Agrobacterium-mediated Cuscuta campestris transformation as a tool for understanding plant-plant interactions.

Autor: Adhikari S; Division of Plant Science and Technology, University of Missouri, Columbia, MO, 65211, USA., Mudalige A; Division of Plant Science and Technology, University of Missouri, Columbia, MO, 65211, USA., Phillips L; Division of Plant Science and Technology, University of Missouri, Columbia, MO, 65211, USA., Lee H; Division of Animal Science, University of Missouri, Columbia, MO, 65211, USA., Bernal-Galeano V; School of Plant and Environmental Sciences, Virginia Tech, Blacksburg, VA, 24061, USA., Gruszewski H; School of Plant and Environmental Sciences, Virginia Tech, Blacksburg, VA, 24061, USA., Westwood JH; School of Plant and Environmental Sciences, Virginia Tech, Blacksburg, VA, 24061, USA., Park SY; Division of Plant Science and Technology, University of Missouri, Columbia, MO, 65211, USA.
Jazyk: angličtina
Zdroj: The New phytologist [New Phytol] 2024 Oct 03. Date of Electronic Publication: 2024 Oct 03.
DOI: 10.1111/nph.20140
Abstrakt: Cuscuta campestris, a stem parasitic plant, has served as a valuable model plant for the exploration of plant-plant interactions and molecular trafficking. However, a major barrier to C. campestris research is that a method to generate stable transgenic plants has not yet been developed. Here, we describe the development of a Cuscuta transformation protocol using various reporter genes (GFP, GUS, or RUBY) and morphogenic genes (CcWUS2 and CcGRF/GIF), leading to a robust protocol for Agrobacterium-mediated C. campestris transformation. The stably transformed and regenerated RUBY C. campestris plants produced haustoria, the signature organ of parasitic plants, and these were functional in forming host attachments. The locations of T-DNA integration in the parasite genome were confirmed through TAIL-PCR. Transformed C. campestris also produced flowers and viable transgenic seeds exhibiting betalain pigment, providing proof of germline transmission of the RUBY transgene. Furthermore, RUBY is not only a useful selectable marker for the Agrobacterium-mediated transformation, but may also provide insight into the movement of molecules from C. campestris to the host during parasitism. Thus, the protocol for transformation of C. campestris reported here overcomes a major obstacle to Cuscuta research and opens new possibilities for studying parasitic plants and their interactions with hosts.
(© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)
Databáze: MEDLINE
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