) expression in Anthurium andreanum Hort. ‘Marian Seefurth’ following microprojectile bombardment. Plasmids containing the Ubiquitin 2, Actin 1, Cytochrome C1 from rice, Ubiquitin 1 from maize and 35 S promoter from Cauliflower Mosaic Virus fused to the GUS-Plus gene were bombarded into in vitro grown anthurium lamina, somatic embryos and roots. The number of GUS foci and the intensity of GUS expression were evaluated from each construct. Ubiquitin promoters from rice and maize resulted in highest number of GUS events and expression in all tissues examined. Due to the slow growth of anthurium plants, development of transgenic anthurium plants may take years. This research should facilitate the development of transformation vectors for expression of desirable traits in anthurium plants. In addition, this research should facilitate the construction of transgenic plants expressing multiple genes under the control of different promoters to avoid potential homology-dependent transcriptional gene silencing.