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Genetic transformation of golden pothos (Epipremnum aureum) mediated by Agrobacterium tumefaciens

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Abstract

To establish a procedure for Agrobacterium tumefaciens-mediated transformation of golden pothos (Epipremnum aureum) plants, the effects of selection antibiotics and the preculture period of stem explants before A. tumefaciens infection were examined. Explants were co-cultivated with A. tumefaciens EHA105, harboring the plasmid pGWB2/cGUS, on a somatic embryo-inducing medium supplemented with acetosyringone. Resulting transgenic somatic embryos were screened on an antibiotic selection medium, and the transgenic pothos plants were regenerated on a germination medium. Hygromycin was the optimum selection antibiotic tested. The preculture period significantly affected the transformation efficiency, with explants precultured for one-day showing the best efficiency (5–30%). Both transformed hygromycin-resistant embryos and regenerated plants showed β-glucuronidase activity. Southern blot analysis confirmed transgene integration into the pothos genome. This reproducible transformation system for golden pothos may enable the molecular breeding of this very common indoor plant.

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Abbreviations

GUS:

β-Glucuronidase

X-gluc:

5-Bromo-4-chloro-3-indolyl-b-d-glucuronide

35S-pro:

Cauliflower mosaic virus 35S promoter

LB:

Luria–Bertani

MS:

Murashige and Skoog

TDZ:

N-phenyl-N′-1,2,3-thiadiazol-5-ylurea

NAA:

α-Naphthalene acetic acid

PCR:

Polymerase chain reaction

NPTII:

Neomycin phosphotransferase

HPT:

Hygromycin phosphotransferase

NOS:

Nopaline synthase

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Acknowledgment

We thank Dr. Shibata for providing the plasmid pGWB2/cGUS.

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  1. School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo, 192-0982, Japan

    Katsutoshi Kotsuka & Yuichi Tada

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  1. Katsutoshi Kotsuka
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  2. Yuichi Tada
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Correspondence to Yuichi Tada.

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Kotsuka, K., Tada, Y. Genetic transformation of golden pothos (Epipremnum aureum) mediated by Agrobacterium tumefaciens . Plant Cell Tiss Organ Cult 95, 305–311 (2008). https://doi.org/10.1007/s11240-008-9444-3

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  • DOI: https://doi.org/10.1007/s11240-008-9444-3

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