Email updates

Keep up to date with the latest news and content from Plant Methods and BioMed Central.

Open Access Methodology

Transformation and regeneration of the holoparasitic plant Phelipanche aegyptiaca

Mónica Fernández-Aparicio12*, Diego Rubiales1, Pradeepa CG Bandaranayake3, John I Yoder3 and James H Westwood2

Author Affiliations

1 Institute for Sustainable Agriculture, IAS-CSIC, Dept. of Plant Breeding, Córdoba, 14080, Spain

2 Virginia Tech, Dept. of Plant Pathology, Physiology and Weed Science, Blacksburg, VA 24061, USA

3 University of California-Davis, Dept. of Plant Sciences, Davis, CA 95616, USA

For all author emails, please log on.

Plant Methods 2011, 7:36  doi:10.1186/1746-4811-7-36

Published: 8 November 2011

Abstract

Background

Transformation and subsequent regeneration of holoparasitic plants has never been reported, in part due to challenges in developing transformation protocols, but also because regeneration of obligate parasites is difficult since their survival depends completely on successful haustorium penetration of a host and the formation of vascular connections. The recent completion of a massive transcriptome sequencing project (the Parasitic Plant Genome Project) will fuel the use of genomic tools for studies on parasitic plants. A reliable system for holoparasite transformation is needed to realize the full value of this resource for reverse genetics and functional genomics studies.

Results

Here we demonstrate that transformation of Phelipanche aegyptiaca is achieved by infection of 3 month-old in vitro grown P. aegyptiaca calli with Agrobacterium rhizogenes harboring the yellow fluorescent protein (YFP). Four months later, YFP-positive regenerated calli were inoculated onto tomato plants growing in a minirhizotron system. Eight days after inoculation, transgenic parasite tissue formed lateral haustoria that penetrated the host and could be visualized under UV illumination through intact host root tissue. YFP-positive shoot buds were observed one month after inoculation.

Conclusions

This work constitutes a breakthrough in holoparasitic plant research methods. The method described here is a robust system for transformation and regeneration of a holoparasitic plant and will facilitate research on unique parasitic plant capabilities such as host plant recognition, haustorial formation, penetration and vascular connection.

Keywords:
holoparasitic plants; gene transformation; haustorium; Phelipanche; Orobanche