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High transformation efficiency in plum (Prunus domestica L.): a new tool for functional genomics studies in Prunus spp.

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Abstract

An improved Agrobacterium-mediated transformation protocol for plum (Prunus domestica L.) hypocotyl slices was developed based on the addition of 2,4-d to the co-cultivation medium. This method increased transformation efficiency up to 10 × (42%) over previous reports with an average efficiency of 25% of hypocotyl slices producing transgenic plants. Timing of each step in the protocol was optimized producing self-rooted transgenic plants in the greenhouse in approximately 6 months. In order to test the system for its utility in functional genomic studies, we developed two hairpin constructs using a fragment of the peach (P. persica) Phytoene desaturase (PDS) gene. When A. tumefaciens with these constructs was used for targeted post-transcriptional gene silencing (PTGS), approximately 50% of the transformed plums were knockout PDS gene plants. The easy and efficient plum transformation protocol that we report here can be readily used for functional genomics studies in Prunus specifically, and Rosaceae and other woody species in general.

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Abbreviations

2,4-D:

2,4- dichlorophenoxy-acetic acid

BAP:

N6-benzylamino-purine

IBA:

Indole-3-butyric acid

ihp:

Intron hairpin

K:

Kinetin

km:

Kanamycin

NAA:

α-Napthaleneacetic acid

PDS :

Phytoene desaturase

PPV-Cp :

Plum Pox Virus coat protein

TDZ:

Thidiazuron

tim:

Timentin

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Acknowledgements

The authors acknowledge the statistical advice provided by John G. Phillips and the technical assistance of Elizabeth Lutton, Ahn Silverstein and Mark Demuth.

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Authors and Affiliations

  1. Department of Genetic Improvement of Fruit Crops, USDA-ARS-Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Cesar Petri, Kevin Webb, Jean-Michel Hily, Chris Dardick & Ralph Scorza

  2. Department of Entomology, Soils and Plant Sciences, Clemson University, Clemson, SC, 29634, USA

    Cesar Petri

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  1. Cesar Petri
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  2. Kevin Webb
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  3. Jean-Michel Hily
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  4. Chris Dardick
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  5. Ralph Scorza
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Correspondence to Ralph Scorza.

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Petri, C., Webb, K., Hily, JM. et al. High transformation efficiency in plum (Prunus domestica L.): a new tool for functional genomics studies in Prunus spp.. Mol Breeding 22, 581–591 (2008). https://doi.org/10.1007/s11032-008-9200-8

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