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Early antibiotic selection and efficient rooting and acclimatization improve the production of transgenic plum plants (Prunus domestica L.)

  • Genetic Transformation and Hybridization
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Abstract

We describe here an improved system for routinely developing transgenic plum plants (Prunus domestica L.) through the use of Agrobacterium tumefaciens. The production of non-transformed "escapes" has been virtually eliminated, and rates of plant establishment in the greenhouse have been dramatically improved. The system is based on the regeneration of shoots from hypocotyls extracted from mature seed. The shoot regeneration medium is Murashige and Skoog (MS) salts and vitamins supplemented with 7.5 μM thidiazuron and 0.25 μM indole-butyric acid. Transferring the explants after co-cultivation to shoot regeneration medium containing 80 mg l-1 of kanamycin and 300 mg l-1 of Timentin reduced the total number of regenerated shoots without affecting the transformation rate. Transformation rates using the described system averaged 1.2% of the hypocotyl slices producing transgenic plants, with a range of 0–4.2%. The transgenic shoots rooted at a rate of 90% on half-strength MS salts and vitamins supplemented with 5 μM α-naphthaleneacetic acid and 0.01 μM kinetin. Plantlets were transferred to a greenhouse directly from culture tubes with a 90% average survival.

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Abbreviations

ACO1 :

Prunus persica 1-aminocyclopropane-1-carboxylate oxidase

CP :

Coat protein

GUS :

β-Glucuronidase

NPTII :

Neomycin phosphotransferase II

PCR :

Polymerase chain reaction

PDV :

Prune dwarf virus

PNRSV :

Prunus necrotic ringspot virus

SGM :

Shoot growth medium

SRM :

Shoot regeneration medium

TIM :

Timentin (SmithKline Beecham, Philadelphia)

TomRSV :

Tomato ringspot virus

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Acknowledgements

The fellowship provided to I.M.G. Padilla by the Spanish Ministry of Education and Culture is gratefully acknowledged.

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Authors and Affiliations

  1. USDA-ARS Appalachian Fruit Research Station, 45 Wiltshire Rd, Kearneysville, WV 25430, USA

    I.M. Gonzalez Padilla, K. Webb & R. Scorza

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  1. I.M. Gonzalez Padilla
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  2. K. Webb
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  3. R. Scorza
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Correspondence to R. Scorza.

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Communicated by K.K. Kamo

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Gonzalez Padilla, I., Webb, K. & Scorza, R. Early antibiotic selection and efficient rooting and acclimatization improve the production of transgenic plum plants (Prunus domestica L.). Plant Cell Rep 22, 38–45 (2003). https://doi.org/10.1007/s00299-003-0648-z

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  • DOI: https://doi.org/10.1007/s00299-003-0648-z

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