AOBPreview published online on July 6, 2007
Annals of Botany, doi:10.1093/aob/mcm118
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Highly Efficient Virus-induced Gene Silencing (VIGS) in California Poppy (Eschscholzia californica): An Evaluation of VIGS as a Strategy to Obtain Functional Data from Non-model Plants
1 Evolutionary Developmental Genetics Group, University of Bremen, Leobener Str., UFT, D-28139 Bremen, Germany
2 Institut für Spezielle Botanik, Johannes-Gutenberg-Universität Mainz, Bentzelweg 9a, D-55099 Mainz, Germany
* For correspondence. E-mail annette.becker{at}uni-bremen.de
Received: 8 January 2007 Returned for revision: 12 March 2007 Accepted: 10 May 2007
Background and Aims Eschscholzia californica: (California poppy) is an emerging model plant for ‘evo–devo’ studies from the basal eudicot clade of Papaveraceae. California poppy has a relatively small genome, a short life cycle and, most importantly, it is amenable for transformation. However, since this transformation protocol is time consuming, virus-induced gene silencing (VIGS) was evaluated as a fast method to obtain functional data for California poppy genes.
Methods: Commercially available California poppy plants were infiltrated with Agrobacterium tumefaciens carrying the tobacco rattle virus plasmids pTRV1 and pTRV2. pTRV2 contained part of the eschscholzia Phytoene Desaturase (EcPDS) gene whose loss of function results in photobleaching of the green parts of the plant and in a lack of floral coloration. The degree and duration of these symptoms was evaluated for vegetative rosettes and plants in flower.
Key Results: It is shown that VIGS is able to effectively down-regulate the EcPDS gene in eschscholzia. Various degrees of silencing were observed starting <2 weeks after infiltration with Agrobacterium tumefaciens in 92 % of the plants. Tissue with silencing symptoms also showed complete or strong reduction of EcPDS transcripts. Strong silencing resulted in almost completely white petals, fruits, shoots and leaves. Plants with a strong degree of silencing will eventually die off; however, others are able to produce EcPDS gene product even after a strong initial silencing and will recover. Silencing was found to be not always systemic, but was often restricted to certain organs or parts of organs.
Conclusions: VIGS is an effective, fast and transient method to down-regulate gene expression in eschscholzia. It serves well to detect prominent phenotypes which may become obvious even if some target gene transcript remains in the plant tissue. However, subtle phenotypes will be more difficult to detect, as extremely strong silencing effects occur in <10 % of all flowers from infected plants.
Key words: Eschscholzia californica, California poppy, Agrobacterium tumefaciens, tobacco rattle virus, Phytoene Desaturase, PDS, virus-induced gene silencing, VIGS, post-transcriptional gene silencing, basal eudicot, EcPDS
These authors contributed equally to this paper.
Present address: Department of Environmental and Plant Biology, Ohio University, Porter Hall, Athens, OH 45701, USA.
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