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A new selection method for pepper transformation: callus-mediated shoot formation

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

We used two genes, TMV-CP and PPI1 (pepper-PMMV interaction 1 transcription factor), to transform commercially important chili pepper (Capsicum annuum) inbred lines (P915, P409) by means of Agrobacterium co-culture. Eighteen independently transformed T0 plants were obtained. The most critical point in the pepper transformation protocol was the selection of shoots growing on calli—referred to as callus-mediated shoot formation (indirect shooting)—because shoots not grown from the callus (direct shooting from the wounded surface) developed into non-transformants. Selection of the correct right callus type also proved to be an important requirement for obtaining transformed peppers. Six different types of callus developed during the selection process. Shoots regenerated from two of these types, while one type regenerated significantly more shoots than the other types, suggesting that the capacity for shoot formation is callus type-specific. Although the transformation rate was low, transformation via callus-mediated shoot formation proved to be reproducible and was confirmed by Southern and Northern blot analyses. Based on the experimental data, we have succeeded in developing a new protocol for the selection and transformation of pepper and expect that it will be used in the future for pepper transformation.

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Abbreviations

AS:

Acetosyringone

CGMMV:

Cucumber green mottle mosaic virus

CMV:

Cucumber mosaic virus

CP:

Coat protein

IAA:

Indole-3-acetic acid

NAA:

α-Naphthaleneacetic acid

PPI1 :

Pepper-PMMV interaction 1 transcription factor gene

TMV:

Tobacco mosaic virus

ToMV:

Tomato mosaic virus

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Acknowledgements

This research was supported by grants to C.H. Harn by the Crop Functional Genomics Center and from the Plant Diversity Research Center funded by the 21st Frontier Research Program of Ministry of Science and Technology of the Korean Government, and by the Center for Plant Molecular Genetics and Breeding Research (CPMGBR) at Seoul National University funded through the Korea Science and Engineering Foundation (KOSEF). We take this opportunity to thank S.H. Lee (Nong Woo Bio Co.) and Dr. H.Y. Lee (Jeju National University) for their technical assistance.

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Authors and Affiliations

  1. Breeding Institute, Nong Woo Bio Co., 537-17, Jeongdan, Yeoju, Gyeonggi, 469-885, South Korea

    Y. H. Lee, H. S. Kim, J. Y. Kim, M. Jung, Y. S. Park, J. S. Lee, S. H. Choi, N. H. Her, J. H. Lee, S. G. Yang & C. H. Harn

  2. Department of Horticulture, Sangmyung University, Cheonan, Chungnam, South Korea

    N. I. Hyung

  3. KT&G Central Research Institute, Yusong, Daejeon, South Korea

    C. H. Lee

Authors
  1. Y. H. Lee
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  2. H. S. Kim
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  3. J. Y. Kim
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  4. M. Jung
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  5. Y. S. Park
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  6. J. S. Lee
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  7. S. H. Choi
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  8. N. H. Her
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  9. J. H. Lee
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  10. N. I. Hyung
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  11. C. H. Lee
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  12. S. G. Yang
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  13. C. H. Harn
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Corresponding author

Correspondence to C. H. Harn.

Additional information

Communicated by I.S. Chung

Y.H. Lee, H.S. Kim, J.Y. Kim and M. Jung contributed equally to this article.

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Lee, Y.H., Kim, H.S., Kim, J.Y. et al. A new selection method for pepper transformation: callus-mediated shoot formation. Plant Cell Rep 23, 50–58 (2004). https://doi.org/10.1007/s00299-004-0791-1

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  • DOI: https://doi.org/10.1007/s00299-004-0791-1

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